Biological Transformation, Libro

Fraunhofer-Forschungsfokus
Reimund Neugebauer Ed.
Biological
Transformation
Biological Transformation
Reimund Neugebauer
Editor
Biological
Transformation
Editor
Reimund Neugebauer
Zentrale der Fraunhofer-Gesellschaft
Munich, Germany
ISBN 978-3-662-59658-6 ISBN 978-3-662-59659-3 (eBook)

https://doi.org/10.1007/978-3-662-59659-3
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Contents
1 From Contrast to Convergence . . . . . . . . . . . . . . . . . . . . . . 1

Reimund Neugebauer and Martin Thum
1.1 Origins: Technology Helps in the Fight for Survival . . . . . . 1
1.2 A New Approach: Nature and Technology to Cooperate and
Converge . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
1.3 Latest: Biological and Digital Transformation Cross-Fertilize
and Enrich Each Other . . . . . . . . . . . . . . . . . . . . . . . . 3
1.4 The Goal: Efficient and Sustainable Value Creation . . . . . . 5
1.5 Biological Transformation at Fraunhofer and Outlook . . . . . 6
2 Biological Transformation . . . . . . . . . . . . . . . . . . . . . . . . . 9
Patrick Dieckhoff, Sophie Hippmann, Raoul Klingner, and Miriam
Leis
2.1 The Meaning of Biological Transformation . . . . . . . . . . . 9
2.2 Nature as Inspiration for Innovations “Made in Germany” . . 10
2.3 The Contribution to Global Challenges and Solutions
for Sustainability . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
2.4 Concepts and Examples from Fraunhofer Research
on Biological Transformation . . . . . . . . . . . . . . . . . . . . 14
2.5 Processes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Sources and Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
3 Biomimetics Research for Medical Engineering . . . . . . . .... 19

Thomas Bauernhansl and Oliver Schwarz
3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . .... 20
3.1.1 Biomimetics as Creative Technology . . . . . .... 20
3.1.2 Biomimetics as a Discipline with Many Sub-
Disciplines . . . . . . . . . . . . . . . . . . . . . . .... 20
v
vi Contents
3.2 Main Strands of Biomimetics . . . . . . . . . . . . . . . . . . .. 21

3.2.1 Biomimetics—Paradigm Shift and Radical
Innovations . . . . . . . . . . . . . . . . . . . . . . . .. 22
3.2.2 Biomimetics Covers Everything Except
Biotechnology . . . . . . . . . . . . . . . . . . . . . . . 23
3.3 The Role of Biomimetics in Biological Transformation . . . . 24
3.4 Biomimetics in Medical Engineering . . . . . . . . . . . . . . . 24
3.4.1 Surgical Bone Punch Modeled on Snake and Cat . . 26
3.4.2 Drilling Modeled on Insects . . . . . . . . . . . . . . . 28
3.4.3 Implants (Endoprostheses) . . . . . . . . . . . . . . . . 30
3.4.4 Exoprostheses . . . . . . . . . . . . . . . . . . . . . . . . 32
4 Innovative Food Products . . . . . . . . . . . . . . . . . . . . . . . . . . 37

Peter Eisner, Ute Weisz, Raffael Osen, and Stephanie Mittermaier
4.1 Introduction and Background . . . . . . . . . . . . . . . . . . . . 38
4.2 Plant-Based Proteins as Functional Food Ingredients . . . . . 40
4.2.1 Extraction Processes for Plant-Based Protein
Ingredients . . . . . . . . . . . . . . . . . . . . . . . . . . 41
4.2.2 Modification of Plant-Based Protein Ingredients . . 44
4.3 Allergenicity of Plant-Based Proteins and Strategies
for Its Reduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
4.3.1 Food Allergies and Food Allergens . . . . . . . . . . . 46
4.3.2 Strategies for Affected Consumers . . . . . . . . . . . 47
4.3.3 Ways to Reduce the Allergenic Potential of Protein
Ingredients . . . . . . . . . . . . . . . . . . . . . . . . . . 47
4.4 Texturizing Plant-Based Protein Ingredients by Extrusion
Cooking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
4.4.1 Extruders and Extrusion Parameters in Food
Processing . . . . . . . . . . . . . . . . . . . . . . . . . . 52
4.4.2 Production of Plant-Based Meat Alternatives . . . . 52
4.5 Further Applications of Plant-Based Proteins . . . . . . . . . . 57
5 Technical Homes for Human Cells . . . . . . . . . . . . . . . . . . . . 63

Christoph Leyens, Udo Klotzbach, Frank Sonntag, Markus
Wolperdinger, Peter Loskill, Thomas Bauernhansl, Andreas Traube,
Christian Brecher, Robert Schmitt, and Niels König
5.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
Contents vii
5.2 Requirements from a Biological Aspect and Modeling

of Biological Processes . . . . . . . . . . . . . . . . . . . . . . . 65
5.3 Microphysiological Organ-on-a-Chip Systems
as an Interdisciplinary Platform . . . . . . . . . . . . . . . . . . 65
5.3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . 67
5.3.2 Specific Organ-on-a-Chip Systems . . . . . . . . . . . 69
5.3.3 Multi-Organ Chips–All-in-One Microfluidics . . . . 71
5.3.4 Outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
5.4 Sensors and Actuators for Automation in Bioanalytics . . . . 76
5.4.1 Automated High-Throughput Microscopy for Cell
Production . . . . . . . . . . . . . . . . . . . . . . . . . . 76
5.4.2 Deep Learning for Classification in Cell Microscopy 78
5.4.3 Optical Coherence Tomography (OCT) as Sensor
Technology in Tissue Engineering . . . . . . . . . . . 80
5.5 Laboratory Automation as the Basis for Process Transfer
in Biological Transformation . . . . . . . . . . . . . . . . . . . . 83
5.6 Conclusion and Outlook . . . . . . . . . . . . . . . . . . . . . . . 88
6 Phenotyping and Genotyping of Plants . . . . . . . . . . . . . . . . . 91

Udo Seiffert and Andreas Herzog
6.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
6.2 Motivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
6.3 General Approach and Examples of Implemented Solutions . 95
6.4 Outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100
7 Cells as Sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105

Stefanie Michaelis and Joachim Wegener
7.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105
7.2 The Importance of Cell-Based Bioanalysis . . . . . . . . . . . 107
7.3 Cell Culture Models for Various Bioanalytical Applications . 109
7.4 Non-Invasive Physical Signal Transducers for Monitoring
Living Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
7.5 Cells as Sensors—Example Applications of Integral Effect-
Directed Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . 117
7.5.1 Impedance-Based Analysis of Changes in Cell Shape
Under the Influence of External Factors . . . . . . . . 118
viii Contents
7.5.2 Analysis of Cytomechanical Changes

with Piezoelectric Resonators . . . . . . . . . . . . . . 121
7.5.3 Quantitative Drug Testing with Sensor Cells and
Evanescent Fields . . . . . . . . . . . . . . . . . . . . . 123
7.6 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124
8 Biopolymers – Function Carriers in Materials Research . . . . . . 129

Alexander Böker
8.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
8.2 Introduction to Biological Building Blocks/Functions . . . . . 131
8.3 Integration of Biological Functions in Materials . . . . . . . . 134
8.3.1 Integration at the Molecular Level . . . . . . . . . . . 134
8.3.2 New Biological Functions in Thermoplastic
Bioplastics . . . . . . . . . . . . . . . . . . . . . . . . . . 142
8.4 Biomaterials for Medicine . . . . . . . . . . . . . . . . . . . . . . 150
9 Biogenic Plastic Additives . . . . . . . . . . . . . . . . . . . . . . . . . . 161

Rudolf Pfaendner and Tobias Melz
9.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
9.2 Plasticizers Based On Renewable Raw Materials . . . . . . . . 163
9.3 Antioxidants from Plant Extracts . . . . . . . . . . . . . . . . . . 166
9.4 Light Stabilizers from Plant Extracts . . . . . . . . . . . . . . . 168
9.5 Flame Retardants from Renewable Raw Materials . . . . . . . 169
9.6 Other Plastic Additives from Renewable Raw Materials . . . 172
9.7 Outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174
10 Organisms as Producers . . . . . . . . . . . . . . . . . . . . . . . . . . . 179

Stefan Rasche, Stefan Schillberg, Felix Derwenskus, Ulrike
Schmid-Staiger, and Ursula Schließmann
10.1 Production of Dietary Proteins in Microbial Cell Factories . . 180
10.1.1 Introduction and Background . . . . . . . . . . . . . . 180
10.1.2 Selection and Optimization of a Suitable
Phenylalanine-Free Dietary Protein . . . . . . . . . . 181
10.1.3 Production and Testing of the Phenylalanine-Free
Protein . . . . . . . . . . . . . . . . . . . . . . . . . . . . 182
10.1.4 Outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
Contents ix
10.2 Plant Stem Cells for the Cosmetic Industry . . . . . . . . . . . 184

10.2.1 Introduction and Background . . . . . . . . . . . . . . 184
10.2.2 Preparation and Production of Plant Stem Cells . . . 184
10.2.3 Optimization of Production Conditions . . . . . . . . 186
10.2.4 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . 187
10.3 Value-Added Compounds from Microalgae—Increased Value
Creation Through Cascading Use and Fractionation . . . . . . 187
10.3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . 188
10.3.2 Microalgal Ingredients and Areas of Application . . 189
10.3.3 Cultivation of Microalgae . . . . . . . . . . . . . . . . 190
10.3.4 Biomass Reprocessing . . . . . . . . . . . . . . . . . . 193
10.3.5 Outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . 195
11 Biologized Robotics and Biomechatronics . . . . . . . . . . . . . . . 199

Norbert Elkmann, Roland Behrens, Martin Hägele, Urs Schneider,
and Susanne Oberer-Treitz
11.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 199
11.2 Human-Robot Collaboration: Overview, Opportunities and
Challenges . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 200
11.3 Potential Applications, Classification of Human-Robot
Collaboration, Standards . . . . . . . . . . . . . . . . . . . . . . . 200
11.3.1 Forms of Interaction . . . . . . . . . . . . . . . . . . . . 201
11.3.2 Hazards and Safeguarding Modes . . . . . . . . . . . 202
11.4 Risk Assessment when Collaborating with Robots . . . . . . . 203
11.4.1 Current Need for Biomechanical Limits . . . . . . . . 204
11.4.2 Differentiation of Biomechanical Limits . . . . . . . 204
11.4.3 Studies on the Development of Load Limits . . . . . 206
11.5 Robotic Applications with HRC: Now and in the Future . . . 208
11.6 Exoprostheses and Exoskeletons . . . . . . . . . . . . . . . . . . 214
11.6.1 From the Mechanical to the Mechatronic Human-
Technology Interface . . . . . . . . . . . . . . . . . . . 214
11.6.2 New Methods of Biosignal Acquisition . . . . . . . . 217
11.7 Summary and Outlook . . . . . . . . . . . . . . . . . . . . . . . . 219
x Contents
12 Future AM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 225
Johannes Henrich Schleifenbaum, Christian Tenbrock, Claus
Emmelmann, Christoph Leyens, Frank Brückner, and Alexander
Michaelis
12.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226
12.2 Deficits and Derived Fields of Action for Metal AM . . . . . . 230
12.3 Objectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232
12.4 State of the Art . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236
12.5 Current Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . 243
12.6 Outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 244
13 Insect Biotechnology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 247

Andreas Vilcinskas
13.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 247
13.2 Antibiotics from Insects . . . . . . . . . . . . . . . . . . . . . . . 248
13.3 Virulence Blockers from Insects . . . . . . . . . . . . . . . . . . 250
13.4 Insect Enzymes for Industrial Biotechnology . . . . . . . . . . 251
13.5 Insects as an Alternative Protein Source . . . . . . . . . . . . . 254
13.6 Insect Biotechnology in Plant Protection . . . . . . . . . . . . . 255
13.6.1 RNA Interference (RNAi) in Crop Protection . . . . 256
13.6.2 Sterile Insect Technique (SIT) . . . . . . . . . . . . . . 257
14 The Resource Principle . . . . . . . . . . . . . . . . . . . . . . . . . . . 261

Bohumil Kasal, Moritz Leschinsky, Christian Oehr, Gerd Unkelbach,
and Markus Wolperdinger
14.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 262
14.2 Wood as a Material . . . . . . . . . . . . . . . . . . . . . . . . . . 264
14.2.1 Structure and Composition . . . . . . . . . . . . . . . . 264
14.2.2 Plant Fibers . . . . . . . . . . . . . . . . . . . . . . . . . 266
14.2.3 Wood and Fiber Composites . . . . . . . . . . . . . . . 268
14.3 Hybrid Materials with Wood and Plant Fibers . . . . . . . . . . 270
14.3.1 Wood-Based Hybrid Materials . . . . . . . . . . . . . 270
14.3.2 Hybrid Materials Made of Plant-Based Fibers . . . . 274
14.4 Hierarchical Wood Structure and Transformation into New
Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278
14.5 Wood as a Source of Chemical Compounds . . . . . . . . . . . 280
14.6 Technology Assessment . . . . . . . . . . . . . . . . . . . . . . . 281
Contents xi
14.7 An Overview of the Digestion Processes . . . . . . . . . . . . . 283

14.7.1 Mechanical Pretreatment (Structure-Preserving
Method) . . . . . . . . . . . . . . . . . . . . . . . . . . . 283
14.7.2 Autohydrolysis/Hydrothermolysis/Vapor Pressure
Digestion (Structure-Preserving Method) . . . . . . . 283
14.7.3 Alkaline Digestion Processes (Structure-Preserving
Processes) . . . . . . . . . . . . . . . . . . . . . . . . . . 286
14.7.4 Sulfite Digestion/Lignin Sulfonation (Structure-
Preserving Process) . . . . . . . . . . . . . . . . . . . . 288
14.7.5 Digestion with Alcohols, Organosolv Process
(Structure-Preserving Process) . . . . . . . . . . . . . 289
14.7.6 Digestion with Organic Acids (Structure-Preserving
Process) . . . . . . . . . . . . . . . . . . . . . . . . . . . 291
14.7.7 Digestion with Ionic Liquids (Structure-Preserving
Process) . . . . . . . . . . . . . . . . . . . . . . . . . . . 292
14.7.8 Digestion by Calcium Oxide (Structure-Preserving
Process) . . . . . . . . . . . . . . . . . . . . . . . . . . . 293
14.7.9 Pretreatments for Subsequent Hydrolysis and
Utilization of the Sugar Fractions (Some Structure-
Degrading Processes) . . . . . . . . . . . . . . . . . . . 293
14.7.10 Utilization of the Lignin Fraction . . . . . . . . . . . . 297
14.8 Pyrolysis to Oil (Non-Structure-Preserving Process) . . . . . . 299
14.9 Pyrolysis with Subsequent Gasification (Non-Structure-
Preserving Process) . . . . . . . . . . . . . . . . . . . . . . . . . . 301
14.10 Fermentation (Non-Structure-Preserving Process) . . . . . . . 302
15 Cognitive Biological Sensors . . . . . . . . . . . . . . . . . . . . . . . . 311

Albert Heuberger, Randolf Hanke, and Claudia Eckert
15.1 Introduction and Motivation . . . . . . . . . . . . . . . . . . . . . 312
15.2 Learning from Biology . . . . . . . . . . . . . . . . . . . . . . . . 314
15.2.1 Insect Swarms as Multi-Modal Sensor Networks . . 314
15.2.2 Beyond Human Vision—Multispectral Sensors and
Polarization . . . . . . . . . . . . . . . . . . . . . . . . . 317
15.3 Learning for Biology . . . . . . . . . . . . . . . . . . . . . . . . . 319
15.3.1 Digitization of Perception—Campus of the Senses . 319
15.3.2 Digitization of Self-Similar Biological Structures . . 323
15.4 The Cognitive Internet . . . . . . . . . . . . . . . . . . . . . . . . 326
xii Contents
16 Prevention of Biofouling . . . . . . . . . . . . . . . . . . . . . . . . . . . 329

Ralf B. Wehrspohn and Ulrike Hirsch
16.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 330
16.2 Formation and Control of Biofouling . . . . . . . . . . . . . . . 331
16.3 Market Development . . . . . . . . . . . . . . . . . . . . . . . . . 337
16.4 Example: Antifouling Through Electrically Conductive
Marine Paints . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 338
16.5 Example: Low-Fouling Membrane Modules for Reverse
Osmosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 342
16.6 Outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 347
17 Urban Agriculture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 351

Eckhard Weidner, Görge Deerberg, and Volkmar Keuter
17.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 352
17.2 New Approaches to Knowledge-Based Urban Agriculture . . 352
17.3 Local Production Sites . . . . . . . . . . . . . . . . . . . . . . . . 353
17.4 Circular Farming . . . . . . . . . . . . . . . . . . . . . . . . . . . 355
17.5 Digital Horticulture . . . . . . . . . . . . . . . . . . . . . . . . . . 358
17.6 Outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 360
18 Digital Villages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 363

Mario Trapp and Steffen Hess
18.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 363
18.2 Smart Ecosystems as the Basis of Digital Ecosystems . . . . . 366
18.2.1 The Platform Strategy—More than the Sum of All
Parts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366
18.2.2 From Platforms to Smart Ecosystems . . . . . . . . . 367
18.3 The Smart Ecosystem of Digital Villages . . . . . . . . . . . . 369
18.4 Success Factors for a Smart Rural Area . . . . . . . . . . . . . 369
18.5 Smart Rural Areas—More than a Theoretical Construct . . . 370
18.6 A Unified Platform with Flexibly Usable Services . . . . . . . 371
18.7 The “Rural Areas” Smart Ecosystem . . . . . . . . . . . . . . . 374
18.8 Outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 377
Contents xiii
19 Alternatives to Growth . . . . . . . . . . . . . . . . . . . . . . . . . . . 379

Christoph Schäfers, Kristina Bette, Florian Herrmann, and Georg
Nawroth
19.1 Introduction: Limits of Growth, Limits of Growth Economy? 380
19.2 The Population Ecology Perspective: Growth Strategy Versus
Capacity Strategy . . . . . . . . . . . . . . . . . . . . . . . . . . . 381
19.2.1 Economic Development from a Resource Perspective 382
19.2.2 Characteristics of Climax Ecosystems . . . . . . . . . 383
19.3 Transfer to the Economy: Climax Economy Characteristics
Such as Niches, Networking, Material Cycles . . . . . . . . . . 385
19.4 Agriculture and Forestry as Pioneers of the Climax Economy 386
19.5 Value Networks for a Climax Economy . . . . . . . . . . . . . 392
19.6 Outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 394
From Contrast to Convergence
Biological Principles Shape Tomorrow’s
1
Technologies
Reimund Neugebauer and Martin Thum
1.1 Origins: Technology Helps in the Fight for Survival
Technology springs from the human mind. Ever since our species has existed on
earth, we have been using our experience and intellectual effort to design and make
things that are to our benefit: tools for making objects, clothing and shelter for
protection against the weather, weapons for hunting and fighting, instruments for
making music, pigments for painting. It took several millennia of development to
reach a level of technology that clearly distinguished humans from animals in their
way of life. The foremost goal in doing so was always to prevail as best possible
against the challenges of nature.
The progress of evolution was originally, broadly speaking, limited to the devel-
opment and propagation of new characteristics via genetic information, and it took
time. Evolution had that time too, because all organisms and species were dancing
to the same tune: changes were recorded in the universal genetic code and trans-
ferred from one generation to the next, with the option of modification. This only
changed fundamentally when the growing human intellect allowed a new platform
to emerge on which innovations could be created, recorded and improved.
Regardless of the duration of a human generation, ideas and expertise could
grow, be passed on from individual to individual and thus spread. Suddenly, the
R. Neugebauer ()
Zentrale der Fraunhofer-Gesellschaft
Munich, Germany
M. Thum
Fraunhofer-Gesellschaft
Munich, Germany
© Springer-Verlag GmbH Germany, part of Springer Nature 2020 1

R. Neugebauer (ed.), Biological Transformation,
https://doi.org/10.1007/978-3-662-59659-3_1
2 R. Neugebauer and M. Thum
pace at which culture and technology were developing increased enormously, and
so did their power and impact.
At first, technology took on the obvious task of conquering nature for the benefit
of humankind; making better use of known resources and finding new sources. The
increasing efficiency with which this happened soon turned nature and technology
into antagonists. Technology in human hands became the means to tame nature.
And nature—forests, lakes, landscapes, animal and plant species—retreated or dis-
appeared.
The effectiveness of this synergy between humans and technology soon re-
vealed that natural resources are finite. Either they eventually run out like raw
materials and forests, or we have to limit ourselves for climatic reasons, as is the
case with fossil fuels. Examples of large-scale environmental damage and health
as well as social consequences for humans increasingly shaped our experience, and
the damage caused by the course of action so far began to make the benefits ques-
tionable. Humankind is about to win the fight against nature—but in doing so, it is
destroying the basis of its own existence.
At the same time, pure research has aroused an interest in biology—far beyond
the circle of researchers themselves. In the search for new avenues, it thus became
evident that extremely interesting solutions can be found in nature as well. The goal
of natural evolution is fundamentally similar to the objectives being addressed by
applied research today, i.e. to achieve optimal functionality with minimal resource
consumption while taking into account adaptability, recycling and the option of
further development. This new perspective on nature thus led to applied research
taking an increasing interest in biological principles and the insights and findings
obtained by the life sciences.
1.2 A New Approach: Nature and Technology to Cooperate

and Converge
There is no point in arguing about the usefulness of the exponentially rising rate of
development of knowledge and technology today: there are so many people living
on earth that acceptable living conditions for all are inconceivable without the use
of technological advances. And the number of people continues to increase, as do
their needs. A global population of 9.8 billion is expected by the year 2050, along
with double the resource consumption and a 1.5-fold increase in greenhouse gas
emissions.
1 From Contrast to Convergence 3
Thus it is clear, with everyone striving for prosperity, that new avenues will need
to be found in the quest for more efficient and especially more resource-efficient
production. Investigating evolution’s ample stock of experience—which has been
producing highly successful results for hundreds of millions of years—is thus be-
coming more necessary than ever. Research to date shows that nature’s inventory,
its products, processes, principles and tools, can be a major source of inspiration
for the development of modern technologies. The approach of treating nature or
biology more as an inspirational partner rather than an exploitable resource is thus
shifting from being an idea to a logical conclusion.
This realization has been growing for years and even decades. Examples from
bionics have contributed to proving the success of natural evolution in the devel-
opment of impressive solutions to technical problems. For example, the complex
microstructuring of surfaces as a means of cleaning was scientifically investigated
in the lotus plant in the 1970s and has been implemented as a technical solution
for the maintenance-free self-cleaning of certain material surfaces since the mid
1990s. Another case is velcro, whose design is based on the barbs found on the
seeds of the burdock plant. It was patented in 1951 and through its variety of appli-
cations it is now familiar to virtually everyone. A drill is currently being introduced
in surgery, which Fraunhofer research modeled on the structure of the ovipositor of
parasitic wasps of the family Ichneumonidae (ichneumonids). It is used to gently
and efficiently prepare a hip bone for the implantation of an artificial hip joint.
Over and above such decisive developments, biological organisms, materials,
structures and processes are being analyzed, evaluated, imitated and used more
and more as sources of inspiration for ideas. This development of technologies,
processes and products based on biological concepts, particularly when the goal
is sustainable value creation, has collectively become known as “biological trans-
formation”. It is not limited to specific areas of technology, but rather describes
a general direction of development, which leads to results that are as fascinating
as they are convincing—and this with increasing speed and a wealth of forms that
would in many cases not be achievable with purely technical research without us-
ing nature as a guide.
1.3 Latest: Biological and Digital Transformation Cross-

Fertilize and Enrich Each Other
The above examples show that the use of biological processes and strategies is ba-
sically not new. However, recognizing the usefulness of processes or structures in
nature is something quite different, in terms of effort, than precisely analyzing their
complexity or even imitating it by means of technology. The current acceleration

of biological transformation in the field of applied research and technology thus
also has to do with another technology, which is itself undergoing a tremendous
spurt in development: digital transformation.
Many of the processes or products of biological transformation presented in this
book can only be realized with the aid of very powerful data processing—a cir-
cumstance which incidentally also finds its analog in nature. In both cases—in the
technological as well as in the biological world—there are highly complex data
storage and data processing systems based on a simply structured universal code:
the genetic code in nature and the digital code in technology.
One example of a digital technology as enabler of a production technology in-
spired by biological systems, is additive manufacturing, also often referred to as
the “printing” of objects. In contrast to the subtractive method where the planned
component is produced, for example, from a block of material by precise removal
of unwanted material parts by means of milling, drilling or grinding technology,
one can produce much more complex structures in the additive process without
generating large amounts of waste. This latter method is also the preferred method
in nature for producing functional structures of almost any size and texture ex-
tremely efficiently. Every tree and every bone, every shell and every organ is built
up additively, cell by cell or layer by layer.
The technological version of additive manufacturing is, however, only econom-
ically possible in the fascinating wealth of options that we know, with the powerful
digital technology that is available today. The digitization of technology is in this
case—as in other cases—a crucial prerequisite for the development of a powerful
biological transformation.
Conversely, biological principles also spur on the performance of digital sys-
tems. Thus, artificial neural networks are modeled on the network of nerve cells
in the brain. They consist of data nodes and weighted connections between them.
Machine learning techniques can be realized by changing various parameters in
this network. Neuromorphic chips are microchips with nerve cell properties and
architecture that are replicated at the hardware level. These components, which are
similar to neurons, simulate the ability of the brain to learn and associate, which
can, for example, accelerate the recognition of patterns in images or in big data
structures. Systems such as these can be far better placed than humans to distin-
guish healthy cells from cancer cells in medical samples. They can also be used
to draw insights from large volumes of data that were not accessible using former
analytical methods.
Also, in the field of sensor technology, adaptations from nature have led to
promising technology trends too. One example is the development of an ultra-thin
optical sensor based on the principle of a compound eye, as seen in insects and
other arthropods. These highly miniaturized optics find application in smartphones,
cars, medical technology or in production facilities, where they help to make the
digital systems more powerful and efficient and result in new product attributes.
Hybrid sensors, which consist of cells and microsystems engineering struc-
tures, are an example of where technological components and living cells interact
directly. They work extremely specifically and may be integrated into analytical
systems. Biological and digital systems are therefore not only becoming more and
more alike, they are also working together more and more, reciprocally enhancing
each other’s performance.
The congruence between materials and biology has long been a concrete re-
search goal in medical applications. Accurate knowledge and the modification of
material properties in the interplay with cells in complex biological systems allows
for targeted control of the interaction with the surrounding tissue in new implants.
Especially when it comes to polymers, biological transformation is in full swing.
This will create pioneering materials that are as functional as they are sustainable.
These materials will improve technical processes and make new therapies or diag-
nostic platforms possible. Synthetic materials that deliberately interact with nature
represent the next evolutionary step in the development of polymeric materials.
1.4 The Goal: Efficient and Sustainable Value Creation
The adoption of biological models, principles and processes rapidly results in

highly functional products with new and striking properties. This aspect is the
primary deciding factor as to whether the innovation can succeed in international
markets. But if we extend our responsibility beyond mere market success, the argu-
ment that products emerging from biological transformation can be produced and
utilized highly efficiently and in particular, resource-efficiently, gains importance.
A classic organizational principle seen in nature is the cycle, and the best exam-
ple of this is the Amazon rainforest. The lushest life in this habitat can be attributed
to the fact that the mineral nutrients remain almost entirely within the ecosystem
and are immediately absorbed and reused after organisms die. If the cycle is bro-
ken through deforestation, the damage to the biosystem is practically irreparable,
because the nutrients are rapidly washed away and lost to the system.
The principle of the intact cycle thus implies the careful use of resources and
thus, in applied research, has long served as an ideal goal for technical systems,
product life cycles, production facilities and ultra-efficient factories. Production
processes that use organisms and their special abilities to produce certain sub-
stances go one step further—nature is no longer just a role model, but now a par-
ticipant. Algae, fungi, bacteria and plants are among these organisms and some
produce substances that are very useful to humans, such as pharmaceuticals, valu-
able chemicals or complex food substances.
The convergence of biology and technology can be followed particularly well
and seen in many places in the production of organic resources: modern agriculture
uses satellite navigation and digital planning tools, highly automated aquaculture
uses microorganisms to produce foodstuffs and other valuable materials in the mid-
dle of a city, biological waste is converted to a source of high quality raw material
in technical plants.
Many such technologies are ongoing developments of already existing pro-
cesses, but some ideas also open up completely new possibilities, especially within
the realm of biological transformation—such as insect biotechnology. Dietary pro-
teins produced in insect bioreactors can be produced with a fraction of the resource
consumption of conventional livestock or crop-based processes. In light of the
world’s food problems, these processes, with their eminent ecological and eco-
nomic advantages, are a promising field of research with enormous potential.
1.5 Biological Transformation at Fraunhofer and Outlook
The Fraunhofer-Gesellschaft has recognized the developmental technological po-

tential of biological transformation. Within the framework of the Fraunhofer 2022
Agenda, which aims to further increase the impact of Fraunhofer research on the
economy and society, Fraunhofer has defined seven Key Strategic Initiatives. Bi-
ological transformation is one of them, and Fraunhofer sees its task not only as
driving the relevant research forward, but also as promoting public awareness of
the topic.
The tremendous dynamization of technological evolution associated with bio-
logical transformation is both a decisive factor for successful international compe-
tition and also a path that we, as the protagonists of applied research, must take in
terms of our responsibility for the wellbeing of all human beings, as mentioned at
the outset. In this way, we follow our fundamental principle: people research for
people.
Biological transformation is such a universal topic, that we have already gone
into and presented developments in the previous volumes of the series “Fraunhofer-
Forschungsfokus” (Fraunhofer Research Focus), which would thematically also fit
the current book: artificial intelligence and additive manufacturing, for example,
were topics in the book “Digital Transformation” and the efficient and environ-
mentally friendly production of natural rubber from dandelions was discussed in

the book “Resource Efficiency”.
The rapid development of medical and health-related research has prompted
us to add another publication in the series “Fraunhofer-Forschungsfokus” on this
topic, to follow on from the current book on biological transformation, in one year.
Other possible areas of research in which Fraunhofer will significantly intensify
its work in the coming years include public safety, quantum technology and the
development of programmable materials.
Biological Transformation
A Research Agenda of the Fraunhofer-Gesellschaft
2
Patrick Dieckhoff, Sophie Hippmann, Raoul Klingner, and
Miriam Leis
Summary
The application of biological principles has resulted in profound changes in sec-

tors as diverse as pharmacy, consumer goods and food production, as well as in
agriculture. Renewable raw materials and the use of production organisms have
long been the emphasis of biotechnology. But the focus is now on linking new
biological processes and findings with other innovations in agricultural science
and in information, medical or manufacturing technology, which also incor-
porate the circular economy. With biological transformation, the Fraunhofer-
Gesellschaft aspires to make a significant contribution to the United Nations’
goals for sustainable development.
2.1 The Meaning of Biological Transformation
The term “biological transformation” refers to the increasing use of materials,

structures and processes of living organisms in technology, with the goal of sustain-
able value creation. As the largest organization for applied research in Europe, the
Fraunhofer-Gesellschaft sees this as a very important field of research and innova-
tion for the future that has the potential to fundamentally transform value creation.
Which materials will we use in the future? How will we manufacture products?
How do we cure diseases and protect ourselves from danger? How do we inte-
grate human economic activity into the cycles of nature? We find answers to these
P. Dieckhoff () S. Hippmann R. Klingner M. Leis

Fraunhofer-Gesellschaft
Munich, Germany

https://doi.org/10.1007/978-3-662-59659-3_2
10 P. Dieckhoff et al.
questions when we make use of life’s innovations and these are integrated into
technology, which is often only made possible through digitization.
Advancing digitization enables us to better understand nature’s optimization
processes, which can be used to design sustainable processes for the economy and
society. This leads to a change process that brings together different disciplines
and economic sectors, particularly digital information technologies, biotechnol-
ogy, neurosciences, materials and nanotechnology, as well as behavioral, social
and economic sciences, and allows novel ideas to emerge at their interfaces. Be-
hind this lies the assumption that digital agendas in the context of Industry 4.0
serve efficiency and networking at their core, but not necessarily sustainability.
The essential needs of society for the protection of public goods such as water, air
and soil, biodiversity, etc. cannot be solved by digitization alone. The Fraunhofer-
Gesellschaft’s concept of biological transformation thus provides a biological par-
allel to digital transformation.
2.2 Nature as Inspiration for Innovations

“Made in Germany”
Using nature as a source of innovation has a long tradition in Germany. The

botanist and microbiologist Raoul Heinrich France received the first German
patent for a bionic invention as early as 1920. He used the seed dispersing capsule
of the opium poppy as a model for a salt shaker that would distribute the small
particles evenly. This paved the way for the general patentability of bioinspired
technological devices, and also made the field of bionics, or rather, “biotechnol-
ogy”, attractive from a commercial point of view. In Germany in particular, the
concept of bioinspired technology is rated as largely positive and considered an
important element for sustainable innovations.
The targeted utilization and adaptation of biological processes to the needs of
humans already has a long history to look back upon. It is believed that some
10,000 years ago, humans started to selectively cultivate plants and perhaps even
use microorganisms to brew alcoholic beverages. Leonardo Da Vinci developed
bioinspired machines in the 15th century that were conceptually way ahead of
his time. In 1943, the concept of artificial neural networks emerged as a simpli-
fied and abstract mathematical formulation of biological neurons, which serves as
a basis for machine learning applications today. In 1958, J.E. Steele coined the
term “bionics” in the English-speaking world, defined as “learning from nature for
2 Biological Transformation 11
technology”, although in the USA in particular, “bionics” is largely understood as

the field of prosthetics and “cybernetic organisms”.
In the 1970s, the process of systematically modifying microorganisms, bacte-
ria and cells in order to use them specifically for the large-scale production of
complex pharmaceutical and chemical components became prevalent. The two
pioneers, Stanley Cohen and Herbert Boyer, turned nature into a production as-
sistant. By learning to multiply the DNA in cells, they invented biotechnology.
Their venture was to apply technical knowledge to living organisms. Today, a wide
range of goods are produced with the help of biological organisms. Bacteria, fungi
and other microorganisms are efficient producers of vitamins, specialty chemicals,
biofuels and humanized implants. Modern laundry detergents now use active wash-
ing substances generated in this way to clean laundry as effectively at a washing
temperature of 30 °C as they used to do at 90 °C, thereby saving energy. The ap-
plication of biological principles has resulted in profound changes in sectors as
diverse as pharmacy, consumer goods and food production, as well as agriculture.
In the 1990s, the technological application of the lotus effect in particular was
used in a variety of products, which also reached the end consumer market. The
first successful demonstrations of neuroprosthetics in humans were also demon-
strated. The brain and nervous system were connected using digital technologies,
for example to control computers and prostheses by electronically reading and
processing neural signals. The German federal government also began to promote
biotechnology in the mid-1990s. This was first sparked off by the BioRegio com-
petition of the German Federal Ministry of Education and Research (BMBF) [1].
Social and political discussions about the concept of bioeconomy also intensified
during this period.
In 2002, the US National Science Foundation (NSF) and the US Department of
Commerce issued a report entitled “Converging Technologies for Improving Hu-
man Performance”. It describes a vision of a development in which the scientific
fields and technologies of nanotechnology, biotechnology, information technology
and cognitive science (NBIC) converge. The EU was skeptical of the highly tech-
nology-driven and individualistic outlook of the ideas coming from the USA and
responded with a counterdraft “Converging Technologies—Shaping the Future of
European Societies” which involved the humanities and social sciences to a far
greater extent.
In Germany, the BMBF provided a further two billion euros for biotechnol-
ogy research between 2001 and 2011. In addition to that, during the German EU
Council Presidency in 2007, a groundbreaking paper was adopted to promote the
bioeconomy: “En Route to the Knowledge-based Bio-economy” [2]. The inter-
ministerial “National Research Strategy Bioeconomy 2030” was published two
years later, with a volume of around 2.4 billion euros [3]. The bio-based economy
became the central issue of this high-tech strategy. The coalition treaty agreed be-
tween the CDU, CSU and SPD in spring 2018 will take the promotion of bio-based
innovations to a new level: “We will drive forward the use of natural principles
and, together with industry, science and civil society, develop an interministerial
agenda, extending ‘from biology to innovation’” [4]. As the BMBF has to date
mainly promoted renewable raw materials or the use of production organisms in
biotechnology, the perspective has now been broadened to include the principles of
nature. The bioeconomy council sees the combination of new biological processes
and findings with other innovations in agricultural science and in information, med-
ical or manufacturing technology as particularly future-oriented.
The technological developments that have a critical impact on the agenda are
advances in information and communication technology (e.g. microelectronics,
sensors, computing and storage capacity, imaging, data transmission) and the re-
sulting findings in life sciences (e.g. through improved capabilities in genomics
and neuroscience as well as general progress in the analysis of complex biological
and ecological systems). With the current Industry 4.0 trend, Germany is suc-
cessfully expanding its expertise in industrial production with the help of digital
technologies. In a similar way, biological transformation offers the opportunity to
take innovation to a new level, shoulder-to-shoulder with the important issue of
sustainability.
2.3 The Contribution to Global Challenges and

Solutions for Sustainability
There are currently around 7.5 billion people on Earth, a figure that may rise to
9.8 billion by 2050. Given the finite nature of available resources—water, food, raw
materials, energy sources—and the pursuit of prosperity for all, serious challenges
arise in the question as to how humanity’s increasing needs can be covered in the
longer term. A continuation of the status quo is untenable, both ecologically and
economically.
One measure of sustainability is the ecological footprint. It measures the en-
vironmental values that a particular region needs to produce the natural resources
it consumes (including food, wood and fibers, as well as space for urban infras-
tructure) and to absorb its waste (including CO2 emissions). The “ecological over-
shoot” describes the situation that is already occurring, that the annual consump-
tion of biological resources exceeds what the earth can regenerate every year.
Currently humanity uses, on average, an equivalent of “1.7 Earths” for resource

replenishment and waste processing. This means that the Earth needs one year and
eight months to replenish what we consume in one year [5].
1.1 billion people are already affected by a shortage of drinking water. By 2025,
up to two-thirds of humanity could, at least periodically, suffer from water short-
ages [6]. The way we deal with waste and our consumption have also become
a problem. Two billion metric tons of waste is generated each year. Packed on
freight trains, this would stretch around the world 24 times. An increasing problem
in this context is the growing amount of electronic waste. According to the UN
report, 44.7 million metric tons were generated in 2016; this amount could rise to
52 million metric tons in 2021. Only about 20% of this is currently being recycled.
Although, according to one calculation, this waste contains materials worth 55 bil-
lion euros [7]. Another problem that is noticeably coming to the fore is the huge
amount of plastic waste on land and in the ocean. Seven million metric tons of it
end up in the sea every year, which is equivalent to the weight of 100 cruise ships
[8]. These challenges can be addressed with the concept of the circular economy.
The fundamental idea of the circular economy and circular production—another
central aspect of biological transformation—aims to mimic biological processes in
which there is no waste, but only material flows that are utilized at different levels.
Since technical products are usually made of materials such as metals that do not
regrow, do not regenerate themselves and cannot ultimately biodegrade, technical
cycles differ from natural ones. Other strategies such as resilient and durable de-
sign or substitution with renewable materials, repair, re-use, reprocessing and, as
a last step, recycling therefore need to be implemented in order to close the techni-
cal cycle and reduce non-recyclable waste as much as possible. New technologies
such as the use of bio-based and biodegradable materials, self-healing capabili-
ties, lightweight bionic structures for energy conservation, additive manufacturing
to reduce waste, and biologically inspired optimization strategies such as swarm
intelligence and the capacity for collaborative intelligence between humans and
machines can all make important contributions here.
In September 2017, the United Nations adopted 17 sustainable development
goals (SDGs) under the “2030 Agenda for Sustainable Development”. The plan is
to initiate relevant change in the direction of development by 2030. The 17 goals
form a framework of action for all nations to refer to in dealing with major societal
challenges until 2030. A total of 169 sub-goals and a set of indicators concretize
the challenges and the monitoring of goal achievements.
At the national level, the German Sustainability Strategy, which is the responsi-
bility of the Federal Chancellery and based on the SDGs, was updated in December
2016. Not only business and civil society, but even more so science is called upon
to make a contribution to achieving the political goals. In 2017, the German Fed-
eral Government initiated the science platform “Sustainability 2030” in order to
involve science more in the implementation of the goals.
The Fraunhofer-Gesellschaft aspires to make a significant contribution to the
United Nations’ goals for sustainable development through biological transforma-
tion. However, biological transformation is not sustainable per se: the irresponsible
use of wood could for example lead to the destruction of forests. Sustainability can
only succeed in a holistic and transdisciplinary approach if the materials, structures
and processes used, coming from living organisms, enable the technical system to
operate more efficiently—in the sense of the greatest possible benefit with the least
possible use of resources, with regard to ecological, economic and social dimen-
sions.
2.4 Concepts and Examples from Fraunhofer Research

on Biological Transformation
All existing living organisms can be viewed as evolutionary success models. Over
billions of years of evolution, nature has developed many solutions that can serve
as models for efficient and effective technologies. These may be material properties
(e.g. the lots effect, structural colors, biocompatibility or the material composition
of bones, mother-of-pearl or spider silk with extremely stable lightweight proper-
ties), structures and shapes (e.g. for excellent aero- or aquadynamics), but also in-
clude the general processes of nature, such as networked communication (nervous
system, neurons), self-organization, swarm intelligence, self-healing, homeostasis
or circulatory processes. Animals and other organisms have likewise developed
sophisticated survival and adaptation strategies over time, for example to camou-
flage, to manage with as few resources and as little energy as possible, to pro-
tect themselves against attack and to survive under adverse conditions. Biological
transformation is a concept for transferring these strategies to technical systems.
As a first step towards systematization, biological transformation can be divided
into three modes of development: inspiration, integration and interaction. These
emerged at different times, use different tools and proceed to this day and into the
future [10].
Inspiration
Arguably the oldest, yet still a relevant way to learn from nature is through inspira-
tion, as the example of the patent for the bio-inspired salt shaker by Raoul Heinrich
France shows. Even additive manufacturing or 3D printing can be mentioned as
a modern example, a process that, among other things, the Fraunhofer Institutes
for Laser Technology ILT and for Ceramic Technologies and Systems IKTS and
the Fraunhofer Institute for Additive Production Technologies IAPT are pursuing.
In nature, everything is built up additively, cell layer by cell layer, which allows
for extremely resource-efficient production with a great variety of geometries and
different material combinations.
Integration
The inspiration-driven replication of natural structures and processes can quickly
be followed by a further development of ideas to include a targeted adaptation
of the biological model to meet technical requirements. One example of this is
scaling and improving the efficiency of production processes for bio-based raw
materials using bacteria or other microorganisms for industrial-scale applications.
For example, silk proteins from spiders or green lacewings can be produced in
large quantities for the production of high-tech, bioinspired and bio-based mate-
rials using modified microorganisms in order to make the applications technically
and economically feasible. This already brings us to the point of integration, where
biological components or principles become an essential part of a technical system.
For example, the Fraunhofer Institute for Interfacial Engineering and Biotechnol-
ogy IGB is researching the use of fungi, algae, bacteria and plants to produce
reusable materials and active ingredients for pharmaceutical products. Even mod-
ern mini-labs, lab-on-a-chip systems, are now integrating real biological cells that
work like mini-organs, for example. These can be used to test the effect of chem-
icals or drugs on individuals. Such highly complex systems require expertise in
microelectronics, materials research and biology, which is why several research
institutes of the Fraunhofer-Gesellschaft, such as the Fraunhofer Institute for Pro-
duction Technology IPT, for Manufacturing Engineering and Automation IPA, for
Biomedical Engineering IBMT, for Material and Beam Technology IWS and for
Interfacial Engineering and Biotechnology IGB are all collaborating on these de-
velopments.
Interaction
If the biological components are not only used passively, for example for the
production of certain raw materials, but are also in active interplay with the tech-
nical systems, one speaks of an interaction. An example of this would be the
bilateral communication between the nervous system and microelectronics in the
field of high-tech prosthetics, where the goal is to make a near-naturally func-
tioning technology. Another current field of research is biosensor technology, in
which biomolecules interact with electronics, for example to mimic the olfac-
tory function of a nose. Here Fraunhofer EMFT is developing miniaturized hybrid

sensors that consist of living cells and technical components and which can be in-
tegrated into microsystems. These can identify substances with a specificity that
only animals would otherwise achieve. The connection to electronic systems as
an interface makes it easier for people to interpret the data. In a similar way,
modern technology and sensors can be used to enable plants to communicate, so
that, for example, environmental conditions for agriculture in enclosed spaces may
be optimally adapted to the plant. Methods such as these are being developed by
researchers at the Fraunhofer Institutes UMSICHT and IME with the aim of im-
proving decentralized food production through vertical and urban farming.
With every step taken from mere inspiration up to communication between bi-
ology and technology, the complexity of the system and thus also the demand for
the necessary understanding of the underlying biological and technical processes
grows. Biology’s useful tools that can be taken advantage of can be divided into
materials, structures and processes.
Materials
Although biological objects are only composed of sugars, proteins and miner-
als, they have a broad spectrum of material properties—just think how different
the consistencies of bone and liver are. The human body consists mainly of the
elements oxygen, carbon, hydrogen and to a lesser extent, nitrogen. Technical ob-
jects usually consist of a far greater number of individual materials. A modern
smartphone consists of at least 40 different elements, most of which are metals
[9]—a challenge for the separation and the subsequent recycling of valuable sub-
stances.
Increased ease of recycling is therefore both an ecological and economic con-
cern. Petroleum-derived materials and additives are to be replaced with bio-based
and renewable alternatives. At the Fraunhofer Institute for Structural Durability
and System Reliability LBF, projects are being pursued in which plasticizers, light
stabilizers or flame retardants are to be obtained from plant extracts. Research is
also being conducted on new sources of nutritional protein and on active molecules
for pharmaceutical agents. The Fraunhofer Institute for Process Engineering and
Packaging IVV is investigating new raw materials and processing methods for
healthy alternatives to conventional foods and protein sources and the Fraunhofer
Institute for Molecular Biology and Applied Ecology IME is researching insects,
a highly successful group of organisms, to harness their successful molecular
strategies for biotechnology and drug discovery.
Biopolymers harbor enormous potential for sustainable, future-oriented materi-
als. Biofunctionalization furthermore lends them an integrative dimension, as they
are not only based on nature in their structure and function, but can also incor-
porate biological components such as proteins, peptides or sugars as constituents.
Wood is also well-known as a valuable bio-based, albeit extremely complex ma-
terial with potential that is far from exhausted. Modern analytical methods have
made it possible to describe the varied characteristics of different types of wood
in more and more detail, thus allowing the utilization of this versatile material to
be optimized. The Fraunhofer Institutes for Wood Research, Wilhelm Klauditz In-
stitute WKI, and Interfacial Engineering and Biotechnology IGB, as well as the
Fraunhofer Center for Chemical-Biotechnological Processes CBP, have focused
on the intelligent utilization of wood-based materials, natural fibers and organic
residues in order to expand the possibilities for sustainable application of these
fascinating materials.
Structures
Biological objects often have remarkable properties which can be ascribed to their
geometry or structure and the specific arrangement of molecules and atoms. Nano
research and nanotechnology are especially well-equipped to offer better and better
insights into the influence that the structural nature of materials can have on their
properties at the smallest level, for example with regard to aero- or aquadynamics
or water repellence. Often structure and material are jointly responsible for the
specific properties, so that a clear separation is not always possible.
Dirt-repellent surfaces as known in nature, like the lotus leaf, are highly sought-
after in technological applications. One particular challenge is to prevent the for-
mation of organic deposits on ships’ hulls, known as biofouling. The Fraunhofer
Institute for Microstructure of Materials and Systems IMWS has developed a non-
toxic, conductive paint system for ships, which uses electric fields to modify the
surface in such a way that organisms cannot settle on the surface. A process based
on the knowledge of the environmental requirements of fungi, algae and other
species that grow in water.
2.5 Processes
Biological systems are characterized in particular by unique abilities such as adap-

tation, self-healing, self-organization, reproduction or homeostasis, which are re-
alized through processes including neuronal signal transmission, DNA transcrip-
tion, immune responses, circulatory principles, etc. Unlike conventional, technical
machines, which need to be powered, controlled, and maintained by external inter-
vention, biological systems are geared towards self-preservation, self-organization,
adaptability and the efficient use of available resources. While most technological
systems have to be dismantled and recycled when they are no longer able to func-
tion, the components of expired biological organisms flow back into circulation,
serving other organisms as food and thus helping to build up new, complex biolog-
ical structures.
Artificial neural networks, artificial intelligence and cognitive systems are
highly topical examples of how the principles of natural information processing
by sense organs, the nervous system and brain can be reproduced in technical
systems, at a high level of abstraction, by sensors, protected data transmission
and machine learning. The “cognitive internet” of the future could be similar to
a perceptive, communicating organism and the Fraunhofer Cluster of Excellence
“Cognitive Internet Technologies” is currently working on its design. This could
sustainably improve industrial production as well as logistics applications or med-
ical diagnostics and more closely combine technical and biological production and
organizational principles.
Sources and Bibliography

1. Federal Ministry of Education and Research (2010) Biotechnology in Germany – 25
years of business start-ups
2. DECHEMA (2007) En route to the knowledge-based bio-economy
3. Federal Ministry of Education and Research (2010) National Research Strategy Bio-
economy 2030
4. CDU, CSU, SPD (2018) A new departure for Europe. A new dynamic for Germany. A
new cohesion for our country
5. https://www.footprintnetwork.org/our-work/ecological-footprint/ Accessed: 09/19/2018
6. http://www.un.org/waterforlifedecade/scarcity.shtml. Accessed: 09/19/2018
7. https://news.un.org/en/story/2017/12/639312-electronic-waste-poses-growing-risk-
environment-human-health-un-report-warns. Accessed: 09/19/2018
8. https://de.statista.com/infografik/10463/daten-und-fakten-zu-plastikmuell-in-den-welt-
meeren / accessed: 09/19/2018
9. http://www.compoundchem.com/2014/02/19/the-chemical-elements-of-a-smartphone/
Accessed: 09/19/2018
10. Bauernhansl, T.; Brecher, C.; Drossel, W.-G.; Gumbsch, P.; ten Hompel, M.;
Wolperdinger, M. (Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung
e. V.) [Ed.]: Biointelligence – A new perspective for sustainable industrial value creation
– results of the biological transformation preliminary study on the biological transforma-
tion of industrial value creation (BIOTRAIN). Aachen, Dortmund, Dresden, Freiburg,
Stuttgart, 2019, Fraunhofer-Verlag. ISBN 978-3-8396-1433-4
Biomimetics Research for Medical
Engineering 3
Innovative Devices and Processes Modeled
on Nature’s Evolutionary Solutions
Thomas Bauernhansl and Oliver Schwarz
Summary
Biomimetics and biotechnology are the disciplines that bring biological knowl-
edge into biological transformation. The difference is that biotechnology uses
organisms directly for the production, conversion and decomposition of sub-
stances, while biomimetics attempts to implement the derived principles in
abstract form using technological means. The three levels at which we can take
up nature’s ideas for a sustainable economic system are learning (1) from the
results of evolution, (2) from the principles of nature, and (3) from the process
of evolution itself.
The findings thus obtained can be implemented in almost all areas of re-
search and industry. In principle, an increase in material and energy efficiency
of up to 30% is possible. The field of medical engineering is particularly predes-
tined for implementation, because the materials of the body, which nature also
works with, are being manipulated. There is cooperation between biomimetics
and biotechnology when it comes to cell-covered implants (hybrids), in the use
of biogenic material and in the functionalization of implants. Great potential for
further synergy between bioengineering disciplines for many more applications
that fall under biological transformation is anticipated.
T. Bauernhansl () O. Schwarz

Fraunhofer Institute for Manufacturing Engineering and Automation IPA
Stuttgart, Germany

https://doi.org/10.1007/978-3-662-59659-3_3
20 T. Bauernhansl and O. Schwarz
3.1 Introduction
Biomimetics can be understood as a creative technology. Although many people

reduce it to the lotus effect or gecko adhesion, it can also be considered an inde-
pendent discipline.
In discussions on biological transformation it became evident that “old”
biomimetics was spoken of, in contrast to artificial intelligence or nanotechnology.
What lies behind this distinction?
Before we deal with biomimetics in its specific application in medical engineer-
ing in this chapter, we will first discuss its definition, development and significance
for biological transformation.
3.1.1 Biomimetics as Creative Technology
The cooperation of two people generally leads to greater creative output than the
work of two lone wolves. The more heterogeneous their wealth of experience and
disciplinary background, the greater potential there is to creatively combine and
interrelate ideas. Because of its interdisciplinary nature, bridging the natural sci-
ences and engineering, biomimetics intrinsically involves, as it were, working on
the boundaries between disciplines and thus has a high potential for creativity.
Undoubtedly, biomimetics is a creative technology—and indeed the only one that
provides proven solutions. Of course, the trials do not take place in a technological
context. It can also be assumed that the maximum achievable optimum does not
exist in nature, because there are many dependencies and boundary conditions that
an evolutionary process is subordinate to. After all, the ability of an individual or-
ganism to survive—in the special case of eusocial animals, the ability of the colony
or of the queen to survive—is the primary goal and not the singular maximization
of an ability.
3.1.2 Biomimetics as a Discipline with Many Sub-Disciplines
The foundation stone was laid for the creation of a discipline when the first def-
inition (biomimetics = biology and electronics) was provided by Jack E. Steele
in 1960. According to the sociologist, Rudolf Stichweh, a discipline is character-
ized by “the homogeneous communication context, the accepted corpus of sci-
entific knowledge and the set of questions, research methods and paradigmatic
solutions to problems” [11]. Wrangling over generally accepted methods and tools
3 Biomimetics Research for Medical Engineering 21
in the biomimetics community resulted in a series of VDI guidelines1 (Associa-

tion of German Engineers). Some of these have already been carried over as ISO
standards. According to Stichweh, the professional socialization of young scien-
tists also has to take place through their own institutions. In Germany this began
with Werner Nachtigall establishing the degree program “Technical Biology and
Biomimetics” at Saarland University in 1990. He was co-founder of the Society
for Technical Biology and Bionics (GTBB) in 1990, which became the first nu-
cleus for a biomimetics community. Today, there are half a dozen independent
bachelor’s and master’s programs in Germany and Austria. The only specialization
in biomimetics for medical engineering is offered at the University of Stuttgart,
greatly underpinned by the research activities at the Fraunhofer Institute for Man-
ufacturing Engineering and Automation IPA. Biomimetics content is taught under
the banner of biomechatronics in the biomedical engineering program at the TU
Ilmenau, too.
The German biomimetics network, Biokon, comprises ten thematic working
groups2, which reflect the breadth and heterogeneity of this discipline today.
Certain recent manifestations such as molecular biomimetics or economic/
organizational biomimetics are not yet represented there.
3.2 Main Strands of Biomimetics
Man has studied bird flight for many years—for application in avionics, for ex-
ample—and obvious analogies from nature were transferred to technology in the
design of the first aircraft. Similar developments later followed in ship and vehicle
manufacturing. The first applications of biomimetics thus came from functional
morphology and biomechanics. This was followed by model-based biomimetics
from the 1950s onwards. From the 1980s, biomimetics was extended to the mi-
1
Biomimetics has been decisively standardized and made transparent by the now
seven VDI guidelines—“Conception and strategy”, “Biomimetic surfaces”, “Biomimetic
robots”, “Biomimetic materials, structures and components”, “Biomimetic optimiza-
tion—Application of evolutionary algorithms”, “Integrated product development process
for biomimetic optimisation”, “Bioimimetic information processing”, “Architecture, civil
engineering, industrial design—Basic principles”, which facilitate and enable the imple-
mentation of bionic developments in technical application Architecture, civil engineering,
industrial design—Basic principles.
2
“Architecture and design”, “lightweight construction and materials”, “surfaces and inter-
faces”, “fluid dynamics”, “robotics and manufacturing engineering”, “sensors and infor-
mation processing”, “biomimetic optimization methods”, “organization and management”,
“education and further training”, “biomimetic medical engineering”.
cro and nanoscale; the lotus effect and gecko adhesion are well-known examples
thereof. In the 1990s, biomimetics was spurred on by innovations in information
technology, nanotechnology and biotechnology.
By now an established scientific discipline, biomimetics generates numerous
innovations today. One particularly significant driver of Biological Transformation
is molecular biomimetics.
Medical biomimetics—and likewise almost all areas where biomimetics is ap-
plied—makes use of all these main strands and constantly develops them further.
If you look at the research topics of the first “Bionic Symposium” in Dayton,
Ohio in 1960, the kick-off congress for the field, you will already find state-of-
the-art terms such as “neural processing, bio-computers, sensor technology, the
improvement of radar through the biological sonar of bats” [1]. The use of the term
“old biomimetics” to differentiate from the bionic use of artificial intelligence, for
example, thus does not correspond to the actual developments.
3.2.1 Biomimetics—Paradigm Shift and Radical Innovations
Generally speaking, paradigm shifts are previous worldviews that undergo funda-
mental change as a result of scientific knowledge. In biomimetics this applies to
at least two discoveries: the “lotus effect” and the “boxfish”. “The smoother, the
cleaner” was the accepted theory before Wilhelm Barthlott discovered the highly
regular, microscopic knobbly structure of lotus leaves under the scanning electron
microscope, which make the adhesion of water impossible. “Dirt repellence is not
achieved by ultra-smooth, but rather by rough surfaces” was the revolutionary in-
sight that could then be technically implemented in Lotusan® facade paint.
While searching for fish with a shape that could inspire ideas for a new class of
car with a steeply rising tail (A-class), Daimler’s attention was drawn to a fist-sized,
shoebox-shaped fish by the Stuttgart zoological-botanical garden, the Wilhelma.
A model of the “boxfish”, as it is officially called in English, was built and tested
in a wind tunnel. Contrary to the school of thought, the model had a cw value that
was only slightly worse than the ideal shaft, which experts initially interpreted as
a measurement error. Daimler developed the bionic car based on the boxfish, with
the design remaining fairly close to the original. Even though it is a family car, it
has a significantly better cw value than well-known sports cars with their classic
streamlined shape.
Based on insights derived from nature, biomimetics offers an extraordinarily
high potential for disruptive innovations. An evaluation of all Fraunhofer patents
and consultation with the patent officers of the institutes revealed that 64 tech-
insects optics
plants robotics/drive
technology
reptiles medical (engineering)
mammals materials
solar technology
fluid technology
other
Fig. 3.1 Assignment of patents identified as biomimetics-related to their sources of inspira-

tion and fields of technology [10]
nologies or inventions of the Fraunhofer-Gesellschaft with more than 250 patents

issued in the years from 2004 onwards, are based on transferred biological con-
cepts. This equates to about three percent (Fig. 3.1).
Of the 54 inventions with a biomimetics background, an above-average number
of inventions won innovation awards: the Bionic Handling Assistant (IPA) won the
German Federal President’s Prize in 2010; FracTherm (ISE), SirexTM (IPA) and di-
rected fluid transport modeled on the horned lizard (IPT) won at the International
Bionic Award in 2008, 2014 and 2016 respectively. The compound eye camera
(IOF) also received several awards. The evaluation of the patents revealed that five
years after filing the initial application, more than 90% still have at least one ac-
tive proprietary right. In other words, the invention is still classified as potentially
usable. This is significantly longer than is the case for conventional inventions and
an indication that more radical innovations are generated by biomimetic inven-
tions.
3.2.2 Biomimetics Covers Everything Except Biotechnology
It is easier to name what does not belong to biomimetics than what can be sub-
sumed under it. The scientific construct developed and publicized by Werner
Nachtigall, addresses three learning levels at which we as humans derive ideas
from nature:
Learning from the results of evolution, in other words, everything we can grasp
or see with a microscope
Learning from the principles of nature—e.g. solar economics, circular economy

or resource efficiency
Learning from the process of evolution itself (e.g. optimization algorithms; [2,
8]).
Key aspects of bioeconomy are thus based on biomimetics. The fields of appli-
cation for this knowledge are almost unlimited. However, the concrete use of living
organisms for the production, transformation or the decomposition of substances
is not one of them—that is the conventional field of work of biotechnology. What
these disciplines have in common, however, is that they have both emerged at the
boundaries of biology and technology and can only be researched on an interdisci-
plinary basis.
3.3 The Role of Biomimetics in Biological Transformation
The sister disciplines of biomimetics and biotechnology, collectively referred to

as bioengineering, are responsible for the transfer of scientific knowledge gained
in biology or its sub-disciplines of microbiology, genetics, zoology, etc. They thus
represent the basic disciplines. Bioeconomy is not classed at the same level. It is an
expression of socio-political will for a more sustainable economic system in which
products, processes and services can be offered in all economic sectors through the
creation and use of biological resources and biological knowledge (see Key Issues
Paper of the Bioeconomy Council [6]).
As indicated in Fig. 3.2, a range of disciplines, most notably information tech-
nology, material science and manufacturing science, underpin the realization of
biomimetic and biotechnological products and processes. In some cases, they en-
able a full implementation for the first time. For example, 3D printing technology
allows the manufacture of lightweight biomimetic structures, and sensor-based
control with real-time capability makes the continuous management of fermen-
tation processes possible.
3.4 Biomimetics in Medical Engineering
The almost unlimited applicability of biomimetics holds true especially in the

field of medicine and medical engineering. According to Ingo Rechenberg [21]
the transfer of biological principles to biomimetic applications makes sense if the
Basic Biomimetics Biotechnology

scientific - Learning from the results of evolution - green biotechnology
disciplines - Learning from the principles of nature (agriculture)
- Learning from the process of evolution - red biotechnology
Biocybernetics (medicine)
- white biotechnology
(industry)
- blue biotechnology
(aquatic organisms)
Enabler IT / digitization Material science Manufacturing
science
Process Biologization
Economic Bioeconomy
system
Result Bioanalogous / biointelligent
production, mobility, communication, etc. systems
Fields of Medical Automotive Logistics Energy Food
application engineering engineering generation production
Architecture Automation Infrastructure … Forestry
Human Housing Mobility Health Security Environment Creating

meaning
needs
Fig. 3.2 A representation of the complex biologization of technology. (Fraunhofer IPA,

Oliver Schwarz)
functional principle, the boundary conditions and the evaluation criterion are simi-
lar in the biological example and in the technical abstraction. This is comparatively
easy in medical engineering owing to the material-related and functional similar-
ity of the example and target application. If we consider the example of a surgical
instrument where the effector has been biomimetically optimized, it is easy to see
that all the technical and functional principles of surgical manipulation—incision,
piercing, suction, holding, etc.—can be found in a multitude of applications in the
animal kingdom. The boundary condition of “the material to be manipulated”—be
it flesh, cartilage or bone—is similar to the boundary conditions for which the ef-
fector organs have been optimized for piercing, gripping, suction, etc. It may be
more divergent with regard to the evaluation criterion. Depending on whether the
selection criterion is effectiveness, quality, speed, pain activation or the compact
design, some biological examples are more suitable for transfer, others less.
In general, an increase in efficiency of between ten and thirty percent can be
assumed for a biomimetically optimized technology, compared to the established
a b
Fig. 3.3 a An anaconda jaw; b re-engineered bone punch. (Fraunhofer IPA, Oliver Schwarz)
state of the art. These figures are not surprising if one considers that material and/or
energy efficiency are the key factors in nature for a species to be competitive and
to ensure its survival.
3.4.1 Surgical Bone Punch Modeled on Snake and Cat
Bone punches have been in use for over 90 years as instruments for exposing sur-
gical sites, especially on the spine. The downside is that after each piece of bone,
cartilage or flesh has been cut out (resected tissue), the instrument has to be with-
drawn and emptied or cleaned. Without changing the external appearance of the
instrument, the Fraunhofer Institute for Manufacturing Engineering and Automa-
tion IPA was to conduct an industrial project to insert a collection receptacle into
the 5 mm wide shaft where the resected material could be collected (Fig. 3.3).
Everyday experience shows that inserting material into a very narrow container
results in blockage of the inlet. A transport mechanism (function) was therefore
sought in nature that achieves the quick, effective and linear transport (evalua-
tion criterion) of bone, cartilage or flesh (boundary condition). The swallowing
mechanism of snakes (“anaconda principle”) fulfilled the requirements: the prey is
transported safely to the stomach via the alternating movement of the upper and
lower jaw with its backward-pointing teeth (Fig. 3.3). The technological analogy
was realized by using a sheet steel cartridge with an inward-pointing row of teeth
and a second, movable row of teeth (Figs. 3.4, 3.5, 3.6). The alternating movement
takes place through operating the grip synchronously with the movement of the
effector.
Fig. 3.4 “Anaconda Principle”: Transport mechanism of resected tissue into the disposable
cartridge of a bone punch. The ball represents the resectate. (Fraunhofer IPA, Miroslav Mik-
losovic)
a b c
Fig. 3.5 Technical implementation of the transport mechanism in the biomimetic bone
punch. a cartridge sheath with internal teeth, b toothed rack with one row of teeth, c as-
sembled resectate cartridge. (Fraunhofer IPA, Miroslav Miklosovic)
Fig. 3.6 Distal end of the biomimetic bone punch with resectate uptake mechanism. (Fraun-
hofer IPA, Miroslav Miklosovic)
The result: up to 40 resectates can now be stored in the 45 mm long cartridge

because it is filled effectively and tightly packed from back to front. This reduces
not only the operation time, but also the risk of infection due to transfers into and
out of the body.
Biomimetic re-engineering of the separation mechanism resulted in an up to

50% reduction of the force required to operate the punch. During punching the
blade is pressed onto an anvil, whereby the material is severed (punching princi-
ple). The blade in a bone punch consists of a semicircle that is pressed onto an
anvil. It was observed that the teeth of a typical carnivore never function according
to the anvil principle of a pair of anvil scissors. A fang at the rear of the dental
arch in dogs and cats severs tendons, flesh, etc. effectively. The teeth in the upper
and lower jaws interlock as they pass each other, severing the material cleanly ac-
cording to the scissor cutting principle [12]. By reducing the anvil to the slightly
smaller size of the semicircular blade, scissor cutting was made possible.
The result: no more last tearing is required to sever the last fibrous components
and the cutting force that needs to be applied could be reduced by up to 50%. The
punch, or actually scissors, is currently undergoing FDA approval.
3.4.2 Drilling Modeled on Insects
In Germany alone, about 220,000 hip replacements are performed each year.
The conically tapered implant shaft must be anchored rigidly in the femur and
not be able to turn (Fig. 3.7). A cavity must therefore be created which is congruent
to the shape of the shaft. In 90% of cases, this is done manually with rasps from
cancellous bone
femur
cortical bone
a b 1 2 3
Fig. 3.7 a Sequence of movements in insect and drill, b Hip implantation using SirexTM .
(Fraunhofer IPA, Kiyoharu Nakajima)
3
4
1 gutter with two hollow channels

2 dovetail guideway
3 channel for executing eggs
a b 4 bristles with two hollow channel
Fig. 3.8 a A hymenopteran drilling with its ovipositor (Fraunhofer IPA, Oliver Schwarz),
b Ovipositor in side view and as cross section. Legend: 1. Sting stylus (saw), 2. Dovetail
guide, 3. Egg exit passage 4. Barbed lancets (shovels). (Fraunhofer IPA, Kiyoharu Nakajima)
a multi-part rasp set, which are driven into the bone with a hammer. Rotary drilling
is not possible because this would only produce round cavities.
The fact that in the whole of human history only one method, rotary drilling,
has been invented for drilling a hole, and that all present-day machining methods
are based on it, required alternatives. A solution was found in the hymenoptera
order of insects, which includes bees and wasps. Their stinging apparatus is a sec-
ondary development of the original egg laying apparatus (ovipositor) for stinging
purposes. This is also the reason only females are able to sting. Other groups, such
as wood wasps or parasitic ichneumons, use the ovipositor to lay eggs in deeper
layers of a substrate, where the hatching larvae feed on fungi or host animals that
live there. This sting is however by no means just a cannula-like tube. The three-
part ovipositor is based on a complex mechanism in which the three lancets are
flexibly connected to each other by dovetail guides (Fig. 3.8).
Unlike the only other drilling principle known to date, rotary drilling, the
Sirex™ drill that this inspired, uses the pendulum stroke principle, in which three
drilling lancets are alternately lifted and lowered (Fig. 3.9).
The pendulum stroke principle opens up completely new possibilities for the
creation of holes and for the design of anchoring systems. Not only round holes,
but also non-circular holes of any cross-sectional geometry can be generated. Hip
a b
Fig. 3.9 a Three-piece effector of the SirexTM in the shape of an implant, b Ergonomic
handling of SirexTM. (Fraunhofer IPA, Kiyoharu Nakajima)
implants usually have a rectangular cross-section and are conically tapered, which
can be easily achieved with the pendulum stroke principle. In addition, no torque
is applied during drilling, which is why drilling may be done directly through the
skin. The multi-part drilling lances can be spread apart or wedged after drilling
and even remain in the drilled hole to act like dowl, for example, when pedicle
screws are inserted into the vertebral body. When the drills are made of flexible
material, they follow the path of least resistance. This allows for following the
curved long bones, for example, so that intramedullary nails may subsequently be
inserted, which are used to temporarily treat fractures of long bones.
The advantages of this biomimetic drilling procedure for hip implants are:
Surgical time saved through the motorization of manual handling, as the final
shape and size of the cavity are produced in one step.
An improved rasping result through the more precise cavity for holding the
shaft of the prosthesis and thus better anchoring in the bone
This results in better and faster healing of uncemented implantations, which
account for about 80% of all implanted total hip endoprostheses.
3.4.3 Implants (Endoprostheses)
In the specific case of implants, the human body itself is the biological model. The
history of medicine shows how long the learning curve has been: from a simple
mechanistic understanding of function and its replication to increasingly differ-
entiated design, that takes increasingly complex biological interdependencies and
boundary conditions into account. In addition to functional morphology, which
Caged- Unicuspid Bicuspid Heart valves Tricuspidpolymer Hybrid Heart

ball valve valve of animal valve valves valves
prosthesis tissue created in
vitro
The first Metallic body Combination Animal Polymers can be Polymer Created by
artificial provided with of polymer transplants processed in a valves tissue
heart a polymer and metallic Advantage: variety of ways colonized engineering
valve in structure valve No lifelong Advantage: with Advantage:
the form Advantage: Advantage: anticoagulant No shear stress, patient’s endogenous
of a ball Long implant Long implant drugs needed optimal flow; own cells;
prosthesis lifespan lifespan Disadvantage: can be tailor- (autologous) grows with
(USA) Disadvantage: Disadvantage: Not made; cells the patient;
Shear stress Shear stress standardized; biomimetic Advantage: minimal
for red blood for red blood limited surface coating High rejection
cells; lifelong cells; lifelong lifespan stability, probability
anticoagulant anticoagulant surface
drugs drugs protection
required from
formation
of deposits
Timeline
1952 1968 1977 2012 Currently in development at Fraunhofer
institutes
Fig. 3.10 Evolution of artificial heart valves. (From left to right: [14–20])
is applied mainly in instrument development, implants also incorporate nano and

molecular biomimetic aspects, which are needed to ensure biostability and bio-
compatibility, antimicrobial properties, or antithrombogenicity.
The example of developments in artificial heart valves is meant to demonstrate
the progress in understanding and in the product (Fig. 3.10). It began with me-
chanical ball valves made of metal, moved on to metallic and then polymeric heart
valves and cell-free animal valves (current gold standard) to elastic polymer valves
developed in cooperation with the Fraunhofer Institute for Manufacturing Engi-
neering and Automation IPA. These include three biomimetic improvements:
A parameterized shape that mimics the tricuspid mitral valve in shape and clo-
sure mechanism;
graded layer thickness and elasticity distribution, which mimic nature and allow
the cusps to move in a flow-optimized way;
a surface coating with carbohydrate chains that mimics the surface of vascular
cells and prevents the formation of blood clots [3, 12].
Using this as a foundation, hybrid valves would be conceivable as an intermedi-

ate step to fully biological valves, which consist of a patient’s own cell material and
can grow along with them (field of research at the Fraunhofer Institute for Interfa-
cial Engineering and Biotechnology IGB). This could combine the reproducibility
and biostability of polymer valves with the patient’s own cell growth to prevent
blood clots.
Implant products are becoming increasingly similar to the imitated body part in
their material properties and structures. This reduces side effects and extends the
life span.
The biogenic development of materials also involves extensive cooperation and
partial overlapping of the disciplines of biomimetics and biotechnology. Bioin-
spired and biogenic materials such as foamed metals used as bone substitute (re-
search field at the Fraunhofer Institute for Manufacturing Technology and Ad-
vanced Materials IFAM), spider and green lacewing silk (research field at the
Fraunhofer Institute for Applied Polymer Research IAP), bioadhesives such as
mussel glue (research field at the Fraunhofer Institute for Manufacturing Tech-
nology and Advanced Materials IFAM) or chitin (research field at IPA and IGB)
will play an increasingly important role in medical engineering. Antimicrobial and
healing properties add a new quality and multi-functionality to wound dressings
(e.g. treatment of burn victims), surgical sutures, or implant materials.
3.4.4 Exoprostheses
Exoprostheses are body replacement parts that fulfill a bodily function from out-
side the body. These include prosthetic limb replacements. Even in ancient Egypt
the desire for mobility and participation in normal life spurred people to develop
artificial replacements. At first clumsy assistive devices were created of wood and
iron, but later attempts were made to mimic more and more features of the lost
limbs, in order to regain body feeling and improve control over the replacement
part. Work was therefore done on the material and structural imitation of the elas-
ticity, energy absorption and release in bones, tendons and joints. These ultimately
determine the perceived physical strain. Oskar Pistorius’ Cheetah racing prosthesis
effectively demonstrated to the public how a technological replacement could be
on a par with a healthy limb [4, 5].
Biomimetics was able to introduce additional efficiency principles from the an-
imal world here. Using the fastest biped endurance runner, the ostrich, which is
similar in size and weight to humans, the Fraunhofer IPA was able to transfer two
biological principles to the design of a racing prosthesis. The movement of the
Fig. 3.11 Sequence of movement of the ostrich prosthesis. (Fraunhofer IPA, Moritz Irion)
ankle joint is coupled to the knee movement in the ostrich prosthesis, as it is in

the ostrich as it runs. The prosthetic foot can thus be pushed off the ground more
strongly by the thigh muscles and also pulled up in the air. To prevent unwanted
buckling, a joint, which is constructed like a four-link gear coupling, guides the
prosthesis into a stable position. This is achieved by using the change in force di-
rection acting on the prosthesis and the movement of the instant center of rotation
of the prosthetic foot part ([7]; Fig. 3.11).
Fraunhofer IPA is also working on the possibility of integrating sensory recep-
tors on the prostheses. This is where electronics comes into play. In English, the
substitution of a body part with the aid of electronic components is known as “bion-
ics”, a special case of the general field of “biomimetics”. In German, no distinction
is made by using “Bionik”.
Particularly for the integration of haptics, temperature and pressure sensors or
the implementation of different grips in prosthetic hands, the natural model is an-
alyzed very precisely and often transferred in broad analogy to the technological
application. The discrepancy between the high density and diversity of skin sen-
sors in the biological hand and a technical hand prosthesis suggests that the current
modest capacity to implement technical sensors will probably lead to the devel-
opment of new, perhaps multimodal types of sensors, which will have to be based
on the biological model. One way to produce such high densities is to use self-
organization processes following nature’s example.
Great progress has been made in the realization of myoelectric prostheses,
which respond to the respective muscle signals of the wearer. The biological inspi-
ration lies in the use of natural control signals and signaling pathways that are re-
tained in the stump. The effectiveness of the myoelectric prosthesis, to specifically
and independently tighten muscle groups, is however influenced by the sequential

control, the number of control signals available, and the abilities remaining after
amputation.
Ideas drawn from nature have great efficacy in medical engineering. The field
will continue to grow within the framework of biological transformation thanks to
the synergy of biomimetics and biotechnology and enable increasingly compatible
and reliable medical technology products to be made.

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cessed: 26.06.2020
Innovative Food Products
New Methods of Preparing Plant-Based Raw
4
Materials Lead to Healthy Alternatives
for Conventional Foods and Protein Sources
Peter Eisner, Ute Weisz, Raffael Osen, and Stephanie

Mittermaier
Summary
The expected increase of the global population to over 9.5 billion by the middle
of this century and the rising consumption of animal food products present one
of the greatest global challenges—securing humanity’s food supply. The use
of new plant-based protein ingredients instead of animal protein preparations
can be an important part of the solution, as the production of animal proteins
requires around five times the area of plant protein production. The follow-
ing article provides an overview of the state of the art production, processing
and application of plant-based proteins in the European food industry. Not only
are the opportunities and advantages presented, but also the shortcomings that
plant-based proteins have had until now, and strategies for optimization. Fur-
thermore, current project results (Fraunhofer Future Foundation) are reported,
in the scope of which new methods for the reduction of the allergenic potential
of plant proteins have been developed. The article is rounded off with a dis-
cussion of technical approaches to the optimization of the taste, texture and
mouthfeel of plant-based foods and examples of the successful implementation
of research results by Fraunhofer spin-offs.
P. Eisner () U. Weisz R. Osen S. Mittermaier

Fraunhofer Institute for Process Engineering and Packaging IVV
Freising, Germany

https://doi.org/10.1007/978-3-662-59659-3_4
38 P. Eisner et al.
4.1 Introduction and Background
Supplying the steadily growing global population with high-quality and safe food
is undoubtedly one of the greatest challenges of the 21st century. Rising prosperity
in many countries is leading to an ever-increasing demand for high-quality animal
foods such as meat, eggs and milk. The production of animal proteins, however,
requires many times more plant proteins for the animal feed, which consequently
increases the land requirement for feed crops. In 2016, over 60% of the world’s
agricultural land was already being used for animal production and cultivating fod-
der. If such growth continues, it is to be feared that the available space will soon
be insufficient [23].
The production of technical raw materials, fine chemicals and fuels from re-
newable raw materials will also increase considerably, for reasons of climate pro-
tection, and in order to realize the United Nations’ global sustainability goals. The
competition for the available land between cultivation of food, animal feed and
technically and energetically used biomass will therefore continue to intensify [11,
29].
As a result of these developments, prices for raw materials have increased
significantly over the past 15 years along with significant price fluctuations. The
assumption that meat consumption will continue to increase is a crucial trigger for
the rising prices. Since the production of animal proteins takes place via the cir-
cuitous route of plant and animal-based feed, significantly more area is required
for the production of animal protein than for the production of vegetable protein.
An area of 5.5 ha is needed, for example, to grow feed for the production of one ton
of protein in the form of pork [69, 83]. By comparison, the production of one ton
of protein in the form of wheat, corn or potatoes has an average area requirement
of 1.0 to 1.2 ha in Germany [24].
Since many agricultural raw materials can also be used as energy sources, the
temporal variation of agricultural raw material prices—whose volatility is deter-
mined by both economic factors and speculation—is similar to fossil fuel price
trends (Fig. 4.1). These increasingly unstable price trends have equal impact on all
important staple foods, such as rice, vegetable oil, cereals or corn [33].
In order to overcome these challenges, it seems urgently necessary to signifi-
cantly reduce the consumption of animal-based food—at least in industrialized
nations—and increase consumption of plant-based alternatives. However, a change
in consumer behavior can only be achieved if it is possible to provide plant-based
foods that are comparable regarding flavor, texture, mouthfeel and enjoyment fac-
tor to traditional, animal-based food products. The plant-derived ingredients re-
4 Innovative Food Products 39
350 140
300 120
Price for Crude Oil US $ per barrel

250 100
Price for Corn US $ per Ton
200 80
150 60
100 40
Corn
50 20
Crude Oil
0 0
Jan 06
Jan 07
Jan 08
Jan 09
Jan 10
Jan 11
Jan 12
Jan 13
Jan 14
Jan 15
Jan 16
Jan 17
Jan 18
Fig. 4.1 Price trends for corn and crude oil since 2006 [33]
quired for this, especially proteins, must therefore have very specific technofunc-
tional properties. The technofunctional profile required ranges from the stabiliza-
tion of emulsions and foams to the formation of gels as well as combining water
and oil in various food matrices [4, 37, 88].
In order to meet the needs of industry for plant-based ingredients, they should
enable the full substitution of animal protein. Only then strictly vegan formulations
can be developed. Protein preparations are thus needed which have a technofunc-
tional profile that comes very close to that of conventional ingredients such as
egg yolk, egg white, casein or gelatine. However, since most plant proteins dif-
fer significantly from the above-mentioned proteins in terms of their molecular
structure, a direct substitution is not possible. The challenge remains: it is not yet
possible to simply exchange the animal proteins in the formulations.
Moreover, the typical aroma and taste sensations of plant-based ingredients
should be minimized as far as possible, as these may be incompatible with the
taste expectations of Western consumers. Many of the vegetable protein ingredi-
ents from legumes, such as soybeans or peas, which are currently available, show
considerable deficits on account of their grassy-green or bean-like aromas as well
40 P. Eisner et al.
as bitter taste and astringency. For this reason, the Fraunhofer Institute for Process
Engineering and Packaging IVV has been conducting intensive research for many
years to be able to provide the food industry with plant protein preparations which
have the functional properties of egg, meat and milk proteins and a neutral taste,
and can thus be used for many applications.
In addition to increases in functionality and consumer acceptance, the nutri-
tional optimization of proteins will become more and more important in the future.
Thus work is underway within the scope of the Fraunhofer Future Foundation’s
“Food Allergen” project to reduce the allergenicity of plant proteins by enzymatic
and fermentative processes and thus to increase the acceptability of these proteins
for consumers who are allergic or allergy-prone. This work and the development
of food products based on plant-based proteins as alternatives to dairy, for exam-
ple, or extruded vegetable proteins as meat substitutes is discussed in this article.
To begin with, however, the extraction processes and the functional properties of
plant-based proteins will be introduced.
4.2 Plant-Based Proteins as Functional Food Ingredients
Plant-based proteins provide a variety of different functions in food. These range

from protein enrichment for increasing nutritional value, to the formation of emul-
sions (e.g. mayonnaise) and foaming (e.g. marshmallows), as well as to the forma-
tion of specific textures or gel structures [4, 37, 88]. The commercially available
protein ingredients used are as diverse as the applications. Alongside well-known
animal proteins such as gelatin and those found in egg, milk, or meat, the use of
vegetable proteins in food products is becoming increasingly important. Plant pro-
tein ingredients are currently mainly produced from soybeans and wheat [27]. The
success of soybean proteins can be attributed not only to high yields per hectare,
low costs and a comparatively high nutritional quality, but above all to the ongoing
research and development of methods for the production and modification of soy
proteins [14].
In recent years, however, also other sources of vegetable proteins have gained
importance. Examples are peas, lupines, rapeseed, sunflowers, rice and potatoes.
In addition to protein preparations that are already available on the market, cur-
rent research is also targeting other raw materials such as quinoa, broad beans and
lentils. In order to fully exploit the potential of plant-based raw materials, and in
particular the potential of legumes and oilseeds, extensive process development is
required also in the future, to enable manufacturers to efficiently extract proteins
of high sensory, functional and nutritional quality. This includes new deoiling pro-
cesses for the simultaneous extraction of oil and native proteins, the development
of extraction and isolation processes, the modification of the properties of plant
proteins (foaming, emulsification, gelation) so that they are comparable to those of
animal origin, and the preparation of modified proteins that are easily digestible by
humans [14]. Further diversification of the market for plant-based proteins is thus
to be expected in the coming years.
4.2.1 Extraction Processes for Plant-Based Protein Ingredients
Proteins are generally divided into classes according to their function in the plant.
A distinction is thus made between storage proteins, structural proteins and bio-
logically active proteins such as enzymes, enzyme inhibitors and lectins. Storage
proteins from plant seeds are best suited for extraction as plant-based protein ingre-
dients. After the cells have been ruptured, the proteins are extracted using a variety
of solvents, and then concentrated. The storage proteins are stored in the seeds in
intracellular “protein bodies”. Table 4.1 provides an overview of the total protein
content as well as the proportions of the different protein fractions in legumes,
oilseeds and cereals according to Osborne.
Based on the proportions of these different protein fractions, it is necessary
to pursue different strategies for protein extraction processes. These are shown in
Fig. 4.2 exemplified by legumes and oilseeds, which are characterized by high
levels of albumins and globulins. Other treatment and extraction procedures are
necessary for cereals, since the proportion of water-soluble proteins is rather low.
They often have a low total protein content and primarily contain protein fractions
which can be dissolved or suspended in ethanol-water mixtures or in alkaline aque-
ous solutions. Although these methods have already been described in literature,
there have been very few studies to date where the potential use of these fractions
in food products is being investigated. Thus, extensive research is still needed in
this area towards achieving the holistic use of cereals instead of simply focusing
on extracting the starch from raw materials such as wheat, rice or corn.
The protein ingredients shown in Table 4.2 are obtained by different manufac-
turing processes that may be summarized in the following groups:
Thermally treated or untreated flours with a protein content that closely corre-
sponds to the basic raw material or hulled basic raw material
Protein concentrates: increased protein content compared to flours. Protein con-
tents of between 65 and 90% are achieved for soy protein concentrates
42 P. Eisner et al.
Table 4.1 Protein content and protein fractions of legumes, oilseeds and cereals. [14, 19,
44, 63, 75]
Total protein Albumins Globulins Prolamins Glutelins
content (%) (%) (%) (%) (%)
Soluble in Water Salt solu- Water/ Alkali
tions ethanol (pH > 11)
Function in the Biological Storage proteins
plant activity
Proportions in
Soybeans 35–40 10 90 Traces Traces
Peas 20–30 15–25 50–65 Traces 15
Broad beans 25 55 Traces 20
Sunflower 20–40 20 60 5 15
seeds (hulled)
Rapeseeds 17–26 20 60 2–5 15–20
Lupine seeds 35–40 25 75 Traces Traces
Wheat 8–15 5–10 5–10 35–45 40
Rice 7–9 2–6 12 4 80
Corn 9–12 4 4 60 26
Table 4.2 Overview of functional and sensory properties, technical production complexity
and achievable market price. [21, 56]
Ingredient Functional Sensory prop- Technical com- Price
(protein content) properties erties plexity
Flours and coarse o Low Low
meals (30–65%)
Protein concen- o/+ o Medium Medium
trates (> 65%)
Protein isolates +/++ + High Medium to
(> 90%) high
Modified protein ++ ++ Very high High
isolates (> 90%)
–: very poor/very low; -: poor/low; o: moderate; +: good; ++: very good
Protein isolates: purified proteins with low content of minor adhering compo-
nents. Protein content over 90%.
The different protein preparations differ both in their production as well as in

their functional and sensory properties. Flours can be applied successfully in sev-
Fig. 4.2 Compilation of manufacturing processes for protein concentrates and isolates.
(Data from [11, 12, 17, 21, 45, 56, 59, 85])
eral applications such as baked goods, dry extrudates (minced meat substitutes) and
as raw material for soymilk production [21]. However, these present plant-specific
“off” taste impressions even at relatively low concentrations, meaning that only
small amounts can be used to support functional properties. By contrast, protein
concentrates and isolates can be used in a much broader product portfolio (e.g. for
the formation and stabilization of emulsions) and also in higher concentrations for
protein enrichment in food products. However, these are also significantly more
expensive than flours. Hydrolysed protein isolates, which have the highest mar-
ket price, are particularly suitable for use in specialty products such as beverages,
sports nutrition, baby food (formula) and balanced diets.
44 P. Eisner et al.
The processes listed in Fig. 4.2 are used for the extraction of protein concen-
trates and isolates from legumes such as soybeans, peas, lentils, lupines, broad
beans and oilseeds (e.g. rapeseed, sunflower). Depending on the objective and ap-
plication, the extraction methods are adjusted with regard to extraction conditions
(pH, temperature, amount of salt, number of stages, etc.) and concentration, in or-
der to achieve specific functional properties. For example, isoelectric precipitation
and ultrafiltration of proteins are suitable methods for the production of protein
isolates with moderate to good protein solubilities [11, 12, 21, 45, 77]. Thermal
precipitation is used primarily for the extraction of (partially) denatured proteins
with a high water binding capacity. One special case is the extraction of proteins
by means of dilution precipitation, whereby isolates with fat-like properties can be
manufactured [59, 82]. Other process options include combinations of processes
for concentrates and isolates, as in the extraction of lupine protein isolates, which
uses an acidic pre-extraction to separate low molecular weight components and
subsequent protein extraction in the neutral to slightly alkaline range [17, 85].
4.2.2 Modification of Plant-Based Protein Ingredients
Since plant-based protein ingredients in their native form often only have some
of the desired application properties, a variety of methods are applied to change
the functional and sensory properties as well as to improved digestibility (Ta-
ble 4.3). The choice of modification conditions (temperature, pH value, added salt
or sugar) allow the functional and sensory properties as well as the digestibility of
the proteins to be adjusted in a targeted manner. As this includes a wide variety of
processing options, it is possible to prepare customized protein isolates for differ-
ent food applications. This diversification results in a broad portfolio of plant-based
proteins with different properties. However, this is only commercially available for
soy protein isolates thus far. There are only a few studies available on further pro-
tein isolates from legumes and oilseeds, so no suitable database for plant-based
protein sources is available as yet. Further investigation is required here in future.
Table 4.3 shows the possibilities described in literature for modifying plant
proteins and their effects on functional and sensory properties as well as on di-
gestibility. As part of the Fraunhofer Future Foundation’s “Food Allergen” project,
the influence of the modification not only on sensory and functional properties, but
also on the allergenicity of the proteins was studied. This project is presented in
the section below.
Table 4.3 The influence of different modification techniques on protein functionality, sen-
sation and digestibility. [7, 9, 21, 43, 45, 49, 50, 51, 56, 77]
Modifica- Technique Influence on
tion Functionality Sensation Digestibility
Physical High pressure Improved gelation Minimal Minimal
treatment
High pressure Increased protein Minimal Minimal
homogenization solubility; improved
gelation
Thermal Steam injection Increased water bind- Reduced Improved
ing; increased fat volatile com- digestibility
absorption; improved pounds
dispersibility
Chemical Acid/alkaline No further/hardly any Flavor en- Presence of
hydrolysis (full) functionality hancers amino acids
and short-
chain peptides
Partial Increased foaming; Formation of Improved
acid/alkaline improved solubility bitter peptides digestibility
hydrolysis in acids; improved
gelation; improved
emulsifying properties
Glycosylation Improved protein Not described Not described
solubility; improved
gelation; improved
emulsifying properties
Succinylation/ Improved protein Not described Not described
acetylationa solubility; improved
emulsifying proper-
ties; improved foaming
properties
Enzymatic Addition of pro- Increased foaming; Formation of Improved
teolytic enzymes improved solubility in bitter peptides; digestibility
(e.g. Pepsin, pa- acids reduction of
pain, bromelain) off-flavors
Fermen- Fermentation Improvement Improved
tative with lactic acid of sensory digestibility
bacteria properties
Combi- Depends on combined procedures
nations
46 P. Eisner et al.
4.3 Allergenicity of Plant-Based Proteins and Strategies

for Its Reduction
4.3.1 Food Allergies and Food Allergens
An allergy is a defense reaction of the body’s immune system to usually harm-

less substances, causing common symptoms such as skin rash, difficulty breathing
or gastrointestinal discomfort. During this process, the antibodies produced by
the body trigger hypersensitivity reactions and therefore fail to provide a protec-
tive immune response against pathogens. In the case of food allergy, considered
a type I allergy, the body activates an immune reaction caused by a specific food
protein mistakenly recognized as a threat. As a result of the allergenic protein con-
sumption, antibodies of immunoglobulin class E (IgE) are formed, which bind, via
receptors, to basophilic granulocytes and mast cells in the blood and tissue, respec-
tively. Frequent ingestion of foods containing the allergenic protein, recognized
by cell-bound IgE antibodies, causes various inflammatory mediators, particularly
histamine and heparin, to be released. This results in a fast and sudden onset of the
symptoms. The parts of the protein that are recognized by the antibodies are called
epitopes. To set off an allergic reaction, two antibodies must generally bind to two
epitopes of the protein, which then trigger the allergic reaction via cross-linking
(Fig. 4.3) [47, 80].
Fig. 4.3 Binding of two antibodies to two epitopes of a tetrameric protein (cross-linking)
and triggering of allergic reaction
Strictly speaking, any food protein could trigger an allergic reaction. However,
a limited number of proteins account for most food allergies. Soy, milk, wheat,
egg, shellfish and peanut products are among the “big eight” food allergens that
are responsible for approximately 90% of all food allergies [16, 86]. In 2007, the
list of allergens was expanded to include fish, crustaceans, lupines, celery, mustard,
sesame, sulfur dioxide and molluscs, thus bringing the total to 14 allergens, which
must be declared in the EU (EU Regulation 1169/2011).
4.3.2 Strategies for Affected Consumers
Resulting from chronic exposure to potentially allergenic proteins, 5 to 7% of the

population is now affected by food allergies. It is expected that these numbers
continue to rise. Nowadays, the only protection for allergy sufferers is consistent
avoidance of allergenic foods, which, however, results in a significant loss in qual-
ity of life.
The food industry is facing a growing demand for allergen-reduced food prod-
ucts in order to avoid potentially allergenic ingredients. While for years the supply
of hypoallergenic food for infants has been vast, for older children and adults, there
are hardly any food products of this kind. One reason for this is that heretofore no
tasty and simultaneously allergen-reduced protein ingredients have been available.
This is exactly where the Fraunhofer Future Foundation project “Food Allergen”
comes into play. One goal of the project is the development and market introduc-
tion of allergen-reduced products, with both a pleasant taste and good functional
characteristics.
4.3.3 Ways to Reduce the Allergenic Potential of Protein

Ingredients
Owing to the increasing significance of food allergies, science and industry are
seeking technologies to reduce the allergenic potential of food proteins. An
overview of the technologies that have been investigated to date is shown in
Fig. 4.4.
Thermal processes are based on the use of dry or moist heat, as is the case for
roasting, pasteurizing and sterilizing. Non-thermal methods include biological and
chemical methods such as fermentation and enzymatic hydrolysis. Of which some
are new methods, for instance the use of ionizing radiation, high-pressure treat-
ment, pulsed UV light and non-thermal plasma. However, these have not yet been
48 P. Eisner et al.
Fig. 4.4 Thermal and non-thermal methods for reducing the allergenicity of food proteins.
[49]
approved for food products, so that thermal, enzymatic or fermentative methods

allow for faster implementation.
Many possible processes may be used to reduce the allergenicity of proteins.
These procedures structurally modify the proteins in such a way that IgE antibod-
ies can no longer bind to the allergenic epitopes. This can be achieved either by
destroying or masking the epitopes. Some studies have shown, however, that the
use of certain techniques may expose epitopes that were originally hidden, possibly
increasing the allergenic potential [68]. Thus, should be avoided at all costs.
Thermal Treatment
Heat is used in the industry for preservation, altering the structure of food prod-
ucts and modifying flavor profiles through induced Maillard reactions. Moreover,
thermal treatment can influence the allergenicity of proteins by causing changes

in their tertiary and secondary structure, namely denaturation and aggregation, as
well as cross-linking with subsequent masking of epitope structures.
The influence of thermal treatment on the allergenicity of food proteins has
been widely described in literature, but with somewhat contradictory results. For
example, the allergenicity of soy protein could be reduced by boiling, microwave
heating or extrusion [25, 58, 61, 72, 84]. In contrast to these results, Shibasaki
et al. [76] and Yamanishi et al. [87] observed an increase in allergenicity following
heating in an autoclave, which was justified by the formation of so-called “neoanti-
gens”. It is assumed that hidden epitopes were uncovered as the protein structure
was altered through thermal refolding [15, 18].
Although thermal treatment certainly has the potential to reduce the allergenic-
ity of proteins, there are limits to this process: the nutritional value decreases,
organoleptic properties change and functionality deteriorates.
Non-Thermal Treatment
In this group, the influence of various physical and chemical methods (such as
ultracentrifugation, ultrafiltration, transglutaminase cross-linking and Maillard-in-
duced carbohydrate conjugation) on the allergenic potential of proteins has been
studied [3, 6, 73, 81, 82]. However, all these technologies were shown to have
only a minor effect on allergenicity. Biological methods such as genetic modifica-
tions are more effective in this regard and can be applied very successfully for the
elimination of allergenic protein fractions [29, 58, 73]. The negative image con-
sumers have of the genetic modification of food has prevented this process from
gaining traction. Methods involving enzymatic treatment, which have long been
widespread in the food industry, appear more interesting from a consumer point of
view.
Enzymatic Hydrolysis
For over 50 years, proteins have been hydrolyzed for a wide variety of food appli-
cations [1]. The most common form of hydrolysis, enzymatic hydrolysis, depends
essentially on the type of protease used (specificity and activity) and the hydrol-
ysis conditions (temperature, pH, ionic strength). The proteins are cleaved at the
peptide bonds, breaking into peptides and amino acids (Fig. 4.5).
A wide range of proteolytic enzymes with different activities may be isolated
and purified from a variety of natural sources. The enzymes are classified into
exo- and endopeptidases according to their mode of activity. Exopeptidases cleave
from the end of an amino acid chain, whereas endopeptidases act on the interior of
a protein molecule.
50 P. Eisner et al.
Fig. 4.5 Enzymatic hydrolysis of a peptide bond. R1, R2, R3 – amino acid side groups;
E – proteolytically active enzymes [50, 78]
Hypoallergenic food, which is currently available on the market, is often pro-

duced by enzymatic hydrolysis, as the epitopes can be cleaved effectively by this
method [25]. A comprehensive screening of enzymes of microbial and plant ori-
gin was carried out to investigate their influence on the allergenic potential of soy
proteins. The results showed that the allergenicity of the main allergens could be
reduced very effectively, while at the same time the technofunctional properties of
the products, such as emulsifying behavior or foaming properties, were retained
or even improved. However, proteins treated in this way often have a bitter taste,
which is more or less pronounced depending on the type of enzyme preparation
used [48, 50]. The emergence of the bitter taste can be accounted for by the release
of bitter peptides, which are formed during enzymatic hydrolysis. These bitter pep-
tides contain hydrophobic amino acids, e.g. leucine, proline or phenylalanine [34].
Numerous methods have been tested to reduce the bitter taste. These range from se-
lective removal of bitter peptides to treatment with activated charcoal or extraction
with alcohol [71]. Another promising way to improve taste lies in the fermentation
of protein hydrolysates using lactic acid bacteria. The bitter taste of soy protein hy-
drolysates can be significantly reduced through the use of Lactobacillus perolens
or Lactobacillus helveticus [51, 52].
High Pressure Treatment and Other Non-Thermal Processes

High-pressure treatment of food products can change the protein structure both
reversibly and irreversibly. This leads to protein denaturation, aggregation or for-
mation of gel-like structures [55]. High pressure-assisted enzymatic hydrolysis of
soy protein at pressures of 400 and 500 MPa before and during enzymatic hy-
drolysis reduced the allergenic potential to almost zero. It is surmised that the
conformation of the proteins is changed by the high pressure treatment in such
a way that they can be attacked by the proteolytic enzymes much more easily. Fur-
thermore, the beany, bitter and green off-flavors of soy protein were reduced to
a great extent. The hydrolysates were shown to have improved protein solubility,
foaming activity as well as fat binding and emulsifying capacity [54].
The potential of other methods such as pulsed UV light, gamma radiation and
direct and indirect low-temperature plasma for reducing the allergenic potential of
soybeans was also investigated. Maximum reduction of the allergenic potential,
from 91 to 100%, was achieved by direct plasma, pulsed UV light and gamma
irradiation. Indirect plasma also resulted in a decrease in antibody binding of up to
89% [53].
All in all it should be noted that there are a number of promising methods and
approaches for producing allergen-reduced food ingredients. However, the taste
and functional properties must always be taken into account in order to make a truly
sustainable contribution to food safety.
4.4 Texturizing Plant-Based Protein Ingredients

by Extrusion Cooking
The high rate of meat consumption is a significant factor that must be reduced if
a sustainable food supply is to be achieved. Traditional plant-based protein prod-
ucts from soy or wheat protein such as tofu, tempeh or seitan, which form an
integral part of the diet in many Asian countries, are hardly accepted as meat sub-
stitutes in most western countries. This is due to their gel-like consistency and
bland taste which is substantially different to most meat products. Hence, the con-
sumer expectations of a juicy, fibrous texture as found in naturally grown meat is
far from being satisfied.
Advances in extrusion technology led to the introduction of textured soybean
flour in the early 1960s. These textured products (TVP = Textured Vegetable Pro-
tein) are prepared using an extrusion cooking process and are now widely available
in the food retail industry. The dried products have a sponge-like consistency, have
a long shelf-life and, once rehydrated, are mainly used as minced meat substitutes
or to provide structure in formed products such as meatballs and burger patties [28].
The market for plant-based meat replacement products has grown significantly
in Germany in recent years. In 2012, turnover was around 156 million euros and
by 2015 sales had doubled (Statista, 2017). Recent developments in the extrusion
of vegetable proteins now allow plant-based meat alternatives with meat-like prop-
erties to be produced from local raw materials.
52 P. Eisner et al.
4.4.1 Extruders and Extrusion Parameters in Food Processing
Extrusion (Latin: extrudere = push out, drive out) refers to a continuous process,
in which viscous, curable materials are passed through a shaping die by means of
screw presses. The first extruded foods were developed in the middle of the 20th
century and can since be found in many product categories. These include starch or
sugar-based foods such as confectionery (licorice, ice cream), snacks (corn curls),
cereals (cornflakes), pasta and baked goods (flat bread), as well as protein-based
food products such as surimi or meat alternatives [54].
Depending on the number of screws in the extruder barrel, a distinction is made
between single, twin and multi-screw extruders (e.g. planetary roller extruder). For
food production, mostly twin screw extruders are being used due to their versa-
tile application, better mixing effect and pumping power, self-cleaning properties
and effective degassing ability compared to simpler single screw extruder systems.
Planetary roller extruders are occasionally used for processing thermally sensitive
formulations such as cocoa-sugar dispersions, ice cream or specialty malts [70].
Extrusion is becoming more and more important in food production due to
the versatile, continuous process design, in which the various components can be
mixed, degassed, thermally and mechanically heated, pasteurized, molded and ex-
panded. The fully automatic operation and high throughput rates, together with
low space and personnel requirements, result in efficient production with low op-
erational costs [28].
Physicochemical reactions take place in the extruder processing zone during
the extrusion of biopolymers whereby the basic raw materials can be modified to
create new textures and flavor profiles. As a prerequisite for targeted product de-
velopment, it is essential to understand the interactions between different extrusion
parameters. These can be divided into groups as illustrated in Fig. 4.6.
The material, machine and process parameters influence the system’s thermal
and mechanical profile and are responsible for molecular and microstructural
changes, which in turn influence the sensory product properties such as texture,
taste, color and shape of the extruded products [74].
4.4.2 Production of Plant-Based Meat Alternatives
The urgent requirement for meat-like, plant-based food products in recent years
has led to new developments in the field of extrusion cooking of plant-based pro-
tein ingredients. The focus is on creating the same visual appearance and sensory
profile during the whole preparation experience as when cooking animal meat.
Fig. 4.6 Selection of extrusion parameters and their interactions
A distinction is made between low-moisture and high-moisture extrusion cooking

according to the water content of the material to be extruded (Table 4.4).
Low-Moisture Extrusion Cooking

In low-moisture extrusion cooking, the highly viscous material can be cooked and
expanded directly on account of the high mechanical shear. If expansion of the
product is desired, a short die is used at the end of the processing zone, which
54 P. Eisner et al.
Table 4.4 Differentiation between low-moisture and high-moisture extrusion cooking

Process control Water Energy Product prop- Intermediate product
content dissipation erties
[%] supply
Low-moisture < 40 Mechanical Porous, ex-
extrusion cook- panded, dry
ing
High-moisture > 40 Thermal Compact,

extrusion cook- fibrous, moist
ing
narrows the flow cross section of the viscous material and results in high pres-
sure infront of the die exit. The hot material relaxes abruptly at the die outlet and
expands due to the evaporation of superheated water. As it expands, the product
is pre-dried, reducing the water content by about 10%. Depending on the choice
of extrusion parameters and recipe formulation, the process can produce extru-
dates ranging in form from expanded to finely fibrous, which may be used as
minced meat substitutes or “meat extenders” in formed products such as meatballs
or burger patties [28, 40].
High-Moisture Extrusion Cooking

During high-moisture extrusion cooking (Fig. 4.7), the feed mixture of plant pro-
tein, water and optionally other ingredients are separately fed to the extruder barrel
and adjusted to moisture contents of usually 60–80%. In the cooking zone of the
extruder, high temperatures of 130 to 170 °C are applied to plastify the protein
mass under high shear and pressure conditions. At the end of the extruder barrel,
a long cooling die is attached to prevent expansion by reducing the temperature of
the hot mass below 100 °C [66]. The cooled die channel further enables the stream
alignment of the viscous protein melt until it finally solidifies which facilitates the
formation of the fibrous structure.
These harsh process conditions are required to induce molecular and mi-
crostructural reactions that result in the formation of tightly adhering protein fibers
that provide the eating sensation of animal meat products.
Protein and other ingredients
Water
Cooling die
Twin screw extruder
Texturized product
Fig. 4.7 Representation of a high-moisture extrusion cooking process for texturizing plant-
based proteins to produce wet-textured meat alternatives
The complex steps involved in the formation of protein fiber structures in this
process are not yet sufficiently understood and have been the subject of numerous
research projects [10]. Globular proteins which are found in seeds as fractions of
storage proteins, such as legumin in peas and conglutins in lupines, are thought
to aggregate at a molecular level. This is achieved by means of high temperatures
inside the extruder, which causes the molecules to partially dissociate and unfold.
This can be observed when a plasticized mass forms in the extruder’s cooking zone
at temperatures above 120 °C. As the process continues, the unfolded proteins in
the hot material are aligned with the flow direction by shear forces between the
extruder screws and through tapering the flow cross section as the mass enters the
cooling die channel. Material-specific differences between the individual material
fractions cause phase separation processes in the shear zone of the die channel.
The fibrous texturate gradually becomes fixed through the protein-protein cross-
linking of disulfide and hydrogen bonds, leaving the die already mostly hardened
as a continuous elastic strand. The geometry of the cooling die and the temperature
gradients chosen in the cross section of the cooling channel allow for the targeted
formation of finer or coarser fiber structures. This knowledge of the interactions
between raw material properties, process parameters and system geometry allows
the typical characteristics of chicken, pork or beef to be reproduced with regard to
their specific fiber structure [21, 67].
56 P. Eisner et al.
Fig. 4.8 Example applications of pea-based texturates made by Amidori
In addition to the requirements for the product quality of meat alternatives, the
origin of the raw materials gains an ever-increasing influence on the purchasing de-
cision. Common formulations consist of a vegetable protein concentrate or isolate,
insoluble fiber, starch as well asother flavor and texture ingredients. Early studies
on the extrusion of defatted soybean flour with high moisture content date back to
the 1980s [35, 38, 60]. In later studies, combinations of soy protein isolate, wheat
gluten and wheat starch were used [13, 41, 42, 45]. These raw materials that are
currently used in most meat alternatives are facing increasing criticism for their al-
lergenic potential, negative associations with genetically modified seeds and long
transportation from their countries of origin.
Whether an alternative protein ingredient can be used in meat substitutes de-
pends on a variety of raw material properties such as composition, functionality
and sensory properties, and usually requires the adjustment of extrusion parame-
ters. Pea protein has recently received increasing attention as an alternative protein
source in meat substitutes due to its nutritional properties and its low potential for
allergic reactions [8, 64, 65]. The extrusion conditions and complex fiber-form-
ing mechanisms of pea protein were further elucidated within the scope of various
research projects at Fraunhofer IVV. On the basis of these studies, the company
Amidori has produced and marketed the first wet-textured meat substitutes made
from regional raw materials based of pea protein (Fig. 4.8).
In the future, other legumes and oilseeds such as beans, lentils, rapeseed or sun-
flower as well as proteins from micro/macroalgae or insects could also be used
as protein sources. Novel taste profiles and particularly favorable amino acid pro-
files could thus be achieved through the targeted combination of different protein
sources.
4.5 Further Applications of Plant-Based Proteins
In addition to working on the extrusion of plant-based proteins to produce meat

substitutes, successful studies on plant-based dairy alternatives to ice cream,
drinks or yoghurt have been carried out at Fraunhofer IVV for more than ten years
[20, 36]. Particular advantages in terms of sensory properties can be achieved
through fermentation with lactic acid bacteria. If suitable strains are used, it is
possible to produce significant amounts of diacetyl during fermentation. These
aromatic metabolic products from microorganisms lend a butter-like aroma to the
plant-based milk alternatives and thus increase consumer acceptance considerably.
Within the scope of extensive scientific studies, market-ready formulations for
cream cheese and yoghurt alternatives could thus be prepared from lupine protein
isolates, which have a sensory profile that is close to that of conventional dairy
products [30, 32]. These products are currently being successfully manufactured
and distributed by the Fraunhofer spin-off Prolupin GmbH and have become
increasingly popular among consumers since 2015. Fig. 4.9 shows a selection
of the products by Prolupin GmbH, which produces cream cheese and yoghurt
alternatives as well as lupine drinks, desserts, pasta and ice cream.
Despite the satisfactory market entry of meat and dairy alternatives, vegan foods
continue to be a niche product in food retail. To increase market share, it will be im-
portant to continue to optimize the sensory properties of the plant-based products.
Lupine dessert
vanilla, Lupine spread
chocolate plain, herb, bruschetta, bell
Lupine alternatives pepper-chili, chocolate
plain, raspbery, mango,
blueberry-blackcurrant, stracciatella
Lupine ice cream

chocolate, N ew !
vanilla, strawberry, Organic lupine pasta
*also av cookies + cocoa, cellentani, tagliatelle, fusilli
aila cookies + caramel
in 0. 5 lit ble
er
Lupine drink Lupine
plain, chocolate, salad cream
vanilla*, chufa* completely egg free
Fig. 4.9 Product portfolio of Prolupin GmbH

58 P. Eisner et al.
In addition to improvements in texture and mouthfeel, the focus is on neutralizing

the typical plant aroma and taste as much as possible. Up to now, most ingredi-
ents still feature many plant-based impressions. Current approaches are aimed at
finding solutions for overcoming existing deficiencies through improved separa-
tion processes and the combination of different raw materials in order to provide
sensorially neutral, nutritionally balanced and highly functional ingredients for the
broad mass market.

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Technical Homes for Human Cells
Micro-Physiological Organ-on-a-Chip Systems
5
as Interdisciplinary Platforms for SMEs,
the Pharmaceutical Industry, Medical Doctors
and Technologists
Christoph Leyens, Udo Klotzbach, Frank Sonntag, Markus

Wolperdinger, Peter Loskill, Thomas Bauernhansl, Andreas
Traube, Christian Brecher, Robert Schmitt, and Niels König
Summary
Life Science Engineering (LSE) focuses on technologies at the interface be-

tween the life sciences and engineering. It covers a very broad product spec-
trum, from the pharmaceutical industry to biotechnology and medical technol-
ogy. An essential element of LSE research is the high degree of interdisciplinary
cooperation. Numerous individual technologies are introduced in this chapter
that have been explored, developed and optimized separately so far. At the
interfaces of these domains, there is great potential for connecting the two disci-
plines, but its realization is still a distant goal. These interfaces are the urgently
C. Leyens () U. Klotzbach F. Sonntag

Fraunhofer Institute for Material and Beam Technology IWS
Dresden, Germany
M. Wolperdinger P. Loskill
Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB
Stuttgart, Germany
T. Bauernhansl A. Traube
Stuttgart, Germany
C. Brecher R. Schmitt N. König
Fraunhofer Institute for Production Technology IPT
Aachen, Germany

https://doi.org/10.1007/978-3-662-59659-3_5
64 C. Leyens et al.
required enablers of biological transformation, which permit the initial inter-

connection of the different domains. Standardized interfaces between biology
and technology must therefore be developed.
5.1 Introduction
Life Science Engineering (LSE) covers a very broad spectrum, from the pharma-
ceutical industry to biotechnology and medical technology. LSE can basically be
viewed from two perspectives that stem from different value chains (Fig. 5.1).
On the one hand, the engineering sciences provide the necessary technical pre-
requisites for producing biological systems. To achieve this, the biological system
first needs to be abstracted and modelled (Sect. 5.2), suitable sensor technology is
required for monitoring conditions and the production process must be controllable
(Sect. 5.3). An essential element for controllability is the ability to align processes
to the specific characteristics and variability of the biological system. Furthermore,
appropriate actuator technology is required to manufacture the biological systems
(Sect. 5.4). The value creation that is thus made possible benefits the life sciences
and, in particular, research in the life sciences.
The second perspective has its origin in the life sciences. Knowledge of engi-
neering applications allows for the implementation of biological systems that may
be integrated in the form of products or into production processes within the frame-
work of biological transformation (Sect. 5.5). Merging these two points of view
Production of
biological system
Engineering sciences Biological systems Life sciences
Integration of
biological
systems
Fig. 5.1 Explanatory model for Life Science Engineering. The two perspectives on biologi-
cal systems from the points of view of both engineering and the life sciences are significant.
(Fraunhofer IPT)
5 Technical Homes for Human Cells 65
under the banner of “Life Science Engineering” lays the foundation for biological
transformation.
5.2 Requirements from a Biological Aspect and Modeling

of Biological Processes
In contrast to conventional rigid processes, modeled in the field of production

technology, cells are inherently dynamic, which affects how the process control is
designed. The complexity of the processes increases further when using cells spe-
cific to an individual person. Patient cells possessing unique properties are used,
for example, for the production of biohybrid implants. In order to master the often
complex relationships, the main influencing factors are required, as well as de-
fined parameters for the characterization of the cell state. If these are determined
through measurements, they allow certain cell properties to be quantified and thus
enable appropriate recommendations for action to be made for the process. Thus,
no rigid modeling of biological processes is carried out; but rather, subprocesses
are selected based on the cell state after a given culture period, and brought to
execution.
This happens analogously in laboratory practice in the sense of laboratory pro-
tocols. These provide an abstract process description and use specific defined val-
ues in a similar way for decision making. The transfer of biological processes to
laboratory protocols is based primarily on empirical experimental data that defines
possible ranges of values as well as specific target values for various process pa-
rameters. These protocols must be re-established with every new deployment in
a laboratory. In doing so, exact ranges of values are defined for the available pro-
tocols by carrying out experiments, and target values are specified so that it is
possible to apply the respective protocol in the laboratory. Sophisticated modeling
of biological processes supports the establishment of these protocols and allows
for their optimization.
5.3 Microphysiological Organ-on-a-Chip Systems

as an Interdisciplinary Platform
There has been a dramatic increase in allergies in the population and serious in-
cidents during clinical trials have taken place world-wide. Further, costs and time
required for the development of a single drug are immense. There is also an in-
creasingly heated discussion on potential health hazards of nanodusts in recent
66 C. Leyens et al.
years. All these are examples bringing our attention to the fact that humankind
does not yet have any adequate methods available for predicting the complex inter-
actions of the human organism in its typical environment with, for example, drugs
or air pollutants that are harmful to the environment and our health. The study of
these interactions has been based on the culture of cell lines in petri dishes and
in vivo experiments on animal models for many decades. The results obtained in
these models can often not be extrapolated to humans, or only to a limited extent.
The reason for this deficiency is, in our opinion, less related to the level of
knowledge that the individual disciplines have attained in the process of learn-
ing about such interactions, but rather to the lack of interdisciplinary solutions.
Convergent technologies are needed in order to be able to model the complex in-
teractions between substances and human organs, or the whole organism under real
conditions and prior to human exposure.
Microphysiological organ-on-a-chip systems allow the production of func-
tional human tissue in a controlled environment and have the potential to lead to
a paradigm shift in preclinical research.
At the Fraunhofer Institutes for Interfacial Engineering and Biotechnology IGB
and for Material and Beam Technology IWS, microphysiological organ-on-a-chip
and miniaturized multi-organ chip technologies are being developed and evaluated
for, among other things, their applicability to substance testing. These systems
open up possibilities for understanding the effects of complex cell-cell and cell-
matrix interactions and generating data on the sum of all processes, which a drug,
for example, may be subject to in the body. These include the absorption of the drug
(resorption), its distribution in the body, biochemical transformation and degrada-
tion (metabolism) and elimination (excretion).
The complex structure of microphysiological organ-on-a-chip systems means
that their production and application require a wide range of technologies from
very diverse industries, including mechatronics, biotechnology, medicine, cell
culture technology, construction and mechanical engineering, electronics and
data/information processing. Close cooperation is thus required between differ-
ent branches of industry and scientific disciplines. The convergence of cutting-
edge technologies and sciences—biotechnology, medicine, computer science,
information technology, microtechnology—combined with innovative, applica-
tion-oriented networking make it possible to gain new insights and develop new
applications that were not possible within individual disciplines.
Fig. 5.2 An entire laboratory on a chip
5.3.1 Introduction
Owing to unforeseeable risks, in many cases it is not initially possible to directly

investigate the therapeutic efficacy of a drug, the toxicity of a molecule (chemical,
pharmaceutical, etc.) or the validity of a mechanistic hypothesis in humans. In or-
der to obtain a first assessment in such cases, preliminary tests must first be carried
out in vitro with (immortalized) cell lines and complex in vivo animal experiments.
Immortalized cell lines are often either of non-human origin or originally derived
from cancerous tissue and are traditionally cultured as 2D monocultures. Animal
models, despite extensive research, are incapable of replicating the complex human
body, and specifically human diseases, as they have a variety of species-specific
68 C. Leyens et al.
physiological differences to humans due to divergent evolutionary developments.

The use of animal models in general is also ethically questionable.
As drug development is built on population-based studies nowadays, medical
treatments can be very different for each individual. What heals one does not work
for the other and may even lead to serious side effects or deterioration in the health
of a third person. The field of medicine is trying to address this problem through
the use of individually customized therapies. “Personalized medicine” is opening
up a new dimension of tailor-made therapies through novel detection methods.
Molecular biology examinations often serve as the basis for medical decisions. For
example, the presence of certain biomarkers or the concentration of certain proteins
can provide information on whether a drug is effective or whether cancer patients
can forgo chemotherapy. Personalized medicine aims to harness knowledge about
the molecular processes of life and to use it for customized prevention, diagnosis
and therapy.
Microphysiological organ-on-a-chip systems can be used to help make predic-
tions for individual patients. Organ-on-a-chip systems are complex, miniaturized,
perfused cell culture systems that simulate medically or pharmacologically rele-
vant functional mechanisms of the human body. The basic principle of organ-on-
a-chip systems is to create a controlled, micrometer-sized environment for the cul-
ture of human organ tissue. It uses methods from the semiconductor industry as
well as laser structuring technologies and combines them with tissue engineering
approaches. Particular emphasis is placed on generating defined physiological cell-
cell and cell-matrix interactions, and guaranteeing specific organ functions. These
include, for example, barrier functions (among others, in blood vessels, intestines,
lungs or on the skin), metabolic or storage functions (including in the liver, kidney
or adipose tissue) or mechanical and supporting functions (including the heart, in
bone or cartilage tissue or skeletal muscle). In addition, the systems are usually
equipped with blood vessel-like perfusion, which allows nutrients, preparations
and cytokines to be transported to the tissue and released and metabolized (waste)
substances away from the tissue, with great temporal and local precision.
In combination with iPS cell technology (induced pluripotent stem cells) a per-
sonalized chip can be built for each patient based on their own cell material, making
it possible to already study their body’s reactions to a substance in the laboratory.
This allows possible serious side effects to be excluded and optimal treatment op-
tions to be identified ahead of the planned treatment.
5.3.2 Specific Organ-on-a-Chip Systems
By using human cell material in conventional cell culture, species-specific dif-

ferences between animal and human studies as well as the killing of laboratory
animals can essentially be avoided. However, in conventional cell cultures the
structural arrangement of different, sometimes highly specialized cell types within
an organ is usually not considered. Defined cell-cell and cell-matrix interactions
are of paramount importance for replicating a physiological cellular function. For
example, mechanostimulation in respiration and intestinal peristalsis, electrophys-
iological stimuli in cardiac muscle contraction and neuronal activity, as well as
light in retinal functionality (retina) have a decisive influence on the formation
and regulation of a microphysiological environment. Therefore disruption of these
microphysiological conditions is the basis of almost all non-hereditary diseases.
Organ-on-a-chip models aim to reproduce these microphysiological conditions as
realistically as possible in vitro, allowing us to leave the typical disadvantages of
conventional cell cultures behind us. One major challenge when culturing com-
plex, three-dimensional organ models under physiologically relevant conditions is
maintaining their function over extended periods of time. Microfluidics and the
associated microfluidic perfusion provide the basis for the standardized, long-term
supply of such artificial tissue.
A diverse array of microphysiological organ-on-a-chip systems is being devel-
oped, manufactured and characterized at the Fraunhofer Institute for Interfacial
Engineering and Biotechnology IGB. The focus of this work is on the reproduc-
tion of tissue-specific in vivo structures, dimensions and environmental conditions
using microfluidic systems.
Heart-on-a-Chip Systems
Heart-on-a-chip systems integrate, for example, anisotropic 3-D cardiac muscle
fibers based on human iPS cells (Fig. 5.3a). As the most common reason for
drug candidate failure is cardiotoxicity or cardiac tissue dysfunction, hiPS cell-
based in vitro models of cardiac and especially myocardial tissues with physiolog-
ical function are of particular importance. The heart-on-a-chip models consist of
microfluidic modules that integrate individual myocardial fibers in special tissue
chambers, thereby culturing them in a physiological environment that corresponds
to the in vivo structure in dimensions and geometry. Experiments with drug prepa-
rations have already revealed that these tissues have advantages over the classically
used cell assays or rodent models in terms of pharmacological prediction. The
physiology of the heart model bears a close resemblance to that of large animals
or humans. A computer-based analysis method was developed to ensure the paral-
70 C. Leyens et al.
Fig. 5.3 a Heart-on-a chip

system, b fat-on-a-chip
system, c retina-on-a-chip
system
lelization of these systems, which allows the automated reading of the heart-on-a-
chip systems (Fig. 5.3b).
Fat-on-a-Chip Systems
Fat-on-a-chip systems integrate functional human adipose tissue in a physiologi-
cal microenvironment in a similar way. Adipose tissue is often overlooked as an
organ, although it accounts for about 20 or 25% of the body weight of healthy
men and women, respectively, and even more than 50% in cases of illness. White
adipose tissue (WAT) is a highly specialized connective tissue with a simple struc-
ture and histology. It has long been perceived to be an organ that serves only to
store and supply energy. Now, however, it is recognized as an important endocrine
organ that releases a variety of cytokines, so-called adipocytokines or adipokines.
The fat-on-a-chip system is a platform designed specifically for the integration of
white adipose tissue with blood vessel-like supply channels, separated from the
3-D adipose tissue by an artificial endothelial barrier (Fig. 5.3b). This allows for
continuous supply and at the same time protection against non-physiological shear
forces. Further, the platform consists of a multilayer PET/PDMS hybrid chip with
tissue chambers of several hundred micrometers in diameter. The fat-on-a-chip sys-
tem allows human adipocytes to be cultured for several weeks while maintaining
the vitality and functionality of the adipose tissue. In addition, the supply channels
could already be coated with endothelial cells in order to generate an even better
recapitulation of the endothelial barrier.
Retina-on-a-Chip System
Another example of microphysiological organ-on-a-chip systems is the retina-on-
a-chip system. Degenerative diseases of the retina, such as age-related macular
degeneration or retinitis pigmentosa, are the most common causes of vision loss.
Existing in vitro or in vivo model systems for the testing of new pharmaceutical
agents as well as mechanistic research are only able to represent the structure and
complexity of the human retina to a very limited extent. The novel microphysio-
logical retina-on-a-chip system combines human iPS cell-based retina organoids
with epithelial and endothelial layers in a microphysiological platform (Fig. 5.3c).
The in vitro model is thus able to allow the physiological interaction of photore-
ceptors with the surrounding retinal pigment epithelium. This interaction, which
is crucial to the functionality of the retina, has not been demonstrable in vitro till
now. The retina-on-a-chip system that has been developed therefore has great po-
tential as a physiologically relevant test system for the development and testing of
new pharmaceutical agents as well as for investigations into the causes of disease.
5.3.3 Multi-Organ Chips–All-in-One Microfluidics
In addition to the development of individual microphysiological organ-on-a-chip

systems, research is being conducted on how a whole organism could be modelled
in a microfluidic network in the future (human-on-a-chip). The multi-organ chips
developed for this purpose provide a small, technical model of the basic functions
of the human body. Specific cells or tissues of individual organs or organ networks
are cultured in them. In order for the cells to fulfill their function as they do in
the human body, the conditions that exist in the body are recreated technically.
This relates, for example, to temperature, pressures and flows that must prevail at
different points, and also the supply of oxygen and nutrients.
72 C. Leyens et al.
In recent years, various design concepts have been introduced for multi-organ-
chips, which may be subdivided into static, semi-static and flexible systems. Static
and semi-static concepts both have a number of limitations that can be avoided by
providing the individual tissues with a need-based supply.
The human body has a complex blood circulatory system, which is able to
achieve the need-based supply of individual organs and tissues by means of nu-
merous complex control mechanisms. This system needs to be reconstructed tech-
nically. A universal plug-and-play kit was developed for this purpose at the Fraun-
hofer Institute for Material and Beam Technology IWS, by means of which it
is possible to quickly and easily assemble customized multi-organ systems. The
kit provides a variety of ultra-compact cell culture modules, micropumps, valves,
reservoirs, mass exchangers and sensors that can be freely combined with one
another. It is therefore possible to individually control the flow and nutrient concen-
trations for each tissue within the multi-organ systems, and thus to ensure a need-
based supply. The combination of microfluidic base chips with customized, ul-
tra-compact, microphysiological modules for different micro-tissues has permitted
a need-based supply of the individual tissues within complex, individually config-
ured multi-organ chips for the first time.
The Multi-Organ Chip

Base chips with integrated micropumps, valves, reservoirs, mass exchangers and
sensors form the core of the IWS multi-organ systems. Over and above the closed
technology chain for the production of base chips and microphysiological mod-
ules, a universal control platform has also been developed and established. The
base chips are assembled from laser-structured individual layers using multilayer
technology. The use of modern laser sources and cost-effective polymer materials
makes the production of these systems flexible, inexpensive and time-saving. Pro-
totypes of microphysiological systems can be put into practice within a few days
thanks to these flexible technologies and the use of laser micromachining technol-
ogy.
The exploded view of a multi-organ chip in Fig. 5.4 serves as an example to ex-
plain the structure and functions of a system of this kind. The system shown here is
produced by multilayer lamination and 3D printing. The functional structure of the
system is divided into layers and incorporates a pneumatic part [1] for distributing
gases, an elastic membrane [2] as a functional element of the integrated pumps and
valves, a microfluidic section [3] for the distribution of liquids and as a cell culture
compartment, and a bottom film [4] that acts as a seal and interface for microscopy
and sensor integration. The system’s fluidic circuit [7] emulates the cardiovascular
system of the human body. For this purpose it is covered with endothelial cells, like
Fig. 5.4 Exploded view of a multi-organ chip
the veins in the human body, and supplies the replicated organs in the cell culture
compartments [10] with nutrients and oxygen. The transport of fluids is facilitated
by an integrated micropump that imitates the heart [6]. Fresh nutrients in the form
of cell a culture medium can be supplied via a fluidic port [9]. A gas exchanger [5]
serves as an integrated lung and regulates the oxygen input in the system.
Pneumatic ports [8] connect the multi-organ system to a controller. This reg-
ulates the pumping speed (in analogy to the heart rate) and the gas volume flow
to the gas exchanger (in analogy to breathing) (Fig. 5.5). As a result, essential
body functions can be carried out in a technical medium, thus providing an ideal
micromileu for the emulation of cell-based questions.
Moreover, various disease models, such as oxygen deficiency or hypertension
can be emulated by varying the technical parameters. By adding stimulants or for-
eign substances such as medications via the fluidic port, the questions posed by
pure medical research as well as the effectiveness of new drugs can be investigated
further.
The associated control platform is based on compact single-board computers,
such as those used in mobile phones and tablets. In addition to customer-specific
sensor and actuator interfaces with hard, real-time requirements, the Linux oper-
ating system provides comprehensive administration and monitoring functions up
74 C. Leyens et al.
Fig. 5.5 a Modular control unit for microphysiological systems, b Organ-on-a-chip system
for emulation of kidney functions
to and including cloud interfaces. Additional functionalities such as self-optimiza-

tion, self-configuration and self-diagnostics can also be implemented to support
and assist users in their increasingly complex work.
5.3.4 Outlook
The cost of introducing a new drug from development to market launch is several
billion euros. In a multi-stage development cycle, out of the approximately 10,000
substances tested in the course of drug development in the preclinical and clin-
ical test phase, on average only one active substance is found, which ultimately
obtains the regulatory approval required for marketability. A development cycle
such as this typically takes over 10 years and the associated costs have increased
by more than 14 times in the last 20 years. Despite steadily rising investment in
research and development, the number of approved drugs has remained relatively
constant. Before they may receive approval, new medicines must be more effec-
tive than substances already on the market and should cause fewer side effects.
Depending on the mechanism of action and target organ, this poses a range of dif-
ferent challenges for the development of new drugs. One of the greatest hurdles is
the transfer of preclinical research results from cell cultures and animal models to
humans. Very often, lack of efficacy or unforeseen toxicity in humans lead to the
rejection of a drug, which was initially successful in animal models.
Microphysiological organ-on-a-chip systems provide a justifiably ethical alter-
native to animal testing and simultaneously promises improved transferability to
humans. Organ-on-a-chip technology, voted among the “Top 10 Emerging Tech-
nologies in 2016” by the World Economic Forum, combines the characteristics of
classical cell line assays (including human genes and cheap, easy handling) with
those of animal models (including multicellular 3D tissues and complex circula-
tion). While there is still a long way to go before traditional animal models can be
replaced, rapid progress in both the development of organ-on-a-chip systems and
in regenerative medicine and tissue engineering has given organ-on-a-chip systems
the potential to become a key technology of the 21st century.
In order for the micro-physiological organ-on-a-chip systems to be made avail-
able quickly and cost-effectively to the pharmaceutical industry and cosmetics
companies, medical and academic researchers, specialized yet flexible manufac-
turing technologies are needed for the production of the systems as well as stan-
dardized and automated processes for obtaining each of the required cell types. In
addition to the established manufacturing processes such as injection molding and
multilayer lamination, additive manufacturing is playing an increasingly important
76 C. Leyens et al.
role in the manufacture of these complex systems. At the same time, innovative
roll-to-roll technologies are now available for the automated serial production of
such systems, which enable different production technologies to be combined in
a series process. In this way, established manufacturing methods such as laser
structuring, hot stamping, lamination and inkjet printing can be combined with
novel technologies such as laser-based surface modifications.
5.4 Sensors and Actuators for Automation in Bioanalytics
As described in Sect. 5.1, the manufacture of biological systems is an essential

element of biological transformation. The complexity of a biological system such
as a stem cell requires appropriate sensor technology to collect the relevant data
needed to make the production process controllable. The sensors that are used
therefore primarily serve to detect the state of the biological product rather than
the state of the production system, which sets them apart from sensors that are
conventionally used in manufacturing technology.
5.4.1 Automated High-Throughput Microscopy for Cell

Production
The use of microscopic imaging methods is standard in the field of cell culture,
especially when it comes to manual cell culture. Confluence is an especially impor-
tant parameter for monitoring progress in cell growth. This is typically determined
by means of phase-contrast microscopy and subsequent image processing. In in-
dustrial, automated cell production with high throughput, microscopes of this type
are a bottleneck due to the relatively long acquisition times. The acquisition times
are also critical given the fact that the cell culture is removed from the incubator
and therefore not in a controlled environment while the image is being captured.
In order to obtain a comprehensive picture of the cells in the cell culture vessel,
the entire surface must be microscopically examined. With a microscope objec-
tive’s small field of view of a few millimeters and the comparatively large surface
area of a microtiter plate (MTP) of around 8 × 13 cm, thousands of individual im-
ages are required to comprehensively examine an entire plate under a microscope
and capture all cells. Nearly 19,000 individual images are required to examine all
wells with a 10X objective, which then have to be stitched together into overview
images. In light of the above, accelerated, high-throughput, automated microscopy
Measurement
LED Flash
Trig
ger Hardware focus
l Controller with measurement
ro
nt path-
Co synchronous
trigger
Control
High-speed camera
Motorized stage
Piezo z-stage
Fig. 5.6 Components for microscopy with a moving object. A central controller ensures
the synchronization of movement, lighting and image acquisition. Focusing takes place via
a hardware-based focus position measurement and a z-axis elevation stage. (Fraunhofer IPT)
systems are of high relevance for cell production. Current systems for cell cul-
ture vessels such as microtiter plates are limited with respect to acquisition time
because of the “stop-and-go” imaging mode, where the MTP must be accelerated
and decelerated for each individual image. The acceleration and speed of the stage
can only be increased to a limited degree, since abrupt starting and stopping causes
fluctuations of the liquid nutrient medium, which affects the exposure of the indi-
vidual images. Depending on the magnification, a microscope image of an entire
plate can therefore easily take several hours.
Significantly shorter acquisition times can be achieved by using an “on the fly”
approach with continuous movement of the cell culture vessel during imaging.
This is made possible by hard synchronization of the traversing motion with the
lighting and image acquisition. Special shutter modes ensure that images are free of
moving blur, and a focus position measurement based on optical coherence tomog-
raphy (OCT) guarantees that they are sharp [1]. Fig. 5.6 shows the arrangement of
the individual components used for the high-speed microscopy approach. The mi-
croscope stand into which the components are integrated is not depicted here. In
the case of cell microscopy, inverse microscopy is usually used.
The acquisition is carried out by pre-measuring the position of the bottom of
the cell culture vessel via the co-axially integrated OCT system. This is followed
by image acquisition with simultaneous meandering movement of the sample in
78 C. Leyens et al.
Constant velocity – Trigger aktive

Acceleration
120 mm
a b
Fig. 5.7 a Meandering scanning strategy for high-speed microscope imaging; b Full-field
phase contrast image of one MTP with 4X objective and an image size of 40 GB. (Fraunhofer
IPT)
the X, Y and Z directions. All images are captured with an overlap because the
trigger signals contain jitters and gaps arising between the individual images are
undesirable.
Thanks to the optimized management of the shutter mode and image storage,
the camera can be read out at the full frame rate of 100 images per second at
5.5 megapixels and 16 bits. On completion of the image acquisition, the individ-
ual images are collated using special algorithms (so-called stitching). The image
acquisition process takes about 1 min for a 4X microscope objective, which re-
sults in an approximately 40 GB image file (Fig. 5.7) At low magnifications, the
acquisition time is limited by the travel speed of the microscope stage.
5.4.2 Deep Learning for Classification in Cell Microscopy
High-speed microscopy provides a method for quickly recording quality parame-

ters during automated cell culture. Confluence is the main quality parameter, which
provides relevant information about cell growth and must therefore be monitored at
regular intervals. Other factors, which are of interest, include the size and shape of
cell colonies and the cell density. In the production of stem cells, the recognition of
hiPS clones (human induced pluripotent stem cells), differentiated and dead cells
is important. In manual cell culture, the classification is carried out using the expert
a Phase contrasst area of an image Segmented image Segmented image (zoom)
Single Cellsover Differentiated

Background iPS cell iPS cell colony monolayer cells Dead cells
Fig. 5.8 a Section of a microscope image of a single well in phase contrast with segmenta-
tion; b Class definitions with sample images from the training dataset. (Fraunhofer IPT)
knowledge of a biologist. The automated process requires appropriate algorithms,

that perform this classification efficiently and reproducibly.
Image processing algorithms based on deep-learning approaches were thus de-
veloped for the extraction of quality parameters from large-scale microscope im-
ages. The motivation for this was, that classic algorithms such as PHANTAST are
clearly too slow to process large area images of MTPs in a reasonable time of less
than one minute per well for a 6-well MTP. Furthermore, there was a need to dif-
ferentiate between significantly more than two classes (foreground, background).
In order to distinguish between six classes in this specific case, a classification was
developed based on a CNN (convolutional neural network), which determines the
classification of one well in a 6-well MTP in just 34 s using GPU computing.
The following classes have been defined (Fig. 5.8): Background, iPS cell (sin-
gle), iPS cell colony, differentiated cells, dead cells and cells that have outgrown
the monolayer. The CNN was trained using 40 images which included all the
classes. Data augmentation was used to increase the training data set. Filters were
thus employed that added modifications in, for example, rotation, translation,
Gaussian blur and brightness. The robustness of the classification, particularly
with respect to variations in image brightness and sharpness, could be increased
with the last two filters.
80 C. Leyens et al.
Fig. 5.8 shows a section of the phase contrast micrograph of a single well with
hiPSC and the segmented image thereof. Differentiated cells can be reliably dis-
tinguished from the iPS cell colonies. Moreover, isolated iPS cells and dead cells
can be detected.
Parameters such as the confluence of each class may be calculated on the ba-
sis of the image classification. Furthermore, colonies can be detected by means of
an additional blob detection method and position, area, edge and roundness ex-
tracted for this purpose. These then also permit analyses of the growth behavior of
individual colonies. The parameters in turn flow into automated decision-making
processes in the automated cell production.
5.4.3 Optical Coherence Tomography (OCT) as Sensor

Technology in Tissue Engineering
The products of tissue engineering have become increasingly important for

biotechnology and regenerative medicine in recent years. Cell microscopy as
described in Sect. 5.3 has only limited suitability for the monitoring of production
processes and quality assurance and is therefore mainly applied in the imaging
of adherent cells such as iPS or mSC (mesenchymal stem cells). The selection
of imaging options in tissue engineering of 3D cell cultures such as organoids or
micro- to macroscopic tissues is reduced to fluorescence-based methods such as
confocal microscopy or its special form, light sheet microscopy. These methods
have the disadvantage that the samples must be prepared with markers and also the
penetration depth is limited to a few 100 µm.
In contrast to that optical coherence tomography (OCT), an imaging technique
based on low-coherence interferometry, functions without the additional use of
markers and is contact-free. By using OCT it is possible to acquire tomographic
sectional images of biological samples, similar to ultrasound imaging. Since the
principle was discovered in the early 1990s, OCT rapidly became the gold standard
for retinal diagnostics in ophthalmology [2].
OCT systems are constructed similarly to a Michelson interferometer (Fig. 5.9).
Light from a broadband superluminescent diode is coupled into a fiber optic system
and split into two parts with a fused fiber coupler. One part of the light is directed
into a reference arm, where it is reflected by a mirror. The other part is directed
into a scanning device, by means of which the measuring beam scans across the
sample using Galvo scanners and/or motorized axes in a raster pattern.
Fig. 5.9 Schematic structure of a Fourier domain OCT system. (Fraunhofer IPT)
The measuring beam penetrates biological tissue and is backscattered as a func-

tion of the density variation—similar to an acoustic wave in ultrasound imaging.
The backscattered light from scattering centers located at different depths is fed
back into the fiber and superimposed on the light in the reference arm from the
fused fiber coupler onwards, where the two light components interfere with each
other. The light interference is recorded in a spectrometer and evaluated by a com-
puter.
The subsequent signal processing produces a depth-resolved density profile in
the form of an A-scan for each point. By scanning with a mirror, so-called B-scans,
tomographic sectional images can be acquired. With a second mirror volumetric C-
scans can be acquired.
A fully automated OCT system was developed for the automated production of
artificial skin models within the scope of the Fraunhofer foundation project “Au-
tomated Tissue Engineering on Demand”. By means of this system, the quality of
the skin models can be characterized after growth is complete. Topography, homo-
geneity and defect formation are automatically analyzed to detect defective skin
models [3–5].
Fig. 5.10a shows the OCT system, which automatically positions the skin sam-
ples in the individual wells below the measuring head, focuses the sample in the
Z-direction and can record B-scans. For quality assurance, an image processing
of a B-scan is carried out to correct defects such as air bubbles and topography
(Fig. 5.10b).
82 C. Leyens et al.
a b
Fig. 5.10 a Fully automated OCT system for the measurement of skin samples in MTP;
b B-scan of an artificial skin sample captured using OCT. (Fraunhofer IPT)
Fig. 5.11 Images of a microfluidic chip taken by light microscope and OCT. The structural
width of the microfluidic channels is 100 µm and can be clearly visualized by OCT. (Fraun-
hofer IPT)
Since OCT can generally be used with all materials that the light of the near-
infrared spectrum that is used can penetrate, it is also suitable for quality assurance
in the production of microfluidic chips (Fig. 5.11) as well as for the visualization
of tissue/organ-on-a-chip applications.
5.5 Laboratory Automation as the Basis for Process Transfer

in Biological Transformation
Material substances and samples are transformed into data and information in labo-
ratories. This means that laboratories are basically knowledge-producing data fac-
tories where decisions are made. Laboratories are thus an integral part of modern
value creation structures and should therefore be operated efficiently, economically
and in a controlled manner. Improved methods of extracting information from sam-
ples and material substances have enabled the life sciences to dramatically improve
their performance over the last 30 years. As a result, we can now offer therapies that
are customized and personalized for a specific patient, in the context of personal-
ized medicine. The development of sequencing technology has made a significant
contribution to this.
The human genome was fully sequenced for the first time during the Human
Genome Project in the late 1990s and published in 2001. The project took several
years and cost US$2.7 billion. Sequencing technology has since made tremendous
progress. Full genome sequencing can now be done in just a few days for a few
thousand euros. There are already concepts for reducing costs to less than 100
euros.
Sequencing technology is just one example of the development of analytical
techniques for the accurate and comprehensive characterization of biological sam-
ples. As a result of the knowledge thus gained about biological functions and
processes, these technologies have now been incorporated into direct application
areas such as diagnostics. Individual analysis and characterization also offer the
opportunity to develop much more individualized measures to combat diseases. At
the same time, this knowledge permits targeted modification of biological systems
to cover certain functions.
As a result of the comparatively late development of this analytical technique
in relation to industrial production, many laboratories today are still not automated
and work processes are structured in a factory-like way. Due to the ever-increasing
demand for conclusive analytical results, there is a need for flexible automation of
work processes. Automation is mainly used in the laboratory environment today
84 C. Leyens et al.
Fig. 5.12 Impressions of working environments in industrial production and in life science
laboratories [6, 7, 8, 9, 10, 11]. (Fraunhofer IPA)
for the development of pharmaceutical agents, where the effects of compounds

from substance libraries on a disease-dependent target are investigated.
Here, automation allows for a huge increase in the number of measurement
points through the multitude of combination options with different concentrations
of substances, and thus increases the productivity of the laboratory.
Laboratory methods and analytical technologies usually require targeted and
often lengthy preparation of the samples before the actual measurement technique
is used. These tasks can be performed by laboratory machines today. Owing to the
great number of different measurement techniques, the individual characteristics
of methods and variety of sample preparation options, it is not enough that modern
laboratory machines simply increase the throughput. It is far more about the quality
of samples, reproducibility through the use of secure and standardized procedures
and the safety of the laboratory users.
Biological samples are usually primarily one or more portions of tissue that
have been taken from an organism. They may be liquid (blood, urine, etc.) or solid
(biopsies, smears, etc.). The tissue consists of a composite of extracellular matrix
and cells as the smallest building block of life with its own metabolism. Informa-
Fig. 5.13 Tree structure for carrying out laboratory analyses. (Fraunhofer IPA)
tion areas can be differentiated within the cells, for which the analytical methods
differ significantly. The analysis of the cells themselves is based on cell analysis
methods and gathers information of the cells’ metabolome. In this process the cells
remain vital. Proteomics is a further area, whereby the protein structure of the cells
is analyzed. The third area is the analysis of the genome and transcriptome, which
takes the form of sequencing studies for example. The structure of the laboratory
analyses is shown in Fig. 5.13.
86 C. Leyens et al.
Fig. 5.14 Full automation of all areas of biotechnological analysis according to the tree
structure for carrying out laboratory analyses. (Fraunhofer IPA)
The extracted cells form the starting material for any laboratory analysis. Since
the isolation of cells from tissue matrices is highly complex or even impossible,
sometimes if there is a lack of available donor material, “cell stocks” are often
used for the investigations. These cell lines have the advantage that they provide
comparable material for later experiments. These cell lines can also be propagated
outside the organism to provide a practically infinite pool of cell samples. Artificial
tissue production has also taken off recently because cells in a three-dimensional
tissue structure demonstrate a more natural behavior.
Automation can also be structured according to the structure of the analyti-
cal areas in life science laboratories. As a result, the individual measures have
significantly improved reproducibility. A wholly automated laboratory consists of
functional modules which, when flexibly combined, result in the tree structure
shown above. Each functional module represents an actuator or sensor, each of
which carries out individual process steps in the laboratory’s work process.
Fig. 5.15 Cocoon system for individualized cell culture [12]. (2018 © Lonza)
Since at the core of the procedures is a variable biological sample, the process
parameters must be largely prevalidated. Ideally, process standards can even be
generated in this way, which can be used as a reference by an automated process.
In general, it is important to monitor both instrumentation and biological samples
by means of integrated sensors, so that is possible to respond to dynamic changes
in the biological samples. Digital process models can be created with which the
real data may be compared. This offers real-time transparency over the process
workflow and device functions and thus creates an interface between biology and
technology. Fig. 5.14 shows the conceptual structure of a fully automated process
solution.
A distinction must be made between the functional levels when laboratory pro-
cedures are automated. The so-called assay function characterizes the sample’s
biochemical processes in the absence of external influences. The primary functions
are all the actuator and sensor processes that result in direct manipulation of the
biological samples. There are additional secondary functions where the biological
sample is indirectly manipulated, by transporting the sample vessel, for example.
Finally, auxiliary functions round off the entire system, although these functions
no longer have any influence on the actual biological sample.
This structure is not recognizable in many laboratory machines today, because
many functions are already integrated into an automated system. Such solutions
are thus correspondingly inflexible. In order to do justice to the unique character
of a biological sample, it is advantageous if individualized process solutions are
designed for the biological sample in the form of functional modules, provided
they are primary functions.
88 C. Leyens et al.
An example of this is the Octane Cocoon system, which was designed for indi-
vidualized cell culture (Fig. 5.15). All primary functions are represented in a single
unit, the Cocoon.
Many existing laboratory automation solutions are being transferred to new ar-
eas in the context of biological transformation. As laboratory automation continues
to develop, process sensors and control software that makes use of artificial intel-
ligence are of vital importance. As these are integrated, it will be possible to fulfil
new functions in the future, and tasks which are currently performed manually
at a great cost will be taken over by machines. The integration of artificial intel-
ligence in particular allows for direct interaction between a robotic solution and
a biological sample.
Additional potential lies in the progressive miniaturization of primary functions
in smart lab-on-a-chip systems. They too can be automated via interfaces.
5.6 Conclusion and Outlook
Generally speaking, biological transformation has the potential to revolutionize in-

dustrial value creation by gradually blending biological and technical approaches
to produce biointelligent systems. These interfaces are the essential enablers of bi-
ological transformation, which permit the intermeshing of the different domains.
Standardized interfaces between biology and technology therefore need to be de-
veloped.
A multimodal biointerface system (MMBIS) designed at the Fraunhofer Insti-
tute for Production Technology IPT could serve as an example of such an approach
(Fig. 5.16).
Signal
Stimulus
Maintain vital functions
Fig. 5.16 Schematic structure and main functions of a multimodal biointerface system
(MMBIS). In addition to maintaining the vital functions of the biological system, stimuli
and signals are transmitted to and from the biological system. (Fraunhofer IPT)
The MMBIS is a deliberate combination of different individual technologies,

some of which are already available, which represent the following elementary
functions:
1. Maintaining the vital functions of the biological system: This includes both
nutrient transport (e.g. via microfluidics) and temperature control.
2. Transmission of stimuli: It must be possible to transfer information to the
biological system if biointelligent systems are to be realized. The usable infor-
mation channels such as optical, chemical, electrical or mechanical interfaces
must be technically integrated into the system. They enter the MMBIS as inputs
along with the vital functions.
3. Reading out signals: The functions of the biological system need to be made
accessible via integrated sensors or technical interfaces with external sensors
(similar to those listed under point 2). The reading is either taken in response
to a stimulus or at any other point when, for example, state variables of the bio-
logical system need to be monitored. Stimulus and response do not necessarily
have to use the same information channel e.g. electrical stimulus of a “muscle-
on-a-chip” and mechanically picking up the muscular contraction.
It would be ideal to implement the MMBIS in miniaturized form, in order to

achieve a high integration density and have the ability to run the three elemen-
tary functions mentioned above in parallel. Platform technology of this kind can
be built based on production technologies used for microfluidic chips, where ad-
ditional optical conductor paths are included by plastic replication, and electrical
conductor paths are added by inkjet printing or pick-and-place machines for more
complex circuits. This could also be implemented very cost-efficiently in roll-to-
roll processes with potential for high-throughput.
With the approach outlined here, a wide variety of bio-intelligent systems could
be realized using a uniform basic layout for the MMBIS, from photo-biosensors
to biological energy stores in cellular mitochondria to micro-bioactuators. All in
all, this would provide a universal, microscale platform technology, a micro-mul-
timodal biointerface system (MMBIS) that offers a toolbox for various types of
interfaces between biology and technology.
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Phenotyping and Genotyping of Plants
Phenotyping of Crop Plants Using Spectral
6
Sensors and Artificial Intelligence
Udo Seiffert and Andreas Herzog
Summary
The digitization of the economy and society naturally does not fail to include
agriculture. Having a plant-based bioeconomy as primary production for many
downstream industries is a key component of digital transformation. The bi-
ological transformation of the economy, or rather of the world, derived from
the much-vaunted trend of digitization, is thereby extended by another very
interesting component. This is ultimately the optimization—at least at spe-
cific points—of biology by biology. In other words, the optimization of plants
as biological systems is carried out using technological processes and meth-
ods, the approach and design of which have been modeled on or inspired by
various biological principles. On the technological side, bio-inspired systems
include spectral sensors and, in particular, artificial intelligence as the central
component required for the phenotyping needed to achieve the aforementioned
optimization.
6.1 Introduction
According to statements by the German Federal Government’s Bioeconomy Coun-

cil, it can be assumed that global demand for biomass (approx. 14.4 billion metric
tons in 2012) will rise significantly in the coming decades [1]. This demand is
driven primarily by a growing global population (9.6 billion people by 2050 [2]).
U. Seiffert () A. Herzog

Fraunhofer Institute for Factory Operation and Automation IFF
Magdeburg, Germany

https://doi.org/10.1007/978-3-662-59659-3_6
92 U. Seiffert and A. Herzog
Africa
Australia Wine
ASEAN
Middle East
South America Fruit
North America
Europe
Vegetables
Specialized crops
Field crops
Nutrion
Plant protecon
Variety recognion
Seed
Crops
Soil
Fig. 6.1 Representation of the area expanded upon in the introduction in the context of
plant-based issues (horizontal), the crops considered (vertical) and corresponding geographic
components (depth axis). (Fraunhofer IFF)
Plant-based products are used not only for pure human nutrition (food) and
to feed a likewise growing livestock population (feed)—together about 11 billion
metric tons—but also increasingly as a renewable raw material for energy genera-
tion (fuel)—about 2.4 billion metric tons—and about 1 billion metric tons are used
as raw material (fiber). Together they are often referred to as “The Four Fs”.
This trend is overshadowed by a change in climatic conditions in many areas of
the world, which is causing traditional cultivation regions to shift and necessitating
new plant varieties for existing sites. As a result, the global goal for plant cultiva-
tion and agriculture is to increase productivity and efficiency while at the same time
optimizing plant traits (stress tolerance, nutritional or application-relevant ingredi-
ents, phosphorus efficiency, etc.). In addition, efforts in respect to sustainability are
becoming ever more important, particularly in terms of the utilization and mainte-
nance of biodiversity. This is an approach at the genotype level. Appropriate tools
6 Phenotyping and Genotyping of Plants 93
and methods for plant phenotyping are required in order to support plant cultiva-
tion (e.g. in the form of support systems for selection) and agriculture through the
use of technology (e.g. through on-site sensing) in achieving these objectives.
This goes far beyond the approach of precision agriculture that has been applied
till now. New challenges arise in the close combination of genotypic optimization
in the form of new varieties, their phenotypic assessment during plant breeding and
productive cultivation. Moreover, this market can by no means be considered lo-
cally anymore, but must be regarded as a complex global structure. Fig. 6.1 shows
the connection between plant-based issues, the cultures being considered and geo-
graphic components in the form of a three-dimensional cube structure.
6.2 Motivation
As mentioned in the introduction, the optimization of crop plant characteristics is

becoming increasingly important. Applications range from utilization of the plant
as a resource for human and animal nutrition, as a biological raw material and
energy generation. The identification and optimization of characteristic ingredients
of the plant as a whole or of certain parts of the plant play a central role in this
context.
Biological or chemical wet laboratories employ a number of analytical methods
for this purpose (e.g. chromatographic separation methods such as liquid chro-
matography (HPLC) or gas chromatography (GC), coupled with various detection
methods, such as enzyme-based or immunological detection methods). These usu-
ally “use up” the samples, in other words they are destructive methods [3]. Making
the results from plant analyses available in real time using the aforementioned
procedures is typically impossible. In many cases this is disadvantageous or un-
acceptable, for example in high-throughput screening or in automated production
operation.
By contrast, multispectral and hyperspectral imaging are powerful analytical
methods, which provide spectral data that reflect, for example, the concentrations
of various organic or inorganic ingredients. In order to be able to use this spectral
data for the non-contact, non-destructive identification of plant-based ingredients,
the challenge lies in assigning these measured values to the abovementioned es-
tablished methods—this is referred to as transformation. This transformation is
highly complex when a great number of ingredients of interest are considered, and
analytical methods are not yet available which severely restricts the application of
statistical methods. Furthermore, a high level of (biological) expertise is required
here and needs to be taken into account accordingly [4, 5]. Machine learning meth-
Fig. 6.2 Using selected reference plants, two methods of data collection and processing
are brought together in a mathematical calibration model. Hyperspectral imaging is non-
invasive and conceptually real-time capable, while the bottom path represents, so to say, the
gold standard. (Fraunhofer IFF)
ods such as artificial neural networks or fuzzy techniques allow support systems
to be constructed that are based on the working methods of the human expert [6]
through
their adaptability and trainability,

taking cognitive relationships into account,
the introduction of a certain degree of self-organization and
the ability to process fuzzy, incomplete (expert) knowledge.
This transformation can therefore be identified on the basis of characteristic

relationships and subsequently generalized.
Fig. 6.2 shows the system model, the fundamental relationship between spec-
tral and wet laboratory data and how they correlate. The mathematical calibration
model is always the critical component here. This incorporates the relationship
between the characteristic properties of the spectral data and the parts of the cor-
responding wet laboratory analysis that are relevant to the application [7, 8]. The
existence of suitable calibration models ultimately opens up a path to numerous
concrete applications.
The system model illustrated here represents the generation of a model. The key
advantage in terms of practical application is that, following the successful creation
and validation of the calibration model, the bottom pathway of the wet laboratory
analysis is theoretically no longer required.
Fig. 6.3 Data cube with spatial (x, y) components and spectral component (). (Fraunhofer
IFF)
The potential applicability of this methodology is huge, ranging from pure

biological research to plant breeding and diverse applications in agriculture and
forestry.
6.3 General Approach and Examples of Implemented

Solutions
Building on the system model depicted in Fig. 6.2, in this section the general tech-
nological approach is explained and illustrated by means of several examples.
Hyperspectral imaging forms the sensory core, which depending on the appli-
cation may be reduced with respect to the actual local and spectral resolution used.
The physical measurement principle used is molecule-specific absorption or
reflection across the electromagnetic spectrum. The typical result of hyperspec-
tral imaging is a so-called data cube, which features the two spatial components
(x and y) plus the spectral components () available for each of these spatial posi-
tions (Fig. 6.3, 6.4).
Fig. 6.4 Laboratory setup of a hyperspectral line scan camera with linear translation unit,
artificial broadband illumination and data acquisition computer. (Fraunhofer IFF)
Fig. 6.5 A typical spectral signature for plants in the wavelength range of about 400 to
2500 nm. This is an example of the exact data vector which results for a specific spatial
position x and y. (Fraunhofer IFF)
Machine learning methods have proven to be particularly advantageous for

processing the raw recorded spectral data in the context of the concrete applica-
tion [9]. Over and above the implementation of the actual task—usually in the
form of a classification (qualitative analysis) or regression (quantitative analy-
sis)—questions also arise about the relevance of the individually recorded spec-
tral bands in the context of current scientific studies, especially with a view to
a low-priced, multi-spectral approach tailored to the concrete application. More-
over, questions concerning the transferability of the mathematical calibration mod-
els that have been created to different sensor hardware and application scenarios
as well as their interpretability in the respective application context are of partic-
ular interest [10, 11]. This spans a vast field which is beyond the scope of this
contribution. For additional information, reference can be made to a range of other
publications.
The technological approach described is characterized by a number of proper-
ties that are typically positively rated both versus alternative approaches and by the
market.
Direct measurement of the condition of a plant via hyperspectral imaging in

the IR, UV and visible range (Fig. 6.5) and offline calibration of the system via
suitable biochemical analytical methods (e.g. mass spectrometry, liquid chro-
matography),
Use of a variety of spectral bands ! high-dimensional image stack,
Identification of a variety of plant ingredients ! high-dimensional target vector.
In order to realize the full potential of this approach the use of machine learning
methods for the analysis and nonlinear modeling of this data is recommended and
essential in a range of applications.
On the plus side, there are, among others, the following possibilities for con-
crete applications [12, 13, 14, 15]:
Instantaneous quantification of plant-based ingredients and diseases—non-con-

tact, immediate, no laboratory testing,
Significant extension beyond systems that have been available for a long time,
such as GPS control of agricultural machines based on soil models, weather and
environmental models.
as well as direct (mostly single-channel) measurement of one or a few parameters,

e.g. chlorophyll (UV range), nitrate content, water (via thermal imaging cameras).
The main areas of application pertaining to plants that have been identified so
far are listed in detail and graphically illustrated below.
1. Agriculture
Environmentally friendly and resource efficient use of operating supplies
(water, fertilizers, pesticides, etc.),
Qualitative (e.g. protein content, starch content, etc.) harvest forecasting,
not only quantitative (harvest quantity in tons),
Analysis of disease infestation before it visibly breaks out,
Analysis of storage condition.
2. Commercial plant breeding and fundamental biological research
Extension of the automated greenhouse (precision phenotyping),
Non-contact, direct measurement of suitable markers in the plant,
Both for traditional plant breeding and green genetic engineering (indepen-
dent of regional political or social consensus on green genetic engineering).
3. Traceability of the product chain
Seed supply, protection of breeders against illegal reproduction of improved
and optimized varieties,
Transport routes and storage of harvested crops,
Identification of origin (e.g. country or region) using biochemical markers
occurring naturally in the plants,
Consumer-centered sensors for determining freshness, lack of handling, ori-
gin, etc.
4. Optimization of agricultural logistics as well as wholesale and retail trade
Planning of agricultural machinery as well as transport, storage and process-
ing capacities through sensor-based extrapolation of harvest time, quantity
and quality,
Continuous supply of downstream units in the value chain with plant-de-
rived products,
Optimized delivery of seasonal plant-derived products.
5. Safety in genetic engineering
Evidence of genetically modified organisms from the laboratory in regards
to open-field cultivation and harvest (‘genetic’ contamination by outcrossing
or from the previous year’s planting) up to the processed product (food and
feed area, biological materials, bioenergy).
Application Scenario in Agriculture

The available data and/or information on, among other things, weather, genetic
variation and soil condition is expanded by a hyperspectral measuring system.
The data thereby collected is processed online using a ready-made mathemati-
cal calibration model combined with the farmer’s expert knowledge. This allows
Fig. 6.6 Application scenario in agriculture: Location adaptive application of operating sup-
plies in relation to plant nutrition and phytosanitary measures. (Fraunhofer IFF)
Fig. 6.7 Application scenario in commercial plant breeding and fundamental biological re-
search. (Fraunhofer IFF)
for, among other things, the location adaptive distribution of operating supplies in
relation to plant nutrition and phytosanitary measures (Fig. 6.6).
Application Scenario in Commercial Plant Breeding and Pure Biological

Research
The typical, existing sensor technology based on simple color images (RGB data)
is extended by the use of appropriate hyperspectral sensors. The added value for
the application is often that all existing data sources are aggregated for processing
and evaluation (Fig. 6.7).
6.4 Outlook
An important aspect relating to the phenotyping of crop plants is the way the
newly-developed sensors are presented. This ultimately determines whether a tech-
nology is adopted by the market. In addition to aspects such as the precision and
robustness of the collected data and of course the price, straightforward, flexible us-
age is also important. The more traditional applications for achieving high area out-
put e.g. as a handheld device, mounted on a farm machine, or even manned aerial
photography are increasingly being complemented by the use of multicopters for
example (Fig. 6.8). These are ideal for systematically sampling small to medium
areas with a high level of detail for pure research or on yield testing plots for plant
breeding purposes.
With the increasing availability and degression in price of multicopters as well
as the miniaturization of spectral sensors, this will become a common form of
spectral-based phenotyping for small to medium areas. This technology represents
a practicable compromise between flexibility and handling, spectral and spatial
resolution as well as price (investment or service).
Fig. 6.9 illustrates a practical application scenario from which the handling and
currently achievable spatial resolution may be derived. A section of an agricultural
test field (large picture, left) is shown with marked areas. These show the opera-
tors of the system at the edge of the field (top), the calibration standard used for
the spectral white balance (center) and the plant population of one plot (bottom).
Depending on the altitude and the cameras or lenses used, the proportions shown
can be adjusted upwards and downwards, at the expense or in favor of the resulting
area output. The picture below shows a brightness code (a color code in the orig-
inal), which shows different areas based on chemical composition such as paths,
borders, plant population. Using an application-dependent calibration model, the
phenotypes of the relevant crop plants in the plots can be biochemically resolved.
Fig. 6.8 Example implementation of a six-armed multicopter with mounted hyperspectral

camera operating in the VNIR wavelength range for flexible aerial recording of the health
and nutritional status of crop plant field populations. (Fraunhofer IFF)
Another interesting aspect that goes beyond the abovementioned inclusion of

spectral data for phenotyping concerns the presentation of the processed data as
information for the user in the context of the specific application. In this regard,
prototypes of technical systems have already been developed based on augmented
reality (AR).
Fig. 6.10 shows an example of a pair of AR glasses, which feeds the sen-
sor data processed in the context of the application into the user’s field of view
and superimposes it on their natural vision. A color code in the original image
indicates the existence of two different nutritional states (red—undersupplied,
blue—oversupplied). Optimal nutrition is typically not displayed, as it represents
the normal state.
In the example application shown in Fig. 6.10, data acquisition and visualiza-
tion take place at different times. With the increasing availability of miniaturized
spectral sensors and mobile computing power and broadband data supply in rural
areas, e.g. 5G, online methods that integrate spectral image acquisition, evaluation
by a pre-trained calibration model and visualization by means of AR glasses or
Fig. 6.9 a Agricultural test field and inset images: system operator (red), the calibration
standard used for the spectral white balance (yellow) and plant population of a plot (blue).
b The brightness code shows different areas based on chemical composition, such as paths,
margins and plant population. (Fraunhofer IFF)
Fig. 6.10 Using augmented reality (AR) to visualize processed spectral sensor data. (Fraun-
hofer IFF)
helmet are theoretically also conceivable. Again, this could be combined with the
use of multicopters. The user thereby acquires a flying spectral eye—another im-
plementation of the biologization of agriculture that is nothing short of spectacular.

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Cells as Sensors
Effect-Directed Rather than Concentration
7
Analysis
Stefanie Michaelis and Joachim Wegener
Summary
Cells are the fundamental base units of life and can be used as living sensors,
if the reaction of the cells to an external chemical, biological or even physi-
cal stimulus can be sensitively detected and converted into an electrical signal.
Physical signal transducers such as transistors, electrodes or optrodes are partic-
ularly suitable for this, because they allow cells to be examined non-invasively
and without labeling, even over long observation periods. The quantification of
biological effects is thus made possible through such biotechnological hybrids
of living cells and technical components, without having to rely on the use of
experimental animals. This article provides an overview of the possibilities for
culturing animal cells in a laboratory environment, describes the state of the art
with regard to signal transducers being used for analysis and concludes with
selected examples of the use of cellular sensors in drug testing and risk assess-
ment.
7.1 Introduction
The isolation and in vitro cultivation of cells from animal organisms in the labo-
ratory is a scientific method that can be traced back to the beginning of the 20th
century [1].
S. Michaelis () J. Wegener

Regensburg Branch Lab, Fraunhofer Research Institution for Microsystems and Solid State
Technologies EMFT
Regensburg, Germany

https://doi.org/10.1007/978-3-662-59659-3_7
106 S. Michaelis and J. Wegener
At that time, the motivation to do so arose (as it still does today) from endeav-
oring to investigate the properties of certain cells or cell aggregates under defined
external conditions, without being subject to the diverse and not very reproducible
variances within an intact organism. A deeper understanding of cellular physiolog-
ical events at a molecular level can usually only be obtained if it is possible to study
the cell species of interest in isolation from the complexity of its original biological
environment. The question of whether the knowledge gained from cells cultured in
the laboratory (in vitro) is transferable to their behavior in the organism (in vivo)
is the subject of a wide-ranging, fundamental debate to which there is no uni-
versally applicable answer. However, the proliferation and variety of applications
of cell culture technology and the resulting advances in technology and medicine
demonstrate the tremendous value of such in vitro studies, without wanting to ig-
nore the limitations of the method. Initially, primarily cells or tissue fragments of
cold-blooded donor organisms were used. Knowledge about the composition of
the media necessary for the sustenance of the cells was still so limited that cul-
tures were only successful in drops of blood. However, isolation and cultivation
techniques rapidly developed further. In 1951, the long-term culture and prolifer-
ation of human cells in the laboratory was possible for the first time, after cells
were taken from an aggressively growing cervical tumor [2]. The cells isolated
then were named “HeLa cells” after the initials of the donor, Henrietta Lacks, and
are still being used today, almost 70 years later, as a simple cell model in research.
Today, cell culture models have been developed for almost all animal tissues and
these are often, but not always, derived from tumor tissue. The unrestrained, long-
term and robust growth of tumor cells makes them particularly suitable as a starting
material for establishing cell culture models that are to be preserved and studied
under laboratory conditions. As has been impressively documented in the example
of HeLa cells, these tumor-derived cell lines are able to live and divide for many
years in the laboratory under suitable culture conditions—without necessarily fully
shedding the characteristic properties of the tissue of origin. Moreover, they can be
stockpiled practically by means of cryopreservation and stored almost indefinitely,
but also recultured as needed at any time, even at short notice. The different types
of cultured cells and their significance in certain areas of biomedical research are
systematically summarized in the following section.
7 Cells as Sensors 107
7.2 The Importance of Cell-Based Bioanalysis
The range of biomedical and biotechnological applications of cultured cells is

diverse and ever expanding, as isolation techniques and culture conditions are
constantly improved. Cultured cells serve as a model for detailed research on the
structure and functioning of the animal organism and its developmental biology
and pathologies. There is also just as much potential in application-oriented fields
such as drug discovery, risk assessment and chemical sensor technology. In the
context of the new and further development of pharmaceuticals, the extremely
productive methods of combinatorial chemistry and modern molecular biology
make large numbers of new potential drugs available every day. But their bio-
logical activity and effectiveness are questionable at first. Although initial basic
physicochemical assays can be used to screen out substances, an extraordinarily
large number of candidates still have to undergo a systematic activity test. While
the use of experimental animals may be indispensable for the final assessment of
a substance, in the early stages of development it is neither useful nor practicable,
particularly on ethical grounds, but also for reasons of time as well as economic
and bureaucratic factors. Moreover, the biochemical effects of an active substance
are very hard to study at a molecular level in the complex and often variable en-
vironment of a living animal. In view of the above, numerous in vitro assays have
been developed that do not require the use of animal models and can be performed
with high throughput (high throughput screening, HTS) under the reproducible
conditions of a laboratory environment. The biological system is frequently substi-
tuted by individual, isolated and purified biomolecules (proteins, nucleic acids) in
this test procedure in order to keep the number of molecular components involved
manageable. However, this reduction of the test system to the molecules directly
involved brings a disadvantage with it: Although an activation or inhibition of the
immediate biological target structure is determined, the triggering of the intended
physiological response of the cell is often dependent on a subsequent multilaterally
controllable signal transduction, and may not be observable in some circumstances.
Thus, for example, by isolating and reconstituting a cell surface receptor (a large
proportion of today’s drugs target cell surface receptors), the binding of a poten-
tial drug to the receptor can be detected and physicochemically characterized, but
whether or not the biological activity associated with the receptor is achieved or
effectively prevented can often not be answered by such assays.
The use of cultured cells as sensory elements in such assays (cell-based assays)
represents a kind of intermediate level between experimental animals and purified
receptors. The existence of cell culture models of the respective target tissue and
furthermore the availability of bioanalytical methods with which the response of

the cells to the administration of a particular active substance may be sensitively
registered are prerequisites for the successful application of cell cultures in phar-
maceutical drug and cytotoxicity screening. In many of the current biochemical
methods that are optimized for high throughput, the cells are often killed at a given
time during the experiment and the entire cell contents examined with regard to the
intracellular concentration of those metabolites, proteins or nucleic acids which are
presumed to relate to the cell function altered by the drug. An unexpected cell re-
sponse may remain undetected in this type of assay, which has such a precisely
defined specificity. In addition, these studies only allow the progression of the cell
reaction over time to be mapped to a very limited extent. As a rule, the cell reaction
is only analyzed after a predetermined exposure time (endpoint assays), since the
cells can thereafter no longer be reused. From a bioanalytical point of view, this has
the disadvantage that the temporal component of the cell reaction is not adequately
captured, even though this may often reveal informative details about the cellular
reaction. For example, the duration of exposure is a key criterion for potential toxi-
city in the risk assessment of a chemical. However, this criterion is only obtainable
in an end-point assay by tedious repetition of the experiments at different exposure
times, while continuous observation of the cells studied reveals the temporal evo-
lution in a single experiment. In order to continuously observe the cell reaction,
non-invasive and preferably label-free assays need to be used, which can function
without influencing the cells through the measurement itself, or introducing a re-
porter substance into the cell culture. In light of this, physical analysis methods,
which can detect the reaction of a cell population to a given stimulus in a time-
resolved manner without the contaminating influence of a reporter substance, are
receiving increasing attention. In this regard, microscopic assays as well as several
other non-imaging optical technologies have emerged in recent years, which have
the potential to become very important in the future. In all of these techniques the
cells are cultured directly on a physical signal transducer (noble metal electrode,
transistor, quartz oscillator, etc.), which is, on account of its functional principle,
able to non-invasively register certain functional or structural changes of the cells
by altering physical parameters (conductivity, refractive index, viscoelasticity etc.).
The most advanced signal transducers developed for this purpose are summarized
in Sect. 7.4. The combination of an appropriate cell culture model and a physical
signal transducer is what lends these biotechnological hybrids their unique and oth-
erwise almost unattainable level of performance, and adds another very promising
component to the portfolio of bioanalytical methods available.
7.3 Cell Culture Models for Various Bioanalytical

Applications
The cells used in cell-based assays or cell-based sensors in the sense described
above are by nature the most sensitive and at the same time most critical component
of the hybrid structure. The vitality and physiology of the cells must remain unaf-
fected by being cultured on the signal transducer and by the measurement process
itself, so that the results of the study remain relevant. This statement may seem triv-
ial, but it implies that the stringent cell culture conditions (37 °C, physiological pH,
incubation in isotonic fluids, adequate oxygenation, sterility) are maintained at all
times. Even if these basic conditions are met, the changes in the chemical environ-
ment associated with culture conditions in vitro and loss of the three-dimensional
architecture of the tissue almost invariably lead to changes in the phenotype and
differentiation of isolated cells. If the goal is to model the in vivo situation as pre-
cisely as possible, primary cell cultures are thus usually used (Fig. 7.1), which are,
as a rule, maintained in culture for only a short time after their mechanical or en-
zymatic isolation from the tissue of origin [3]. Cells in primary cultures display
the closest similarity to the corresponding cells in vivo, with regard to their dif-
ferentiation [3] when they are used immediately after isolation and not propagated
or stored. Longer-term culture and propagation ex vivo leads to an increasing loss
in their differentiation and to selection of those subtypes that can adapt to ex vivo
Fig. 7.1 Overview of the different cell types that can be used in cell-based sensors or cell-
based assays. (Fraunhofer EMFT)
laboratory conditions the quickest. The need to isolate these primary cells anew
for each study makes their use less attractive and more expensive, even though
their features with respect to physiology, enzymes and receptors resemble those
of cells in the tissue of origin the most. For example, primary cultured liver cells
(hepatocytes) are extremely valuable sensor cells for toxicological studies. How-
ever, published studies show that the freshly isolated hepatocytes begin to lose their
characteristic phenotype after just 24 h ex vivo. The time window for the advanta-
geous use of these cells is therefore very limited [4].
In the case of highly specialized research questions so-called co-cultures have
started to be used, in which such cell types are cultured together that are also in
contact with each other in vivo [5]. Constructs of this type can also be used as
sensor systems if the cell-cell interaction is able to remain unaffected by the signal
transducer [6].
If the most important goal is not the precise modeling of a specific anatomical
structure, but rather general cellular characteristics, then cell lines are often used
(Fig. 7.1). These can be cultured in vitro for a long time, continue to multiply,
and are generally easy to handle. They are derived from primary culture cells by
continuous culture and propagation in vitro. Initially, such cells have a finite life-
time limited to about 65 cell divisions (Hayflick limit) before they show signs of
cell senescence and eventually die off. On closer inspection, they are therefore re-
ferred to as a finite cell line. When spontaneous or experimentally induced genetic
transformation occurs in the course of culture caused by viruses or carcinogens,
these cells lose their growth limitation and their finite lifespan [4]. From this point
on, they can be cultured in the laboratory with virtually no time limitation, are
available in any desired quantity and are therefore also known as immortalized cell
lines.
Apart from their elemental metabolism and cell structure, these cells inevitably
show little similarity to their precursors in the body. The ability to store such cells
in the long term by cryopreservation and to only culture them when needed [3]
make them a highly attractive and versatile alternative to primary cell cultures.
A great variety of both finite and immortalized cell lines from virtually any tissue
of the human body is commercially available from cell depositories, so their use
does not require prior isolation from a donor organism.
In addition to cells such as these, isolated and derived directly from a living
organism, modern molecular biology can also customize cells for a sensory task
through genetic manipulation. This method has become extremely widespread in
modern biomedicine. These cells produce one or more surface receptors or en-
zymes in significantly higher copy numbers than is natural, so they can be used as
particularly sensitive sensors for the molecules that interact with these receptors
and enzymes. Concepts of this nature play a prominent role in the development of
pharmaceutical agents and have become indispensable in the repertoire of exper-
imental possibilities. An alternative approach is to equip wild-type (natural) cells
with a so-called reporter gene. The design is such that the corresponding protein
is only produced in the presence of the substance that is to be detected or in the
event of cell stress. The resulting proteins (e.g. fluorescent proteins or enzymes
that specifically activate a dye) can be quantified optically, so that the analyte to
be detected or cell stress is indirectly identified [7, 8]. Such reporter cells, which
are products of genetic engineering, have already been used to successfully detect
nanomaterials [9], heavy metals [7] or skin irritants [10]. The possibility of cryo-
preservation is a very important aspect for the use of these genetically modified cell
lines, since targeted genetic manipulation that aims to produce customized sensor
cells involves a considerable amount of work and can only realize its full potential
through being able to preserve the cells.
Stem cells have a special role to play as potential sensor cells because they can
be used to investigate differentiation processes and how they may be influenced
by external factors [11]. Studies of this kind are of especially great importance
in chemical risk assessments, particularly with regard to biological development
processes. Breakthroughs in the production of human induced pluripotent stem
cells (hiPS), produced by genetically reprogramming adult human cells, have again
greatly expanded the potential applications, by using complex differentiation pro-
tocols to generate human cell types that are usually difficult to access [11]. Thus,
it is already possible, and this trend will increase in the future, to use human cells
(from all tissues) as sensors, which by their nature have the greatest relevance for
biomedical research questions.
7.4 Non-Invasive Physical Signal Transducers

for Monitoring Living Cells
Physical signal transducers can detect the response of living cells to an external
stimulus of chemical, biological or physical nature without the addition of dyes,
fluorescent probes or other auxiliary reagents. Consequently, they have no molec-
ular specificity, but integrate individually across the different regions of the cell
body, and report on the integral refractive index, the integral electrical resistance,
changes in cell shape, or cellular micromechanics. This very pronounced integra-
tion across many individual cell structures makes these measurement principles
referred to as “holistic”, because they capture the cell as a whole or in large part.
Holistic methods are completely unbiased as a result of this distinctive feature that
they integrate over the entire cell body. Such a bias is inherent to many label-
dependent methods, however, because the need to choose a molecular probe (chro-
mophore, antibody) before conducting the experiment establishes the cell property
to be investigated a priori. All other cell properties or parameters are disregarded.
Data acquisition using holistic techniques is carried out non-invasively with very
low amplitude physical disturbance factors (current, voltage, mechanical deforma-
tion, light intensity), so that continuous observation of the cells is possible even
over very long periods of days or weeks, without the cells being affected. The du-
ration of the experiments is not determined by the readout in these methods, but by
the cellular sensors under the given experimental conditions. The great, frequently
underestimated advantage of this continuous data acquisition is the detailed infor-
mation it provides on the dynamics of the cellular response, which can otherwise
only be obtained with great effort.
Fig. 7.2 outlines the most sophisticated signal transducers for label-free, non-
invasive monitoring of cell-based assays. In all the illustrated methodological ap-
proaches, the cells grow directly on the surface of the signal transducer, which may
be the gate of a transistor, a planar gold film electrode or an optical waveguide. For
adherent cell cultures, a distance of only 30 to 200 nm separates the lower cell
membrane, adjacent to the substrate, and the surface of the signal transducer. This
very close spatial interaction between the cell and the technical surface is essen-
tial for the sensitivity of the methods displayed, but can only be influenced by the
experimenter to a very limited degree. The cells are initially suspended and then
sediment on the surface in a spherical form where they actively spread out if the
surface is coated with adhesion promoting proteins. After a few hours, an equilib-
rium distance to the surface is established, which can only be altered within certain
limits by a change in the protein coating of the technical surface. There are sev-
eral more measurement concepts over and above those illustrated in Fig. 7.2, but
these have not yet made it into routine use. The signal transducers illustrated can
be subdivided into those that allow the central energy metabolism of the cells to be
analyzed (a and b) and those that detect changes in cell shape, micromechanics or
the integral refractive index of the cell body. The latter type indicates a mass shift
within the cells and in the space between cell and surface (c to f). They all reflect
the integrated response of the whole cell and each is holistic in its own way, and
thus applicable to many different experimental scenarios.
Fig. 7.2a outlines the concept of using a transistor with an ion-sensitive gate
(ion-sensitive field-effect transistor, ISFET) as a signal transducer. The electri-
cal signal of the transistor therefore becomes dependent on the concentration on
the gate surface of the ion for which the ISFET was designed, thereby permitting
its integral quantification. Of particular importance in the context of cell-based
a d
b e
c f
Fig. 7.2 Non-invasive physical signal transducers for the monitoring of adherent cells in
cell-based assays. a ion-sensitive field-effect transistor (ISFET), b fluorophore-doped poly-
mer matrix, c impedance analysis of cell-covered film electrodes, d piezoelectric shear wave
resonator, e surface plasmon resonance, f optical waveguide. (Fraunhofer EMFT, according
to [26])
assays is the use of a proton-sensitive ISFET, which measures changes in pH be-

neath the cells located on the gate [12, 13]. The background to this approach is
that the metabolic activity of cells is inextricably linked to the secretion of pro-
tons, which result in extracellular acidification. Owing to the tiny volume between
the cell membrane and the gate surface (see above), even minute changes in pro-
ton secretion are sufficient to register changes in the central energy metabolism.
Light addressable potentiometric sensors (LAPS) were groundbreaking for the de-
velopment of metabolic sensors. In these, a photocurrent induced by point-like
illumination from the rear side is measured by a light-sensitive semiconductor if
cells have grown on the front side of the semiconductor [14–17]. The potential
of the surface facing the cells is again pH dependent and alters the induced pho-
tocurrent. In this way, it is also possible to detect a change in metabolic activity
of the cells on the front side with LAPS. In contrast to the ISFET transducers, the
measurement can be carried out in a spatially-resolved manner as the laser beam
responsible for the rear illumination is scanned across the surface pixel by pixel and
the corresponding local photocurrent is registered. ISFET measurements are how-
ever integral for all cells growing on the gate. A comparative study has confirmed
very similar analytical characteristics such as sensitivity, drift and reaction time for
both methods [18]. What the two methods have in common is that they both have
to be operated under pulsating flow in order to repeatedly dissipate the released
protons and to guarantee a constant chemical milieu for the cells over time. The
analytical readout is therefore the extracellular acidification rate (EAR) rather than
an equilibrium pH value. Alternatively, the oxygen consumption rate (OCR) of
the cells can be determined by amperometric methods using a miniaturized Clark
electrode or similar approaches based on ISFETs. In contrast to measuring the oxy-
gen consumption of living cells, extracellular acidification provides information
on both the anaerobic and aerobic metabolism. Devices that allow measurement of
both parameters (EAR and OCR) for a cell population and thus provide a differen-
tiated picture of the metabolic situation at any time of exposure have the greatest
information content.
A noteworthy alternative to the electrochemical oxygen and pH determina-
tion described above are technologies based on luminescence (Fig. 7.2b). These
approaches use luminophores whose luminescence is shifted in wavelength as
a function of pH value or quenched in its intensity as a function of oxygen concen-
tration. The luminophores do not come into direct contact with the cells, but are
embedded in a hydrogel (pH) or an oxygen-permeable polymer. These methods
are therefore also regarded as label-free approaches [19].
In addition to the quantification of extracellular pH, which is applicable to all
cell types, field-effect transistors (FETs) [20–22] and planar noble metal electrodes
[23] play a major role in recording the action potentials of electrically active cells
such as neurons or heart muscle cells. Action potentials are waves of membrane
depolarization that surround the whole cell and can be detected as extracellular
changes in potential. The cells are not punctured with a microelectrode as they
are in patch clamping, an established method for deriving such action potentials,
but can spread undisturbed on the gate of the FET or the metal electrode. Planar
FETs are therefore eminently suitable for non-invasively registering the effects of
all types of stress on neuronal activity and conducting a quantitative analysis. Of
particular interest are arrays of FETs or noble metal electrodes that capture the
electrical activity in a culture with spatial and temporal resolution and thereby al-
low cell communication to be mapped over extended areas of a neuron population.
The approach illustrated in Fig. 7.2c is also based on planar noble metal films.
This was originally referred to as electric cell-substrate impedance sensing (ECIS)
and has a widespread commercial distribution nowadays [24, 25]. When ECIS is
used, the cells to be examined are cultured on planar gold film electrodes that are
less than 100 nm thick and are applied to the bottom of a conventional petri dish or
a multiwell plate using established thin-film technology. A small current of a few
microamps is used to measure the AC resistance (impedance) of the electrodes. As
cells adhere to and spread out on the electrodes, the current is forced to flow around
the dielectric, poorly conductive cell bodies on extracellular pathways—below and
between the cells. The measured impedance of an electrode that is completely
covered with cells is thus determined by the dimensions of these electrolyte-filled
interstices and thus by the cell shape. However, unlike in microscopy, changes in
cell shape are not seen in an image, but are recognized by changes in the measured
impedance during an ongoing experiment. In this process, the observed impedance
changes integrate over the entire cell body, including cell-cell contacts, the cell-
surface contacts and the cell membrane. In this way, changes in cell shape of the
order of less than 50 nm can be detected, which is an order of magnitude less than
the resolution limit of conventional optical microscopes.
As described in the following application examples, the synchronous contrac-
tions of beating cardiac muscle cells can be registered just as well as the stochastic,
uncoordinated shape fluctuations of non-beating cells. In doing so, the temporal
resolution ranges from milliseconds to hours. These specifications make ECIS
a very versatile signal transducer for the continuous monitoring of adherent cell
cultures while they are exposed, for example, to drugs, toxins, nanomaterials or
xenobiotics in general [27–30]. The need for a high sample throughput in drug or
risk assessments has been addressed by the development of 96- or 384-well elec-
trode arrays.
Piezoelectric signal transducers are also used when cell responses are regis-
tered in cell-based assays, as outlined in Fig. 7.2d. The most advanced of these
is the quartz crystal microbalance (QCM), which was originally developed for the
mass-sensitive analysis of adsorption reactions at solid-liquid interfaces [31]. The
core component is a thin, disk-shaped crystal oscillator made of ˛-quartz crystal
that is covered on both sides with thin gold film electrodes for making contact to
electronic drivers.
If an alternating voltage is applied to these surface electrodes, a resonant shear

oscillation is produced in the quartz parallel to its planar surfaces, due to the piezo-
electric coupling. This mechanical oscillation of the quartz disk is highly sensitive
to all adsorption and desorption processes on the surface, which can be recognized
by a corresponding change in the resonance frequency [31]. It is also possible to
extract material properties of a material adsorbed on the quartz surface, such as its
viscosity or elasticity [32, 33] from the measured data. In this way, changes in the
micromechanics of the cells adhering to the quartz can be measured quantitatively
via this signal transducer and made accessible in a time-resolved and integrated
manner for a population of several thousand cells. The ability to observe mechan-
ical changes in a cell assembly has resulted in a multitude of applications [34, 35]
including the observation of cells under the influence of toxins [36] and nanoma-
terials [38, 39]. Despite the shear oscillation necessary for the measurement, the
QCM method is considered to be non-invasive, since the oscillation amplitude is
a maximum of one nanometer in the center of the quartz, under the usual experi-
mental conditions, and falls off towards the edges in the form of a Gaussian profile.
The quartz disks have very high resonant frequencies of several megahertz, so the
combination of this high-frequency deflection with only small amplitude excludes
the possibility of the cells being affected. The temporal resolution of the QCM
method can be reduced to just a few milliseconds, which is more than adequate
for the vast majority of biomedical exposure scenarios associated with a change in
cellular micromechanics.
Surface plasmon resonance (SPR) spectroscopy is the most widely used label-
free method in molecular bioanalysis. Similar to the QCM technique described
above, SPR permits the quantitative analysis of adsorption processes at the solid-
liquid interface. Typically, an interaction partner is immobilized on the surface of
an SPR chip while the potential binding partner is made available in the supernatant
solution. If binding occurs at the interface, the refractive index in the vicinity of
the surface changes, which can be sensitively read out. As indicated in Fig. 7.2e, it
is possible to measure the tiniest changes in the refractive index on the surface of
a conductive material. In the classic Kretschmann configuration, the base of a glass
prism is coated with gold by vapor deposition. This thin gold layer serves as the
mass-sensitive signal transducer. If monochromatic light strikes the interface at an
angle greater than the critical angle of total reflection, quasi total reflection of the
incident light occurs. A small portion of the electric field strength passes beyond
the interface however and generates an evanescent electric field in the vicinity of
the surface that causes the mobile electrons in the conduction band of the gold
to oscillate. The resonance condition for these density fluctuations of the surface
electrons (plasmon) depends on the refractive index in the adjacent medium, which
can therefore be quantified very sensitively based on the intensity of the reflected
light. In response to an external stimulus, cells grown on such an SPR chip pro-
duce a measurable change in refractive index, which is attributable to a dynamic
mass redistribution (DMR) in the cell [40–42]. The SPR readout is also integral
and without molecular specificity in the context of cell-based assays and sensors,
but for this reason it has a similarly wide range of applications to the previously
described ECIS method. Typical areas of application include preclinical drug test-
ing [43, 44] as well as risk assessment of xenobiotics [45]. For data analysis and
interpretation, it is of great importance that the penetration depth of the SPR signal
for commonly used measuring devices is only 100 to 200 nm, so that the cell body
can only be “looked into” up to this depth. The experimentally measurable dy-
namic mass redistribution is therefore limited to the lower portion of the cell body
and the gap between cell membrane and the surface. Processes taking place on the
upper cell membrane are not detectable by this method under standard conditions.
A closely related technique that also determines changes in adherent cells based
on refractive indices is based on the use of resonant waveguide gratings (RWG).
As shown in Fig. 7.2f, a waveguide of this kind behaves like an optical filter. Only
one wavelength of the polychromatic, white excitation light is propagated through
the waveguide and decoupled. The wavelength of the transmitted light is deter-
mined by the refractive index of the sample that is in contact with the waveguide.
Similarly to SPR, this technique can be used to detect dynamic mass shifts in the
cells with extremely high sensitivity. This technique, which is commercially avail-
able in 384-well format, has made significant contributions to the understanding
of pharmacological problems in recent years. The RWG technique does not only
resemble SPR in the fact that changes in refractive index are quantified, but also in
its limited sample penetration depth of 100 to 200 nm.
7.5 Cells as Sensors—Example Applications of Integral

Effect-Directed Analysis
The operating principles of cell-based sensors are elucidated by means of three
examples in the sections that follow. Unlike typical analytical questions, these
methods do not seek to quantify an analyte in concentration units, but rather to
quantify its effect on the sensor cells. In bioanalysis, the term “effect-directed
analysis” (eda) has become established for this analytical strategy. For complex
samples, such as environmental samples, the sample is first broken down into frac-
tions using conventional separation techniques, each of which is then analyzed for
its effect on the sensor cells.
Further fractionation of the first positive fractions progressively isolates the

toxic substance and ultimately allows it to be quantified by chemical methods. This
type of sample analysis is very goal-oriented in that it isolates a bioactive compo-
nent in a sample due to its functional significance, and the complex methods of
chemical analysis thus need only be applied to this subset of the whole sample.
7.5.1 Impedance-Based Analysis of Changes in Cell Shape

Under the Influence of External Factors
When studying the toxicity of heavy metals, it is preferable to use kidney cells as
sensors, because these cells are involved in detoxification following heavy metal
exposure, in vivo. The signal transducer used in this case is the ECIS method
(Fig. 7.2c), where the cells grow on planar gold film electrodes and any cell shape
changes during exposure are detected by measuring the impedance. Fig. 7.3a shows
a cell-covered gold film electrode with a diameter of 250 micrometers, as is typi-
cally used. Cell death due to toxic exposure always results in cell shape changes,
but the mechanism of cell death determines whether the cells shrink or swell.
a b
Fig. 7.3 a Cell-covered gold film electrode for the impedimetric analysis of changes in cell
shape. The circular electrode has a diameter of 250 µm. b Variation of impedance |Z| over
time at a measurement frequency of 4 kHz when densely grown layers of kidney cells (NRK)
on the ECIS electrodes are brought into contact with increasing concentrations of cadmium
chloride. For lower concentrations the toxic reaction only sets in after several days. (Fraun-
hofer EMFT)
If cells start a genetically programmed self-destruction process (apoptosis) un-

der the influence of a toxic substance, the cell body will shrink and be split into
fractions. If the cells simply die under the influence of external toxic stress without
their own intervention (necrosis), the cells swell and finally burst. Fig. 7.3b shows
the variation of the impedance over time for an electrode completely covered with
rat kidney cells (normal rat kidney, NRK; finite cell line) under the influence of
increasing concentrations of the heavy metal salt, cadmium chloride.
The increasing concentrations of cadmium chloride (CdCl2 ) are added after
about 12 h. The initially high impedance of the cell-covered electrodes decreases
dose-dependently, as a result of the onset of cell poisoning, since the bodies of the
kidney cells contract under the influence of the toxic heavy metal. The contraction
of the cell bodies and loss of attachment to the culture substrate are typical cell
responses to an incipient poisoning. The illustrated toxicity study shows that the
cells can be monitored continuously with the ECIS method over several days, and
the long-term toxicity of smaller cadmium concentrations (30 µM) can also be re-
vealed. A similarly detailed analysis using endpoint assays can only be achieved
with an enormous amount of work and material input.
Fig. 7.4 shows a related example of a toxicity study in which the same kidney
cells as shown in Fig. 7.3 were exposed to increasing concentrations of carbon
a b
Fig. 7.4 a Time course of the normalized impedance at a measuring frequency of 4 kHz
for gold film electrodes covered with a continuous layer of NRK cells. The cells were
incubated with increasing concentrations of carbon nanoparticles at time t = 1 h. b Classi-
cal biochemical cytotoxicity assay after 12 h exposure time. The quantitative analysis gives
a value of (0.48 ˙ 0.02) mg/mL for the half-maximal effective concentration EC50. (Fraun-
hofer EMFT)
nanoparticles. The risk assessment of nanomaterials has become a highly regarded

and much-addressed field of research because of their widespread distribution. In
this example, the impedance was normalized, that is, the first point measured after
the addition of the nanoparticles is taken as a reference point. At the smallest doses
of up to 0.3 mg/mL, no influence on the cells can be detected over the entire mea-
suring period of 24 h. The first cell reactions to a concentration of 0.4 mg/mL are
detectable after an exposure time of 12 h. Higher concentrations of carbon nanopar-
ticles reduce the impedance to the value of a cell-free electrode within ten hours,
indicating either a complete cell rounding or permeabilization of the cell mem-
brane.
By comparison, Fig. 7.4b shows the dose-response curve of a classic biochem-
ical vitality test for a 12-hour exposure time. The assay yields a concentration of
approximately 0.5 mg/mL as the concentration of half-maximal cell impact, which
is very consistent with the impedimetric data. The advantage of continuous cell ob-
servation, however, is that based on the dataset shown in Fig. 7.4a a dose-response
relationship can be drawn up for any exposure time between zero and 24 h, with-
out having to conduct a further experiment. When a biochemical endpoint assay is
used, the entire experiment must be repeated for any other exposure time.
Fig. 7.5 illustrates impedimetric cell observation particularly clearly using the
example of cardiac muscle cells cultured in the laboratory on ECIS electrodes. Af-
ter a continuous cell layer has been established on the electrodes, these cells begin
to contract synchronously and rhythmically in much the same way as in the intact
cardiac muscle. This periodic cell shape change manifests itself in the recorded
impedance profile through corresponding peaks, from which both the frequency of
the “heartbeat” and its amplitude may be quantified.
Fig. 7.5 High temporal

resolution impedance pro-
file for a population of
rhythmically beating heart
muscle cells on an ECIS
gold film electrode. (Fraun-
hofer EMFT)
The combination of beating heart muscle cells (cardiomyocytes) and an im-

pedimetric signal transducer is currently widely used to detect cardiotoxicities, for
example in connection with drug side effects, without having to resort to the use
of experimental animals. The possibilities offered by stem cell technology allow
human heart muscle cells that are obtained by reprogramming adult cells from
other human tissues to be used in these experiments. In this way, the otherwise
commonly observed differences between different species with regard to their sen-
sitivity to drugs or toxins may safely be circumvented, which would otherwise lead
to a residual uncertainty in the assessment of the study results.
7.5.2 Analysis of Cytomechanical Changes with Piezoelectric

Resonators
The micromechanics of animal cells are primarily determined by a system of in-

tracellular protein filaments. This cytoskeleton within the cells is crucial for their
mechanical stability, since animal cells, in contrast to plant cells, do not have a me-
chanically resilient cell wall. Further, micromechanical changes of the cell body
are also involved in many physiological and pathophysiological processes. In light
of this, techniques for studying cytomechanics from a biomedical point of view are
very important and in great demand. The gold standard for such investigations is
scanning force microscopy (SFM) whereby individual cells are mechanically de-
formed and the force required to do so is registered. The offsetting of deformation
and force results in descriptive quantities such as Young’s modulus for character-
izing the cell stiffness. SFM provides a subcellular lateral resolution that allows
for accurate mapping of cytomechanics, even in subregions of a single cell. How-
ever, studies carried out on a cell ensemble are very time consuming and can only
be conducted under physiological conditions to a limited degree. Furthermore, as
part of the measurement process, the cells are subjected to considerable mechan-
ical strain. An alternative, integral approach is based on the QCM technique, in
which the cells are cultured on the surface of piezoelectric quartz resonators. The
acoustic impedance of the cells at the surface and thus cytomechanical changes
can be retrieved by analyzing the shear oscillations with amplitudes of less than
one nanometer. The readout integrates over the entire cell population on the quartz
surface and thus captures tens of thousands of cells in one measurement process.
The temporal resolution can be optimized down to the range of seconds. This non-
invasive measurement takes place in the environment of a regular cell culture in-
cubator and thus under physiological conditions.
a b
Fig. 7.6 a Fluorescence micrographs of the actin cytoskeleton in a population of canine

kidney epithelial cells (MDCK) before (A) and after (B) exposure to 5 µM cytochalasin D,
which is known for the breakdown of filaments. b Progression of load impedance of a quartz
resonator continuously covered with MDCK cells under the same experimental conditions
over time. The change in the load impedance |ZL | is proportional to the change in acoustic
impedance. (Fraunhofer EMFT)
Fig. 7.6 shows fluorescence micrographs of a population of canine kidney cells

in which a portion of the intracellular cytoskeleton has been stained. In Fig. 7.6a
the undisturbed organization of the filaments can be seen in a thick filament belt
and nearly parallel aligned intracellular filaments. Fig. 7.6b depicts their state after
exposure to the fungal toxin cytochalasin D, which completely breaks down this
part of the cytoskeleton to structureless aggregates, within 50 min [35].
By following the action of cytochalasin on the cells by means of piezoelectric
resonators, the progression of the load impedance over time as shown in Fig. 7.6b
is obtained, which is proportional to the acoustic impedance of the cells. The time
course of the load impedance |ZL | documents the time-resolved “softening” of
the cells under the influence of 5 µM cytochalasin D until a new equilibrium state
is reached after about 50 min. The QCM technology can potentially permit paral-
lel operation, so that in future a moderately high sample throughput will also be
possible with this micromechanical sensor.
7.5.3 Quantitative Drug Testing with Sensor Cells and

Evanescent Fields
Drug development often focuses on the potential stimulation of cell surface recep-
tors, including the activation of subsequent signaling cascades. The largest class
of cell surface receptors is the G-protein coupled receptors (GPCR). These are the
target of more than 40% of prescription drugs on the market today and they are
therefore receiving a lot of attention in current drug development. The histamine
receptor, involved in many allergic reactions in the body, belongs to this class of
receptors.
Fig. 7.7a shows the culture of human U373 cells (finite cell line) on SPR sub-
strates presenting the histamine H1 receptor on their surface. If these cells are
brought into contact with histamine, then the time curve of the reflectivity R shown
in Fig. 7.7b is observed, which indicates a dynamic mass redistribution in the cells
as a result of the receptor activation in very precise temporal detail.
a b
Fig. 7.7 a Stimulation of cell surface receptors with activating ligands triggers signal cas-
cades in the cell, which bring about a dynamic mass shift. This can be analyzed by means
of time-resolved SPR measurements. b The sensograms show the reflectivity over time dur-
ing incubation with the agonist histamine, with or without preincubation with an antagonist
(blocker) such as mepyramine, using human U373 cells as an example. (Fraunhofer EMFT)
If, however, the cells are previously incubated with a histamine antago-
nist—which binds to the receptor but does not activate it and subsequently blocks
it (Fig. 7.7)—then the cell response is not triggered even if histamine is added. It
is important to emphasize that also the antagonist—in the example shown above,
the H1 antagonist mepyramine—binds to the receptor, but this binding does not
trigger a cell reaction. Only when using an activating ligand (agonist) such as his-
tamine itself, does a cell reaction occur. This example clearly demonstrates that the
hybrids of cellular sensors and physical signal transducers described here allow for
an effect-directed analysis that is not usually used to quantify an analyte, but rather
to analyze its influence on a living biological system. Dose-response relationships
of agonists and the inhibitory potency of antagonists can be quantitatively deduced
from raw data of this type.
7.6 Conclusion
Hybrid sensors made up of living cells and physical signal transducers are char-
acterized by their wide range of applications and their high level of automation
and medium to high throughput. The effectiveness of the different approaches
demonstrated here in just a few examples can be transferred to many drug and
cytotoxicity studies. The systems described here stand out for the non-invasive na-
ture of the measurement (which also allows long-term effects to be registered), the
independence from analytical labels and the temporal resolution of the experiment,
which allows for the uninterrupted documentation and analysis of a dynamic cell
reaction. Creative developments of new or optimized signal transducers, innovative
data acquisition and analysis routines as well as the combination of different signal
transducers in a single measurement setup—also including electrical manipulation
methods—are continuously extending the field of application and will in future
allow further fundamental parameters of cell physiology to be quantitatively deter-
mined in a wide range of experimental scenarios.

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Biopolymers – Function Carriers
in Materials Research 8
Polymeric Materials with Biological Functions and
Biomaterials for Medicine
Alexander Böker
Summary
Bionic systems that follow the approach of “learning from nature” have been
around for many years. However, the fusion of biology and materials is cur-
rently being completely rethought at the Fraunhofer Institute for Applied Poly-
mer Research IAP. Modern methods of molecular biology, biotechnology, poly-
mer chemistry and materials science are enabling the development of innova-
tive functional materials with outstanding properties and functions in interdis-
ciplinary research projects. For this purpose, biomolecules such as proteins,
peptides or carbohydrates are incorporated directly into polymers, thus trans-
forming their natural function into a material. Filtration through protein pores,
biocatalysis in thin films and sugar-mediated diagnostics become possible. The
next step is implemented based on these advances at the molecular level: Strate-
gies are developed for bringing together labile biomolecules and thermoplastic
polymer processing—seemingly irreconcilable opposites—to produce biofunc-
tional plastics. For medical applications, the need for congruence between ma-
terials and biology has been known for a long time. New implants allow even
more precise control of the interaction with tissues, made possible by accurate
knowledge and modification of the material properties in interplay with cells in
complex biological systems.
The biological transformation of polymers is in full swing, leading not only
to future-oriented materials that are sustainable and functional, but also to ma-
terials that enhance engineering processes and facilitate new therapies or di-
A. Böker ()
Fraunhofer Institute for Applied Polymer Research IAP
Potsdam, Germany
https://doi.org/10.1007/978-3-662-59659-3_8
130 A. Böker
agnostic platforms. Plastic material designers can not only learn from nature,
they also need to develop plastics that use nature and deliberately interact with
it—this is the next step in the evolution of polymer materials.
8.1 Introduction
The term biopolymers in its narrower sense is applied to polymers which are
of direct biological origin—especially cellulose, starch, chitin or proteins—but it
also refers to polymers whose monomer units were manufactured based on re-
newable raw materials (e.g. cellulose, starch or sugar). Under older definitions,
biodegradable polymers (such as polylactic acid/PLA or Ecoflex made by BASF)
were generally classified as biopolymers and bioplastics, whether based on fossil or
natural resources. The Fraunhofer Institute for Applied Polymer Research IAP has
longstanding key competencies in the development of materials based on the afore-
mentioned biopolymers and now defines the term biopolymer even more broadly
to include polymers that are used as functionalized biomaterials in a biological en-
vironment or additionally possess certain biological functions. Fraunhofer IAP is
breaking new ground with this approach, especially in terms of biological func-
tionalities, which go far beyond the familiar bionic approaches. The biofunctional
polymers are not only modeled on nature in terms of their structure and functional-
ity, but also receive their function through direct incorporation or other integration
of biological building blocks from nature (e.g. proteins, peptides or glycopoly-
mers/sugars) into the respective polymeric material. This has the advantage that in
some cases multifunctional systems are used that have been optimized over mil-
lions of years, whose highly specific functions can hardly be optimized further. In
nature, proteins act, among other things, as catalysts, recognition motifs, enzymes,
transporters and nanochannels. However, the fact that the proper folding of the
protein structure is essential for these functions presents a great challenge. In non-
natural environments, the stability of proteins is a critical factor. One objective is
therefore to protect the biomolecules from denaturation when they are embedded
in a polymer material. The situation is more favorable for antimicrobial peptides
or for glycopolymers/sugars. Another approach follows the adaptation of artificial
materials to natural environments for use as medical materials for implants.
In this chapter, we describe current studies being conducted at Fraunhofer IAP.
To begin with, building blocks and features of interest are introduced, thereafter
possibilities for integrating biological functions at the molecular level, on polymer
surfaces, or in corresponding bulk materials are demonstrated. This is followed by
8 Biopolymers – Function Carriers in Materials Research 131
an overview of work being done at IAP on biofunctionalized plastics as biomateri-

als for medicine, using the example of a keratoprosthesis in ophthalmology.
8.2 Introduction to Biological Building Blocks/Functions
In the past decades, globular, soluble proteins were modified in multiple ways and
incorporated into synthetic hybrid materials. Over the last few years, another class
has been targeted, namely the transmembrane proteins. Here, the task is com-
plicated by the fact that membrane proteins occur in lipid membranes in natural
systems and therefore have pronounced hydrophilic and hydrophobic sections. The
hydrophobic region of a transmembrane protein must be especially protected by
stabilizing agents so that the proteins are soluble in water at all. Certainly, the mim-
icking of cell membranes by producing synthetic analogues represents a promising
research goal, so a variety of strategies have been developed for embedding mem-
brane proteins in a polymer matrix. One of the largest transmembrane proteins is
FhuA (ferric hydroxamate uptake protein component A). It is a monomeric channel
protein composed of 22 antiparallel ˇ-sheets, which is found in the outer mem-
brane of E. coli bacteria [1] where it acts, inter alia, as a transporter for an iron-
peptide complex. FhuA can be easily engineered using biotechnological methods.
For example, variants have been produced that differ from the wild-type protein
by having a protein channel that is completely open or one that is enlarged by the
insertion of further ˇ-sheets [2, 3]. Fig. 8.1 shows the structure and dimensions of
FhuA and the distribution of the hydrophilic and hydrophobic sections.
The equally interesting class of hydrophobins includes proteins with a pro-
nounced, chemically-stabilized amphiphile that therefore behaves similarly to sur-
factants [5, 6]. Their ability to assemble on surfaces in thin, structured monolayers
can be used to reverse the polarity of polymeric surfaces. Their immobilization
on, for example, polar polyamide or cotton makes the surface of the latter hy-
drophobic, i.e. water-repellent, and thus results in increased dirt repellency similar
to the lotus effect. Non-polar polymer materials such as polyesters can similarly
be equipped in such a way as to make them either oil-repellent or water-compati-
ble, thereby improving the water absorption capacity of synthetic fibers and hence
also the wearing comfort. Hydrophobin coatings can be applied, for example, by
spraying and do not require complex preparation of the polymeric surface.
Surfaces that are functionalized with enzymes and thus enzymatically active
can be obtained by applying preparations of the enzyme with polymeric matrix
material and/or crosslinking agents to the selected surface by means of various
coating methods [7, 8]. For this, spray application or knife coating are examples
132 A. Böker
Fig. 8.1 Structure of FhuA wild-type and the ACVFtev variant with fully opened protein
channel in side and top view. [4]
that come into question. Alternatively, appropriate layers can also be produced
at a water-air interface and then transferred to the substrate by means of suitable
transfer techniques. Enzymatically active surfaces can be used for materials with
antimicrobial properties, for biocatalysis and for sensor technology.
Short peptides (e.g. antimicrobial peptides) usually have no tertiary structure
and therefore display greater stability under the usual polymer processing condi-
tions such as high temperatures, or can even be used as biomaterials themselves
[9, 10]. It is therefore also possible to process these together with bulk materials
following conventional injection molding or extrusion methods. Loading a poly-
meric molded part with peptides over its entire volume ensures that even when the
workpiece is subjected to abrasive stress, a functional surface is always available.
In doing so, even low peptide concentrations are sufficient to achieve an effect,
which underscores the cost effectiveness of their application.
Sugars, also known as glycans, play an essential role in numerous biological
processes over and above their role as an energy source and supplier. In this way
they are involved in friend-enemy recognition and the immune response, mediate
the formation and maintenance of cell aggregates, support the viscoelasticity of
joints or act as a protective layer on cells against environmental influences [11,
12, 13]. In addition, they act as supporting substances, for example in plants (cel-
Fig. 8.2 Glyco-polymer coated surfaces (treated) following incubation with eGFP-express-
ing E. coli compared with uncoated surface (untreated). The diminished fluorescence shows
that hardly any E. coli adhere to the treated surface. (Fraunhofer IAP)
lulose) or insects (chitin). Particularly the latter is already frequently found in its
deacetylated form as chitosan in biomaterials for the medical sector.
Introducing glycans into materials as functional agents can lead, for exam-
ple, to materials, which microorganisms such as bacteria can no longer adhere
to (Fig. 8.2), or which can be completely resterilized by wiping with water. A con-
ceivable application in this regard is the development of paint for biocompatible
antifouling.
In order to avoid depletion of functionality however, sugars can also be incor-
porated directly into the workpiece in the form of glycopolymers, since these can
be processed at higher temperatures. When skillfully linked to biocompatible poly-
mers, new bio-based adhesives that can be controlled with regard to their durability
are just as easy to obtain as new water-based lubricants. The incorporation of gly-
cans into suitable bulk materials can be used for the production of cost-effective
filtration systems for particles, heavy metals, micro- and nanoplastics and even for
microorganisms, and thereby harness some of the biological versatility of sugars
in plastics.
The use of highly controlled sugar structures opens up the field of medical tech-
nology and diagnostics [14, 15]. Thus, new biosensors are becoming available for
the rapid diagnosis of bacterial infections or intelligent implant coatings. Such im-
plant coatings can act as cell recruiters and thus improve the implant’s ingrowth,
or they may have a cell-repelling effect and thus prevent implant ingrowth, even
if only partially. The biocompatibility of the polymers means that neither of these
options has a negative effect on the immune system.
134 A. Böker
8.3 Integration of Biological Functions in Materials
8.3.1 Integration at the Molecular Level
Stefan Reinicke and Ulrich Glebe
If the aforementioned proteins with their multitude of functions are incorporated

into different materials, they can be combined with or interact with the material
surrounding them in different ways at the molecular level. The protein is not only
simply embedded in a macromolecular network, but may also have fixed chem-
ical linkages to a substrate via its own reactive groups. In this process, the type
of linkage as well as the structural characteristics of the material are of crucial
importance for the application that is ultimately targeted. Usually, proteins can
be attached to the surrounding material via a chemical bond or through attractive
interactions. These interactions can be so strong that, despite the lack of cova-
lent attachment, a firm, lasting bond between protein and surrounding material is
ensured. This applies, for example, to electrostatic interactions that utilize the pro-
tein’s surface charges. Then again, proteins usually have hydrophobic structural
elements on their surface, which in turn allow corresponding hydrophobic interac-
tions with a suitable material. One much-noted example of a hybrid material based
on such physical interactions and that consists of protein and a synthetic compo-
nent would be enzyme conjugates with graphene-like materials [16]. Graphene has
outstanding electromechanical properties and serves as a conducting medium for
an electrical signal that is generated by the enzymatic conversion of a specific an-
alyte. The most prominent example of such an enzyme-based redox system is the
oxidation of glucose by glucose oxidase. Systems based on the inclusion of a re-
dox-active enzyme in a thin, electrically conductive polymer layer on an electrode
surface by electropolymerization [17] will be aimed at similar applications. The
minimal thickness of the resulting enzymatically active layer, as well as its elec-
trical conductivity are ideal for the construction of fast-reacting, highly selective
sensors for bioanalytics.
Many examples of the non-covalent incorporation of proteins into a polymer
matrix are also to be found for the insertion of membrane proteins into polymer-
some membranes [18]. Polymersomes are vesicles where an amphiphilic block
copolymer forms a membrane in an aqueous medium. The insertion of protein
channels into the polymer membrane allows for selective transport into or out of
the vesicles. Reaction spaces are thus created in the interior of the polymersome.
For example, enzymes can be entrapped in the polymer vesicles and the substrate
Fig. 8.3 A selection of various chemical reactions for the covalent bonding of proteins to
synthetic materials. (Fraunhofer IAP)
can enter the vesicles through the protein pores. In this way FhuA variants with
open protein channels (Fig. 8.1) were inserted into polymersome membranes to fa-
cilitate the transport of DNA [19]. The protein channel can also be blocked initially
and only opened to allow transport when triggered by a reduction reaction [20] or
light [21]. Polymersomes with inserted aquaporin Z (AqpZ) could be spread on
porous substrates to obtain a planar membrane with incorporated protein channels
[22].
However, physical interactions are not always suitable as a principle for the
binding between protein and polymer matrix, because they are not always strong
enough or they may occasionally be significantly weakened by changes in envi-
ronmental parameters. Too much disruption of the structure of the protein by such
interactions may also entail a significant loss of protein functionality. For this rea-
son, in many cases researchers revert to the covalent attachment of the protein.
Functional groups of the protein which are easily chemically addressable are used
for this purpose. These include, above all, the primary amino groups of the ly-
sine residues and the thiols of the cysteine residues, since these are available for
a large number of chemical reactions [23] (Fig. 8.3). Other amino acid residues as
well as the N- and C-terminus are in principle also addressable. Moreover, biolog-
ical modification at the molecular level also makes it possible to incorporate non-
136 A. Böker
Fig. 8.4 Overview of the different approaches to the synthesis of protein/polymer conju-
gates: a “grafting-to”, b “grafting-from”. [29]
canonical amino acids into the protein sequence, which facilitate further linking
reactions that may be partly orthogonal [24]. The most prominent example of this
is the incorporation of reactive groups into the protein, which are suitable for the
highly efficient and selective azide-alkyne “click” coupling [25].
Nanoscale protein/polymer conjugates are a special case of protein hybrid ma-
terials. Here individual, mostly linear polymeric chains are covalently tethered to
the protein. This linkage gives the protein a number of new and improved proper-
ties, or makes it accessible for further processing into the desired material in the
first place. For example, protein-based drugs can be protected in this way from
premature degradation in the human body [26]. Switchable polymers as the syn-
thetic component in a conjugate on the other hand allow for the targeted control of
enzymatic activity [27, 28]. A third application, described below, exploits the in-
creased surface activity of the conjugate. The covalent binding of polymer chains
to proteins is usually achieved via two different mechanisms. When a prefabri-
cated polymer chain is attached to the protein, it is called a “grafting-to” approach,
whereas the growthbuild-up of the polymer chain away from the protein surface
is called “grafting-from” (Fig. 8.4). In “grafting-from” approaches, it is usually
possible to reach a higher density of polymer chains on the protein, but the pro-
tein needs to be sufficiently tolerant towards the polymerization conditions. The

isolation of the conjugates is usually easier with the “grafting-from” approach, as
only small molecules like monomers and possibly a catalyst must be removed. In
certain isolated cases, a “grafting-through” approach comes into question. Here,
the chemically modified protein acts as a macromonomer, which is incorporated
into the growing polymer chain. Which approach will ultimately promise success
varies from case to case.
Adsorption and self-assembly of the protein/polymer conjugates at interfaces
can produce thin, proteinaceous films that can be used for a variety of applica-
tions. Although water-soluble proteins usually already have some surface activity
on their own, it often appears to occur at a much lesser degree. On top of that,
adsorption at the interface usually involves a loss of protein structure thereby de-
stroying the functionality of the protein. Conjugates often prove to be much more
stable in this respect. In addition to their increased surface activity along with the
stabilization of the protein, the conjugates provide their own “immobilization ma-
trix” in the form of the attached polymer chains in the course of self-assembly.
Although the polymer matrix can also be created at the interface beforehand and
the protein subsequently attached [30–32], the self-assembly of the conjugates al-
lows a certain flexibility in the generation of the films due to the fact that a wide
variety of conjugate building blocks can be used. The co-assembly of different con-
jugates makes it possible to generate materials that combine different functions, for
example, different enzymes that can represent entire reaction cascades.
Looking at this in perspective, it is of great importance, especially with regard
to the creation of complex chemical structures for the pharmaceutical sector. Fur-
thermore, the self-assembly approach generally ensures a high protein density and
thus also functionality, while simultaneously maintaining the homogeneity of the
film.
A prominent example of the concept described is the self-assembly of fer-
ritin/polymer conjugates. Here initiator groups were attached to the lysine residues
of the protein for a polymerization reaction whereafter N-isopropylacrylamide
(NIPAAm) was polymerized according to the “grafting-from” strategy. Fer-
ritin/PNIPAAm conjugates show significantly greater surface activity than the
protein or polymer alone [33]. The conjugates were consequently assembled at
water/oil interfaces and formed “Pickering” emulsions. “Pickering” emulsions are
those that are stabilized by large, soft particles, as opposed to simple surfactant-
stabilized emulsions. Stable capsules were obtained by assembling conjugates with
a UV-crosslinkable copolymer and then covalently linking them (Fig. 8.5a) [34,
35, 36]. The ferritin/polymer conjugates were not only crosslinked at emulsion
interfaces, but also on planar surfaces. Ferritin is an iron-containing protein com-
138 A. Böker
a b
Fig. 8.5 a Synthetic scheme for ferritin/polymer conjugates which were subsequently used
to stabilize “Pickering” emulsions. b Overview of the production of protein-polymer hybrid
membranes using the example of the protein ferritin [34, 39]
plex consisting of 24 subunits [37], which after self-assembly of the conjugates

served as a placeholder for pores to be subsequently generated (Fig. 8.5b). These
pores were formed in a post-treatment step by denaturation of the ferritin [38]. The
film thus ultimately obtained has uniform pores, due to the use of monodisperse
proteins and can therefore be used as a highly selective membrane for separations
based on size exclusion.
In addition to globular proteins such as ferritin, which serve only as place-
holders for pores, membrane proteins can also be used, which can act directly
as nanopores within a membrane on account of their channel-like structure. Cur-
rent studies are concerned, for example, with the incorporation of the membrane
protein FhuA into appropriate films.
The synthesis of FhuA/polymer conjugates is the first example of the mod-
ification of a membrane protein in this way (Fig. 8.6a) [2, 3]. FhuA has been
biotechnologically altered in such a way that not only is an open protein channel
formed, but several amino acids in the protein were also exchanged. Analogous to
the example of the ferritin conjugates, polymerization initiators were attached to
the lysine residues. In order to avoid polymer chains to grow inside the channel
and block it again, the lysines were only positioned on the outer protein surface
and above the hydrophobic region of the protein (Fig. 8.6b). FhuA/polymer con-
jugates also stabilized “Pickering” emulsions, which were still visible even after
a b
Fig. 8.6 Overview of a the synthesis of FhuA/polymer conjugates and b the FhuA vari-
ants used; Lysine residues are shown in magenta. c “Pickering” emulsions stabilized by the
conjugates are still stable after 40 days (left: emulsion, right: two-phase mixture without con-
jugates); the cryo-SEM image shows a capsule after the conjugates have been crosslinked.
[2, 4]
40 days (Fig. 8.6c) [4]. After crosslinking the polymer chains, a capsule could be
visualized using cryo-scanning electron microscopy (cryo-SEM).
However, membranes for complex separation problems represent only one case
where the self-assembly of conjugates may be applied. Biocatalytically active films
can also be produced in this way. For example, in one current study, the enzyme 2-
deoxy-D-ribose 5-phosphate aldolase (DERA) has been similarly incorporated into
a thin film [39]. This film allows the enzyme to be used in a continuous synthesis
process for statin-like drug motifs (Fig. 8.7) to make the hitherto relatively mate-
rial-intensive and ineffective batch process more effective and to render it a real
alternative to conventional syntheses.
140 A. Böker
2-Deoxy-D-ribose-5-
phosphate aldolase (DERA)
Fig. 8.7 Immobilization of the enzyme 2-deoxy-D-ribose-5-phosphate aldolase (DERA) in

an ultrathin, polymer-based film to generate a biocatalytically active material for the syn-
thesis of enantiopure hydroxyaldehydes. The use of a thin film such as this instead of using
the enzyme in solution allows for continuous process control, which makes for a much more
efficient synthesis process. (Fraunhofer IAP)
Another approach, similar to protein/polymer conjugate stabilized “Pickering”

emulsions is to covalently bind the conjugates together at the emulsion interface
using crosslinking agents. It was shown that these structures, known as pro-
teinosomes, can be used as reaction spaces [40], can catalyze cascade reactions
when several enzymes are used [41] and can produce hierarchical structures with
multiple proteinosome layers [42].
These examples of self-assembling conjugates can be grouped into the class of
ultrathin films in terms of the nature of the final proteinaceous material [43]. The
particular advantages of such films make it worthwhile to consider them separately.
There are numerous other methods for the generation of ultrathin, proteinaceous
films besides the self-assembly of suitable conjugates mentioned above. These
include not only the electropolymerization on electrodes in the presence of the
protein already discussed, but above all the Langmuir technique. This technique
creates thin films on water surfaces by dispersing amphiphilic substances directly
on the surface. Movable barriers, which allow almost any variation of the available
surface, allow films of different densities to be produced (Fig. 8.8). Due to the fact
that a protein is usually bonded out of the subphase, it is possible to use the variable
density of the film to ultimately control its loading density. Another advantage is
the extremely small thickness of a single monolayer at the interface of only a few
nanometers. Films with variable layer thicknesses in the lower nanometer range
can be formed in a controlled manner by multiple transfers from the interface to
Film Compression
Protein Adsorpon from the Subphase
Protein Matrix forming Polymer
Fig. 8.8 Functional diagram of the structure of a proteinaceous thin film via Langmuir tech-
nique. The dispersion of matrix material on the water surface is restricted by the movement
of the barriers to any desired degree before the protein binds out of the subphase. (Fraunhofer
IAP)
a substrate (Langmuir-Blodgett technique). Under appropriate conditions, proteins

can also be incorporated into preformed polymer films at the interface. Following
dispersion of the amphiphilic polymers, the insertion of proteins such as bacteri-
orhodopsin, cytochrome C oxidase and OmpF (outer membrane protein F) could
be demonstrated by the increase in surface pressure [44, 45]. Alternatively, the
polymer films can also first be transferred to a substrate and then the protein incor-
porated into the film [46] or adsorbed on to the film [30].
Of course, one of the greatest advantages of ultrathin films is the absence of dif-
fusion limitations in the transport of substances with which the embedded protein
142 A. Böker
is intended to interact. At the same time, however, the amount of active protein is
limited by the minimal amount of material. Consequently, such films are mainly
used in biosensors, where fast reaction times but no high throughput rates are re-
quired [47]. Glucose detection by means of glucose oxidase bonded to conductive
polymer structures is an example of this [48, 49]. As already stated, ultrathin pro-
teinaceous films may also be used as membranes. Here, they usually constitute
the active separation layer in a membrane module, where above all their extreme
thinness makes it possible to work with lower pressure gradients, which ultimately
leads to more economical separation processes. The ferritin membranes previously
discussed are a good example of this.
A further special class of molecular, biofunctionally integrated polymers are
the so-called glycopolymers. These consist of a synthetic polymer backbone with
attached glycan chains and therefore act as glycoprotein mimics.
These hyperbranched polymers may, for example, be presented on silicon sur-
faces and after enzymatic modification be built up to develop a consecutive assay
for the identification of ligands for specific interactions (Fig. 8.9) [50]. Alterna-
tively, bio-based proteinaceous scaffolds such as albumins can also be used as
fully biocompatible multivalent carriers [51, 53]. Possible applications of these
structures include using them as capture molecules or as innovative agents in
biomedicine that target lectins.
On top of that, glycans can be used to adjust physical properties such as the
deformability or hydrophilicity of bioplastics. Surface-bound glycans furthermore
offer the possibility of adjusting lubricity, and even specific interactions or the
repulsion of microorganisms that interact with plastic surfaces.
Modern synthetic methods enable the otherwise complex, regioselective modi-
fication of sugars with polymerizable groups to be carried out in a short space of
time and with high yield [53]. Thus, this class of polymer, which could also display
thermoplastic properties depending on the backbone, may also be used as a base
polymer in future.
8.3.2 New Biological Functions in Thermoplastic Bioplastics
Ruben R. Rosencrantz and Jens Balko
Biomolecules carrying a variety of complex functions allow to create plastics that

have specifically adapted physical and biological parameters. And even catalyt-
ically active products are now possible. The classes of biomolecules described
b c
Fig. 8.9 a Synthetic scheme for glycopolymer brush gradients on silicon surfaces The
preparation of the glycomonomer takes place without the use of protective groups in only two
steps by means of MWI (microwave radiation) based synthesis. b Glycopolymers produced
by conjugation of lactose to polyethyleneimine. c Multivalent glycan presentation on an al-
bumin molecule leads to strong interaction with lectins (orange) and their multimerization.
(Fraunhofer IAP)
144 A. Böker
above are available for this purpose, although their integration into thermoplas-
tic (bio)plastics, involve numerous challenges. For the successful integration of
biomolecules it is not only essential to know the structural properties of the
molecule, but also how to maintain its stability and suitability for the desired
functionality.
The central research and development goal is to implement the biological func-
tions carried by the bio-building blocks into thermoplastic bioplastics or to apply
them to surfaces, by means of practical processing methods. The consistent use
of bioplastics is especially important in view of their biodegradability, should the
plastic products end up in the environment after being used. The technical and
material challenge is the following: the temperature-sensitive biological building
blocks, which lose their functionality at the denaturation temperature TDen, are pro-
cessed at the processing temperature TP for thermoplastics. When the processing
is carried out in solution, this step can be bypassed.
The currently unresolved issues and their technological implementation are ad-
dressed by the “BioPol” project group, established in 2018 by Fraunhofer IAP
at the Brandenburg University of Technology (BTU) Cottbus-Senftenberg, with
degressive initial financing from the state of Brandenburg, and operated in close
cooperation with the university’s biotechnology professorships.
Peptides
Peptides, some of which exhibit surprising temperature stability, are predestined
to withstand processing at elevated temperatures while maintaining functional in-
tegrity [54, 55]. Functions that are generated in this way can be found primarily in
the area of antimicrobially active plastics. Antimicrobial peptides can be used for
this purpose, which require, for example, different degrees of freedom in terms of
the self-assembly of pore structures, depending on the mechanism of action. This
needs to be taken into consideration with regard to the type of processing.
Enzymes
Enzymes for the production of catalytically active materials represent the greatest
challenge being integrated into thermoplastic materials. Although heat-stable en-
zymes that can withstand temperatures close to 100 °C have been known for a long
time, the majority of biocatalysts are designed to work at lower temperatures in na-
ture [56, 57]. Therefore, modifications must be made to retain processability while
preserving the biocatalytic function. Besides genetical engineering that allow more
temperature-stable variants of proteins to be produced [48], another noteworthy
alternative is stabilization through the use of polymeric or low molecular weight
additives [59, 60, 61]. There are numerous approaches available (Sect. 8.3.1) ready
to be transferred to production scale. Possible options include the direct conjuga-

tion of polymer chains to proteins, the use of saccharides to increase the stability in
dry state or the production of CLEAs (chemically linked enzyme aggregates) [62].
The latter are dense, active protein aggregates, which are intrinsically stabilized
by cross-linking. The synthesis of CLEAs is still process in a laboratory scale, but
it ought to be possible to pursue an approach for the production on a commer-
cial scale, too. In particular, non-toxic cross-linking methods which are suitable
for a variety of biomolecules need to be developed here. Modern technologies
such as microwave-assisted syntheses have already been used to achieve non-toxic
crosslinking of biomolecules at low temperatures in the past [63].
Glycans
At Fraunhofer IAP, research is already being conducted on how glycans (polysac-
charides) can be incorporated into polyesters with retaining their functional in-
tegrity (hydrophilicity). However, processing options have not yet been considered.
The main advantage of bulk incorporation is to maintain the function towards
abrasion. At the same time, however, a disadvantage of the minimal mobility of
the biomolecules in the material arises, which could, for example, significantly
reduce enzymatic function. There are examples of the incorporation of enzymes
into epoxy resins where activity is maintained [64, 65], as well as more tempera-
ture stable systems with glycans or peptides, which should permit extrusion while
maintaining functional integrity. Here again, the modification of the biomolecules
plays a crucial role. Amines, for example, interfere in the polyester synthesis pro-
cess and must be removed from the starting material. However, since amines can be
essential for the activity of peptides, it may be necessary to use a different synthesis
to obtain suitable polymers. Another way is to mask interfering chemical groups
without reducing the activity of the biomolecule. Being incorporated into bulk ma-
terials, numerous functions result: controlled hydrophilicity and hydrophobicity,
biocatalytically active components, antimicrobial properties, controlled mechan-
ical properties, triggered degradation, etc. The advantage of greater functional
stability is noteworthy here, since no loss of function can arise due to wear.
Thermoplastic Polymers
The typical processing temperature TP of thermoplastic polymers ranges between
100 and 250 °C, with retention times of around 3 to 5 min and pressures of 20 to
1000 bar. In the forming steps (compound, semi-finished product, product), these
process windows can be repeated, which always implies increased material stress.
The most well-known bioplastic available on the market is polylactic acid PLA
(TP = 150–200 °C). The bio-based and readily biodegradable polybutylene succi-
146 A. Böker
nate PBS (150–180 °C) resembles much more the commodity plastic polyethylene
PE with regard to its thermo-mechanical properties. It is therefore more suitable
for flexible film applications. Optimized blends of PLA and PBS make it pos-
sible to achieve high thermostability, as it is required during foil printing or for
some food packaging. During processing, the melting temperature of the material
is almost completely achieved by the dissipated energy and through shearing. The
continuous service temperature of a plastic semi-finished product or component
are directly related to the processing temperature. If high thermostability is desired,
naturally a plastic material with high processing temperatures will be selected.
By contrast, bio-building blocks denature under influences such as shearing and
high temperatures. The denaturation of proteins starts already at TDen of 40 to
60 °C, for glycans only at about 150 °C [66]. The rapid onset of loss of protein
function (short-chain peptides are more stable) is due to the altered or disrupted
three-dimensional molecular structure, which is based on non-covalent bonds. Gly-
cans are thermally more stable because their spatial molecular structure plays only
a minor role for their functionality. Therefore, the central requirement for the bio-
building blocks is that they retain their function despite all thermoplastic process-
ing and forming steps. The formulation, often started in aqueous solution, must be
customized, so that the aqueous phase may be transferred to the surface or into the
bulk workably and efficiently. If successful, the bio-building blocks are homoge-
neously distributed and immobilized in sufficient concentration. Biodegradability
is ideally sought after as an inherent material function in that biodegradable plas-
tics are predominantly used as matrix materials.
The biodegradability is tested at the Processing Pilot Plant for Biopolymers in
Schwarzheide using a respirometer based on conventional industry standards.
The compounding of petroleum-based plastics and bio-building blocks cur-
rently appears able to deal with the problem that the majority of the plastic
materials used today are not biodegradable. For example, enzyme-based master-
batches were introduced that can be compounded into polyethylene (PE) to make
PE biodegradable. While this system aims at the use of PE, numerous other en-
zymes are known which could be used in a concerted cascade reaction sequence of
esterases, lipases, laccases etc. to enable the biodegradation of, for example, PET
and other plastics.
Processing Technology
The processing technology is selected according to the intended functional use,
either as a functionalized interface or functionalized bulk material. The Processing
Pilot Plant for Biopolymers at Fraunhofer IAP offers a modern and versatile fleet
of machines (Fig. 8.10a) with which compounds are produced from bioplastics
Fig. 8.10 Processing Pilot Plant for Biopolymers in Schwarzheide. a Modular twin-screw
extruder with granulation line for compound production. b Schematic outline of extrusion
foam production. (Fraunhofer IAP)
and bio-building blocks such as glycans, and subsequently processed into semi-
finished products such as injection-molded parts or blown films. The extrusion
foaming plant can produce high to medium density foams with densities ranging
from 1.0 to 0.3 g/cm3 (Fig. 8.10b). When open-celled foams are used as filters,
glycans on the inner surfaces of the foam cells can thus be used for the removal of
heavy metals, microplastic particles or microorganisms.
148 A. Böker
Peptides are used to equip surfaces with, for example, antimicrobial properties.
In existing co-extrusion processes such as 3-layered flat film and blow molding
machines, the peptide can be embedded into the functional layer in a bioplastic
matrix such as PCL at a low processing temperature (TP = 100–120 °C). The carrier
layer may then be another bioplastic with a higher melt temperature, such as PBS
or PLA.
The layer of bio-building blocks can however also be applied from solution
in order to avoid any influence of temperature. High-density engineering compo-
nents produced by injection molding or filament-based 3D printing (fused filament
fabrication), are treated by dipping or spraying for this purpose. The technology
for layer-by-layer deposition, yet to be tested on a pilot plant scale, will in future
allow the deposition of thin, precisely adjustable layers with layer thicknesses of
approximately 100 to 500 nm [67].
Relevant biofunctions that may be added to plastic coatings include, for exam-
ple, glycans to achieve a microorganism-repellent coating, but also antibacterial
peptides or even enzymes for biocatalytically active layers (Sect. 8.2). No mat-
ter which coating technology is specifically followed, the biofunctional building
blocks must be incorporated into a formulation. One conceivable example is the
use of chitinases as a fungicidal or insecticidal surface coating. Simple saccha-
rides, such as lactose, are a very robust class of biomolecules and can already be
used to make surfaces hydrophilic or to adjust their lubricity. Similarly robust,
despite their lower temperature resistance, are antimicrobial peptides that lead to
correspondingly equipped packaging [68, 69].
The formulation must be adapted depending on the biomolecule, since an en-
zyme, for example, requires significantly more degrees of freedom than a glycan
or peptide in order to exert its effect. It is possible to modify glycans, peptides and
enzymes with polymerizable groups. This allows for direct incorporation of the
components. In terms of functions, hygienic surfaces are a primary target that can
be worked towards as these have been identified as a large market with relevant
opportunities for utilization. In the widest sense, this also includes anti-fouling
applications.
Likewise, biosensory applications for diagnostics are being investigated, as new
approaches are needed especially in the field of bacterial infections.
There are some initial findings on the use of glycopolymers as biosensors on
interfaces and surfaces [70, 71]. A first step towards a cheap, simple and minia-
turized biosensor was the use of perforated, gold-coated films as sensor material
for a surface plasmon resonance system. This was used to analyze bacterial entero-
toxins and substantiated the remarkable anti-adhesiveness of the glycopolymers
(Fig. 8.11) [72].
a b c
Fig. 8.11 LSPR system for analyzing the binding of bacterial enterotoxins. a Schematic lay-
out of the system with a gold-plated, perforated gold film and grafted glycopolymer brushes
presenting a specific glycan ligand (Gal˛3Galˇ4GlcNAc) of the toxin TcdA. The flow-
through geometry minimizes mass transport limitations. b Binding curve of TcdA on the
specific ligand. c Negative control TcdA on glycopolymer brushes that do not present a lig-
and. (Based on [72])
Fig. 8.12 a Sketch of a glycopolymer-based micelle. Dark blue: glycan. Black: polymer
backbone and hydrophobic part. Red: dye. Light blue: lectins interacting with the micelle. b
Dye- or drug-loaded glycopolymer micelles interact with microorganisms for diagnosis and
therapy. (Fraunhofer IAP)
Furthermore, the self-assembly of glycopolymers in aqueous solutions to form

micellar structures that serve as a basis for novel drug encapsulations is also of in-
terest (Fig. 8.12) [73]. These can specifically dock on to microorganisms, directed
by the glycans, and thus be applied in diagnostics. The species-specific adminis-
tration of drugs is also conceivable as a future development.
150 A. Böker
8.4 Biomaterials for Medicine
Christian Schmidt, Olivia Mauger, and Joachim Storsberg
Biomaterials are synthetic or non-living natural materials that are used in medical
therapy and diagnostics thanks to their biocompatibility and relevant functionality.
Most consist of plastics, ceramics or metals, but also of biopolymers. They en-
ter into chemical, physical and biological interactions with the biological systems
through their medical functions. Biomaterials are designed and rigorously tested
for targeted applications, especially with a view to the replacement or support
of functions in the human body. The ISO 10993 standard assesses their biocom-
patibility and tests their physicochemical interactions with the body. Because the
chemical and biological interaction with the organism takes place on the surface of
the biomaterial, the coating of materials is a way to increase their biocompatibility.
A bio-inert material can thus be made bioactive. The molecular functionalization
of materials for clinical applications is described below, using an example from the
field of ophthalmology.
Ophthalmological implants are a special example of the use of polymeric bio-
materials in medicine. Corneal diseases arising due to infection, injury or burns can
result in irreversible damage. In severe cases of disease progression, drug therapies
are usually insufficient, which is why transplants of parts of the cornea or entire
corneas play an important role. Religious or cultural factors regarding the origin
of the material contribute to the scarcity of corneas available for transplantation,
and risk factors also exist in connection with the donor’s medical history [74, 75,
76]. Although the technology for transplantation of segments of the cornea has
been significantly improved in recent years [77, 78], a large number of people face
a long wait for the necessary transplantation [79]. Therefore, for some patients
only artificial corneas, so-called keratoprostheses, come into question as corneal
replacements. Several keratoprostheses are indicated for ultima ratio patients. The
Potsdam-Halle Keratoprosthesis or MIRO-KPro has been available since 2009.
Historical Background
The first written description of an artificial cornea as a substitute for a cloudy
cornea stems from Guillaume Pellier de Quengsy (1750–1835). In 1789/90 the
French doctor and eye surgeon published a two-volume textbook for prospective
ophthalmologists on the state of 18th century ophthalmic surgery [80]. In his trea-
tise on various diseases of the cornea in the first volume, the author described
an artificial cornea and recommended indications and surgical techniques. Differ-
ent corneal opacities or spots and their causes were described and discussed in
the first article (pp. 92–109). Here the author mentioned the usual treatment for
cloudy corneas, in his time, with ammonia water (then known as spirit of hartshorn
[81]), which he nevertheless rejected as ineffective for severely “stained” or scarred
corneas and therefore proposed a new operation ([80], p. 94 ff.). This was followed
by a detailed description of the tailor-made prosthesis with illustrations of the ker-
atoprosthesis and the surgical instruments required for its implantation. For this
purpose, thin, polished, clear, convexo-concave glass with the diameter of the ex-
tracted cornea was enclosed in a thin silver ring. A shallow groove was produced
at the outer edge of this ring of the thickness of the original natural cornea to al-
low for anchoring to the tissue. The author recommended that the surgeon review
the ingrowth of the keratoprosthesis. However, in case the artificial cornea was not
sufficiently integrated into the tissue of the eye, the author furnished the prosthesis
with two to three cotton threads which were attached between the ring and the lens
to sew the prosthesis into the tissue with the help of the described aids.
Pellier de Quengsy also took a critical view of the proposed procedure in his
review. The comments he listed are still guidelines for both ophthalmologists and
materials scientists today, as can be seen from the selected citations below:
(1) A foreign body such as this must surely cause significant disturbance or even
damage to the patient. The discomfort would however only be felt alike the one
following the insertion of an artificial eye and would diminish over time. Pellier
de Quengsy also recognized the brittleness of the lens material as a limiting
factor, although he considered the likelihood of glass breakage to be very low.
As an alternative, he proposed the development of a keratoprosthesis from some
other, non-brittle, transparent material. He also recommended the fabrication of
the prosthesis in one piece. These statements have not lost their relevance even
today, because true matching of biomaterials with natural tissue has still not
been achieved [82–84].
(2) Secretions may be retained in the eye due to the lack of natural pores on the
prosthesis. This could be remedied by having a small separation between the
ring and the sclera or, better still, having several small holes punched into the
silver ring, which are made during production. These holes would thus act as
artificial pores, allowing all discharges to flow through them. In the end, he sug-
gested making small holes around the silver ring so that the various secretions
of the eye would not pollute or degrade the artificial lens, leaving the lens clear
and transparent. Even if Pellier de Quengsy had wanted to put the small holes
on the edge of the artificial cornea for another reason, he formulated, already
in his time, the thesis, still held today, that for the adaptation of a biomaterial
152 A. Böker
it is necessary to take into account the interactions between tissue and artificial
material. These problems are still not fully resolved today [85]. In this context,
diversity within cohorts makes it difficult to formulate overarching principles
[86, 87] and significantly increases the complexity of the questions to be an-
swered.
In summary, one can see how modern Pellier de Quengsy’s concept was, despite
the fact that the anatomy of the eye was not yet fully understood in the 18th century.
In his book, Pellier de Quengsy expressed that he placed great hope in progress
and the ingenuity of scientists to optimize his idea and find a remedy for people
suffering in this area of eye medicine. Unfortunately, it is not known whether he
ever performed this operation on a patient. It was not until many years later that
his idea was taken up again, although without reference to his writing.
Within the definitions for devices for clinical applications, there is harmony
between the wording of US and EU descriptions [88, 89] with regard to the ‘in-
tended use’, which is in turn in agreement with Pellier de Quengsy’s standpoints
formulated in the 18th century.
Modern Keratoprostheses
The Boston keratoprosthesis (B-KPro, also referred to as the successor to the
Dohlman-Doane Kpro) [90] was approved by the US Food and Drug Adminis-
tration (FDA) in 1992 and its optimization is ongoing [91]. It consists of a donor
cornea attached to a cylinder made of polymethyl methacrylate and a titanium ring
for its fixation. This keratoprosthesis is well tolerated by more than 80% of patients
over a period of up to 17 months after implantation [92]. In long-term observations
of patients with a BKPro among other things, increased intraocular pressure with
concomitant optic nerve damage is described [93].
The osteo-odonto keratoprosthesis (OOKP) as well as the tibial bone Kpro [94–
96] both contain plexiglas lenses. A complex, multi-stage surgical procedure is
necessary for the implantation of the OOKP [97]. The optic mount in this case
consists of the root of a tooth previously removed from the patient, including sur-
rounding bone. The prosthesis is sutured into the patient’s cornea and covered
with oral mucosa. This prosthesis remained firmly anchored in more than 80%
of patients for over 19 years [98]. For both the B-KPro and OOKP, increasing
intraocular pressure is one of the common late complications [99].
The Aachen keratoprothesis [100–102] consists chiefly of a silicone optic and
a polyvinylidene difluoride textile with functionalized surface as the haptic (etym.
from Greek haptein = to fasten, touch). The textile gives the prosthesis increased
flexibility, while its mesh size favors the epithelialization of the haptic and the
nutritive supply of the eye.
The Potsdam-Halle keratoprosthesis (MIRO-KPro) [103] was developed in ac-
cordance with the interdisciplinary strategy for biomaterial production: a high level
of biocompatibility and functionality of the product is sought right from its de-
sign, through its material synthesis and material processing up to its adaptation via
surface modifications. The keratoprosthesis was designed from a piece of highly
transparent plastic, which is given a delicately optimized shape through precise
milling. The optic consists of a cylinder, which is surrounded by a slightly con-
cave border, the so-called haptic, at about one third of its height. This edge is
furnished with fine holes for surgical suturing. The porosity of the haptic favors
cell growth around the optic and its flexibility counteracts the risks of torsional
and compressive forces in ocular tissue. The MIRO-KPro consists of a hydropho-
bic polymer with good biocompatibility [104]. The requirements demanded of the
prosthesis arise from the natural surrounding milieu of the eye that interacts with
its surfaces oriented in different directions. While the haptic of the implant should
grow together well with the cells of the surrounding tissue, no cells should settle
on the optic of the keratoprosthesis. This would severely impact the transparency
and translucency of the material. The front, outer side of the optical area must also
be wettable to ensure a stable tear film. The surfaces of the biomaterial are there-
fore modified in different ways to achieve these desired implant properties [81, 89,
105]. The surface of the optic should remain cell-free and its outer surface must be
compatible with the tear film. In order to make it wettable and thus induce adhe-
sion of the tear film the surface is hydrophilized. Practically, this is achieved by the
photochemical polymerization of a hydrophilic monomer directly on the surface.
A monomer solution is applied anteriorly, exclusively on the optic and irradiated
with UV light of a suitable wavelength, whereby a thin hydrophilic polymer layer
is formed on the surface. This critical change in polarity results in improved wetta-
bility and thus allows the formation and dispersion of the tear film on this surface.
The haptic surface should, by contrast, be cell-friendly. The aim is to achieve
increased cell adhesion and deposition of extracellular matrix both anteriorly and
posteriorly, so that the prosthesis grows into the surrounding tissue well. It is there-
fore subjected to a completely different treatment. The surface of the rim is first
activated by nitrogen atmospheric plasma [106] during which both sides of the
optic (anterior and posterior) must be covered. The plasma treatment ionizes the
material surface and causes an increase in the surface tension, thus improving the
adhesion of the subsequent coatings. The plasma-activated haptic is then alter-
nately drizzled with aqueous solutions of the biocompatible, charged biopolymers,
chitosan (polycation) and heparin (polyanion). This layer-by-layer technique [106]
154 A. Böker
Fig. 8.13 Keratoprosthesis

for ultima ratio patients.
(Fraunhofer IAP)
Fig. 8.14 Implanted ker-

atoprosthesis. [79]
is used to build up layers of polyelectrolytes that serve as an adhesive primer for

an additional protein layer. The surrounding natural cells recognize the growth
factor of the RGD sequences bound to the haptic and are stimulated to colonize
its surface. This specific interaction with the cells thus induces their adhesion and
proliferation.
Biocompatibility has been verified by in vitro tests with primary porcine corneal
epithelial cells [89, 104, 106]. While few cells were visible on the anterior and
posterior sides of the optic, prolific cell growth was observed on the haptic. This
speaks for the tolerability of the material, as no cytotoxic effects were detected.
The keratoprosthesis was tested on rabbits as an animal model at the Halle Uni-
versity Clinic for Ophthalmology. No significant complications arose in the course
of the experiments. The implant was firmly anchored in the surrounding tissue, the
optic remained transparent and permeable to visible light and overgrowth with cell
material was successfully prevented [88]. The keratoprosthesis has already been
implanted in ultima ratio patients (Figs. 8.13 and 8.14; [103, 107]). The coating of
the haptic proved to be beneficial for growth into the surrounding tissue. In con-
clusion, the results to date demonstrate that a well-tolerated keratoprosthesis has
been developed, for which further improvement is envisaged. At present, this pros-
thesis is indicated for targeted use as an ultima ratio remedy for patients where
conventional therapeutic approaches have failed.

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Biogenic Plastic Additives
High-Quality Plastic Additives Made from Natural
9
Raw Materials Benefit the Circular Economy
Rudolf Pfaendner and Tobias Melz
Summary
Additives for plastics based on natural raw materials (“bio-additives”) are well-
known substances that have been used as such or in chemically modified form
in the plastics industry for many years. However, interest in new bioadditives
is increasing in line with the goal of replacing petrochemical raw materials
and promoting a circular economy, as well as with the increasing demand for
biopolymers. Since practically all polymers require additives to guarantee their
properties, processing and application, it is logical for biopolymer formulations
to also be developed entirely on the basis of renewable raw materials, i.e. both
the polymer and the additive. The most important additives include plasticizers,
antioxidants and flame retardants.
9.1 Introduction
Plastic additives, formerly referred to somewhat deprecatingly as auxiliary

substances, have now become essential components in all applications of plas-
tics—from synthetic fibers to coatings, to molded parts of all kinds. For example,
additives ensure the gentle processing of plastics during thermal processing, in-
fluence functional properties such as flame retardance and guarantee long-term
use even under demanding conditions. As very few new plastics are entering the
market anymore, additives are the real drivers of innovation in the design of plastic
properties to enable new areas of application. Additives are as varied as their
R. Pfaendner () T. Melz

Fraunhofer Institute for Structural Durability and System Reliability LBF
Darmstadt, Germany

https://doi.org/10.1007/978-3-662-59659-3_9
162 R. Pfaendner and T. Melz
2TGUGTXCVKQPQHRQN[OGTRTQRGTVKGU < INQUUGPJCPEGTU

< RTQEGUUKPIUVCDKNK\GTU < OCVVKPICIGPVU
< CPVKQZKFCPVU 'ZVGPUKQPQHCRRNKECVKQPUUGTXKEGNKHG
< CPVKQ\QPCPVU < 78CDUQTDGTUNKIJVUVCDKNK\GTU
< JGCVUVCDKNK\GTU < CPVKQZKFCPVU
< NWDTKECPVU < HNCOGTGVCTFCPVU
< CEKFUECXGPIGTU < KORCEVOQFKHKGTU
< QRVKECNDTKIJVGPGTU
/QFKHKECVKQPQHRQN[OGTUVTWEVWTG
< DKQEKFGUCPVKOKETQDKCNCIGPVU
< EJCKPGZVGPFGTU
< HQCOKPIDNQYKPICIGPVU
< ETQUUNKPMKPIEQWRNKPICIGPVU
< FGHQCOGTU
< CPVKETQUUNKPMKPICIGPVU
< TGRGNNGPVU
< EQORCVKDKNK\GTU
< OCTMGTU
< FGITCFCVKQPCFFKVKXGU
< RKIOGPVU
< TJGQNQI[OQFKHKGTUVJKZQVTQRKECIGPVU
< HKNNGTU
/QFKHKECVKQPQHUWDUVCPEGUWTHCEGRTQRGTVKGU < HKNNGTFGCEVKXCVQTU
< CPVKUVCVKECIGPVU < TGKPHQTEKPIOCVGTKCNU INCUUECTDQPHKDGTU
< PWENGCVKPICIGPVU < TGE[ENKPICFFKVKXGU
< CPVKPWENGCVKPICIGPVU < CEGVCNFGJ[FGHQTOCNFGJ[FGUECXGPIGTU
< VTCPURCTGPE[GPJCPEGTU ENCTKHKGTU < QZ[IGPUECXGPIGTU
< RNCUVKEK\GTU < FGQFQTK\GTU
< UWTHCEGOQFKHKGTU < EQPFWEVKXGCFFKVKXGU
< UNKRCPVKDNQEMKPICIGPVU < J[FTQRJKNK\KPIJ[FTQRJQDKPICIGPVU
< CPVKHQIIKPICIGPVU < CFJGUKQPRTQOQVGTU
< FKURGTUCPVU
Fig. 9.1 The world of plastic additives. (Fraunhofer LBF)
applications (Fig. 9.1) and can be classified according to their functions. In the
broadest sense, they either preserve polymer properties (e.g. processing stabilizers
and antioxidants) or extend polymer properties. The latter group includes addi-
tives that chemically modify the polymer structure (e.g. through chain extenders,
crosslinking agents), those that modify the physical structure including the surface
properties (e.g. through nucleating agents) and above all those that extend the
applications and service life (e.g. flame retardants, light stabilizers).
The global market value for plastic additives is estimated to be around US $58
billion by 2020 (excluding fillers and reinforcing materials such as glass fibers)
[1], which is equivalent to 17 million metric tons. The growth of additives in terms
of quantity and value is thus logically following the growth of plastics [2]. The
main factors for growth in the additives market, in addition to the overall increase
in plastics, are lightweight plastics and environmental and toxicity aspects [1].
Today, most additives are still produced from petrochemical raw materials via
chemical processes. However, with the growth of biopolymers, additives based on
renewable raw materials are increasingly in demand. Certain natural products, once
9 Biogenic Plastic Additives 163
isolated and if necessary purified, can either be used directly as additives (e.g. some
antioxidants) or in the form of “building blocks”, i.e. synthesis components for the
production of additives. Some additives produced from renewable raw materials
have already been in use for many years, e.g. epoxidized soybean oil as a plasticizer
and heat stabilizer in PVC applications. Interestingly, the first thermoplastically
processable plastic based on cellulose nitrate (“celluloid”) was supplemented by
the natural product camphor [3], which, according to current terminology, fulfills
the function of a plasticizer. This made it possible for the first time to mold a non-
deformable material into a desired shape in a simple thermal processing step.
Plasticizers already contain a high proportion of renewable raw materials. Other
additives that will be discussed in more detail below are antioxidants, light stabi-
lizers, and flame retardants. This is rounded off by a brief discussion of lubricants,
transparency enhancers, biocides/antimicrobial agents, repellents and deodorizers.
In each case, the functions of the additives are described, as well as the state of the
art with respect to additives from renewable raw materials (“bioadditives”), their
use in current petrochemical-based polymers, which still accounts for more than
98% of the market, and the use of bioadditives in polymers made from renewable
raw materials (“biopolymers”). The combination of biopolymers and bioadditives
allows formulations to be developed that are based entirely on renewable raw ma-
terials.
9.2 Plasticizers Based On Renewable Raw Materials
Plasticizers in the form of additives (so-called external plasticizers) act as process-

ing aids by reducing the melting temperature and glass transition temperature of
a polymer. At the same time they modify the polymer properties. For example, they
may lower the elastic modulus. The plastic becomes more elastic and softer. Plas-
ticizers constitute the majority of all plastic additives in terms of volume and value
(US $21.4 billion (2013) [1]), although their prices per kilogram are also in the
lowest category. More than 80% of all plasticizers produced are used in polyvinyl
chloride mainly for soft PVC (P-PVC) [4]. Other polymers of industrial importance
to the plasticizer market are polyvinyl butyral (PVB), polyvinyl acetate (PVAc),
polyvinylidene chloride (PVDC), cellulose esters and polyacrylates. Application
concentrations are usually between 10 and 50% by mass. Important criteria in the
selection of plasticizers are compatibility with the polymers, low volatility, low
solubility in water, high extraction stability, resistance to aging, lightfastness, resis-
tance to microbial attack, environmental compatibility, toxicity including possible
degradation products as well as expense.
a b
Fig. 9.2 Plasticizers based on epoxidized vegetable oils: a epoxidized soybean oil, b epox-
idized castor oil, c epoxidized linseed oil (each major component). (Fraunhofer LBF)
From a chemical perspective, plasticizers are weak solvents for polymers. As

a result, the range of possible basic chemical structures is relatively extensive.
Major classes include phthalates, phosphates, adipates, citrates, cyclohexane-1,2-
dicarboxylates, terephthalates and trimellitates, i.e. esters formed with long-chain
alcohols. The inevitable substitution of phthalates, which were the main plasticiz-
ers used for many decades (e.g. dioctyl phthalate, DOP) due to their toxicity and
for environmental reasons, has also accelerated the development of new plasticiz-
ers from renewable raw materials.
Plasticizers derived from renewable raw materials are however by no means
new, since epoxidized vegetable oils have been in use for many decades [5]. Es-
sentially, these are triglyceride vegetable oils based on soybean oil, linseed oil,
castor oil or sunflower oil. They contain several unsaturated double bonds, which
are epoxidized by means of peroxide or peracetic acid (Fig. 9.2). In halogen-con-
taining polymers such as PVC, the epoxy group also acts as a heat stabilizer during
processing, as it can react with HCl, which is formed in the thermal degradation of
PVC in the first step.
Furthermore, it is possible to use the reactive epoxide group for subsequent
reactions when the ring is opened and for the synthesis of, for example, the corre-
sponding acetates [6]. More recently, the principle of epoxidized plasticizers has
been transferred to other raw materials, e.g. cardanol, which is obtained from the
shells of cashew nuts [7].
Fig. 9.3 Plasticizers based on a isosorbide esters and b, c citric acid esters. (Fraunhofer
LBF)
Isosorbide can be obtained from glucose by hydrogenation and subsequent de-

hydration. The (oligo) esters with dicarboxylic acids or fatty acids, which are in
turn obtained in this way, are alternatives to conventional phthalates (Fig. 9.3). Low
volatility, good compatibility with PVC as the main application and biodegradabil-
ity are advantages, whereas the increased water absorption is considered a disad-
vantage [6, 8]. In particularly sensitive applications such as medicine, toys and
food packaging, esters of citric acid are now used, which are obtained from cit-
rus fruits and cane sugar, but still, above all, via chemical synthesis (Fig. 9.3).
These plasticizers are also good alternatives to phthalates, but they are much more
costly [9]. In addition to these, plasticizers have also been studied that are obtained
via chemical processes from vegetable oils and waste materials. Examples include
the acetic acid esters of hydrogenated castor oil [10], fatty acid esters of camphor
derivatives [11] and wood liquefied by means of polyols [12].
On the research front, plasticizers from renewable raw materials have since
also been introduced into biopolymers (e.g. polylactic acid (PLA), polysaccha-
rides such as starch polymers), in order to increase the flexibility of polymers in
film applications. Plasticizing effects are reported for polyols such as glycerol, for
fatty acid esters, but also with citrates and soybean oil [4]. Isosorbide dioctoate is
particularly effective in PLA due to its increased miscibility, which is also reflected
in the improved flexibility and transparency of the polymer formulation [13].
9.3 Antioxidants from Plant Extracts
Plastics are essentially organic materials. They age through oxidation and photoox-
idation, which lead to changes in the surface, to discoloration and ultimately to the
mechanical failure of plastic parts. The process known as autoxidation leads to
changes in the polymer chain via radical reactions, e.g. to a reduction in molecular
mass or to crosslinking of the polymer chains at a molecular level. Stabilizers are
introduced to delay aging and to guarantee the useful lifetime.
Antioxidants secure the processing of plastics and their long-term use by
interrupting autoxidation. Without these additives, the diverse applications of
polypropylene, other polyolefins and plastics in general seen today would be un-
thinkable. Antioxidants are used in concentrations of between 0.05 and 1% (higher
for elastomers), depending on the polymer and the application. Important criteria
in the selection of antioxidants are high efficacy at low concentrations, compati-
bility with the polymer, low volatility, low migration, high extraction stability, no
contribution to discoloration, environmental compatibility, low toxicity including
potential degradation products and cost.
A distinction is made between primary antioxidants, which act as radical scav-
engers, and secondary antioxidants, which react with the hydroperoxide intermedi-
ates formed in the degradation process. Primary antioxidants are, from a chemical
perspective, sterically hindered phenols or (semi) aromatic amines. Secondary an-
tioxidants are mostly phosphites or phosphonites and less commonly, sulfides.
The combination of primary and secondary antioxidants leads to synergistic ef-
fects. Sterically hindered phenols mainly feature di-tert-butylphenol groups (or
tert-butyl-methylphenol groups), which contain chemical groups of the polymer to
be stabilized as further structural units in order to ensure good compatibility. Other
elements include a comparatively high molecular mass to guarantee low volatil-
ity during processing and in application. A certain mobility (migration) within the
polymer is however advantageous for efficacy, so that as a rule very high molec-
ular masses are avoided. The phosphites are usually sterically hindered phosphite
ester structures. Sulfides are mainly long-chain esters of thioglycolic acid. The ef-
fectiveness of the antioxidants is generally assessed by drawing on the processing
stability of plastics (e.g. by multiple extrusion) and the long-term thermal stability
(e.g. by accelerated aging at elevated temperatures).
Phenol structures often occur in nature, but phosphites/phosphonites or (semi)
aromatic amines do not. Studies on the use of antioxidants from renewable raw
materials in plastics have therefore focused on phenolic antioxidants from various
sources.
b c
d e f
Fig. 9.4 Examples of antioxidants from renewable raw materials: a tocopherol; b quercetin;
c curcumin; d sylimarin; e rosmarinic acid; f ferulic acid. (Fraunhofer LBF)
A phenolic antioxidant from nature that has been known for a long time and
commercially used (to a lesser extent) in plastics is vitamin E (tocopherol and its
isomers, Fig. 9.4) [14, 15]. Vitamin E is a relatively good processing stabilizer, but
its contribution to long term thermostability at elevated temperatures is quite weak.
Owing to its high price, vitamin E is preferably used as a booster in combination
with traditional stabilizers [16] or for particularly sensitive applications such as
ultra high molecular weight PE (UHMWPE) implants [17].
Phenol-containing plants and their extracts, from green tea to red grapes to
gingko leaves, have cell-protecting, antiviral and anti-inflammatory properties and
are available as dietary supplements and medicines.
It is therefore not surprising that these substances have also been tested as sta-
bilizers in plastics. Well-known examples are extracts of green and black tea [18],
rosemary [19], red wine extract [20] or tomatoes [21]. The flavonols contained in
the plants and their glycosides such as quercetin, hyperin, rutin [22], dihydromyre-
quetin [23] or sylimarin are responsible for the effect [24, 25]. Other phenol-
containing phytonutrients such as curcumin [26] or synthetic derivatives such as
esters of rosmarinic acid, polymers of caffeic acid [27], derivatives of ferulic acid
[28] and cardanol, a monophenol distillate from cashew nutshells [29] were inves-
tigated (Fig. 9.4). Even lignin displays a (weak) antioxidant effect in polyolefins
[30, 31].
The pure phenol-containing substances demonstrate a stabilizing effect in poly-
mers which is quite comparable to that of synthetic antioxidants. This has been
seen, for example, in the polyolefins, polypropylene (PP) or polyethylene (PE),
which are most often investigated owing to straightforward detection methods.
Combinations with secondary antioxidants also result in a synergistic effect. For
example, it has been shown that a combination of curcumin and a phosphonite
in LDPE is superior to an analogous synthetic antioxidant combination in terms
of processing stabilization (multiple extrusion) [26]. The effect of antioxidants is
present in all polymers that undergo oxidative degradation via radical mechanisms.
Hence applications of antioxidants from renewable raw materials have also been
investigated in biopolymers, for example in polybutylene succinate [28] or in starch
polymers [32].
The stabilizing effect of the natural antioxidants essentially depends on the con-
centration of phenol groups and their chemical environment. This can be quite
high. However, the natural antioxidants are often colored or prone to discoloration,
which is undesirable in most plastics e.g. for packaging or film applications. Fur-
thermore, the solubility in the polymers is often low, thus limiting their compati-
bility. This can lead to the formation of deposits during processing and plate-out
during use. The thermal stability (volatility and degradation) of phenols from re-
newable raw materials is often lower than that of synthetic antioxidants, which
means that their use is limited or impossible at high processing temperatures, as
required for engineering plastics, for example. Furthermore, there are very few
studies to date on how the bio-antioxidants contribute to the long-term thermal sta-
bility of polymers and to their possible synergistic effect with classic long-term
stabilizers such as hindered amines (hindered amine light stabilizer, HALS).
9.4 Light Stabilizers from Plant Extracts
Most polymers in outdoor applications require light stabilizers in addition to ag-

ing stabilizers (antioxidants). This is the only way a photooxidative degradation
can be prevented or at least delayed. The light stabilizers of today are both UV
absorbers and radical scavengers based on hindered amines, so-called HALS com-
pounds. UV absorbers (e.g. o-hydroxybenzophenones, o-hydroxyphenyltriazines,
o-hydroxyphenylbenzotriazoles, cinnamates, oxanilides) absorb the UV light, con-
vert the energy into heat in a non-radiative transition and thereby protect the poly-
mer from the UV radiation. HALS light stabilizers are chiefly based on tetram-
ethylpiperidine derivatives and act as radical scavengers, similar to phenolic an-

tioxidants, interrupting the degradation process, thereby allowing the active struc-
ture to regenerate. Pigmented plastics generally demonstrate higher UV stability
than non-colored polymers; e.g. carbon black exerts a very good light protection
effect in black-colored polymers. A wide range of standard and specialized com-
mercial products are available on the market as UV absorbers and HALS.
Chemical structures that correspond to industrial UV absorbers or HALS com-
pounds are rarely found in nature, with the exception of cinnamic acid esters. No
comparably effective light stabilizer based on renewable raw materials has there-
fore been reported to date. However, to protect against UV radiation, plants in
arid and alpine areas produce flavones and flavonols, which are deposited on the
plant surface and thus protect the underlying plant cells [33]. The few studies on
light stabilization by means of stabilizers from renewable raw materials refer to
polyphenols with flavonol structures, which act as antioxidants. These studies thus
prove the familiar effect that polymers stabilized by antioxidants are more light-
stable than unstabilized plastics. These investigations of the photo-oxidative stabi-
lizing effect of natural antioxidants were thus carried out on typical polymers such
as polypropylene [34], bio-polyethylene [35] or biopolymers made from renewable
raw materials, e.g. quercetin in PLA [36, 37] and in starch polymers [38].
However, the effect of phenols as light stabilizers is weak, at least in comparison
to synthetic light stabilizers. A prodegradant effect was established for quercetin
and other compounds at high concentrations, which is explained by the presence
of transition metals in the natural product extracts [36, 39]. However, with accel-
erated biodegradability being desirable in biopolymers such as PLA rather than
a disadvantage (at least for packaging materials), the question of improved light
stability has hardly been raised to date.
9.5 Flame Retardants from Renewable Raw Materials
Conventional plastics as organic materials consist mainly of carbon, hydrogen and

oxygen and are therefore—with the exception of a few inherently flame-resistant
high-performance polymers—relatively highly flammable. In addition, they have
a high energy content and therefore release a considerable amount of energy when
combusted. In order to reduce or eliminate the fire hazard of plastics, it is there-
fore necessary to minimize the flammability of many plastics and to use flame-
retardant plastic compositions. To achieve this, additives are usually introduced
into the plastic in the form of flame retardants that prevent ignition for a certain
time or significantly delay the spread of fire. This allows affected people to es-
cape and material assets to be protected. Flame retardants are the second largest
class of plastic additives in terms of volume and value, demonstrating the highest
growth rate of 5.4% a year [1]. The main factors here are increased requirements
for fire protection, increasing safety awareness and the replacement of traditional
flame retardants for environmental and toxicity reasons. The required proportion
of flame retardants in the plastic can be very high, up to 80% by mass. A mass
percent of 15 to 30 is most commonly used, depending on the particular polymer
and application.
From an environmental point of view, certain brominated flame retardants have
been banned. The call for halogen-free flame retardant solutions is thus becoming
ever louder. It is therefore not surprising that flame retardants are also being sought
from renewable raw materials. The basis for halogen-free flame retardants is either
inorganic compounds—e.g. aluminium hydroxide or magnesium hydroxide—or
they are based on organic nitrogen compounds such as melamine or on organic
phosphorus compounds such as ammonium polyphosphate. There are therefore
two strategies for flame retardants made from renewable raw materials: either us-
ing naturally occurring phosphorus compounds or equipping natural products with
flame retardant functional groups. The naturally available phosphorus compounds
are limited to phytic acid (Fig. 9.5), which is found in maize, soybeans and grains
or their derivatives. Phytic acid itself is water-soluble, hygroscopic and only ther-
mally stable to a degree, which limits its incorporation into plastics. Furthermore,
hydrolyzable plastics such as PLA are substantially damaged during processing
with phytic acid. Synergistic approaches such as metal salts (zinc, aluminum) or
nitrogen-rich bases like melamine cannot improve the thermal properties to any
significant extent. However, flame retardant properties can often be demonstrated
in combination with other flame retardants.
Both oil-based plastics such as PVC [40] or polypropylene [41] and biopoly-
mers such as PLA [42] were tested in this regard. Another possibility is to combine
different nature-based flame retardants, e.g. phytic acid with lignin [43] or phytic
acid with cellulose [44]. However, even these synergistic combinations cannot
solve the lack of thermal stability. Mechanistically, phytic acid, its derivatives and
combinations lead to crust formation (“char”) in the event of fire. This barrier pro-
tects the polymer from further combustion. A total flame retardant concentration
of about 30% is needed to achieve an adequate effect.
Another example of flame retardants from renewable raw materials that has
been published is the use of casein. Casein is the protein portion of milk. PLA con-
taining 15% casein by mass achieved a very good rating according to the UL 94 V-
0 standard. Here again, the thermal stability during processing is a challenge [45].
a b
Fig. 9.5 Examples of flame retardants from renewable raw materials. a phytic acid, b car-
danol phosphate, c tannin. (Fraunhofer LBF)
As an alternative to the natural product phytic acid, which contains phosphorus,

phosphoryl groups can be incorporated into natural products via phosphorylation
reactions to produce flame retardants [46]. The phosphorylation of lignin by means
of phosphorus pentoxide deserves particular mention here. The lignin contains
phosphorus and thus leads to a certain flame retardancy, for example in ABS (acry-
lonitrile butadiene styrene) [47]. In principle, all other bio-phenol derivatives, as
described in Sect. 9.3 can also be converted into potential flame retardants via phos-
phorylation. Cardanol [48] is a representative example worth mentioning here. For
other polyphenols, such as tannin, [49] it is assumed that the aromatic structures
will lead to char formation when used as potential flame retardant. Precondensa-
tion and the formation of crosslinked structures thus weaken the already somewhat
inadequate thermal stability.
9.6 Other Plastic Additives from Renewable Raw Materials
In addition to the important plastic additive classes already mentioned, there are
a number of other additives based on renewable raw materials that are of lesser
importance in the overall context. These include nucleating agents, biocides, re-
pellents and deodorizers. Furthermore, several additives such as lubricants and
antistatic agents are based on simple natural raw materials or their reaction prod-
ucts.
Nucleating agents are used to control the morphology of semi-crystalline poly-
mers. In this way the crystallinity or the crystallization rate can, for example, be
increased. Transparency enhancers, or clarifiers are a special subclass of nucleating
agents, which are used in polypropylene [50].
The bulk of the market is based on sorbitol derivatives (Fig. 9.6), where the
starting material is sorbitol, a sugar molecule that can be obtained from glucose
via enzymatic reactions. Sorbitol acetals such as bis(p-methylbenzylidene)sorbitol
have also been described as nucleating agents in PLA. The low crystallization ten-
dency of PLA often requires an additive to achieve adequate mechanical properties
[51]. Another natural product, orotic acid, found in dairy products and mold fungi,
is also effective for this purpose [52].
Biocides or antimicrobial agents are an important market in the hygiene sector.
The aim is to achieve a surface effect in plastics, which protects the user from harm
caused by microorganisms. Moreover, there are applications that prevent microor-
ganisms from causing damage to the plastic itself. Such damage can lead to visual
deficiencies such as stains on the surface, contamination and deterioration of me-
chanical properties. Soft PVC is a particularly susceptible substrate in this respect.
Some natural products feature biocidal properties, such as many vegetable oils,
which contain phenols or sulfides [53] as active components. Chitosan is a biocidal
Fig. 9.6 Nucleating agents based on natural products. a bis(p-methylbenzylidene)sorbitol,

b orotic acid. (Fraunhofer LBF)
Fig. 9.7 Examples of biocides: a chitosan, repellents, b capsaicin, c deodorizer ricinoleic

acid salt. (Fraunhofer LBF)
polymer that is worth mentioning (Fig. 9.7). It is a polyaminosaccharide produced

from the chitin of crustacean shells [54].
Repellents prevent the destruction of plastics (e.g. cables) by rodents or insects.
Additives of choice here are substances with a strong bitter or pungent taste, such
as capsaicin, an ingredient found in pepper [55].
Natural fragrances such as pine scent are used as deodorizers in recycled plas-
tics, for example. Alternatively, reactive substances can be used to improve odor
by absorbing odorous substances, e.g. those containing sulfur or amines. Salts of
ricinoleic acid, the main component of castor oil [56], and cyclodextrins, which are
obtained enzymatically from starch, are examples of commercial products based
on natural products [57].
A simple class of natural products that will not be discussed in more detail
here are fatty acids and their derivatives such as salts, esters and amides. These
are used in large quantities as lubricants, PVC heat stabilizers, acid scavengers or
simple antistatic agents. Fatty acids are derived from both plant and animal sources.
Depending on the origin, in some cases different degrees of efficacy or tendencies
towards discoloration can be seen [58].
9.7 Outlook
In general, additives have the advantage over polymers that they are generally used
in smaller amounts and possess a higher value. In principle, the same additives can
be used for biopolymers and petrochemical polymers. A substantial market share
can thus be more easily realized which is independent of the commercial growth
of the actual biopolymers.
Plasticizers are the class of additives which already offer a broad and very ef-
fective commercial range today. These can compete quantitatively with the current
overall biopolymer market. The next few years will show to what extent the market
will implement this.
Antioxidants occur in various forms in nature, but often present deficiencies
compared to synthetic products, for example, in terms of temperature stability
and color consistency. Without taking the required cost optimizations into ac-
count, potential areas of application are rather to be found for polymers with lower
processing temperatures or less critical color requirements, e.g. biopolymers and
recycled plastics. Engineering plastics such as polyamides have been excluded so
far, mainly for reasons of temperature. Substances from natural resources are also
only likely to be introduced into the field of light stabilizers to a limited extent.
Intensive research is being done on flame retardants from renewable raw mate-
rials, but here too many approaches fail because of the comparatively low thermal
stability of natural products. This means that important fields of application such
as engineering plastics are to a large extent excluded. It remains to be seen to what
extent chemical conversions of natural products, such as condensation reactions
and crosslinking reactions, will reduce this shortcoming.
With additive manufacturers’ increasing awareness of the need to use renew-
able raw materials and/or to increase the biological share in their portfolio, even
more additive classes will become important in the future. New additives from re-
newable raw materials should thus be able to achieve a substantial market potential
relatively quickly. A proportion in the order of 10% of the additive market, corre-
sponding to five billion euros, should be accessible within the next few years. There
is, however, still a considerable need for research and data in many areas pertain-
ing to additives before the potential applications of additives from renewable raw
materials in comparison to synthetic products may be secured.

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Organisms as Producers
Production of Value-Added Compounds Using
10
Microorganisms, Algae and Plant Cells
Stefan Rasche, Stefan Schillberg, Felix Derwenskus, Ulrike

Schmid-Staiger, and Ursula Schließmann
Summary
One aspect of bioeconomy is the use of biological resources such as plants,

animals and microorganisms to produce value-added compounds and active in-
gredients. Thus, this biomass not only serves as fodder and foodstuff and as
a source of energy, but also as a supplier of important bio-based industrial
products such as specialty chemicals, bio-based plastics, surfactants, colorant
or pharmaceuticals. Furthermore, individual biological systems such as animal
or microbial cells and even plants can be optimized or genetically modified to
produce proteins, oils or metabolites for different industrial applications.
In this chapter, the efficiency of biological production systems is illustrated
by means of three examples: the production of a dietary protein in genetically
modified bacteria, the preparation of plant stem cells for the cosmetic industry
and the production of proteins and valuable lipid fractions such as carotenoids
from microalgae. These three scenarios demonstrate the efficiency of biological
systems in general. This approach can be applied to a variety of other classes of
products, placing biological production at the heart of bioeconomy.
S. Rasche () S. Schillberg

Fraunhofer Institute for Molecular Biology and Applied Ecology IME
Aachen, Germany
F. Derwenskus U. Schmid-Staiger U. Schließmann
Stuttgart, Germany

https://doi.org/10.1007/978-3-662-59659-3_10
180 S. Rasche et al.
10.1 Production of Dietary Proteins in Microbial Cell

Factories
Stefan Schillberg and Stefan Rasche
Project Title:
Production of dietary proteins in microbial cell factories
Aim:
Production of a phenylalanine-free dietary protein to support dietary treatment of phenylke-
tonuria
Co-Operating Partners:
Fraunhofer Institute for Molecular Biology and Applied Ecology IME, metaX GmbH
Research Plan, Funding Volume:

Funding period 6 years, 955,082 C (Federal Ministry of Education and Research, BMBF)
Contact:
Stefan Rasche, Stefan Schillberg (Fraunhofer IME)
10.1.1 Introduction and Background
Phenylketonuria (PKU) is one of the most common congenital metabolic disorders

in humans, occurring in Germany with an incidence of 1 : 8000. Affected patients
cannot break down the amino acid phenylalanine, because, due to a genetic defect,
they either lack the enzyme phenylalanine hydroxylase or it is present, but less ac-
tive. As a result, phenylalanine accumulates in the body causing phenylpyruvate,
phenylacetate or phenyllactate to be formed, which if untreated leads to severe
brain damage with associated mental retardation, seizures and spasticity. The ef-
fective treatment for PKU that has been successful for about 45 years is a special
low-protein diet that should ideally be followed for life. In this diet, the amount of
phenylalanine consumed is strictly controlled and limited to the essential amount.
Since common foodstuffs contain a lot of protein and thus also phenylalanine, PKU
patients are reliant on a low-protein diet. This is supplemented with phenylalanine-
free, synthetic amino acid mixtures to ensure an adequate supply of essential amino
acids that cannot be produced by the human body. However, the poor organolep-
tic properties of these amino acid mixtures as well as their limited solubility and
suspendability make it difficult to adhere to the diet. Dietetics research and the nu-
trition industry are therefore constantly on the lookout for new dietary approaches
10 Organisms as Producers 181
for PKU patients, which promise an improvement in flavor and use in food prepa-
ration.
Low phenylalanine or phenylalanine-free proteins offer an alternative to syn-
thetic amino acid mixtures as their functional properties make them easier to pro-
cess, thereby also contributing to an improved flavor. In collaboration with the
company, metaX, Fraunhofer IME has identified a protein with an optimal amino
acid composition for nutrition. The protein was optimized using genetic engi-
neering methods, so that it no longer contains any phenylalanine, yet remains
structurally intact and is suitable for microbial production. This protein, called
GSP105, can be produced in the range of tens of grams per liter in the Pseu-
domonas fluorescens production system and then purified by standard chromato-
graphic techniques. Initial experiments on mice have shown that a diet based on
GSP105 can significantly reduce phenylalanine levels in the blood.
10.1.2 Selection and Optimization of a Suitable Phenylalanine-

Free Dietary Protein
In order to meet the requirements of a protein for human nutrition, the phenylala-
nine-free proteins should demonstrate a balanced amino acid profile with a high
biological value, as such proteins are converted into endogenous proteins partic-
ularly well. The biological value of a protein is usually defined by the so-called
potato-egg principle (Kartoffel-Ei-Standard, KES), which should ideally lie above
100, the reference value for chicken egg protein. In addition, the ideal protein can-
didate should have low allergenic potential, not be too large (less than 50 kilodal-
tons, kDa) to facilitate production in the bacterial production strain, and preferably
not be patented.
With the aid of an algorithm that takes the dietary protein requirements outlined
above into account, the UniProt protein database has been analyzed for more than
830,000 proteins from animal, plant and microbial organisms that are used in hu-
man nutrition or used to produce such products (e.g. lactic acid bacteria for yogurt
production). In doing so a protein candidate from Bacillus subtilis could be identi-
fied, which met the above requirements, but still contained a single phenylalanine
residue. Since this 19 kDa GSP protein did not contain the essential amino acid
tryptophan, the phenylalanine residue was replaced with tryptophan using genetic
engineering methods. The phenylalanine-free protein, now designated as GSP105,
thus had a particularly high biological value (KES = 137) and also did not dis-
play any allergenic potential, as verified by an in silico analysis using the database
FARRP (farrp.unl.edu). The use of the GSP105 protein for the treatment of PKU
has been filed as a patent [2].
10.1.3 Production and Testing of the Phenylalanine-Free

Protein
Large-scale production of the phenylalanine-free GSP105 protein must be possi-

ble before it is introduced into the diet of PKU patients, since the protein will be
required on a ton scale if it is successfully commercialized. The industrial produc-
tion of proteins for various applications in the pharmaceutical, nutrition and animal
feed industries as well as in the chemical industry is mainly accomplished by an-
imal and microbial cells such as yeasts or bacteria. In the food industry, mainly
bacteria are used that are genetically modified and thereby produce a new, so-called
recombinant protein. A large proportion of the enzymes used for the production of
foodstuffs (e.g. in bread and baked goods, cheese, fruit juices, confectionery, meat
and sausage products, noodles, etc.) are thus produced with the aid of genetically
modified bacteria [3]. A particularly efficient platform for the production of re-
combinant proteins is the bacterium, P. fluorescens, which allows yields of up to
tens of grams per liter of culture volume [1]. Annual production capacities on a ton
scale can thus be achieved by using large bioreactors (> 20,000 liters) to cultivate
the bacteria.
The corresponding gene was introduced into the bacterium to produce the
GSP105 protein. Initial cultures of the genetically modified bacterium in shaking
flasks (2.5 liters with 0.5 to 1 liter of culture volume) have already resulted in
yields of 2.5 g per liter. Culturing in bioreactors (5 to 350 liters working volume)
increased productivity to an average of 20 g per liter. In this way 3.5 kilograms
of GSP105 could be produced within just a few months. The protein was puri-
fied with high yield by means of immobilized metal ion affinity chromatography.
Excess salts were then removed by hollow fiber filtration and the protein was
dried by freeze and/or spray drying. After drying (Fig. 10.1), various experiments
were carried out to characterize the protein (determination of purity, phenylalanine
content, detection of DNA contamination).
The purified and dried protein has a neutral odor and a pleasant, almost neutral
taste. It tastes neither sweet nor sour or bitter and these flavor characteristics thus
make it ideal for the preparation of dietary foods. Initial experiments with PKU
mouse models, which were given the phenylalanine-free GSP105 protein produced
Fig. 10.1 Efficient production of the phenylalanine-free GSP105 protein is carried out in the
bacterium P. fluorescens. The protein can be produced and purified in large quantities and,
having been dried by freeze drying (a), it may be aliquoted and shipped for different analyses
(b). (Fraunhofer IME)
in P. fluorescens, showed a reduced phenylalanine value that was well below the
required limit.
10.1.4 Outlook
The preparation of GSP105 has made a protein available that fulfills all the char-
acteristics of a dietary protein for the treatment of PKU patients and which can
be produced in sufficient quantities in the P. fluorescens bacterial production plat-
form. Following further animal experiments approval of the protein as a novel food
will be applied for. Since the GSP105 protein has not yet been used for nutritional
purposes, extensive food safety analyses are required.
The bacterium P. fluorescens has proven to be a very effective platform for
the production of recombinant proteins. The production system was in-licensed
from Dow AgroSciences. As part of this research license, other dietary, agricultural
and industrial proteins are also produced in this bacterium at Fraunhofer IME.
The aim of these projects is to establish economic and sustainable production and
purification processes, so that industrial partners may prepare protein candidates
for various application markets.
10.2 Plant Stem Cells for the Cosmetic Industry
Stefan Rasche and Stefan Schillberg
Project Name:
Plant stem cells for the cosmetic industry
Aim:
Production of plant stem cells and optimization of production conditions
Fraunhofer IME, Dr. BABOR GmbH & Co. KG

Funding period 2 years, 250,000 C (industry)
Contact:
Stefan Rasche, Stefan Schillberg (Fraunhofer IME)
10.2.1 Introduction and Background
Plant extracts are an important raw material for the ever growing cosmetic industry
on account of the tremendous diversity in their active ingredients and properties.
Attention has recently shifted from plant extracts grown outdoors to preparations
from tissue or cell cultures. In contrast to extracts from whole plants or fruits, the
use of cell cultures can ensure a consistent product quality, especially with regard
to the relevant ingredients. The cell cultures are cultivated under sterile condi-
tions, which precludes the possibility of exposure to microbial contaminants such
as endotoxins. The use of pesticides can also be avoided, which benefits both the
product quality and the environment. Furthermore, the production is independent
of climatic conditions, seasonal restrictions or the geopolitical situation, which
guarantees customers a continuous supply.
10.2.2 Preparation and Production of Plant Stem Cells
On behalf of the Aachen cosmetics company Dr. Babor GmbH & Co. KG, plant
cell cultures were to be established from the fruit tissue of the pear cultivar “Cham-
pagner Bratbirne” (Pyrus communis cv. Champagner Bratbirne) as well as the wild
Fig. 10.2 Callus cultures (left) displaying undifferentiated cell growth are created from the
fruits of the Champagner Bratbirne (upper panel) and the wild service tree (lower panel).
These plant stem cells are grown in culture vessels of up to five liters with 2.5 liters of
culture medium to provide biomass (right). (Fraunhofer IME)
service tree (Sorbus torminalis)—a tree with 1 to 2 cm large, red-brown fruits. For
this purpose, the fruits of both plants were surface-sterilized to remove unwanted
germs and the fruit tissues were cultured on nutrient media. In addition to sugar,
these media also contain plant hormones that induce undifferentiated cell growth
(Fig. 10.2). The resulting cell clusters can then be further cultured in liquid medium
where they continue to grow in suspension as single cells or small cell aggregates.
Since these undifferentiated cells can theoretically be regenerated to intact plants
when other plant hormones are added, the cosmetic industry also refers to these
cell cultures as plant stem cells.
In order for larger quantities of pear and service berry stem cells to be prepared,
they are shaken at 28 °C in 5-liter containers with 2.5 liter filling volume for uni-
form nutrient supply. Thus, without complex pre-culturing, tens of kilograms of
cell biomass can be produced within seven to 14 days. After harvesting the cells
by filtration, they are disrupted by means of a homogenizer, and insoluble con-
stituents are separated by continuous centrifugation. The clear cell extract is then
mixed with alcohol and can be added in this form to cosmetic products e.g. creams.
10.2.3 Optimization of Production Conditions
Once the plant stem cell cultures were established, the proportion of plant cell
biomass per liter of culture volume was about 150 g. However, the cells were still
secreting a great quantity of unwanted polysaccharides and lipids. The culture con-
ditions such as inoculation volume, temperature and culture time were optimized in
order to both reduce the polysaccharides and lipids in the culture and also increase
the cell biomass. In the past, one factor at a time was changed when optimizing
processes. For example, the optimum temperature was first determined and then
the culture time was optimized at that temperature. However, this procedure does
not produce an optimal test result since the individual effects and interactions of the
various influencing factors in the process are not recognized. By contrast, the statis-
tical planning of experiments, known as Design of Experiments (DoE), allows for
the systematic planning and statistical evaluation of experiments. The functional
relationship between process parameters and the derived results can thus be de-
termined and mathematically described with little manpower and methodological
effort required [6, 7].
Statistical experimental design helped to reduce the polysaccharide and lipid
content of Champagner Bratbirne and wild service berry cultures to less than 5%,
and increased cell biomass yields by nearly 300%. At the same time production
costs could be reduced by a factor of 3 [4]. This demonstrates the great potential of
statistical experimental design to improve production processes. In addition to the
environmental conditions, various culture vessels were also evaluated with regard
to yield, production capacity and cost/benefit analysis. There was a choice between
1 liter Erlenmeyer flasks, 5 liter disposable bioreactors, 10 liter wave bags and
a 200 liter disposable bioreactor [5]. The 5 liter disposable bioreactors performed
best in comparative studies due to their low acquisition and operating costs and
their ease and flexibility in handling.
10.2.4 Conclusion
Plant cell cultures present an attractive alternative to the use of whole plants or
fruits, which may have fluctuating crop yields and quality depending on the seasons
and environmental conditions or be contaminated with harmful substances. The
growth conditions of the stem cell cultures could be optimized and a significant
reduction in production costs could be achieved by using statistical experimental
design. The plant-based extracts from the Champagner Bratbirne and wild service
berry cultures have been used in selected products for the cosmetics industry since
late 2013.
10.3 Value-Added Compounds from Microalgae—Increased

Value Creation Through Cascading Use and
Fractionation
Felix Derwenskus, Ulrike Schmid-Staiger, and Ursula Schließmann
Project Name:
Value-added compounds from microalgae
Aim:
The aim of the subproject was to culture three microalgae strains (Phaeodactylum tricornu-
tum, Chlorella vulgaris and Nannochloropsis sp.) to have the highest possible content of the
target ingredients and to investigate cell disruption and extraction processes with regard to
the holistic value creation of microalgae biomass for the food industry.
Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), the Institute of In-
terfacial Process Engineering and Plasma Technology (IGVP) at the University of Stuttgart,
Max Rubner Institute Karlsruhe, Institute for Clinical Nutrition of the University of Hohen-
heim, University of Freiburg

Project period 01/01/2015–12/31/2017
Funding volume 390,000 C
Contact:
Dr. Ursula Schließmann
Fig. 10.3 Light-microscopic images (resolution 1 : 1000) of various microalgae species.

From left to right: Chlorella sorokiniana (green alga), Phaeodactylum tricornutum (diatom)
and Anabaena species (cyanobacterium) (Fraunhofer IGB)
Table 10.1 Allocation of different growth conditions

Heterotrophic Mixotrophic Photoautotrophic
Light x x
CO2 x x
Organic C source x x
10.3.1 Introduction
Microalgae are one of the world’s most important biomass producers. They do not
only make a major contribution to global oxygen production, but are also able to
produce high quality compounds such as proteins, omega-3 fatty acids or antioxi-
dant dyes, which are of particular interest to the food, animal feed, cosmetics and
pharmaceutical industries. They represent a group of plant-like, unicellular organ-
isms, of which there are an estimated 300,000 different species on earth today.
So far, about 40,000 species have been described and a few analyzed in detail.
The term “microalgae” encompasses both prokaryotic cyanobacteria and eukary-
otic microalgae species (Fig. 10.3), which can grow in a variety of environments
such as seawater, freshwater and on different soil surfaces.
Depending on the particular species, microalgae can grow under heterotrophic,
mixotrophic or photoautotrophic conditions (Table 10.1).
In the project presented here, the microalgae were cultivated photoautotroph-
ically. The focus of the investigations was the production and fractionation of
proteins and valuable lipid fractions from the three microalgae strains Phaeodacty-
lum tricornutum, Chlorella vulgaris and Nannochloropsis sp. Culturing processes
for the target products developed in-house at the Fraunhofer IGB were used, for
example, to guarantee the production of biomass with a high pigment and EPA
content.
Fig. 10.4 Ingredients from microalgae and their target markets. (Fraunhofer IGB)
10.3.2 Microalgal Ingredients and Areas of Application
The composition of the microalgal ingredients depends on the selected strain

and the processing conditions during biomass cultivation. Fig. 10.4 provides an
overview of the ingredients present in microalgae and target markets.
Under optimal culture conditions, proteins can account for up to 60% of the
total dry mass. They are highly suitable for food and animal feed applications,
as the amino acid profile is in line with WHO/FAO recommendations [10]. Both
the techno-functional and the nutritional properties are crucial in this regard. The
amino acid composition of microalgae is such that almost all species have a higher
proportion of essential amino acids than the FAO requires [26].
Some species of microalgae (e.g. Phaeodactylum tricornutum, Pavlova lutheri,
Nannochloropsis oceanica) contain phospholipids and galactolipids in their
chloroplast membranes, which in turn contain polyunsaturated omega-3 fatty
acids, especially eicosapentaenoic acid (EPA, C20:5, !) and docosahexanoic acid
(DHA, C22:6, !). EPA is found mainly in fish oils and can act as a precursor to
prostaglandin-3, which can inhibit platelet aggregation. A specific EPA intake is
also suspected to help reduce inflammation and the symptoms of depression [19].
When microalgae cells are cultivated under conditions of nitrogen or phos-
phorus deficiency, some species (e.g. Chlorella vulgaris) can accumulate large
amounts of triacylglycerides, which consist of glycerol and saturated as well as
monounsaturated fatty acids (mainly C16–C18). Under suitable conditions, these

fatty acids make up more than 60% of the total dry matter [20]. In addition to the
main products, however, microalgal biomass may also contain several high-value
secondary products such as carotenoids (e.g. astaxanthin, ˇ-carotene, fucoxanthin,
lutein) and phytosterols, which are of interest for their antioxidant and anti-inflam-
matory properties [9, 14, 18]. Some carotenoids can be used as natural, healthy
food colorants.
10.3.3 Cultivation of Microalgae
Selection of Microalgae Species

The fatty acid profile of microalgae is species-specific and differs widely depend-
ing on the different cultivation conditions. When conditions for growth prevail,
the diatom P. tricornutum contains a very high level of polyunsaturated eicosapen-
taenoic acid (EPA, C20:5n3). Algae such as C. vulgaris and N. oceanica when
cultivated under conditions of nutrient limitation, have predominantly unsaturated
and monounsaturated C16 and C18 fatty acids, which function primarily as storage
lipids.
Large quantities of pigments such as carotenoids and xanthophylls (oxygenated
carotenoids) are found in the diatom P. tricornutum in particular, which can make
a significant contribution to value creation. Fucoxanthin, one of the main pigments
associated with photosynthesis, is particularly present alongside beta-carotene and
zeaxanthin. From an industrial point of view, fucoxanthin is of special interest
because of its health benefits, which are largely based on the structure and func-
tional groups of the molecule [22]. Commercially available fucoxanthin is cur-
rently obtained predominantly from the processing residues of macroalgae. How-
ever, macroalgae have an approximately tenfold lower fucoxanthin content (in
terms of dry biomass) than microalgae and can only be harvested from the sea
on a seasonal basis [28].
For the initial cultivated biomass, a fatty acid content of 44.3% w/w for C. vul-
garis, 18.7% (of which 3.7% eicosapentaenoic acid, EPA) for N. oceanica and
9.0% (5.1% EPA) for P. tricornutum was established, based on dry biomass. The
protein content was determined by means of IR spectroscopy and was, respec-
tively, 19.5% w/w for C. vulgaris, 38.4% w/w for N. oceanica and 55.5% w/w for
P. tricornutum. While the two green algae C. vulgaris and N. oceanica produced
under nitrogen limitation predominantly demonstrated unsaturated and monosat-
urated C16 and C18 fatty acids, which function primarily as storage lipids, the
a b c
Fig. 10.5 Various bioreactor systems for microalgae cultivation: a Raceway ponds in
Southern California [27], b Tubular reactors (AlgaePARC, Wageningen University, The
Netherlands), c Flat-panel airlift bioreactors. (Fraunhofer IGB)
diatom P. tricornutum showed a very high content of polyunsaturated eicosapen-

taenoic acid (EPA, C20:5n3). Fucoxanthin could be produced in amounts of up to
2.6% w/w of the dry biomass through cultivation.
Influence of the Reactor System

Cultivation in reactor systems makes it possible to influence the constituents of
the microalgal biomass by regulating various process parameters, especially the
nutrient supply and light intensity [20]. Different systems are used depending on
the desired product and the species of algae. The most common systems are “open
ponds”, tubular reactors and flat panel reactors (Fig. 10.5). In the project described,
flat-panel airlift bioreactors (Fig. 10.5c) were used.
These were developed at Fraunhofer IGB and consist of chambers separated
by static mixers. These generate a swirling flow in each chamber of the reactor
(Fig. 10.6a) [11]. The flow pattern constantly brings the algae cells from the dark
side of the reactor to the side facing the light (Fig. 10.6b). This ensures optimal
light distribution, resulting in a very high productivity of up to 2 g L1 d1 dry
biomass (DBM) and a high biomass concentration of up to 20 g DBM L1 .
These reactors are equipped with automation systems that enable full control of
the CO2 level, temperature, pH, and nutrient concentration in the culture [20]. The
reactors can be operated indoors with LEDs (Fig. 10.7) or outdoors using natural
sunlight (Fig. 10.8).
Fig. 10.6 a Side view of a flat-panel airlift bioreactor; b Schematic representation of the
flow pattern in each chamber. The cyclic flow pattern provides for the transport of microalgae
cells from the side of the bioreactor that faces the light to the side facing away from the light.
(Fraunhofer IGB)
Fig. 10.7 Flat-panel airlift bioreactor (FPA) with artificial lighting, pH and temperature con-
trol and automatic feed. (Fraunhofer IGB)
Fig. 10.8 FPA bioreactors in the Fraunhofer CBP pilot plant in Leuna. (Fraunhofer CBP)
10.3.4 Biomass Reprocessing
Cascading Use
Although microalgae can in some cases be used as whole cells for nutritional pur-
poses, it is often worthwhile to fractionate the various components. This increases
the added value and compensates for the relatively high production costs. The de-
cisive factor is the development of suitable cell disruption and extraction methods
under the premise that the resulting fractions maintain their techno-functional or
physiological properties in the subsequent processing steps. By combining suitable
cell disruption methods (e.g. mechanically by means of an agitator bead mill) with
subcritical and supercritical extraction methods, integrated overall processes can
be developed, and with these the biomass can be pre-treated for use in the high-
end sector.
For this purpose, the cells must first be gently disrupted in order to obtain the
lipophilic and hydrophilic fractions in successive extraction and separation steps
(Fig. 10.9).
Harvesting, Cell Disruption and Extraction

The first step required for cell harvesting and biomass concentration is solid-liquid
separation (e.g. by flotation, filtration or centrifugation). This is usually followed
by cell disruption to ensure direct contact between the solvent and the product
compounds that are to be extracted.
Fig. 10.9 Extraction and separation steps for the fractionation of microalgal ingredients
using the example of proteins and lipids. (Fraunhofer IGB)
Up to now, mechanical and chemical or enzymatic methods have been used as

digestion methods—for example high-pressure homogenization, disintegration in
agitator bead mills, application of a pressure gradient, pulsed electric field (PEF)
treatment, heating, osmotic shock treatment or chemical and enzymatic treatment
[13, 16]. In many cases, mechanical cell disruption using an agitator bead mill or
high-pressure homogenizer proved to be the most effective method [25].
Classic solvent extractions are among the methods used for the extraction
of lipophilic ingredients such as triacylglycerides, polar membrane lipids or
carotenoids [13]. New, cascading extractions with high-pressure extraction meth-
ods, however, demonstrate clear advantages, for example, relatively minimal
environmental impact compared to conventional solvent extraction. Processes
such as subcritical, pressurized liquid extraction (PLE) with organic solvents (e.g.
ethanol, ethyl acetate) or supercritical fluid extraction (SFE) with CO2 and polar
co-solvents, if needed, can be applied sequentially to separate products according
to their polarity. Both extraction methods operate at high pressure and moderate
temperature, thereby allowing the nutritional value and techno-functionality of the
extracted compounds to be retained [23]. Furthermore, several suitable solvents
are available that meet the requirements and regulations of the food and animal
feed sector.
In order to obtain additional added value, microalgae proteins are either ex-
tracted from the residual biomass following extraction of the lipid fraction, or are
separated beforehand during cell disruption [24]. The results obtained in the project
show that extractions with subcritical and supercritical fluids (PLE) are suitable
methods for lipid and carotenoid extraction from microalgae and are in principle
very well-suited to the extraction of the fraction of value-added compounds for the
areas of application addressed. The yields, especially those from the subcritical ex-
traction method, are significantly higher than previously published literature data.
It could also be shown, in the case of PLE extraction, that the efficient extraction
of aqueous biomass is possible. This in turn renders a preceding, energy-intensive
drying step superfluous, which is essential in the case of supercritical extraction,
for example.
For the production of ultrapure extracts, e.g. for pharmaceutical or cosmetic ap-
plications, further processing steps are necessary for the lipid fractionation beyond
the simple sequence shown here. For many applications in the food and animal feed
industry, however, further purification of the fractions is not required. It could be
demonstrated that both the extracts obtained and the disrupted microalgal biomass
in its current composition show properties beneficial to health on account of their
ingredients [12, 21] and that they moreover have excellent bioavailability [15].
The high levels of pigments such as fucoxanthin (e.g. in P. tricornutum) can make
a particularly significant contribution to the added value of the microalgal biomass
(Fig. 10.10).
10.3.5 Outlook
The potential added value of the respective ingredients is shown in Fig. 10.10. The
AERTOs community of European research institutes on the strategic topic of bio-
based economies (Fraunhofer together with, among others, SINTEF Norway, SP
Sweden, VTT Finland, TNO Netherlands, VITO Belgium and Tecnalia Spain) also
investigated microalgal biomass reprocessing methods, evaluated them with regard
Fig. 10.10 Value creation potential of microalgal ingredients. [8]
to technological and economic factors and developed value chains which facilitate
their economical production.
Information from the Baden Württemberg bio-economy research association
also flowed into the evaluation. This led to the insight that it is not only the added
value that has to be considered in order for algae biorefineries to be established,
and that the processing costs for algal products on a commercial scale are not the
critical factor for economic success, but that the key factor influencing costs is
algal production in photobioreactors as well as in raceway ponds.

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Biologized Robotics and Biomechatronics
Opportunities and Challenges in Human-Robot
11
Collaboration
Norbert Elkmann, Roland Behrens, Martin Hägele, Urs

Schneider, and Susanne Oberer-Treitz
Summary
In this chapter, after introducing the topic of “biologized robotics and biomecha-
tronics”, the authors begin by discussing the opportunities and challenges of
human-robot collaboration. They consider the potential applications, forms of
interaction as well as hazards and how to avoid them, among others, by defining
load limits. After looking ahead at the future of human-robot collaboration, they
go on to present medical technology applications such as endoprostheses and
exoskeletons. Here they start by describing the transition from the mechanical
to the mechatronic human-technology interface. The chapter is rounded off
by looking at new approaches to recording biosignals, and the combination of
functional electrostimulation with actuators, and concludes by introducing the
concept of hybrid exoskeletons.
11.1 Introduction
Nature has always been a source of inspiration for innovations, especially in

robotics. The vision of the ultimate machine is therefore still that of a humanoid,
which imitates human mobility and agility as well as our perceptive and cognitive
N. Elkmann () R. Behrens

Fraunhofer Institute for Factory Operation and Automation IFF
Magdeburg, Germany
M. Hägele U. Schneider S. Oberer-Treitz
Stuttgart, Germany

https://doi.org/10.1007/978-3-662-59659-3_11
200 N. Elkmann et al.
abilities, and can therefore perform practically any task. Although market-ready
products still seem far removed from this vision today, robotics is increasingly
benefiting from functional principles derived from biological models. The motiva-
tion behind this is to make kinematics more compact, more agile, safer and more
graceful.
The next level is biointelligence, i.e. the fusion of biological structures and pro-
cesses with information processing, as already applied in the latest learning control
systems in robotics or prosthetics. The present chapter on biologized robotics and
biomechatronics focuses on the highly topical subject of safe interaction between
human and machine. On the one hand, it is about the safe division of tasks in pro-
duction, on the other hand, there is also a focus on robots or machines that humans
can carry on their bodies.
11.2 Human-Robot Collaboration: Overview, Opportunities

and Challenges
Industrial production companies are currently undergoing a techno-economic

transformation that is characterized by the progressive automation of manual tasks.
Collaborative robots play a role as an element of this transformation. Thanks to
their integrated safety sensors and specialized design, they can safely work in the
immediate vicinity of humans. Conventional protection measures used in automa-
tion technology, e.g. safety guards, are then no longer mandatory. However, the
greatest advantage of this increasingly widespread type of robot is the ability to
adjust existing means of production to create a smooth, on-demand and cost-effec-
tive automated process [8, 54, 58]. At the same time, collaborative robots open up
new solutions to counter current societal change and its economic consequences
by relieving employees with disabilities of physically demanding work [2, 42, 50]
and thereby reducing the physical or mental overload of employees in general [2,
20, 40, 44, 55].
11.3 Potential Applications, Classification of Human-Robot

Collaboration, Standards
Human-robot collaboration (HRC) encompasses the division of labor between hu-

man and robot. It combines the fine motor skills and cognitive abilities of humans
with the performance capability of a robot to create a flexible and ergonomic work
system [6, 20, 26, 27]. Despite modern safety features, collaborative robots are
11 Biologized Robotics and Biomechatronics 201
subject to the same legal safety requirements as conventional machines. Hence

their distributors must carefully consider the hazards and risks to humans and the
environment that arise from the particular robotic application [58]. As with other
human-machine interfaces, the risks largely depend on the level of interaction be-
tween humans and robots [43].
11.3.1 Forms of Interaction
In an industrial environment the nature of a human-robot interaction can easily be

described by the following features [5, 40]:
Shared workspace: Human and robot share a common workspace (collaborative

space).
Simultaneous work: Human and robot work in the collaborative space simulta-
neously.
Physical contact: Human and robot work “hand in hand” (physical contact is
required to perform the task).
The order of these features follows an implicit logic. For example, humans and
robots can only work simultaneously if there is a common workspace. Likewise,
physical contact can only occur if the robot and the human work in the collabora-
tive space simultaneously, etc. The possible implications result in four forms of
HRC [5, 40, 55]:
Co-existence: Human and robot carry out independent activities in separate

workspaces that are not directly related to each other.
Sequential cooperation: Human and robot work one after the other in the col-
laborative space in order to achieve a common process goal. There is sequential
alternation between manual and automated work.
Parallel cooperation: Human and robot work simultaneously in the collaborative
space in order to achieve a common process goal. Direct contact between the
two cooperating partners is not required for the operational task.
Collaboration: Human and robot work hand in hand to achieve a common pro-
cess goal. Direct contact is required for the operational task.
The four forms are based on the conceptual differentiation provided by Spillner
[55]. Their analysis shows that these forms are in a consistent relationship with the
three features introduced above (Fig. 11.1).
Fig. 11.1 Forms of HRC and standard safety principles. [6]
11.3.2 Hazards and Safeguarding Modes
The planning and implementation of the forms of interaction must guarantee that
human health is not endangered during cooperation with the robot [2]. For hu-
mans, the most common hazard is the mechanical effect of clamping or collision
with the robot [2, 50]. Since enclosure of the robot with safety fences contradicts
the guiding principle of collaborative robotics, appropriate protective measures are
required [21, 29, 30].
To this end, international regulations specify four safeguarding modes in DIN
EN ISO 10218-2 and ISO/TS 15066:
Safety-RATED Monitored stop: The robot stops when a human enters the col-
laborative space and stays in this state for as long as a human remains there.
Speed and Separation Monitoring: Human and robot work simultaneously in
the collaborative space. Human safety is ensured by maintaining a minimum
distance to the robot. The robot stops if the human comes closer than this dis-
tance.
Power and Force Limiting: Direct contact between human and robot is permit-
ted. Human safety is ensured through adherence to biomechanical limits.
Hand Guiding: A control unit at the end effector of the robot converts movement
commands given by the human into movements. Clear workplace design, a safe
speed limit and intuitive operation ensure human safety.
In the Federal Republic of Germany occupational health and safety is cov-

ered by the autonomous law of the German Social Accident Insurance (DGUV),
whereby the four safety principles are mandatory for all operators of collaborative
robots in an industrial production facility [43]. The right hand side of the taxonomy
in Fig. 11.1 indicates which principles are suited to which form of HRC. It is clear
from this that the safeguarding mode Power and Force Limiting is applicable to
every form of HRC. This explains why Power and Force Limiting (PFL) currently
enjoys a high priority among users and robot manufacturers [1, 8].
11.4 Risk Assessment when Collaborating with Robots
How severe the potential hazards of a collaborative robot are depends on which
features underpin the human-robot contact. Table 11.1 introduces three qualitative
features to better differentiate these kinds of contact [11, 23, 24, 31, 38]. Currently,
ISO/TS 15066 prescribes that no surface parts of a collaborative robot may have
sharp points or edges [17]. This stipulation is based on the high hazard potential
that arises from such contours [5]. Thus, two applicable features remain with four
possible combinations, each representing a specific contact case (Fig. 11.2).
Table 11.1 Features of contact between human and robot

Feature Form Brief description
Load curve Quasi-static The contact force rises slowly. Its progress over time
displays no exposed maximum and remains at a constant
value
Dynamic The contact force rises quickly and drops quickly after
reaching its maximum
Spatial Unconstrained The human body part touched by the robot can move out
constraints of the way in the direction of the contact force
Constrained The touched body part is obstructed in the direction of
the contact force and cannot move out of the way
Contour Blunt The surface contour that touches the human tends not to
penetrate the epidermis at moderate forces
Sharp The surface contour tends to penetrate the epidermis
even with minimal force
Fig. 11.2 Differentiation

of dangerous human-robot
contacts and availability of
biomechanical limit values
for their safeguarding (up
to now ISO/TS 15066 has
only listed verified limit
values for clamping)
11.4.1 Current Need for Biomechanical Limits
Fig. 11.2 indicates with a check mark that ISO/TS 15066 currently only lists re-
liable limit values for clamping. For all other cases of contact, it either does not
include any verified limit values (as for push and clamping collision) or the limits
are not yet verified (applies to free collision).
For the case “push”, there is no need for customized load limits as there is no
risk of injury, because the robot is operating at a very low speed [5, 24]. According
to the estimation of the DGUV, load limits are not required for a clamping impact
either, as severe injuries can hardly be avoided if clamping occurs at high robotic
speed. Thus, action still needs to be taken for free collisions, which can occur just
as frequently as clamping in workspaces with collaborative robots. The urgency
of this need is exacerbated by the fact that PFL is currently enjoying the greatest
attention among users of collaborative robots (Sect. 11.3.2).
11.4.2 Differentiation of Biomechanical Limits
ISO/TS 15066 defines the onset of pain as the maximum allowable stress due to
clamping and collision with a collaborative robot. This threshold indicates the
moment at which a feeling of mechanical pressure changes to become a minor
sensation of pain [10, 25]. The onset of pain applies to all contacts that are deliber-
ately made by humans (intentional contact) or which are preceded by foreseeable
misuse according to Machinery Directive 2006/42/EC (unintentional contact).
A pain stimulus is triggered when the intensity of the effect of a force on the
body reaches the range where there is a risk of tissue damage [41, 65]. The force
inducing the pain is thus related to the contact area, which transmits it to the human
[9, 48].
A relationship between the force and surface area results from the normal stress,
which arises on the surface of the impacted body part [49]. From a mechanical
point of view, this is an interfacial phenomenon of the stress-strain state in the af-
fected tissue [67]. This operates in the form of local forces on receptors in the skin,
which become active at a certain threshold of force and trigger the transmission of
pain signals to the spinal cord. Neurological transmission structures in the spinal
cord accumulate the information from the activated receptors. Only when the sum
of all these signals reaches a certain threshold is the stimulus transmitted further to
the brain [60].
Taking the pain biomechanics of the body into account, it is necessary to distin-
guish the type of mechanical contact:
Edged or semi-sharp contact—an increase in tension within the contact area

activates individual receptors and their combined high potentials exceed the
transmission threshold.
Flat or blunt contact—a force evenly distributed over the contact area activates
a large number of receptors and their combined low potentials exceed the trans-
mission threshold.
The need arises from both types of contact and their associated pain transmis-
sion processes to apply normal stress and force-based load limits. A simplified
model of the relationship between the two limit values is illustrated in Fig. 11.3.
It shows the maximum allowable contact force FOC as a function of the contact
area AC . The curve increases linearly up to the area AOC . Its slope corresponds to
a limit value O C based on normal tension. From the area AOC onwards, FOC assumes
a constant value FO0K .
8
<O A AC AOC
C C
FOC .AC / D (11.1)
:FO0K otherwise
Fig. 11.3 Differentiation of limit values according to the type of contact. (Loosely based on
[30])
11.4.3 Studies on the Development of Load Limits
Fraunhofer IFF has been involved in the field of “biomechanical load limits for
safe human-robot collaboration” since 2010 and has carried out numerous studies
and research projects. At the beginning of these activities, a pendulum-like testing
device was developed, which was the first worldwide able to investigate the effect
of collision loads on subjects (Fig. 11.4). Since 2016 the Fraunhofer IFF has also
had an algometer at its disposal from the Institute for Occupational Safety. This is
a testing device that is used to determine load limits for clamping contacts. Dur-
ing the studies, the Fraunhofer IFF collaborated with clinical facilities of the Otto
von Guericke University in Magdeburg. The responsible ethics committee of the
university positively approved the studies.
The shape of the impactor can be varied on both measuring devices. It was
necessary to vary not only the impactor for the collision, but also the pendulum
parameters, collision speed and collision mass. A collision mass of up to 20 kg can
be set, thanks to its robust construction. At full deflection, it reaches a collision
speed of 1.25 m/s.
Fig. 11.4 Collision experiment with a pendulum acting on the forearm muscle of a subject
in the Fraunhofer IFF laboratory. (Fraunhofer IFF)
The algometer can also be equipped with different impactors for investigating
clamping loads. The speed with which the clamping force increases is also freely
adjustable. Both testing devices have various measuring means that record all the
quantities relevant to the load. These include piezoelectric load cells and a system
from TekScan for recording the normal stress, which arises between the impactor
and the impacted part of the body.
Studies on the experimental determination of pain thresholds are well advanced
and have been presented to the German and international standardization bodies.
Supplementary studies are currently being prepared. These include investigations
into the onset of injury, for which Fraunhofer IFF has already generated globally
unique initial findings in two preceding studies.
The results of the Fraunhofer IFF studies form the fundamental basis for the
design of safe workplaces with physical human-robot collaboration today and will
continue to do so in the future. Parallel to the experimental part of the study and
the goal of defining verified limit values, Fraunhofer IFF is working on the devel-
opment of body models that will permit the hazard assessment of robots by means
of simulation in the future. The use of models means it will no longer be necessary
to measure, for example, collision or clamping forces at the real robot setting. In
future, a virtual measurement in a simulated environment will be sufficient. The
results from the subject studies make an important contribution to the development
of the models by helping to verify the models and their quality. The finite element
method (FEM) serves as the basic technology [61]. Initial research has proven this
approach to be both viable and trend-setting.
11.5 Robotic Applications with HRC: Now and in the Future
In industrial robotics, HRC is usually associated with the physical cooperation

between an operator and a robot performing joint production tasks or with co-
existence, i.e. robotic applications where no safety fences are used and without
explicit cooperation between human and robot. In addition, cognitive robotic sys-
tems can be implemented for numerous other applications, such as informational
cooperation or the interpretation of operator actions or environmental conditions
as control signals. The available robotic safety technology, such as safety controls
to limit working and speed ranges, allows for direct interaction between robot and
operator while taking the normative specifications into account.
The influence of the operator can be extended in conjunction with integrated
sensors without limiting the autonomy of the robotic system.
In the following, HRC applications are presented, subdivided according to the
forms of HRC defined in Sect. 11.3.1 and evaluated in terms of areas of application
and productivity.
Operators and robots work in collaboration when, for example, motion and con-
trol signals are transmitted to the robotic system by direct physical cooperation. In
this way, some of today’s robotic systems already integrate force-moment sensors,
or at least provide appropriate interfaces for the input of motion specifications e.g.
in the form of Cartesian position corrections. This allows, for example, the op-
erator’s skills relating to the positioning of tools and products to be used in the
process. These can in turn be superimposed through the analysis of imaging sen-
sors and comparison with computer-aided models of the production data. Fig. 11.5
shows an HRC system for programming welding paths by manual guidance and
recording workpiece positions using a line scanner matched with stored CAD data
[36].
When hand-guided programming—or programming by demonstration—is used
in welding, for example, welding quality can be optimized through path inter-
polation algorithms, and the time required for an inexperienced operator can be
Fig. 11.5 Hand-guided robot programming for small batch production. (Fraunhofer IPA)
reduced many times over compared to conventional programming [39]. Thanks

to integrated joint-torque sensors, KUKA’s LBR iiwa robot can also use physical
cooperation during the process over and above the programming, to make posi-
tion corrections or enter control signals in the process. In the case of the iiwa,
the integrated force sensor system has been implemented with a focus on safety
and therefore also serves to put the safety principle of power and force limitation
described in Sect. 11.3.1 into practice [34].
The HRC system, SILIA, shown in Fig. 11.6, which has been implemented as
a sequential cooperation using the safety principle of speed and separation mon-
itoring, functions without haptic interaction. It is used to perform manual and
automated operations at a cooperative assembly station. 3D cameras track the op-
erator’s movement. On the basis of this information, the workstations are cleared
for the robot’s use. An estimation of sequences in the precedence graph and cor-
responding hypothesis formation are used to anticipate operator movements. The
subtasks of the process assigned to the robot or operator are stored in an intelli-
gent sequence controller. This makes the system productive and at the same time
allows the operators to structure their activities freely. The safety of the physical
cooperation is thus made possible by monitoring a minimum distance between the
Fig. 11.6 HRC assembly cell with time-staggered utilization of cooperative work spaces.
(Fraunhofer IPA)
robot and the operator, where reaction and braking times are taken into account
according to the specifications for functional safety.
In the future it will be essential to not only use lightweight robots, but also to
introduce heavy duty robots into HRC, especially in applications that improve the
ergonomics for the worker.
Fraunhofer IFF has implemented the first instance of human-robot collabora-
tion with a heavy goods transporter on behalf of Adam Opel AG in Rüsselsheim
(Fig. 11.7). In conventional door assembly during continuous flow production, an
employee removes the doors with a hand-guided handling system and brings them
to the mounting position on the vehicle. The door is then mounted on the body.
In the newly developed process, a robot with high load capacity automatically re-
moves the door from the door conveyor and moves it to the installation position.
Only then is the work area released for the employee, who then takes over the
installation.
During installation by the employee, the robot holds the door, keeping it syn-
chronized with the conveyor belt speed, resulting in a spatially close collaboration
between human and robot. Particular attention was paid to safety in implementing
this system, which could be achieved through the use of selected safety sensors, se-
cure grippers and the targeted combination of safe operating modes for the human-
robot collaboration (see Sect. 11.3.2).
Mobile robot systems implemented as highly integrated HRC systems whose
manufacturing abilities can be made available to different workstations on demand,
independently of fixed installations (Fig. 11.8), promise the breakthrough to the
adaptable factory of the future.
Fig. 11.7 Human-robot collaboration with a heavy goods transporter for door assembly.
(Adam Opel AG)
As part of the ARENA20361 research campus, Fraunhofer IPA developed the

concept of a mobile screwing assistant as a surface-movable platform with a sen-
sitive robotic arm equipped with various tools such as grippers or screwdrivers
(Fig. 11.8). By adapting the tools to a workstation laid out for manual work, the
scope of work can be systematically allocated according to the respective strengths
of human and machine and dynamically assigned to the station in line with the
available capacities [59].
In this process it becomes evident that the safety principle implemented signifi-
cantly influences productivity, e.g. through dimensioning a safe speed according to
whether or not a person is present and the collision risk associated with how the
system is currently equipped.
Therefore, in order for it to be possible to implement a robotic system for paral-
lel cooperation and to adjust the productivity for each HRC scenario, the existing
rigid applications of safety technology must be overcome. Only then is dynamic
1
ARENA2036 (Active Research Environment for the Next Generation of Automobiles) is
the largest and leading research platform for mobility in Germany, in which the entire value
creation chain of the fully digitalized vehicle of the future is being rethought and imple-
mented.
1 worker, 2 workers, 1 robot 4 workers at

1 worker 2 workers
1 robot installed at 2 stations 2 stations
Work cycle [s] 107 74 55 37 28

1 station 2 stations more than 2 stations
Fig. 11.8 Mobile door installation station as a highly integrated HRC robotic system in
adaptable production. (ARENA2036/Rainer Bez, Fraunhofer IPA)
configuration possible. For this reason, “plug & safe” concepts are being used
at Fraunhofer IPA, which expand the aspects of dynamic process adaptation by
decentralizing the safety configuration and communication to safety-related com-
ponents.
With the aid of an online risk analysis tool (Fig. 11.9), as developed within
the scope of the LIAA project [37], the specific hazards of an HRC system were
to be evaluated already in the planning phase of an application. An automated risk
identification for an HRC workstation is carried out using a model-based approach,
and the automation planner is supported in the decision-making regarding work
equipment selection for the application. This allows the decision-maker to better
estimate how great the influence of different safety principles and components is
on the efficiency of the application when a particular process is implemented [4].
Fig. 11.9 Online risk analysis with automated hazard identification based on semantic de-
scriptions. (Fraunhofer IPA)
Robotic systems in networked production plants must offer versatile technol-

ogy that can, in real time, evaluate a high volume of product and process data,
which in some cases is marked with uncertainties. Only then can HRC contribute
to the implementation of adaptable production systems, by possessing a flexible
degree of automation. Through the virtualization of production in the context of
Industry 4.0 further requirements are added, over and above compliance with the
basic requirements for physical safety. These concern data security and the pri-
vacy of the production and user data. Safety is therefore a three-fold factor (safety,
security, privacy) and a central criterion when commissioning modern production
plants.
This topic is of high relevance for production based in Germany, which is
also reflected in current funding measures available to companies. Examples in-
clude the projects “Roboshield” (www.roboshield.de) and “CyberProtect”, which
support companies based in Baden-Württemberg in the implementation of safe ap-
plications with automation. In developing software platforms with key integrated
safety-critical technologies, digital tools are being developed that govern access to
the production system, software and data through secure deterministic communica-
tion techniques, pre-operational and operational simulations and risk assessments.
Artificial intelligence and machine learning approaches are thus to be used as tech-
niques to verify the safety of manufacturing equipment, to support manufacturers,
system integrators and end users of industrial automation technologies in tomor-

row’s production facilities.
The robots used in service robotics come even closer to humans. In addition
to the direct evaluation of user input and object localization, the field of semantic
interpretation of language, gestures, or human behavior is already playing a much
greater role in cooperation here. While the human being cooperating with an in-
dustrial robot is regarded as the operator of the robotic system, service robots offer
many more diverse possibilities for cooperation with humans.
11.6 Exoprostheses and Exoskeletons
11.6.1 From the Mechanical to the Mechatronic Human-

Technology Interface
Biomechatronics as a subject notably includes aspects of robotics, measurement

and control technology, biomedical engineering, biomechanics, human-technology
interactions and rehabilitation measures as research topics.2
Among other things, it involves the development of intelligent systems to sup-
port and restore defective functions of the human body [62].
Mechanical Solutions
Exoskeletons have by now reached a product-relevant state of the art as mechanical
human-technology interfaces, as orthoses from technical orthopedics. Both mod-
ern pelvis, leg and foot orthotic devices for the rehabilitation of cerebral palsy
[19] and passive load-bearing pelvis, leg and foot structures such as Noonee’s
ChairlessChair® can absorb and redirect loads into the ground. Such systems are
already medically established in applications for physical relief and support and
in certain cases of incomplete paralysis, they represent the only way patients can
be mobilized to walk. In the field of occupational health and safety applications
for the reduction of physical fatigue and possibly the delay of musculoskeletal
ailments, all passive exoskeleton systems are still in the early testing phase of ini-
tial products and prototypes. It is already becoming apparent that purely passive
exoskeleton structures, while highly relevant in individual factory application sce-
narios, are only capable of variable application to a limited extent, due to their lack
of adaptivity. This will probably limit their areas of application in the future.
2
According to the definition of Biomechatronics given in the Biomechatronics study pro-
gram of the TU Ilmenau, 2018.
Mechatronic Solutions Without Positive Energy Input

Adaptive braking systems have also long been established in technical orthopedics.
These control, for example, the moment of resistance that is generated in hydrauli-
cally braked knee joint modules using motor-controlled valve trains. This provides
a relatively energy-efficient method to influence the movement demands a user
places on a prosthetic joint in terms of stability or lability [63]. Potential benefits
of such microprocessor-controlled prosthetic knee joints include flexible adapta-
tion to the ground when walking, increased walking safety, increased user activity,
mental relief when walking, increased quality of life, a more cost-effective thera-
py—from a holistic point of view—happier patients, less energy required while
walking and a visibly as well as biomechanically measurable more natural gait.
Clinical evidence for these desired functions is still limited, in particular due to the
lack of comparable study designs, but many internationally run clinical studies and
initiatives for the standardization of assessment tools are working on providing this
evidence [51].
With regard to biological-technical assimilation, in the case of such an exopros-
thetic knee joint a technical control of valves is available that is based on the user’s
kinetic and kinematic data.
In addition, the user adjusts neuromuscularly as a biological system to the me-
chanical behavior of the prosthesis. According to the definition, one can certainly
speak of assimilation here. The mechanically communicating interface is either
a prosthetic socket system or, in certain cases, the direct connection of a skin pen-
etrating implant to the bone. Such mechatronic and control solutions are already in
use as products in paralysis orthoses for the knee (C-Brace® , Otto Bock). The chal-
lenging difference in assimilation to the prosthetic knee joint is that the technology
and controller must influence the same natural joint as harmoniously as possible
[28].
Mechatronic Solutions with Positive Energy Input

In the case of muscle weakness, missing musculature following amputation and
active relief during hard physical labor where the musculature is intact, energy
needs to be introduced into the human system. This is already established and has
been implemented for motorized prosthetic hands to a largely satisfactory degree.
The technical solution is much more demanding for high power density actua-
tors worn on the body for the lower extremities, which the user does not find too
heavy. Ossur’s PowerKnee® and later Otto Bock’s BiOM Ankle® were the first
leg prosthesis components able to take over the critically high joint moments and
angular speeds while walking [12]. Whether the solution is a DC motor/gearbox
combination or a spring/DC motor actuator, such systems provide direct controller
Fig. 11.10 Active upper body exoskeleton “Stuttgart Exo-Jacket”. (Rainer Bez/Fraunhofer
IPA)
communication between human and technology, as the prosthesis controller reacts

adaptively to user kinetics and kinematics and the human gait adapts neurologically
to the prosthesis. One could refer to these as mechanically coupled, assimilated
control systems. Such actuator and control strategies are incorporated in various
paralysis exoskeletons for the lower extremities [32]. Active exoskeletons are also
being developed for the upper extremities, for example the “Stuttgart Exo-Jacket”
(Fig. 11.10) at Fraunhofer IPA [47].
Hybrid Controls
Biosignal interfaces with the technical controller of a prosthesis, orthosis or exo-
skeleton represent an extension of the above control strategies, where the aim is
optimal voluntary control by the user. Here too, hand prostheses are early demon-
strators of the technical feasibility. For over 50 years, hand actuators have been
controllable via electrical muscle signals through outlets in the skin. Weak sig-
nals, muscle cross-talk and soft-tissue shifts are all challenges affecting raw data
acquisition. Significant research findings have emerged at the sensor and data pro-
cessing level that have significantly improved the sensitivity and specificity of
the muscle signal for controller applications. New knowledge is also emerging
in neuroscience, which is equally supportive for technical systems in the com-
plex multiscale simulation of muscles. Such biosignal-influenced controllers are
also found in exoskeletons such as Cyberdyne’s HAL® , termed “cybernic volun-
tary control” (CVC) and “cybernic autonomous control” (CAC) [35]. This system,
with its hybrid controllers, is already being used in several countries, especially in
stroke rehabilitation. Here, “hybrid” refers to the combination of recorded kinetic
and kinematic measurement data and biosignals from the user. Therapy robots as
standalone devices also use these signals in some cases [16].
11.6.2 New Methods of Biosignal Acquisition
The weaknesses associated with the muscle signal acquired at the skin surface
have been indicated above. A variety of other methods for biosignal acquisition are
therefore being sought for hybrid controllers in order to assimilate the biological
and technical system in biomechatronics. Some of the research strategies currently
being pursued are mentioned here by way of example.
Implants in the muscle itself can also record the electrical activity in the acti-
vated skeletal muscle [45]. In addition, research is currently being carried out to
determine to what extent muscle movement can be acquired externally and robustly
via small implanted magnets and used for the voluntary control of hand prosthe-
ses [57]. Magnetic navigation is fundamentally considered to be a high-resolution
spatial measurement method in medical technology. The muscle activation itself is
a late signal. It has therefore been interesting for some time now to acquire data
earlier in the signal chain of voluntary motor function, namely in the area of the pri-
mary motor cortex. This is possible to a limited extent with various sensor systems
such as classical electroencephalography (EEG) for the detection of brain waves,
but also, for example, with near-infrared measurement systems [53]. This kind of
measuring system attempts to detect the activation of brain regions via thermal
phenomena. Implant experiments on monkeys have also been successful, as has
been shown at the EPFL in Lausanne [13]. Here, signals from the primary motor
cortex could be used to activate peripheral muscles. Two approaches are mentioned
below, in which nerves and muscles are prepared through surgical intervention for
better data interfaces for biosignal-based controllers in prostheses.
Targeted muscle reinnervation (TMR) follows the approach of providing easily
accessible and trainable muscles with new nerve connections for sensor systems
such as surface EMG. New possibilities for even more intuitive user control of
prosthetic hands have emerged out of this, especially at the Northwestern Uni-
versity in Chicago [14, 33]. End-to-end suturing of the agonist and antagonist in
the amputation stump is another new surgical route to biosignals. Here the vol-
untary contraction of the agonist leads to the stretching of the antagonist and to
the physiological stretch stimulus. Thus, in contrast to amputation techniques with
end fixation of the muscle stumps onto bones or soft tissue, an agonist-antagonist
interaction in the stump area is possible for the first time both in terms of actuator
and thus also sensory activity. This technique is referred to as AMI® and is cur-
rently being researched at MIT and Harvard Medical School and other institutes
[56]. At present, eight patients who have undergone lower leg amputations have
already been treated with this surgical technique and been given multi-axial, con-
trollable prosthetic feet. It is expected that both of these technologies will improve
the voluntary control of prostheses.
Functional Electrostimulation Combined with Actuators

Another dimension may be mentioned in the field of hybrid controllers in hu-
mans: The movement of an anatomical joint through the synchronized actuation
of paralyzed muscles and externally mounted motors [15, 66]. The aim here, de-
spite damage to the nerve, which leads directly to the muscle, is to stimulate the
fundamentally intact, but unstimulated muscle externally [46]. With functional
electrostimulation (FES) two important goals are being pursued in particular: On
the one hand, skeletal muscle has a more favorable power-to-weight ratio than
exoskeletal drives; on the other hand, its activation can return intact sensory in-
formation from the muscle (depending on the defect) to the spinal cord and brain.
This could lead to better rehabilitation success in the understanding of modern in-
tensive neurorehabilitation on account of the sensory feedback. In 2016, several
patients with spinal cord injuries rode in a trike race with externally stimulated
leg musculature in the Cybathlon research olympiad for the first time [3]. Among
them was a patient with a novel thigh nerve implant (First in Human, Cleveland
Clinic), which was able to activate several nerve branches and thereby several mus-
cle groups in the thigh by means of external stimulators [52].
Hybrid Exoskeletons
Hybrid exoskeletons are currently being explored as doubtless the most complex
exoskeletal biology-technology interaction, both for medical rehabilitation and oc-
cupational health and safety applications which guard against overloading. The
HAL® system mentioned above is a pioneering product for exoskeletons with hy-
brid controllers for motors worn on the body. In experiments currently being con-
ducted by M. Goldfarb at Vanderbilt University in Nashville, prospective patients
from three therapy centers are provided with an actuating hybrid of functional elec-
tro-stimulation of their own muscles and power assistance of the limbs [22].
11.7 Summary and Outlook
Fraunhofer institutes have played a key role in the development of technologies for
the safe interaction of humans and machines and their use in robotics, orthoses or
exoskeletons. The realized examples presented here display the basic principles of
biologization: The adaptation of technology to humans—right through to physical
symbiosis—and the use of biologically inspired control technology for recognition
of intent during movement.
The robotics of the future is characterized by imitation of the human or other
biological model. Yang et al. [64] present the scientific and ongoing technical
development of future robots on the basis of ten challenges, which can almost
entirely be classified as biologized robotics. These include the use of bionic de-
sign and functional principles as well as the combination of functionally integrated
materials with biologically inspired manufacturing processes. Miniaturization, ef-
ficient scalability and division of labor are also included, as well as the use of robot
swarms and the comprehensive application of artificial intelligence for the reliable
recognition of people and objects, and for intuitive interaction as well as the con-
stant performance optimization of robotic systems. Systematically tackling these
challenges and transforming them into useful robots and assistance systems are the
subject of extensive research activities.

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Future AM
The Next Generation of Additive Manufacturing
12
Processes
Johannes Henrich Schleifenbaum, Christian Tenbrock, Claus

Emmelmann, Christoph Leyens, Frank Brückner, and Alexander
Michaelis
Summary
Additive manufacturing (AM) is a technology with high disruptive potential that

is currently undergoing heated discussion. The combination of Industry 4.0 and
AM makes it possible to print industrial products directly based on digital data.
This can result in sustainable change in industrial value creation chains through-
out the whole spectrum of manufacturing engineering. Universities, research
institutes and young companies recognized the potential of additive manufac-
turing processes at a very early stage and have developed them into marketable
systems that have found their way from their applications in prototyping into
the manufacture of end products. A new branch of industry has emerged that
radiates throughout the entire value chain—from materials production and ma-
J. H. Schleifenbaum () C. Tenbrock

Fraunhofer Institute for Laser Technology ILT
Aachen, Germany
C. Emmelmann
Fraunhofer Research Institution for Additive Manufacturing Technologies IAPT
Hamburg, Germany
C. Leyens F. Brückner
Fraunhofer Institute for Material and Beam Technology IWS
Dresden, Germany
A. Michaelis
Fraunhofer Institute for Ceramic Technologies and Systems IWU
Hermsdorf, Germany

https://doi.org/10.1007/978-3-662-59659-3_12
226 J. H. Schleifenbaum et al.
chine technology and additive manufacturing processes as a service up to the

integration of additively manufactured components into new products.
There are still various “links” missing along the process chain, however,
before the comprehensive and cross-sector use of additive manufacturing pro-
cesses can occur. These include universal data formats, the uninterrupted link-
ing of digital and real process chains as well as concepts for the scalability of
AM processes with regard to build rate and component size, so that produc-
tion of larger quantities also becomes economically viable. Suitable concepts
are also lacking for the manufacture of multi-material components with AM-
adapted materials or the universal automation of the process chain up to and
including the postprocessing of components.
Numerous initiatives are working on solving these problems. For exam-
ple, in the “futureAM” focus project two strategic goals are being addressed,
namely, securing and expanding Germany’s technological leadership in the area
of metal AM, as well as establishing a comprehensive cooperation platform for
highly integrated collaboration, which makes use of the decentrally distributed
resources of the Fraunhofer-Gesellschaft and interested partners in the field of
AM. Technological leaps are needed to ensure this technological leadership and
any significant further development. These required leaps in technology may
be subdivided into four dimensions. Specifically, these are Industry 4.0 & the
digital process chain, scalable & robust AM processes, materials and system
engineering & automation.
The cooperation platform is not only created through the intensive collabo-
ration within and between the individual fields of action, but especially through
the development of a “Virtual Lab”. Out of this collaboration, with the participa-
tion of all partners and using the newly developed technologies, cross-industry
and cross-sector demonstrators are built, which indeed come from a range of
industrial sectors important to Germany.
12.1 Introduction
“Additive manufacturing is far more than just a promising trend: the applications
seem endless” [1]. This quote is representative of the optimistic spirit in much of
the manufacturing industry in Germany. Especially universities, research institutes
and young companies recognized the potential of additive manufacturing processes
at a very early stage and have developed them into marketable systems, which
12 Future AM 227
Fig. 12.1 Market and competitive environment
have found their way from their applications in prototype construction into the
manufacture of end products.
A new branch of industry has emerged that radiates throughout the entire value
chain—from materials production and system construction and additive manufac-
turing as a service to the integration of additively manufactured components into
new products. Up to now, there have mainly been niche applications, but the enor-
mous growth potential is causing more and more companies from the most various
industries, as well as large companies such as General Electric, Siemens, etc. and
additive manufacturing suppliers to surge onto the market.
The overall situation for Germany can be summarized as follows (Fig. 12.1):
Additive manufacturing represents a key technology for global future markets
with high growth potential, in which Germany has become the world market leader
with regard to metal AM. Despite this great potential, there are increasing signs that
Germany may fall behind internationally when it comes to applications. The recent
acquisitions of Concept Laser, a German manufacturer of 3D metal printers, by
GE Aviation and of the renowned service provider CITIM by the Swiss company
Oerlikon are an indication of this, as are the major AM-specific funding initiatives
of leading industrialized countries such as
USA: “America Makes—National Additive Manufacturing Innovation Insti-

tute” with over US $200 million start-up financing,
China: 3D Printing Alliance is establishing ten 3D institutes with US $245 mil-
lion in six years or
Nanjing plans US $100 million regional development project,
which go far beyond German funding measures in terms of volume.

In light of this, national initiatives for additive manufacturing were also devel-
oped in Germany. In a concerted approach between the Fraunhofer-Gesellschaft
and industry, the activities initiated by Fraunhofer for a research and development
program were put forward to the Federal Ministry of Education and Research
(BMBF) and a program for the technology transfer of additive manufacturing
to SMEs was proposed to the Federal Ministry of Economics and Technology
(BMWi). These initiatives are in line with concepts and recommendations currently
being developed for additive manufacturing by the Leopoldina (German National
Academy of Sciences) and acatech (German Academy of Science and Engineer-
ing). At the same time, the industrial initiative “Digital Engineering and Additive
Manufacturing” (IDEA) was submitted to the BMBF under the management of
Siemens and with the involvement of the VDI Technology Center. The aim of this
initiative is for Germany to become the world market leader in the application of
digital, additive manufacturing. The Fraunhofer-Gesellschaft is also a central re-
search partner to industry in this initiative. Final decisions on the realization of
these programs are still pending, but the chances are high.
A glance at the market trend in additive manufacturing also shows that this
topic is already extremely important to Fraunhofer today, but will be even more
so in the future. In 2015, the global market for additive manufacturing (materials,
facilities and services) amounted to US $5.165 billion. For the year 2021, a market
volume of more than US $26 billion is predicted [2]. The greatest growth potential
is expected in the area of metallic materials. Medium growth rates of 20% are
expected for the metal sector for the years 2015 to 2021, with a market volume of
between US $1.9 and 2.8 billion for the year 2021 [3].
In addition to the market development of AM for metallic materials, the follow-
ing reasons point to a focus on the metal sector:
The rapid development of additive manufacturing and the continuously increas-

ing interest of industry in recent years is based primarily on the development
and provision of processes, materials, facilities and systems engineering for
metallic materials.
12 Future AM 229
Metallic materials and components play a dominant role in Germany’s most

important sectors e.g. the automotive industry, mechanical and plant engineer-
ing, tool and mold making, turbo machinery, the aerospace industry and med-
ical technology, where the need for multifunctional, individualized products is
steadily increasing. AM is an ideal way to address this need.
Germany currently still has an excellent starting position in the additive man-
ufacturing of metallic materials. German companies such as EOS, SLM Solu-
tions, Concept Laser or Trumpf are globally leading providers of the appropriate
facilities and system technology with a global market share of around 70%.
Companies such as Siemens, Airbus, VW, BMW and MTU are among the
world’s first leading industrial users of AM.
The Fraunhofer-Gesellschaft and other German research institutions are among
the world’s most renowned for metal AM. They need to use their knowledge
base collaboratively to expand the leading position of German industrial com-
panies and thus prepare themselves for the next step AM needs to take on the
road to industrialization and broader distribution.
The importance of additive manufacturing processes (AM, 3D printing) to in-

dustrial production as well as the need for action were intensively discussed and
documented within the Fraunhofer-Gesellschaft:
Paper “National Initiative Additive Manufacturing”, Fraunhofer-Gesellschaft,

April 2016
Concept paper “Mittelstand 4.0 Competence Centers for Additive Manufactur-
ing”, Fraunhofer-Gesellschaft, September 2016
Strategic meeting of the board on 11/3/2016 in Leipzig as part of a workshop
on additive manufacturing.
The Fraunhofer-Gesellschaft has taken a leading role in the area of research and
development, especially with regard to metallic materials. Typically, individual in-
stitutes excel in and occupy different key areas, from material research through to
process development and machine technology, quality assurance and data integra-
tion, and develop these in close collaboration with industry.
Institutional research in an international context has seen increasing reinforce-
ment of major national initiatives. These sometimes massive investments fund
individual institutions and in particular create new networks and new market par-
ticipants. These changes represent increasing competition for the “Erfolgsmodell
AM@Fraunhofer” (AM success model@Fraunhofer). The planned project, with
a format that is based on the Fraunhofer lead projects, aims to strategically expand
the competitive position of Fraunhofer in the field of additive manufacturing and

establish new unique selling points.
Within the Fraunhofer-Gesellschaft, the institutes represented in Allianz Gen-
erativ, the Fraunhofer Additive Manufacturing Alliance, cover many of the com-
petencies needed in additive manufacturing across all types of materials as well
as along the entire value chain. However, the most intensive R&D activities in the
Fraunhofer-Gesellschaft with the greatest growth are currently in the area of metal
AM. Furthermore, within the framework of this project, the Fraunhofer Research
Institution for Additive Manufacturing Technologies IAPT (formerly LZN, Laser
Zentrum Nord GmbH) will be integrated into the Fraunhofer-Gesellschaft as an
institute with proven expertise in the field of metal AM.
12.2 Deficits and Derived Fields of Action for Metal AM
The sustainable, broad-based industrial implementation of additive manufacturing

achieves shorter innovation cycles as well as continuous improvements in develop-
ment and production. It therefore requires a holistic view to be taken of the vertical
and horizontal process chains (Fig. 12.2) as well as their integration into estab-
lished production chains. The vertical process chain includes the development and
Fig. 12.2 Process chains in additive manufacturing

12 Future AM 231
Fig. 12.3 State of the art and deficits in the metal AM sector
provision of suitable starting materials for the additive processes (in most cases
powder, as applicable, also wire), the actual AM process (e.g. SLM, LMD)as well
as the implementation of the AM processes with robust AM machines. Other im-
portant aspects of the vertical chain include quality assurance for online monitoring
and error detection during the AM process and the support of process develop-
ment and optimization through the development of simulation tools on different
scales. The horizontal process chain begins with the design of components and
tools suitable for AM, to take advantage of the tremendous degree of freedom of
AM. Following manufacturing by means of various AM processes, a post heat
treatment step is often carried out, both to improve component quality (e.g. HIP)
and to set desired structural properties. The final step of the horizontal chain is
usually the post-processing or finishing of the components. This includes, for ex-
ample, the removal of support structures and the separation of the component from
the substrate plate and the finishing of the component to achieve the required di-
mensional accuracy and surface topography.
The deficits that arise from the state of the art when it comes to metals are
summarized in Fig. 12.3: alongside the horizontal process chain consistent data
formats are lacking, existing simulation tools are not integrated into the process
chains, and there is no uninterrupted link between digital and real process chains.
In the vertical process chain, new concepts are lacking for the scalability of the
AM processes with regard to build rate and component size so that the production
of larger quantities may also become economically viable. Direct error detection
during the manufacturing process is another yet unsolved task.
With regard to materials, the range of materials that can be processed using AM
is still severely limited. The development of AM-adapted materials is still in its
infancy with only a few examples of AM-specific material developments. Also, the
multi-material processing for the optimization of component functions with stress-
resistant materials requires interdisciplinary process and material development and
is still outstanding today. One of the biggest shortcomings of the AM process is the
lack of automated post-processing of components. Numerous manual activities are
still required along the vertical and horizontal process chain, which significantly
impair the profitability of AM.
The objectives, solution approaches and course of action of this project are
derived from these major deficits.
12.3 Objectives
This project aims to achieve two strategic objectives:
Technology leadership: to expand the leading position in additive manufactur-

ing (AM) through new, fundamental technological developments and establish
the conditions for the next generation of additive manufacturing, so that the fu-
ture viability of the German industry is sustainably reinforced, especially in the
area of manufacturing engineering.
Cooperation platform: to create a cooperation platform with outstanding infras-
tructure and tremendous potential for synergy through the merger of excellent
Fraunhofer Institutes and the integration of Fraunhofer IAPT with its likewise
proven expertise in the AM sector into the Fraunhofer-Gesellschaft. This plat-
form ensures highly efficient, coherent cooperation and a strong capacity to
innovate, and guarantees that all the facets of this complex topic are addressed
and covered.
The need for a next generation of metal AM in order to sustainably secure

technology leadership arises from the realization that the developmental steps in
the area of AM processes and technologies are increasingly yielding less marginal
utility (Fig. 12.4, phase III). The scaling of productivity by means of existing laser
12 Future AM 233
Fig. 12.4 R&D expenditure and performance capacity of metal AM
and optical systems can serve as an example here (Sect. 12.4). The current develop-
ment path is aimed at the linear scaling of laser power through the use of multiple
scanning systems. In this process, however, a saturation region is reached regarding
the machine, build volume and process capabilities. Moreover, any further scaling
along this path will increase costs disproportionately, because it would not only
multiply the most expensive components, but also result in increasing complexity
in coordination. In order for significant further development to be possible, a tech-
nological leap analogous to the transition from mobile phone to smartphone is
necessary (Fig. 12.4). The foundations for this technological leap are to be devel-
oped in this project.
In order to achieve the strategic goals mentioned above, the project structure
shown in Fig. 12.5 was developed, which is characterized by two special features:
1. The deficits described in Sect. 12.2 were addressed and four fields of action
derived. As far as possible, these address all important issues along the vertical
and the horizontal process chain.
2. The cooperation platform is not only created through the intensive collabora-
tion within and between the individual fields of action, but most significantly
through the development of a “Virtual Lab”. The demonstrators are to be built
as a product of this collaboration and with the participation of all partners.
Fig. 12.5 Project structure and fields of action
The fields of action “Industry 4.0 & digital process chain” (Field of action 1),
“Scalable & robust AM processes” (Field of action 2), “Materials” (Field of ac-
tion 3) and “Systems engineering & automation” (Field of action 4) are of partic-
ular importance for the next generation of metal AM. The technical and scientific
objectives presented in Fig. 12.6 were therefore defined, which are to be achieved
in each case under the leadership of one institute and with the participation of the
others.
Another unique selling point of this project is the integrative, cross-institute and
cross-platform production of demonstrator components in order to demonstrate the
performance capacity of the technologies that have been developed:
Demonstrator components are to be manufactured from various sectors relevant

to Germany in order to demonstrate the cross-sector significance of the devel-
opments in this project.
The technical and scientific developments in the respective fields of action es-
tablish the basis for production.
12 Future AM 235
Fig. 12.6 Envisaged technical and scientific goals
The data processing and handling and the production of the demonstrators
takes place with the involvement of the four participating institutes in a joint “Vir-
tual Lab”. The complete process chain for the production of a steering knuckle
as a demonstrator from the automotive industry is shown by way of example in
Fig. 12.7. The first step is the design or re-design of the component to,for example,
incorporate grid or hollow structures into the component to reduce the weight. The
software tool developed at IAPT within the first field of action for the design of
AM components will be used for this purpose. The second step is to manufacture
the steering knuckle by means of the SLM process, using the newly developed
scalable AM concept machine (action field 2 at ILT). Thereafter, a reinforcement
structure made of a second high-strength material is attached at IWS (action field 3)
using the LMD method (multi-material AM). The automated removal of the sup-
port structures as well as postprocessing of the component are carried out using the
procedures developed in action field 4 under the lead management of IWU.
The combination of the development of new technologies (“technology leaps”)
in the individual fields of action and integrated processing in the joint Virtual Lab
creates a unique combination that has a positive influence both internally and ex-
ternally.
Fig. 12.7 Collaboration in the “Virtual AM Lab”
12.4 State of the Art
The initiatives mentioned in Sect. 12.1 and the existing funding projects and activ-
ities are aimed primarily at detailed innovations that are to be implemented in the
area of existing process and system technology, and also at the integration of addi-
tive manufacturing into industrial production environments. Various aspects must
be ensured for this: the robust production of larger quantities and reproducibil-
ity, process reliability and reliable system technology and installation engineering,
including control and drive technology, along the horizontal and vertical process
chains. Examples of what is required to achieve this include the optimization of
existing procedures, development of consistent software solutions in the form of
AM-specific CAD/CAM chains and establishment of standard interfaces suitable
for industry. Other approaches to the ongoing development, however, display less
and less marginal utility, so that both the leaps in development and the competitive
advantage gained become smaller. The state of the art and the tasks within the four
fields of action are described below.
12 Future AM 237
Field of Action 1: Industry 4.0 & Digital Process Chain

The digital process chain linked with Industry 4.0 represents a higher-level field of
action. Fundamentally, the following priorities are to be addressed here:
Fully interlinked individual steps of the vertical process chain, from the provi-
sion of the starting materials, to additive processing with materials in the form
of powder or wire and post heat treatment, right up to the finishing step e.g.
by machining processes. The adaptation of the interfaces for communication
between human, machine and component plays a key role here.
The universality of data formats along the entire process chain from the de-
sign of the components e.g. with grid structures right up to finishing is already
the subject of current developments. The development and implementation of
powerful simulation tools e.g. for calculating distortions, even for complex and
larger components, and feeding these back into the AM workflow represent im-
portant tasks. In this way, based on the CAD model and with the aid of the
simulation, the CAD model can in turn be modified (e.g. machining allowance)
and the optimized processing strategy and support structures implemented.
The horizontal process chain (Fig. 12.2) is to be made significantly more flexi-
ble and efficient by networking and digitizing all hardware and software sys-
tems, so that the direct implementation of a complex digital model (design
phase) into a physical component can be carried out “smoothly” along the entire
chain. Throughout all phases of product development, a uniform representation
model is therefore needed that describes the characteristics of the product in
accordance with the requirements. This begins with the resolution-independent
description of the geometry and connects seamlessly with the simulation of the
product properties. This step makes it possible to use locally adapted process
parameters as well as locally adapted inline quality assurance. If product and
process tolerances are exceeded, corrective, inline intervention into the produc-
tion processes is possible using multimodal measurement and control data.
The following deficits exist in this subject area:
Poor planning and coordination of the entire (digital) process chain

Lack of standardization in the transfer of production-relevant data
Resolution-independent representation of local product properties for the gen-
eration of locally adapted manufacturing strategies and quality assurance
Reworking due to inadequate process results in the individual steps
Poor alternating information flow for manufacturing processes from results of

previous process steps
Limited feedback of incidents in the manufacturing procedure
The resulting consequences are significantly increased manufacturing costs, in-

adequate quality, too high throughput times and reduced flexibility and decreasing
productivity.
The step towards a decentralized and dynamically operating system as de-
scribed by Industry 4.0 can be accomplished through the rigorous networking
of all processes involved in the product and manufacture planning, the actual
manufacturing as well as quality assurance.
Field of Action 2: Scalable and Robust AM Processes

The limits to the scalability of the processes are illustrated with the example of
selective laser melting (SLM) as the currently most important process for the pro-
cessing of metallic materials. According to the current state of the art an increase
in the build rate is possible via two approaches: on the one hand several laser beam
sources are applied simultaneously for the SLM process (parallelization), on the
other hand higher laser power is used, up to the multi-KW range. Although both
approaches lead to an increase in the build rate, the limits are reached in terms of
process stability, which decreases with increasing laser power (e.g. spatter forma-
tion, the dynamics of the melt as a function of the melt volume) and the associated
inferior component quality. In addition, machine costs escalate in line with the
available laser power, so unit costs are only marginally reduced or not at all.
In the latest commercial machines, the build rate can be increased by using mul-
tiple laser scanner systems. Currently, up to four such systems are used in parallel
(e.g. EOS M400-4 [4] and SLM Solutions SLM 500 HL [5]). The limitations of
scan field size and requirements for protective gas flow mean that the build volume
of the largest commercial system is 800 × 400 × 500 mm3 (Concept Laser X-Line
2000R [6]) at present. Scaling up through the use of scanner arrays is conceptu-
ally described in the patent literature, however actual proof of concept is not yet
available. The use of diode laser systems for SLM was first described by ILT [7, 8].
Other than a similar publication by the University of Sheffield [9] with significantly
lower overall laser power and melting rate, no further publications exist. It is there-
fore necessary to develop new illumination concepts using new beam sources (e.g.
diode lasers) and novel parallelized beam deflection systems (miniaturized scanner
arrays), which allow build rate and component size to be scaled much more simply
and cost-effectively. These new illumination concepts will be transferred to and
implemented in innovative machine and system technology.
12 Future AM 239
In addition to SLM, laser metal deposition (LMD) is generally also suitable for
additive manufacturing. Unlike SLM, in the case of LMD component size is only
limited by the handling system. Furthermore, LMD is also suitable for repairing or
modifying components or processing different materials in a component. A major
deficit and significant obstacle in additive manufacturing with LMD is the low pre-
cision (allowance 0.5–1 mm) of the applied volumes achievable at high application
rates (> 2 kg/h) so far. For this reason, a new process for the additive manufacture of
3D volumes is being developed, using extreme high-speed laser welding (EHLA).
When processing rotationally symmetrical components, much thinner layers
(25–250 µm) can be produced with EHLA than with laser metal deposition (LMD),
at feed rates of up to 250 times faster while at the same time achieving high build
rates. So far, however, the EHLA process has only been implemented for the pro-
cessing of cylindrical components where the required high feed rates of up to
250 m/min are achieved by rotation of the components. For the high-speed pro-
cessing of flat or three-dimensional surfaces, it is therefore necessary to develop
the system technology (kinematics and suitable powder feed nozzles) and appro-
priate build strategies.
A further example is the topic of robustness and quality assurance. Accord-
ing to the state of the art, measuring methods such as pyrometry, thermography
and videography are applied, whereby measurement signals from the interaction
zone can be indirectly correlated with discontinuities arising in the process [10–
17]. There are many approaches and studies that relate to this. However, none of
the approaches has yielded a quality assurance system yet, which can help detect
typical defects reliably during the course of the additive manufacturing process.
What is needed for the next generation of process monitoring systems, there-
fore, is the development of measuring techniques with which defects such as pores,
cracks and bonding defects can be detected directly and not only a posteriori. One
approach here is measurement by means of ultrasound. A laser is used to generate
ultrasound in the newly manufactured solid body, which is then locally detected
by laser interferometry. Inline measurement of process-inherent ultrasound to de-
tect near-surface defects is also possible. Inline measurement methods offer the
prospect of being able to have a corrective effect on the build process.
Over and above inline measurement and quality assurance, the identification
of a robust process window for various materials, machines, beam sources, etc. is
indispensable for widespread industrial application. To date, no suitable databases
and technology processors are available for this, which could fulfill a manufac-
turer-independent approach. One possible approach here is the coupling of a tech-
nology database system with corresponding CAD/CAM modules in order to secure
universal, robust process windows.
Field of Action 3: Materials

The topic of materials is also of great importance in additive manufacturing. Ac-
cording to the state of the art, the materials used are mainly those which have been
developed and optimized for conventional production routes such as casting. These
materials are not suited, or are not optimally suited, to being processed with ad-
ditive methods. The reason for this is the very short temperature-time cycles and
the associated high rates of cooling. One example is the problem of hot cracking
in nickel or aluminum-based alloys, which is also known in welding technology.
Just by adapting the process parameters, a solution is only possible in very narrow
process windows and impossible for many alloys. Alloy technical measures are
required here to suppress cracking.
However, the combination of high cooling rates and local metallurgy also opens
up the possibility of developing new materials or simplifying existing production
routes. The most prominent example is Al-Sc alloys. Due to the rapid cooling in
additive manufacturing, the Sc can be forcibly dissolved in the Al matrix. Subse-
quent precipitation hardening produces nanoscale Al-Sc particles that increase the
tensile strength to values of up to 400 MPa. In addition, there are new, innovative
materials on the threshold of industrial utilization—e.g. iron aluminides, high en-
tropy materials or metallic glasses—for which additive manufacturing offers ideal
constraints for setting fine (e.g. eutectics) or non-equilibrium microstructures (e.g.
glass).
Other key topics are new approaches to multi-material processing, the pro-
duction of material composites, material gradients or the production of particle-
reinforced materials. It goes without saying that the use of several material com-
ponents in one component presupposes the flawless, separate processing of each
individual material, which poses a major challenge in the case of Ni-based super-
alloys such as MarM247. In addition to the gradients addressed, the combination
of materials can also be used to realize discrete transitions, a sheath-core structure
or hard materials, e.g. through embedding carbides in matrices.
Another important topic in the field of materials is the provision of AM-adapted
powders for the various processes. There are considerable requirements here with
regard to the specifications, their influence on the AM process management and on
the final component properties. Quality assurance of the powder is another impor-
tant task. The topic of powder materials is being addressed in cooperation with the
Fraunhofer Institute IFAM.
Field of Action 4: System Technology and Automation

In this field of action, concepts are still lacking on how additive manufacturing is to
be integrated into existing manufacturing landscapes and systems. Especially the
12 Future AM 241
mutual interdependencies of the technologies have not been adequately described

and tested. One example is the choice of manufacturing process for the (automated)
removal of the support structures. Depending on the capability profile of the pro-
cesses to be used, the support structure used in additive manufacturing is designed
in accordance with the restrictions of the subsequent process.
This may mean that a (thin-walled) support structure might need to be replaced
with solid material further along the process chain due to the subsequent steps,
because this helps to avoid an interrupted cut and chatter marks, surface defects,
etc. arising during machining processes. The mapping of these interdependencies
and their implementation in a fully automated system are still largely unexplored.
In order to exploit the potential of additive manufacturing for the production
of highly integrated functional systems, automated processes are needed for multi-
material processing using established powder bed-based processes. When SLM is
used, material variations have so far primarily been undertaken in the direction
of building. A further powdered material can be introduced by subsequent knife
coating or by interrupting the process, removing the powder and recoating with
the second powder [18–21]. Material combinations have not yet been comprehen-
sively realized at the build level. For the purpose of an automated and economical
production process, material combinations are therefore largely avoided.
Before greater quantities can be reproducibly manufactured and process reli-
ability guaranteed, additive manufacturing must be integrated into industrial pro-
duction environments and process chains. The focus here is on the development
and implementation of concepts for the integration of the SLM process, in partic-
ular into existing manufacturing landscapes and systems. A major obstacle to this
is the removal of the support structures inherent to the SLM process, which is at
present largely performed manually, as well as the required post-processing of the
components. The development of solutions for the linking, automation and auton-
omization of these process steps represents a significant milestone on the road to
the industrial implementation of additive manufacturing.
Although about 70% of the total cost of additively manufactured components
results from post-processing, only a few isolated research activities addressing au-
tomated post-processing are known up to now. Premium AEROTEC, EOS and
Daimler recently agreed to cooperate on the development of an automated pro-
cess chain for additive manufacturing (Fig. 12.8). The process steps to be taken
into account are additive manufacturing, CNC milling, heat treatment, EDM, laser
texturing and robot-based assembly [22, 23].
The state of the art in research and technology relating to the post-processing of
components manufactured by means of laser beam melting amounts, for the most
part, to manual or semi-automated surface processing by sandblasting, plasma pol-
Fig. 12.8 Future manufacturing concept. [23]
ishing, vibratory grinding or grinding [24]. For large surface area components such
as tools and when the surfaces have special requirements (visible and functional
surfaces), the post-processing usually takes place via milling on a CNC machine
[25].
A surface treatment of additively manufactured components, which however
only takes the surface quality (roughness) and not their dimensional accuracy with
regard to shape and position tolerances into account, and also does not consider the
removal of support structures, is a partially automated process and referred to as
“MMP Technology” by BINC Industries. Like manual post-processing, however,
this is time consuming and costly.
Commercial systems are already available for powder removal [5, 28], which
are however not linked to the subsequent mechanical post-processing. An example
of such a system is shown in Fig. 12.9.
Up to now, additive manufacturing has focused on prototypes and special ap-
plications in small batch quantities, which have a high proportion of manual work,
especially in post-processing. Automation of downstream processes has not yet
taken place and additive processes have not been integrated into automated pro-
cess chains. Initial mass production scenarios (e.g. GE Aviation’s fuel injector
pumps [26], borescope bosses made by MTU Aero Engines [23] or acetabular
cups manufactured by electron beam melting [27]) utilize large-scale additive pro-
duction of the component that is always the same. In doing so, the automation
of post-processing is not envisaged or the post-processing is rigidly related to the
same component that is manufactured over and again and is thus manually pro-
12 Future AM 243
Fig. 12.9 SLM 500 with powder supply station and removal station. [5]
grammed in a time-consuming way and permanently linked. The same applies to

the handling (gripping) of the components. Flexible solutions do not exist for the
automatic post-processing of individual component geometries, which are typical
of additive manufacturing and particularly advantageous (e.g. from the point of
view of mass customization).
12.5 Current Results
As planned for the first year of the project, the foundations for cooperation in the
joint Virtual Lab were defined and suitable systems analyzed based on whether
they would meet the requirements for an Industry 4.0-compatible architecture. The
first significant progress according to the work plan could be achieved within the
fields of action. Thus, for example, an initial model is available for the digital twin
that is to be developed, which incorporates the core process chain elements of addi-
tive manufacturing, including component, customer, order, facilities, materials and
building tasks. In the field of process scaling and robustness, the basic machine de-
velopment of the optical and mechatronic system has been prepared. The central el-
ement thereof—the processing head with fiber laser system and five scanners—has
been developed and the necessary vector allocation cohesively implemented using
software engineering. In the field of materials/multi-material production, a graded
material transition from alloy 316L to Inconel 718 could be successfully generated
by means of LMD. In accordance with the work plan, the process chain analy-
sis required for complete automation was concluded and a concept created for the
planned autonomous manufacturing cell.
Synergies and cross-cutting issues that go beyond the objectives and the work
plan have been realized, especially on the side of data processing and software
for machine control. The participating institutes have thus defined a new data for-
mat [*.1FC], which includes all the relevant information for carrying out the SLM
process. In its current version, the data format incorporates the layered processing
paths of the components to be manufactured, the respective associated local pro-
cess parameters and metadata, e.g. information about the processing strategy used.
In doing so, the [*.1FC]-data format is flexibly designed to be able to implement
extensions according to requirements, as the project progresses.
12.6 Outlook
In the further course of the project, different priorities will be set in the various
fields of action. Thus, for example, for the AM component identification and op-
timization, an expansion of the catalog of bionic structures for the substitution of
component elements will take place on the software side. In addition, further man-
ufacturing restrictions for topology optimization and specified tasks and functions
will be implemented in the user interface. On top of that, the first approaches to
error influence simulation are to be created and automated data input generation es-
tablished. In the development of digital twinning, the validation of the requirement
specifications is planned. Furthermore, the complete twinning for all the elements
of the process chain will be continued based on the previously developed model,
so that an expanded and more detailed model is created.
In the area of the scalable SLM system concept, the programs developed for
path planning and scanner control will be developed as a basis for the process
control and extended, closely tied to the hardware. The optics concept and the
flow of protective gas will also be developed further with regard to scalability. In
further studies on direct fault detection, parameterization will be investigated for
ultrasonic measurements, the integration of sensor technology into an experimen-
tal SLM system continued and evaluation procedures established for in-process
measurements. The measuring and process monitoring systems developed for the
LMD will subsequently be tested for their suitability.
Based on the results already achieved in the field of (multi)-material develop-
ment, the focus here is on the precise analysis of the phases that arise. In addition,
a reduction in the number of individual alloy components (e.g. a binary alloy sys-
12 Future AM 245
tem) should allow for better analysis and evaluation of occurring phenomena. The
mechanical characterization of multi-material test specimens that are generated
may also be included within this field of action.
With regard to the automated process chain, the process development for com-
ponent integration will be completed in the next work package, so that the first
RFID-chip identifiable components can be produced.

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08.05.2017
Insect Biotechnology
Insects as a Resource
13
Andreas Vilcinskas
Summary
Insect biotechnology can be described as the development and application of

biotechnological methods to make insects or their derived molecules, cells,
organs or associated microorganisms available as products or services for ap-
plications in medicine, plant protection or industry. This emerging field, also
known as yellow biotechnology, is rigorously pursuing translational research
approaches with considerable value creation potential. The Bioresources Divi-
sion at the Fraunhofer Institute for Molecular Biology and Applied Ecology
(IME) is one of the world’s leading research institutions in insect biotechnol-
ogy. Researchers here are establishing technology platforms that systematically
identify and characterize natural products and enzymes from insects and make
them utilizable. Innovative technologies for the use of insects in the biocon-
version of organic waste into valuable raw materials are also being developed
here. In addition, biological and biotechnical processes for the sustainable and
environmentally friendly control of insect pests and vector insects are being
developed at the Fraunhofer branch in Giessen.
13.1 Introduction
In terms of biodiversity, insects with over one million species described are con-
sidered the most successful group of organisms. Over the course of their evolution,
they have developed a vast arsenal of active ingredients and enzymes with which
they can defend themselves against diseases and enemies or secure their food
A. Vilcinskas ()
Aachen, Germany

https://doi.org/10.1007/978-3-662-59659-3_13
248 A. Vilcinskas
supply. This biodiversity, which is evident in insects at the species level, is also
reflected on the biochemical and molecular level. Insects and the microorgan-
isms associated with them produce an unbelievable variety of natural products that
should be tapped for the benefit of humanity. This is a research objective of insect
biotechnology, which has also become known as yellow biotechnology [1, 2, 3].
However, insect biotechnology extends beyond pure natural product research,
because only those active ingredients that can be produced in sufficiently large
quantities to at least to cover costs can be marketed. Peptides or enzymes synthe-
sized by insects or their symbionts may be produced as synthetic or recombinant
analogues. However, they often have complex folding patterns or are modified after
translation, so they cannot be represented in this form by bacteria or yeasts. In light
of this, expression systems based on insect cells are booming in biotechnology.
These also offer various advantages over vertebrate cell lines. The development of
cost-efficient fermenter technologies based on insect cells is an engineering chal-
lenge being addressed by Prof. Peter Czermak’s department in IME’s Bioresources
Division [4, 5].
13.2 Antibiotics from Insects
New antibiotics are urgently needed due to the increasing prevalence of multidrug-
resistant bacteria. This need is aggravated by the fact that in the last 60 years, no
entirely new structures have been brought to the market for controlling the bac-
teria with the greater potential threat, the Gram-negative bacteria. At least, after
50 years of stagnation, two new active ingredients have been found for Gram-pos-
itive bacteria. Since the screening of chemical substance libraries remained largely
unsuccessful, natural products are once again turned to as a resource for new an-
tibacterial agents.
However, the low-hanging fruits have already been harvested. Extremely time-
consuming research, the low probability of finding new antibiotics, and the high
cost of development have led many large industrial companies to abandon antibi-
otics research. In order to develop new solutions to this dilemma, Prof. Andreas
Vilcinskas, Head of the Bioresources Division in Gießen, and Prof. Peter Ham-
mann of Sanofi founded the Sanofi-Fraunhofer Center of Excellence for Natural
Products Research in 2014. Sanofi brings one of the world’s largest banks of bac-
terial strains, as well as expertise in developing marketable products to this new,
jointly led venture, while Fraunhofer employees develop innovative approaches
for efficiently screening for new natural products against Gram-negative bacteria.
For the further development of the drug—from the scaling up of the substance ex-
13 Insect Biotechnology 249
Fig. 13.1 Life cycle of the common green bottle fly, Lucilia sericata. The eggs of the green
bottle fly are often deposited in wounds of animals and hatch into tiny larvae, which can
be used in wound therapy if they are kept germ-free. After several larval stages, pupation
takes place. The adult flies only visit flowers. Both larvae and pupae play a role in forensic
entomology
traction, to the co-fermentation of the microorganisms with pathogens up to and

including pharmacology and clinical testing—complementary expertise also exists
on both sides.
The Sanofi-Fraunhofer Center of Excellence for Natural Products Research is
one of the three largest research institutions in the world that identifies new active
ingredients for the development of new antibiotics. In order to reduce the costs for
its maintenance, the bacterial strain bank can be used by other industrial partners.
In doing so, the assumption is made that there is no competition between the
respective product ranges. Currently Corteva, for example, is involved. In this con-
stellation, the supernatants of the cultured microorganisms cannot only be screened
against Gram-negative bacteria, but also against plant pathogens and insects. Shar-
ing the costs and risks of modern natural products research between several part-
ners and expanding the possible value creation results in new opportunities to
accelerate the discovery of new antibiotics using creative approaches.
250 A. Vilcinskas
Within the scope of the collaboration with Sanofi, over one hundred different
antimicrobial peptides from insects were investigated for their potential suitabil-
ity for the development of antibiotics. The evaluation of their therapeutic potential
using rigorous industrial evaluation criteria made it possible to bring two antimi-
crobial peptides from the wound maggot, Lucilia sericata (Fig. 13.1) into the
preclinical research stage. These do not only have a potent action against gram-
negative bacteria (including numerous clinical isolates), but also do not show cy-
totoxicity or evidence of any side effects or the development of resistance in the
amounts tested.
13.3 Virulence Blockers from Insects
Among the most common bacteria in hospitals that develop resistance to clini-
cally used antibiotics is the Gram-negative bacterium, Pseudomonas aeruginosa.
This species is one of the so-called ESKAPE pathogens (Enterococcus faecium,
Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseu-
domonas aeruginosa and Enterobacter species), which are considered to be the
most important causes of nosocomial infections. The need for new antibiotics is
most urgent for combatting these bacteria. Lung infections caused by antibiotic-
resistant bacteria are among the most important causes of death, according to the
WHO [19].
However, the identification of new active ingredients with antibacterial activity
and their development into antibiotics is very costly and risky. Treatment strategies
that do not aim at directly killing the pathogens, but rather focus on neutraliz-
ing their pathogenicity or virulence factors, are therefore regarded as a promising
alternative. These include all the substances produced by the pathogens that are es-
sential for the successful infection of the host, for example toxins that are secreted
by the bacteria to, among other things, overcome the host’s immune defenses and
to make its proteins usable for its own metabolism [6].
Among the bacterial exotoxins, the metalloproteases belonging to the M4 fam-
ily play a prominent role, with thermolysin as their prototype. These are not only
essential as virulence factors, they also cause symptoms associated with infections
such as haemorrhagic bleeding, necrosis and sepsis [7]. In light of this, M4 metal-
loproteases such as pseudolysin from P. aeruginosa, vibriolysin from the cholera
pathogen, Vibrio cholerae, or aureolysin from S. aureus have been identified as
target molecules for the development of virulence blockers. The inhibitors of M4
metalloproteases that have been tested so far, such as phosphoramidon, are how-
ever not suitable for the development of new anti-infective agents, because they
also inhibit human matrix metalloproteases and can thus cause serious side ef-
fects [7]. The search for innovative virulence blockers therefore focuses on those
molecules that act highly specifically against thermolysin-like metalloproteases
without inhibiting human enzymes.
The pseudolysin produced by P. aeruginosa is one of the most important vir-
ulence factors produced by pathogens. A novel inhibitor of bacterial metallopro-
teinases (IMPI) discovered in insects inhibits biofilm formation, swarming motil-
ity, virulence factor production and antibiotic resistance in P. aeruginosa in vitro
[7]. In the BMBF-funded project “Inhalable virulence inhibitors from insects for
the treatment of lung infections (4-IN)”, the Fraunhofer Institutes for Molecular Bi-
ology and Applied Ecology IME and for Toxicology and Experimental Medicine
ITEM are working on the preclinical testing of IMPI with regard to its potential as
a novel active agent for the prevention and treatment of pulmonary infections. For
this purpose, GMP-compliant IMPI test specimens are being produced and their
efficacy evaluated in established in vitro and in vivo models of lung infections
caused by P. aeruginosa. The development of IMPI in this project is supported by
regulatory measures. The necessary toxicological and pharmacological safety tests
are being developed in coordination with the regulatory authorities.
13.4 Insect Enzymes for Industrial Biotechnology
In the course of evolution, insects have acquired the ability to use almost any
organic substrate for food, such as wood or carcasses. To make this possible,
they have developed specialized enzymes that are of interest for the bio-economy,
among other things for the bioconversion of organic waste. In the targeted search
for new enzymes to be applied in industrial biotechnology, the IME’s Bioresources
Division lets itself be led by knowledge-based approaches.
As part of the research program “Application of Insect-associated Microorgan-
isms in Industrial Biotechnology” (AIM-Biotech) jointly funded by Fraunhofer
and the Max Planck Society, Fraunhofer IME’s Bioresources Division and the Max
Planck Institute for Chemical Ecology in Jena are examining how the caterpillars
of the clothes moth, Tineola bisselliella, can eat and digest textiles and how the
larvae of the black soldier fly, Hermetia illucens, can transform slurry, used deep-
frying fat or silage waste into fats and protein. The latter could in future be used,
for example, for fish farming or animal nutrition (Fig. 13.2).
A particularly impressive example of the targeted search for new enzymes is
a kind of burying beetle, the common sexton beetle, Nicrophorus vespilloides,
which can locate small mammal or bird carcasses from miles away, which it buries
252 A. Vilcinskas
Fig. 13.2 As part of the “AIM Biotech” project, the Bioresources Division of IME is inves-
tigating the potential applications of symbiotic microorganisms from three species of insects
that have adapted to special ways of feeding: the Nicrophorus vespilloides burying beetle,
the black soldier fly, Hermetia illucens and the clothes moth, Tineola bisselliella
in the ground and then uses as food for itself and its offspring (Fig. 13.3). It is note-
worthy that the burying beetles not only preserve the carcasses chemically, which
are often several hundred times heavier than they are [9], but that they are also able
to digest them fully, in front of their mouths, skin, hair and all.
For this, they must possess enzymes that could be used, among other things,
for the bioconversion of slaughterhouse waste [10]. In the AIM Biotech project,
a multitude of new insect enzymes has been discovered that are being produced
recombinantly to explore their potential for applications in industrial biotechnol-
ogy.
Fig. 13.3 Life cycle and breeding behavior of the common sexton beetle, Nicrophorus
vespilloides. On discovering a carcass, burying beetles of different species fight each other
until the biggest pair wins the carcass and buries it in the ground. They process the carcass
using enzymes from glandular secretions. It soon assumes a roundish shape and is deposited
in a breeding chamber, where mating and egg deposition take place. Once the larvae have
hatched, they are initially fed regurgitated food pulp by the parents until they can eat their
food independently when their mouthparts are mature. After a few weeks, the final larval
stages pupate and then ultimately crawl out of the ground as new beetles
254 A. Vilcinskas
13.5 Insects as an Alternative Protein Source
From a humanitarian point of view, securing global nutrition is a compelling ne-

cessity and a geopolitical challenge that requires massive investment. The world
population is growing steadily and with it the global need for protein. Conventional
methods will not be able to meet this need. In Germany too, meat production is
flourishing in an effort to secure human protein supply. In this way, Germany has
meanwhile even become one of the world’s largest meat exporters. The produc-
tion of meat as a classical protein source has already left far-reaching ecological
traces. The growing livestock numbers are a heavy burden on the environment. The
spread of large quantities of slurry and manure increases the nitrate load of soil and
groundwater.
There is tremendous dormant potential in insects for the bioconversion of bio-
logical waste and as a resource for proteins, fats and other substances of industrial
interest. In this context, insect-based industries can offer solutions to global prob-
lems and opportunities for innovative value creation [11]. IME’s Bioresources
Division is actively engaged in the development of new technologies to exploit
insects for bioconversion and develop them as a resource for the bioeconomy. Her-
metia illucens, the black soldier fly, is of great interest in this respect [12, 13]. It
is particularly well suited to the production of valuable products such as proteins
and fats from organic waste, which in turn can be utilized as animal feed in live-
stock farming and for aquaculture. In this way, insects can facilitate the creation
of a circular bioeconomy. The use of insects to produce animal protein via bio-
conversion is not only cost and energy efficient, but also extremely sustainable. In
“insect farming”, insects can be bred in so-called insect reactors on a ton scale. The
enormous potential has been recognized: the insect business is growing as a future
market around the world.
Insect-based manufacturing processes are more efficient and protect the en-
vironment. In comparison with conventional livestock farming, insects offer nu-
merous economic and ecological advantages, e.g. they require little space. The
production of one metric ton of soy requires about one hectare of arable land. In
the same area up to 150 metric tons of insect protein can be obtained from black
soldier flies. Another advantage is that producing proteins or animal feed from in-
sects is largely freely scalable. The method is just as suitable for small-scale farms
as it is for industrial production plants [14].
The environment and climate are sustainably protected in that the water re-
quirement of insects is significantly lower than that of livestock. In addition, they
produce no harmful methane and up to a thousand times less carbon dioxide than
livestock. Industrial insect breeding can be done without antibiotics and therefore
also reduces the emergence of resistant pathogens. Insect species that are used in-
dustrially do not transmit diseases to humans or livestock or endanger biodiversity
[14].
The methods are particularly sustainable, because insects allow for closed ma-
terial cycles. They efficiently convert organic waste from agriculture or the food
industry into valuable substances. In the process of producing proteins and fats
from insects, other raw materials are also created that are valuable for industrial
applications e.g. chitin and chitosan that are used in the food and cosmetics in-
dustries. Even their excrements can be used as an excellent organic fertilizer or
to operate biogas plants. Thus, the energy-neutral production of insect proteins is
therefore even conceivable. Conventional animal husbandry can also profit from
insect farming as feeding with insect proteins has a positive effect on the health of
livestock.
These arguments above justify the growing worldwide acceptance and increas-
ing investment in the industrial exploitation of insects. Since official EU approval
was granted for the use of insect proteins in aquaculture in 2017, Fraunhofer scien-
tists and Prof. Klaus Eder from the Department of Animal Nutrition at the Justus-
Liebig-Universität in Gießen, together with industrial partners such as YNSECT
and Alternative Protein Corporation, are researching how suitable insect proteins
are as feed for aquaculture and for animal nutrition.
13.6 Insect Biotechnology in Plant Protection
Another research focus of IME’s Bioresources Division is the development of new

biotechnological methods for the sustainable and environmentally friendly control
of insect pests and vectors. The growing world population demands a rapid in-
crease in the industrial production of plants. It is the basis of food and animal feed
production and at the same time also supplies important energy resources. While
food supplies need to be secured for ever more people, at the same time there has
also been a conditional decline in suitable areas for agricultural production as a re-
sult of climate change. In order to compensate for this development, the yields
obtainable from the available agricultural land need to be increased by intensify-
ing agricultural practices. Although biologically and ecologically sustainable plant
cultivation is an alternative, it requires even greater areas. This in turn implies the
loss of structural diversity in landscapes.
The effects on biodiversity are even more serious. If one considers the propor-
tion of agricultural products that are destroyed on areas under cultivation and in
256 A. Vilcinskas
storage warehouses by insect pests, insects can be regarded as our greatest com-
petitors for food. For decades, insecticides have been used to control them. These,
however, result in the well-documented and much-lamented loss of biodiversity of
insects and other animals such as bats and birds that depend on them as a diet.
The decline in bee populations and the associated loss of their beneficial role in
pollination are also causally related to the widespread and long-term use of insec-
ticides. The increased use of pesticides may indeed limit crop losses and losses due
to plant diseases and pests, but as a result, specific selection processes can actually
promote the spread of insecticide resistance in various insect pests.
The increasing resistance to insecticides signals the rising need for action for the
development and approval of new active ingredients for crop protection applica-
tions. Fortunately, environmental and consumer protection is receiving increasing
attention and the development of alternative options for the sustainable control of
insect pests is becoming increasingly important. At the same time, crop protection
is being confronted with new invasive insect species, for which there are no sat-
isfactory control options that can avoid sensitive economic losses. One example
of this is the cherry vinegar fly, Drosophila suzukii, which causes great damage in
fruit cultivation. Its presence in Hessen was first established in 2012 and by 2014
a third of all fruit growers already complained about harvest losses. The official
prognosis is that fruit cultivation will no longer be economically viable if this in-
sect pest continues to spread.
With this in mind, insect biotechnology develops new options for the sustain-
able, environmentally friendly and consumer-friendly control of insect pests and
vectors [15]. This is also aided by the development of new biotechnological meth-
ods that complement and support biological control strategies, making them more
efficient. Various innovative strategies are being developed in insect biotechnol-
ogy that will be applied in crop protection in the future. Two of these are briefly
introduced below: RNA interference (RNAi) and sterile insect technology (SIT).
13.6.1 RNA Interference (RNAi) in Crop Protection
RNA interference (RNAi) is a natural mechanism of eukaryotic cells that has been
conserved by evolution. It is characterized by the targeted shutting down of genes
in order to defend against viruses. In doing so, the immune system recognizes the
double-stranded RNA (dsRNA) and degrades it. RNAi technology is based on the
fact that synthetic dsRNA, which corresponds to the sequence of target genes, can
inhibit their expression and thus also the synthesis of the corresponding protein.
The 2006 Nobel Prize in Medicine was awarded to Craig C. Mello and Andrew Z.
Fire for the discovery of RNAi. RNA interference plays a particularly important
role in the defense against foreign RNA for plants. This technology has therefore
also been successfully adapted for use in crop protection. This is achieved by ge-
netically modifying crops so that they produce a certain species-specific dsRNA,
directed only at the target organism, which turns off an essential gene in the insect
pest, thereby affecting the development or lethality of the insect pest. The species-
specific design of the dsRNA means that it has no harmful effect on non-target
organisms. One problem is that many plant pests such as the western corn root-
worm, Diabrothica virgifera, or the rape beetle, Meligethes aeneus, cannot be bred
and that there are no sequenced genomes or genetic tools available, which makes
it more difficult to identify suitable target genes. Only genes which can be turned
off in the insect by the tiniest amounts of dsRNA after oral ingestion with food are
suitable as target genes for crop protection.
Innovative high-throughput procedures for the cost-effective identification of
target genes have been developed in IME’s Bioresources Division for one of the
most important model insects, the red flour beetle, Tribolium castaneum [16].
Tribolium is particularly suitable for these studies because its genome has been
sequenced and it has a robust systemic RNA interference response that is also trans-
mitted to offspring (parental RNAi). Homologous sequences to the target genes
identified in Tribolium are then sought in the transcriptomes of different insect
pests and new species-specific dsRNAs are constructed. These constructs are ex-
pressed in the crop plant.
One example of the success of this approach is the production of maize plants,
which have become resistant to the corn rootworm through this method [17]. The
RNAi mechanism is now established as a promising and, above all, future-proof
method and has made alternative, insecticide-free approaches possible in plant pro-
tection. Thus, the RNAi concept can be used to supplement or even completely
replace the use of Bt toxins in crop protection. In the meantime, applications have
been made for worldwide approvals for the cultivation of RNAi plants. As the
dsRNA is engineered to be species-specific, no impact on non-target organisms is
expected. Nevertheless, it is important that efficient and safe standards for the fu-
ture use of RNAi technology both in plant research and in crop protection continue
to be developed.
13.6.2 Sterile Insect Technique (SIT)
Large-scale yet environmentally friendly pest control is a global challenge. A key

objective of governments, research institutes and industries is integrated pest con-
258 A. Vilcinskas
trol through the combination of innovative physical, chemical and biological con-
trol solutions. SIT has already been applied successfully for 60 years. The tech-
nique reduces a pest population through the repeated mass release of sterile mem-
bers of the same species. In order to increase the efficiency of this method, large
numbers of the insect species that needs to be controlled are bred, adult males are
separated from the females and exposed to radioactivity to sterilize them. Only
the sterile males are then released, which ultimately compete with the free-living
males for copulation with females, but can produce no offspring themselves.
However, the ionizing radiation used for sterilization impairs the fitness of the
treated insects and thus also reduces the efficiency of SIT. For the control measure
to succeed, the number of released males must be significantly higher than that of
the free-living males (by up to tenfold). New molecular biology solutions are being
developed under the direction of Prof. Marc Schetelig in a Fraunhofer “Attract
Group” at the Bioresources Division of IME, so that the promising SIT may be
applied to further agricultural pests and vectors. The main focus of this research is
on the design of the development processes for three genetic constructs:
1. Marker genes: The clear differentiation of sterile males from free-living males
is indispensable for monitoring the success of a release. Genetic constructs are
being established for this purpose, which entail the stable labeling of males
with fluorescent proteins. These so-called marker genes are also expressed, for
example, cell or organ-specifically and can therefore easily be identified by
fluorescence microscopy.
2. Sexing systems: The release of females should be avoided at all costs in the con-
trol of vector insects, as these hematophagous insects can increase the transmis-
sion of infectious diseases (e.g. malaria, yellow fever, zika or dengue viruses).
For ethical reasons, the release of other vectors is also prohibited. The develop-
ment of sex differentiation systems will increase SIT’s economic efficiency by
eliminating the customary, expensive and labor-intensive insect breeding step,
during which females are initially separated from males and then ultimately de-
stroyed. The development of genetic constructs is thus intended to ensure that
only male embryos are bred on an industrial scale [18].
3. Lethal genes: So-called lethal genes, which are inactive during the breeding
phase and activated after release, are to replace the use of harmful radioactive
radiation. Carriers of lethal genes have greater fitness than radioactively irradi-
ated males and can therefore be released in significantly lower numbers.
SIT represents an innovative and sustainable approach to crop protection. How-

ever, all three genetic constructs will need to be developed over many years of
research for each insect species that is to be controlled. In addition, extensive con-
trol studies are needed to verify that the constructs are stable and species-specific
and that they are not transferable to other insect species.

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19. WHO Fact sheet N°310
The Resource Principle
Utilization and Intelligent Reprocessing Routes
14
for Wood-Based Materials, Natural Fibers and
Organic Residues
Bohumil Kasal, Moritz Leschinsky, Christian Oehr, Gerd

Unkelbach, and Markus Wolperdinger
Summary
From time immemorial wood has been used for a very wide range of appli-
cations on account of its mechanical properties. Its uses range from static ap-
plications in the construction industry and interior design, where for the most
part load-bearing structures are maintained, and extend all the way to energetic
use—in other words, its complete degradation to water, minerals and carbon
dioxide. There are numerous intermediate levels of physical and/or chemical
treatment between these extremes.
In Sects. 14.2 to 14.5, applications are described where all statically signif-
icant structures are retained and combined with other materials such as glass
fiber or even concrete to optimize mechanical properties. Here, chemical pro-
cessing is limited to the bonding of wood components with each other or with
other materials. These diverse combinations allow new mechanical properties to
be achieved. If a hierarchical structure that results in an anisotropic distribution
of mechanical properties is broken down, a near-isotropic distribution profile,
with respect to mechanical properties, can be achieved in composite materials.
B. Kasal ()
Wilhelm-Klauditz-Institut, WKI, Fraunhofer Institute for Wood Research
Braunschweig, Germany
M. Leschinsky C. Oehr G. Unkelbach M. Wolperdinger
Stuttgart, Germany

https://doi.org/10.1007/978-3-662-59659-3_14
262 B. Kasal et al.
If the focus is on the chemical components rather than the mechanical struc-
ture, then wood can be broken down and fractionated using a variety of meth-
ods. Nine of these processes are described and evaluated according to their
respective technical maturity. To this end, it should be noted that there are differ-
ent stages of development: from an established need for pure research through
to industrial applications that have already been implemented. A distinction
should be made between those processes which preserve the chemical struc-
tures—where lignin, hemicellulose and cellulose are regarded as fundamental
structures worthy of preservation—to those which break down these structures.
While many mature applications already exist for cellulose and hemicellulose,
lignin, apart from a few applications, still requires a great deal of research in
order for the synthetic efficiency of nature to be optimally exploited. When
methods are used that degrade the above-mentioned target structures further,
the end products are small molecules, which can be introduced into the gas net-
work to store energy as fuel (bioethanol) or as methane, or can serve as raw
materials for other processes of the chemical industry. An essential criterion for
all these processes is that no residues remain, but rather that residues from other
processes can even be included in the cycle. It is of interest to the chemical in-
dustry that components can be discharged at the different stages of digestion,
which can in turn be used for further production and replace fossil resources.
Should further use no longer be meaningful after various product cycles, then
thermal utilization is still possible and the resulting carbon dioxide can be rein-
troduced into the resource cycle by using catalysts and energy.
14.1 Introduction
Wood and natural fibers of plant or animal origin have been used by humans since
the very beginning. Their importance in society has changed through the course
of history. At first, they were vital for survival and strategically important, they
later fell into oblivion and are now regaining importance. Fig. 14.1 illustrates the
evolution of materials and their relative importance throughout the entire human
history.
As can be seen in Fig. 14.1, plant-based materials are becoming increasingly
important again. The reasons for this shift are manifold, ranging from new tech-
nologies such as bonding and heat treatment to a growing understanding of the
concepts of sustainability. The natural resources that are the result of millions of
years of evolution and continuous adaptation to the environment (in plants and
14 The Resource Principle 263
Fig. 14.1 Relative importance of materials throughout the entire human history. [1]
trees) serve as a model for many man-made materials such as hollow carbon fibers,
nanotubes or for attempts at making adaptable materials with shape-memory abil-
ity. Modern analysis methods such as electron or atomic force microscopy allow
the microstructure of natural materials to be studied and not only contribute to
our understanding of them, but also to discovering new potential for fabricated
or synthetically manufactured materials. New mechanical, thermo-mechanical and
chemical processes for synthetic materials enable new bio-based materials to be de-
veloped that go beyond traditional technologies, e.g. from solid wood to wood fiber
boards or wood-textile hybrids. The challenges lie in the high degree of variability
of this renewable raw material, which makes it extremely difficult to produce ma-
terial with consistent properties, hygroscopicity and biodegradability—which can
also be considered a positive trait.
264 B. Kasal et al.
14.2 Wood as a Material
14.2.1 Structure and Composition
Wood is a complex material that is available in relatively large quantities. The great
number of tree species—current statistics report over 60,000 tree species known to
science today [2]—makes the utilization of wood challenging. Although almost
all species can be used commercially, only a small proportion is used in material
production. If known sustainability concepts are implemented, they can secure an
unlimited supply of wood fibers. However, the global forest area (about 30.6%
of the global land area) is continuously declining and currently amounts to about
4.0 billion hectares [9].
Wood is generally classified into coniferous wood (softwood, gymnosperms)
and deciduous wood (hardwood, angiosperms). The anatomy of these two wood
categories differs considerably. The description of the anatomical characteristics
of the wood goes beyond the scope of this text and the interested reader is invited
to consult the relevant literature (e.g. [4]). The structure of the wood cell has been
extensively studied and the hierarchical structure of wood is schematically illus-
trated in Fig. 14.2.
From Fig. 14.2 it follows that wood retains its fiber structure all the way down
to the polymer chains (cellulose). In addition, the cells themselves are hollow, re-
sulting in an optimal balance of performance (regarding properties) and mass.
Table 14.1 lists the geometric properties of hard and softwood fibers compared
to conventional plant fibers. Often, individual fibers are not separated, but rather
fiber bundles, because fiber bundles are longer than the individual fibers.
Wood fibers are usually short and thick, so they are not suitable for applications
that require, for example, spinning. Chemically, wood consists of three different
polymers: lignin (aromatic), cellulose and hemicelluloses. In addition, wood con-
tains small amounts of other organic and inorganic, see Table 14.2. Lignin and
cellulose are the most prevalent polymers in the world.
The traditional technologies, some of which have been known for centuries,
are sawing, veneer peeling/cutting (veneer-based materials include plywood, lam-
inated veneer lumber, parallel-strand lumber (PSL)), disintegration into particles
(particle boards, OSB = oriented strand board) or thermo-mechanical pulping
(fiberboard of varying density).
The above-mentioned processes are mechanical or thermomechanical and use
adhesion to connectthe particles or fibers together to form a quasi-homogeneous
material. All of these materials are fiber-particle-dominated and the adhesives play
Fig. 14.2 a Hierarchical structure of the wood; b Schematic of the wood cell structure. The
fiber thickness ranges from about 20 to 40 µm
only a minimal role in determining the physical and mechanical properties of the
composite or product. Often, some of these materials are also referred to as wood-
based materials, but they do not behave like conventional composites. The result-
ing properties cannot be derived from the properties of the constituents. Thus, for
example, classical lamination theory is not applicable.
Recent advances in the development of adhesives have allowed the hybridiza-
tion of plant-based materials by combining wood with technical textiles (e.g. glass,
carbon and aramid fibers in combination with solid or laminated wood) [15], met-
als, glass, plastics and other materials. These materials are not always composites
in the true sense and the interaction of the individual materials must be taken into
account, for example with the aid of continuum mechanics. The combination of
wood particles or fibers with an inorganic matrix (e.g. concrete or gypsum) has
been implemented for some time. The challenge here is the alkalinity of the matrix
and the possible deterioration of the lignocellulosic material.
266 B. Kasal et al.
Table 14.1 Geometric properties of cellulosic fibers (single fibers)

Cellulosic Diameter (µm) Length (mm) Density EMC (65% RH at
fibers (kg/m3 ) 20 °C)
Abacá 6–46 2–12 1500 5–14
Bagasse 10–20 0.8–3 900 –
Banana 10–80 1–6 1325 1–15
Bamboo 10–40 1.5–4 1500 –
Coconut fiber 10–24 0.3–3 1250 2–13
Cotton 10–40 10–65 1550 7–8.5
Flax 1.7–76 5–140 1450 7–12
Hemp 3–51 10–55 1200 6–12
Jute 5–30 1–6 1400 10–14
Kenaf 2–50 2–10 1310 7–12
Ramie 4.6–120 40–260 1550 7.5–17
Sisal 5–50 1–8 1400 5–12
Softwood 30–45 3–5 300–500 ~ 12
Hardwood 20–40 1–1.8 150–1000 ~ 12
EMC = equilibrium moisture content, RH = relative humidity of ambient air
Table 14.2 Main components of wood. The proportion of the individual components is very
variable
Cellulose Hemicellulose Lignin Extracts Minerals
40–55% 15–35% Softwood: 28–41% 1–10% 0.2–0.8%
Hardwood: 18–25%
14.2.2 Plant Fibers
Plant fibers represent a significant proportion of natural fibers and are traditionally
used alongside wood fibers for textile and paper production. The production of
paper usually requires delignification unless the original plant has no significant
lignin content, as is the case for cotton. An overview of commercially used plant
fibers is given in Fig. 14.3 and typical fiber properties are listed in Table 14.3.
It can be seen from Table 14.1 that natural fibers vary greatly in their ge-
ometry and properties, which presents a considerable disadvantage compared to
synthetic fibers. All-natural fibers are hygroscopic and absorb water in both liquid
and gaseous form. The fiber saturation point represents the point at which the cell
walls are completely saturated with water and varies between 20 and 35% (based
Fig. 14.3 Classification of natural and synthetic fibers. [28]
on dry weight). This point is reached when the relative humidity of the air is close
to 100%.
Fibers of various origins are compared to manufactured (continuous) fibers and
composite materials in Fig. 14.4 with the aid of Ashby plots. Different forms of
these plots can also be used for the selection of materials. It is important for mate-
rial production, however, that the plant fibers are able to compete with the synthetic
continuous fibers, especially when considering the bulk density and the price to-
gether with the absolute sizes of the technical parameters. Based on the desired
properties such as the elastic limit (Table 14.3), some plant fibers perform better
than synthetic fibers, for example in energy absorption. It should be noted that the
properties of the individual natural fibers are not decisive and should be applied
with caution as the fibers are relatively short and need to be spun into yarns. The
mechanical properties of yarns have lower values than those of single fibers [21].
268 B. Kasal et al.
Table 14.3 Mechanical properties of selected plant fibers compared to typical manufactured
fibers. Values without a specified range represent an average. [28]
Fiber type Density Tensile strength E-modulus Specific mod- Elongation
(g/cm3 ) (MPa) (GPa) ulus at break
(GPa/(g/cm3 )) (%)
Abacá 1.5 15–980 10–70 9 1.0–10
Bagasse 1.25 20–290 3–20 18 ~ 1.0
Bamboo 0.6–1.1 140–800 11–32 25 2.5–3.7
Banana 1.35 400–600 8–20 9 1.5–9
Coir 1.15–1.46 95–230 2.8–6 4 15–50
Cotton 1.5–1.6 287–800 5.5–12.6 6 2–10
Flax 1.4–1.5 343–2000 27.6–103 45 1–4
Hemp 1.4–1.5 270–900 23.5–90 40 1–6
Henequen 1.2 430–570 10–16 11 3–6
Isora 1.2–1.3 500–600 – – 5–6
Jute 1.3–1.49 200–800 10–60 30 0.2–1.8
Kenaf 1.4 200–1000 14.5–53 24 1.5–2.7
PALF 0.8–1.6 180–1627 1.44–82.5 35 1.6–14.5
Ramie 1.0–1.55 300–1000 5–128 60 1.2–4.0
Sisal 1.33–1.5 80–700 9.0–38 17 2.0–7.0
Aramid 1.4 3000–3150 63–67 46.4 3.3–3.7
Carbon 1.4 4000 200–240 157 1.4–1.8
E-glass 2.5 1000–3500 70–76 29 1.8–4.8
S-glass 2.5 4570 86 34.4 2.8
14.2.3 Wood and Fiber Composites
Wood and natural fiber-based composites (WFBC) can be bonded to organic or

inorganic matrices and be either matrix or fiber dominated. Organic matrices are
usually adhesives that are either applied continuously (as with veneer-based ma-
terials) or sparsely (as in particle-based materials). The mechanical properties of
such materials are dominated by the fibers or particles, but the physical properties
such as shrinkage, swelling or water resistance are predominantly influenced or
even controlled by the adhesive properties.
The composites are made from veneer-based materials that only use wood as
a raw material, as well as fiber and particle-based materials that can make use
of virtually any plant containing lignocellulose. Veneer-based materials include
plywood, molded plywood, laminated veneer lumber, parallel strand lumber (PSL)
Fig. 14.4 Properties of a density-specific elastic modulus and b tensile strength. Dashed
lines show a constant material performance for anchor stiffness E/q and strength r/q, beam
stiffness E1/2/q and strength 2/3/q and panel stiffness E1/3/q and strength r1/2/q (GFRF =
glass fiber reinforced plastic, CFRF = carbon fiber reinforced plastic). [1, 28]
and the like. Examples of particle-based products include particle board or OSB
boards. Fiberboards are an example of a fiber-based material. Fiberboards may be
characterized by their low, medium or high density.
The processes for producing veneer or particles are mechanical, whereas the
processes for producing fibers are thermomechanical. Details for this can be found,
270 B. Kasal et al.
for example, in [26]. The adhesives used to bond the particles can be divided into
two basic types depending on their water resistance. These contain phenolic and
urea-formaldehyde adhesives, adhesives based on urethane, melamine or acetate
and combinations thereof. Epoxy-based adhesives are rarely used, e.g. in hybrid
composites such as wood/metal, wood/glass or wood/textile. Almost all WFBCs
are fiber dominated, i.e. their properties are controlled by the properties of the
fibers or particles and not by the adhesives. This is because the adhesives are only
sparsely distributed on the surface of the fibers or particles and account for a mini-
mal proportion of the total composite—usually less than 10% dry weight. For PSL,
the resin content can be up to 15%. Matrix-dominated WFBCs are relatively rare
and examples of these are wood-plastic composites (WPC), usually with a ther-
moplastic matrix such as polypropylene, polyvinyl chloride (PVC), polyethylene
or concrete-wood composite materials. The lignocellulose particles in WPCs take
the form of flour or fibers and make up 40 to 85% of the composite [30]. The
wood-concrete composite materials contain about 30 to 70% wood [30] and the
resulting properties are influenced both by the wood particles and by the cement-
bound matrix. Other inorganic bonding agents such as gypsum or concrete are also
possible.
14.3 Hybrid Materials with Wood and Plant Fibers
14.3.1 Wood-Based Hybrid Materials
The fibrous structure of wood (Fig. 14.2) results in materials with an anisotropic
character. Wood has almost negligible strength in tension perpendicular to fibers
and different strength in tension and compression. Wood is also not conductive; the
specific resistance is about 1.3 × 109 to 1.7 × 107 m. The heat transfer coeffi-
cientis also relatively low at about 0.05 to 0.12 W/mK perpendicular to the fiber
direction and 0.22 to 0.5 W/mK in the fiber direction [20]. The properties differ
greatly depending on the fiber direction, moisture content and temperature. Details
can be found, for example, in [20].
In order to overcome the anisotropic character or to incorporate functionality
such as electrical conductivity, wood can be combined with other materials. If solid
wood is used, the material cannot be treated as a composite material, e.g. by ap-
plying classical lamination theory. Instead a hybrid material is created. A hybrid
foam is depicted in Fig. 14.5, which highlights the disadvantages of the individual
materials.
Fig. 14.5 Example of a hybrid material: metal foam in combination with wood foam.
a Overall structure; b Magnification; c Microscopic image of the metal-wood foam inter-
face; d The fibrous character of wood foam. (a Fraunhofer WKI Manuela Lingnau, b, c,
d Fraunhofer WKI, Bohumil Kasal)
Metal foam provides stiffness and electrical conductivity, but is a poor insulator
and cannot absorb moisture. Wood foam has poor stiffness and strength properties,
but offers thermal insulation properties and serves as a filler material. In addition,
being a hydrophilic material, wood foam allows the hybrid system to absorb water
vapor and liquids. Depending on usage, each of these individual properties may be
rated as either beneficial and positive or harmful and negative.
Wood combined with high-stiffness and high-strength fibers or textile structures
has been used for some time. Fig. 14.6 shows an example of the reinforcement of
laminated wood with a glass fiber textile.
Several statements can be made in this regard. Hybridization can be used to
control material behavior while maintaining many of the original properties. The
textile (or fabric) itself can be used to add a particular functionality (e.g. conduc-
tive fibers, optical fibers, etc. which are integrated directly into the reinforcement
272 B. Kasal et al.
Fig. 14.6 Example of a hybrid material: wood in combination with a glass fiber textile.
a Unreinforced material fails completely due to the tension in the lower fibers. b Reinforced
material shows a pressure failure due to the displacement of the neutral axis. The reinforce-
ment increases ductility in this case. c 5-layer metal-veneer combination. d Cross section of
the metal-veneer hybrid. (Fraunhofer WKI, Manuela Lingnau)
fabric and act as a moisture barrier) or to control the failure mode (e.g. from brittle
to ductile). The material properties of the fabric may be controlled by fiber and/or
yarn material as well as the architecture of the fabric (woven, non-woven fabric,
number of layers, fiber/yarn orientation, etc.). Fig. 14.6 shows a combination of
laminated veneer lumber and steel for the unidirectional forming process. The ve-
neer thickness is 0.5 mm, the steel thickness is 0.53 mm and polyurethane adhesive
(200 g/m2 ) was used.
The hybridization of wood involves a multitude of technical challenges that
need to be addressed. These include differential shrinking and swelling as well
as strain compatibility. For example, the reinforcement in the direction perpen-
dicular to the fibers, as shown in Fig. 14.7, must transfer the total tension at the
tensile strain of 0.2 to 0.4%. This means that the reinforcement stiffness must be
significantly greater than that of wood under tension perpendicularly to the fiber
orientation [14, 15]. Another challenge lies in the different thermal expansion prop-
erties of the reinforcing and reinforced materials.
Fig. 14.7 Example of a hybrid material: radial reinforcement of the wood perpendicular to
the fibers. a Glass fiber rod inserted transversely across the fibers; b Electron microscopic
assimilation graph of GF reinforcement. The GF (white circles show the cross sections of
the individual glass fibers) and epoxy resins are clearly visible. c Normal stress distribution
(positive values are stresses) in the crown of a non-prismatically curved beam that is loaded
by a moment. (Fraunhofer WKI, Bohumil Kasal)
Other combinations may include wood-metal or wood-concrete materials.

Fig. 14.8 shows two forms of wood-concrete combination. Fig. 14.8a shows wood
particles coated with cement paste and Fig. 14.8b the use of adhesive to combine
two solid materials (wood and concrete).
There are fundamental differences between the materials shown in Fig. 14.8.
When the wood particles are combined with the inorganic matrix, in this case
concrete, a new material is created that can be homogenized and macroscopically
even assumed to be isotropic. When the two materials are combined in solid form
(Fig. 14.8b), both materials (wood and concrete) will retain their distinct proper-
ties and the system can be represented using parallel axis theorem as a composite
cross-section (compared to the composite material as in Fig. 14.8a).
274 B. Kasal et al.
Fig. 14.8 a Wood-cement combination in particle shape, density approx. 1400 kg/m3 , wood
particles in cubic form. b Adhesive-bonded wood-concrete composite, e.g. solid wood. The
adhesive allows for shear force transmission. (Fraunhofer WKI, Manuela Lingnau)
Fig. 14.9 shows the cross-sections of the wood-cement composite material with
a large volume fraction of wood (Vw = 0.57). This shows that mixing the hygro-
scopic material with the cementitious materials that require water (in this case
200 kg/m3 ) poses a challenge, because the water is quickly removed from the ce-
ment paste as it is absorbed by the wood.
14.3.2 Hybrid Materials Made of Plant-Based Fibers
Plant-based fibers such as flax, hemp, kenaf, jute, cotton, sisal and others [22] are
usually spun into yarns and then processed into textiles. The fabrics can then be
used like fabrics made from continuous fibers—carbon fibers (CF), glass fibers
(GF), Kevlar. The advantage of plant-based fibers lies in their deformability, recy-
clability and their compatibility with wood. The use of plant-based fibers that have
a similar chemical composition to wood (cellulose, hemicelluloses and lignin) also
expands the range of adhesives beyond just the traditional epoxy resins.
Studies that are still at an early stage of research demonstrate the applicability of
plant fibers in construction [12, 28] or in the automotive industry [29]. Ashby plots
Fig. 14.9 a Wood-cement combination in particle shape: density approx. 1000 kg/m3 ; Vol-
umetric proportion of cement paste = 0.15, wood = 0.57; Wood particles in cubic form.
b Close-up of the interface between the wood (beech wood, density = 880 kg/m3 at 40%
water content). Delamination between wood and concrete is clearly visible after ten wetting
and drying cycles (wetting in water at 20 °C and drying at 70 °C). (Fraunhofer WKI, Norbert
Rüther)
have been developed for various plant fibers to facilitate their selection according
to the number of criteria [25, 28].
Fig. 14.10 shows the failure of steel reinforced concrete, which is further rein-
forced by a flax fabric in the lower (tensile) zone. The use of the flax fabric has
increased the ductility of the beams compared to non-reinforced beams [11].
The plant-based fibers or yarns usually have inferior mechanical properties
compared to manufactured continuous fibers. These yarns can still be combined
into a hybrid fabric with other different fibers and yarns, e.g. thermoplastic fibers,
carbon or glass fibers. This hybrid fabric can then be used in further processing
(e.g. compression molding). Fig. 14.11 shows examples of different combinations
of fabric layers in a composite.
Each layer consists of a single-material fabric. The combination of different
materials within a single layer is shown in Fig. 14.12. Various yarns and fibers can
be incorporated into the textile structure in multiple layers to provide the fabric
with the desired properties and functions.
276 B. Kasal et al.
Fig. 14.10 Steel-reinforced concrete beam with 6-layer flax fiber reinforced plastic (epoxy
matrix). The failure in the concrete shows the expected performance of the adhesive area.
[11]
Fig. 14.11 Examples of hybrid fabrics: a Different combinations of flax yarn and car-
bon fiber fabrics as layering systems. Each layer consists of a single material; from
left to right: 4CF/4FF/4CF, 1FF/8CF/1FF, 8VF/PA11, 8CF/PA11,2FF/8CF/2FF/PA11,
3(FF/CF)/3(CF/FF) PA11, 8FF/PA11. b CF/FF/CF/FF/CF in epoxy matrix: view of the indi-
vidual layers. (c) Side view of the three-layer system. (Fraunhofer WKI, Manuela Lingnau)
Figs. 14.11 and 14.12 show that there are seemingly endless possibilities for the
hybridization of natural fiber textiles, as well as for textiles in general. This requires
analytical methods for pre-planning the textile architecture based on the desired
function and performance. There are extensive studies on textile models [16, 17,
Fig. 14.12 Example of a hybrid material: combination of fibers/yarns from different mate-
rials in a single fabric: a Flax yarn in combination with polyamide fibers; b Magnification;
c Micro-CT image of the textile structure. (a Fraunhofer WKI, Manuela Lingnau, b Fraun-
hofer WKI, Bohumil Kasal, c Fraunhofer WKI, Florian Bittner)
18, 27], but it remains a challenge to link, for example, a structural performance
quality of the textile-reinforced element with the properties of the yarn material and
the fabric architecture. The models must take into account the stochastic properties
of natural fibers, while the fabric geometry can be approximated deterministically
[27].
Fig. 14.13 shows a conductive material (wire, fiber, yarn) which is incorporated
into the flax fabric during the weaving process.
278 B. Kasal et al.
Multi-layer
fabric
Metal
conductor
b
Carbon thread
Metal
contact
Metal
a c contact
Fig. 14.13 Example of a hybrid material: conductive fibers from flax fabric. a Flax fab-
ric with embedded wires b Micro-CT image of the fabric cross-section; c Cross section of
a multilayered flax fabric with embedded, conductive carbon fiber thread. (Fraunhofer WKI,
Florian Bittner)
14.4 Hierarchical Wood Structure and Transformation into

New Materials
Biomimicry is an attempt to imitate plant and animal structures in the development

of new materials, mechanisms and systems. The hierarchical structure of woody
and lignocellulosic plants can be used to design manufactured fiber materials. The
study of wood anatomy shows that wood is based on fibrous building blocks that
extend down to the cellulose chains. The wood cells (tracheids, tracheae, vessels,
etc.) are hollow fibers with cell walls that continue to retain a fibrous character (fib-
rils, microfibrils). Microfibrils result from cellulose chains (Fig. 14.2). This highly
organized structure has developed into an extremely efficient material with highly
specific mechanical properties (Tables 14.1, 14.3, Fig. 14.4). However, the very
variable structure suggests that the wood itself may not be a solution, but rather
model for sophisticated materials [3]. Furthermore, the adaptability and multi-
functionality of plants plants (reaction wood, fluid (water) conduction and stor-
age, self-healing) are desirable in many applications, but this has not yet been fully
Table 14.4 Plants/wood and technical materials and their main differences
Wood/plant Technical materials
Process Biological self-assembly Manufacture
Main compo- Predominantly carbon with H, O and Many different elements: Fe,
nents small amounts of other elements Cu, Si, C, Al, Ni, Cr . . .
Building blocks Organic polymers; lignin, cellulose Organic (synthetic) and inor-
and hemicelluloses dominate (cova- ganic (metals, ceramics . . . )
lent bonding); small units of fat, wax,
turpentine, oils
Structure Hierarchical at all levels Organized, function-driven
Functionality Adaptable, self-healing Limited to the design purpose,
usually not adaptable or only
to a limited extent
Properties Highly variable Organized, relatively low vari-
ability
achieved. The specific properties of natural fibers and wood are clearly able to
compete with other synthetically produced fibers. Table 14.4 summarizes the main
differences between plant-based and engineered materials.
It can be seen from Fig. 14.1 that synthetically produced materials have only
recently overtaken natural materials in significance. If costs are also taken into
account, natural materials such as wood are far better than the newly developed
materials. The geometry and structural hierarchy of lignocellulosic fibers can be
modeled and to some extent copied [23, 24] and wood has been used as a typi-
cal example of a hierarchical, multi-scale structure [3]. Other plants were used as
a model for optimized structures [19]. Micro-CT technique, combined with numer-
ical modeling and 3-D weaving techniques, can be used to develop preforms with
the aim of maximizing the load capacity-to-mass ratio [19]. The fluid flow analogy
can be used to simulate the grain alignment around the knots [10] where the lami-
nar flow passes through the sample, with the knots shown as obstacles (Fig. 14.14).
The micro CT scan is used to obtain the optimized fiber orientation for similar en-
gineering objects. The high strength of the transition zones (tree–trunk–branch) is
achieved by the orientation of the wood fibers and the chemical structure of the
cell walls, which are either rich in lignin (compression wood) or cellulose (tension
wood).
The adaptability of lignocellulosic materials to changing environmental condi-
tions has been studied, but cannot yet be fully exploited. The chemical differences
between tension and compression wood are an example of this [6]. The forma-
280 B. Kasal et al.
Fig. 14.14 Fiber orientation in the branching of a tree: micro-CT images. Modeling of knots
and fiber orientation by 3-D flow analogy. (a, b Fraunhofer WKI, Florian Bittner, c [10])
tion of reaction wood indicates that plants are able to generate a different chemical
composition and structure in their cell walls, based on the dominant load—in this
case compression or tension—to exploit the relationship between the chemical
structure and mechanical properties of the resulting system. Although it is known
that cellulose primarily provides the tensile properties, the exact relationship be-
tween, for example, the degree of polymerization and the mechanical properties of
the material is not yet clear [31]. It appears that a purely observational study of
biological materials is not sufficient and that relationships between chemical com-
position, structure and properties must be investigated. Only then the evolutionary
development of biomaterials be fully exploited and used to develop new generation
materials.
14.5 Wood as a Source of Chemical Compounds

In addition to the development of novel wood-based materials, which preserve
the essential structural features of the starting material (such as fibers or cellular
structures), the processing of lignocelluloses (predominantly wood and agricul-
tural residues) is also of great importance to the chemical industry. The goal is to
obtain smaller, chemically defined units down to molecule level. In doing so, the
focus is on whether and to what extent wood can be used to sustainably produce
basic, fine and specialty chemicals that can become an alternative to substances
produced from fossil materials. According to this approach, it is logical to put so-
called “biorefineries” on a level with those of the petroleum-based industry (i.e.
the conventional refineries) and to partially replace petroleum with renewable raw
materials such as wood. In addition to the broadening of the raw material base and
thus reducing the politically significant competition for fossil raw materials, there
are also advantages for the environment: no additional carbon is introduced into the
biosphere in the form of CO2 and, moreover, an important step is taken towards the
goal of a circular economy.
The processing of wood can be carried out with varying degrees of differentia-
tion. The first step is to split lignocellulose into the three main components already
mentioned: lignin, hemicellulose and cellulose. The digestion methods used for
this purpose are explained below, following a short assessment of the technology.
14.6 Technology Assessment
Digestion processes can basically be divided into those that mostly preserve struc-
tures and those that do not preserve structures. In the former, pure component
separation allows the basic structure of the main constituents of lignocellulose to
Mechanical pretreatment
structure-preserving
(chopping, cutting, grinding)
Autohydrolysis/hydrothermolysis
(Aquasolv hydrothermolysis, prehydrolysis processes, steam explosion)
Digestion with organic solvents - Organosolv processes

(Alcell, Organocell, MEA method, acetone)
Digestion with ionic liquids

Alkaline digestion procedures
(Soda, kraft (sulfate), digestion by calcium oxide, AFEX)
Acid digestion processes with organic or mineral acids

(Sulfite, Bergius, Scholler, Milox, Formacell, Acetosolv, Acetocell)
Pretreatment with fungi

structure-degrading or
(brown and white rot fungi)

structure-changing
Oxidative digestion processes

(Wet oxidation (O2 or H2O2), ozone)
Pyrolysis
Fermentation
Gasification
Fig. 14.15 Digestion processes from structure-preserving to structure-degrading

282 B. Kasal et al.
Table 14.5 Advantages and disadvantages of structure-preserving and structure-degrading

processes
Structure-preserving process Structure-degrading process
+ Use of the synthetic power of nature + Fast implementation
+ Preservation of the biopolymer structure + Economy of scale more possible for
larger facilities
+ Production of sugar as a raw material for + Gaseous products easier to handle in the
fermentation process
+ Decoupling of primary and secondary + Easy integration into existing chemical
refining processes
Challenging component separation Loss of the synthetic power of nature
Slow processes, low implementation rate High energy expenditure (p, T)
No “drop-in” products for the chemical Large facilities required for cost-efficient
industry after the primary step implementation
be completely retained or at least preserved to a significant degree. In methods

where the structure is not preserved, the basic structure of the major components
does not remain intact because it is degraded to a greater or lesser extent by, for
example, a thermochemical or biochemical process.
A variety of processes can be graded appropriately from “structure preserv-
ing” to “structurally degrading” within the digestion processes described below
(Fig. 14.15).
Table 14.5 compares the advantages and disadvantages of a milder, structure-
preserving process to harsher, structurally degrading processing.
The above-mentioned digestion processes are at different stages in their tech-
nological development. The processes are listed according to the so-called Tech-
nology Readiness Level (TRL) in Table 14.6.
An investigation of the technology readiness of various biorefinery concepts
in Europe was presented, among other things, in the Biorefinery Roadmap [32].
References [34, 35, 36, 37, 38, 39, 40, 41] provide further illustrations of various
digestion processes with regard to technology readiness, integrability, efficiency
and detailed process control. Not all of the methods presented here are forward-
looking, only a few have been proven in practice or have the potential to be used
in industrially relevant dimensions. This overview is not exhaustive. The goal is
rather to present the most important and most technologically promising methods.
Table 14.6 Technology readiness [33]

Technology readiness level (TRL) Phase
1 Description of the operating principle
2 Description of the technology concept Laboratory scale
3 Proof of concept
4 Validation in the laboratory
5 Technical validation in a relevant application environ- Pilot/demo/reference phase
ment
6 Demonstration in a relevant application environment
7 Demonstration of application
8 Qualified and complete system (reference) First-of-its-kind facility
9 System successfully operating Commercial phase
14.7 An Overview of the Digestion Processes
14.7.1 Mechanical Pretreatment (Structure-Preserving Method)
Mechanical shredding of lignocellulose can be carried out by means of cutting,

grinding, sanding and other processes. The aim is to increase the surface area and
reduce the degree of polymerization of the incorporated material. A disadvantage
of the mechanical pretreatment is that the fractions of lignocellulose are not present
separately, but rather in one phase. The high energy input, which is up to 70%
higher than that of other processes, depending on the desired particle size, does not
make mechanical pretreatment appear as an attractive option for digesting ligno-
cellulose at present [54].
Another less energy-intensive option for mechanical pretreatment is extrusion.
Here the lignocellulose is defibrillated under heating by the shear forces arising
in the extruder, which shortens the fibers [54]. Both methods are mainly used to
improve the hydrolyzability of lignocellulose, for example via enzymatic action
(TRL 9).
14.7.2 Autohydrolysis/Hydrothermolysis/Vapor Pressure

Digestion (Structure-Preserving Method)
There is a variety of processes that operate on the principle of autohydrolysis. One

of them is the hydrothermal fractionation of lignocellulose at elevated temperature
284 B. Kasal et al.
and high pressure in an aqueous medium. During autohydrolysis, acetyl groups are
split off from the (hemi)celluloses by an autocatalytic reaction. The concentration
of acetic acid in the liquid phase increases with increasing reaction time, resulting
in a decrease in pH. After a certain reaction time, the pH reaches a minimum of
about 3 and does not go below that [42]. In this process, acetic acid is released
from the biomass. The released acetic acid in turn catalyzes the cleavage of the
ˇ-(1-4) glycosidic bonds of the hemicelluloses, which leads to a reduction in the
molecular weight of the hemicelluloses and ultimately to the formation of water-
soluble oligosaccharides and monosaccharides. In the further course of the auto-
hydrolysis, the monosaccharides also undergo dehydration reactions, which leads
to the formation of condensation and degradation products [43].
The processes of Aquasolv® hydrothermolysis, prehydrolysis in the kraft pulp
process and steam explosion proceed according to the principle of autohydrolysis.
Wet oxidation can also be included here, but it is a rather exotic process. Products
of these digestion processes are a solid fraction with cellulose fibers and lignin,
a dissolved fraction including hemicelluloses and soluble extracts, lignin residues
and acetic acid as well as steam-volatile products such as furfural.
In Aquasolv® hydrothermolysis, 30 to 60% of the biomass used is dissolved in
water at 120 to 180 °C (maximum 220 °C) and a relatively high pressure of 40 bar.
The hemicelluloses discharged in this way and now found in solution then undergo
a conditioning process (purification and hydrolysis of the remaining polysaccha-
rides to monosaccharides) and can, for example, be directly further fermented. The
fibers also remain largely unaffected due to the exclusive use of water as a solvent.
The process, which is being developed as an alternative digestion process for pulp
production, is currently still being tested [34, 35]. The technology readiness can be
estimated at four to five (Fig. 14.16).
During prehydrolysis in the kraft pulp process, the hemicelluloses are dissolved
out of the cell structure at 170 °C over 90 min in a pressurized atmosphere of 8 bar.
After appropriate conditioning, here too the dissolved sugars can be fermented to
form various products such as carboxylic acids [46]. Moreover, the process can also
be carried out in two stages in order to achieve a higher conversion rate or a higher
degree of saccharification of cellulose and hemicelluloses [47]. The prehydrolysis
has recently been used in industrial batch or continuous processes, although there
are only a few successful examples [48]. The level of technology readiness is thus
nine.
For the steam explosion digestion process, the biomass is exposed to hot steam
at 200 to 360 °C with relatively high pressure applied for 5 to 20 s. Through the
subsequent sudden release of pressure, the vaporizing water contained in the cells
causes the lignocellulose to disintegrate into fibers, releasing the hemicelluloses
Aquasolv reactor Gas connection (nitrogen/compressed air)
Heating block
Sampling
device with
Pressure compensation
water cooling Water
tank
reservoir
Circulator pump High-pressure pump
Fig. 14.16 Operating principle of Aquasolv® hydrothermolysis
according to the principle of autohydrolysis [49]. The process has already been
applied several times in industry (TRL 9).
For wet oxidation, an oxidizing agent is added to the aqueous digestion con-
ditions under pressure and at elevated temperatures. This may be atmospheric
oxygen, oxygen or hydrogen peroxide. The latter is currently not economically
feasible due to the high chemical costs. Most of the hemicelluloses go into solu-
tion and the relatively strong oxidation process and high degree of delignification
result in low molecular weight carboxylic acids and alcohols being formed (mainly
from the hemicelluloses and lignin). The addition of alkaline substances reduces
the temperature and thus the amount of oxidized hemicellulose. The advantage of
the process is that fewer by-products such as 5-hydroxymethylfurfural (5-HMF)
are formed in comparison to other processes, thereby facilitating the use of the
sugar fraction in fermentation processes. Owing to the strong oxidation during the
digestion process, however, the hemicellulose yield is low. The high temperatures
that are generated during the process and the associated effort in designing the sys-
tem mean that much more development work is required before the process can be
286 B. Kasal et al.
viably applied in industry [50, 51]. The technology readiness is estimated at three
to five.
The process of autohydrolysis or hydrothermolysis is simple, robust and the use
of chemicals is not necessary. However, clean fractionation of lignocellulosic com-
ponents is only possible under great effort and expense. The material utilization of
lignin, for example, is therefore difficult to implement at present owing to the high
level of impurities, or the sugar fraction of C5 and C6 sugar molecules can only be
metabolized in parallel by certain microorganisms.
There is potential for further development of the methods described, especially
with regard to the extraction and value-adding further processing of the hemicellu-
loses, which consist mainly of C5 sugars.
These accumulate in large quantities for example in the prehydrolysis step prior
to the kraft digestion process in the production of chemical pulps, where high lev-
els of cellulosic purity are aimed for. The process of autohydrolysis also plays an
important economic role in the production of bioethanol, where steam explosion
is widely used as a pretreatment step or digestion process prior to the hydrolysis
of the cellulose (Sect. 14.9). The technology readiness of the processes, which are
estimated to be at level 6 to 7, can be illustrated using the example of a process
developed by the company Abengoa (Fig. 14.17). Abengoa has taken a demon-
stration scale plant into operation in Spain that has the capacity to pretreat 70,000
metric tons of straw per year for ethanol production [52]. A pilot plant of the same
size was commissioned by the company logen in Birch Hill, Ottawa, Canada in
2011 [53]. Abengoa also set up a pilot plant with a capacity of 34,000 metric tons
of wood per year in Hugoton, Kansas, USA in 2011. From a current perspective, it
should be noted that Abengoa‘s efforts were not successful and the facilities have
now changed ownership.
14.7.3 Alkaline Digestion Processes (Structure-Preserving

Processes)
There are also a variety of methods for the alkaline digestion of lignocellulose,
the most important of which are the kraft digestion (sulfate digestion) and soda
digestion processes. Basically, the raw material is put into a reactor together with
chemicals and water and digested at 140 to 180 °C, a pressure of 6 to 12 bar and
a pH of 14 for 0.5 to 6 h. The products are pulp consisting of cellulose and hemicel-
luloses and black liquor. The lignin contained there can either be used materially
or as an energy source after the digestion chemicals have been recovered. At this
Fig. 14.17 Abengoa demonstration plant. (Source: Press release from June 26, 2013; “Aben-
goa inaugurates its first demonstration plant using Waste-to-Biofuels (W2B) technology”)
point, it should be mentioned that the method is not limited to wood as a starting
material, but that other plant components that contain less lignin, such as straw
and grass and cotton remnants can be utilized in the respective areas where they
are cultivated while maintaining the fiber structure. The use of grass is particularly
interesting for economic reasons, although the proportion of grass used can only
be increased up to about 50% without causing an unacceptable reduction in quality
in paper manufacture [117].
The kraft digestion takes place at temperatures between 160 and 175 °C for
0.5 to 2 h at 9 bar. The active chemicals are NaOH and Na2 S. The soda process
runs at 160 to 170 °C for 5 to 6 h and is applied in the production of paper pulp
from deciduous wood or annual plants according to the state of the art. The active
digestion chemical is NaOH.
Alkaline digestion, despite the complex chemical recovery that is usually sul-
fur-based, is an established and efficient process for pulp production. An integral
part of the process is the chemical recovery, during which a large part of the lignin
must be used thermally. There is clear potential for development here with regard
288 B. Kasal et al.
to the utilization of secondary streams. This includes the pre-extraction of hemi-

celluloses and their material utilization, gasification of the black liquor for material
and energetic use or the extraction of lignin for material utilization. The process
is already being applied in industrial practice within the scope of the kraft process
for the production of pulp with plant sizes of over two million metric tons of wood
consumption per year. Nearly 80% of the pulp produced worldwide is made ac-
cording to this process, amounting to 128 million metric tons in 2011 [55]. Thus,
conventional pulp plants that operate according to the kraft process have a technol-
ogy readiness level of nine.
For the extraction of additional bio-based chemicals through discharge and fur-
ther chemical or microbial processing of various biomass components according to
the concept of the “Integrated Forest Products Biorefinery”, the technology readi-
ness level is highly process-dependent.
14.7.4 Sulfite Digestion/Lignin Sulfonation (Structure-

Preserving Process)
Sulfite digestion also has a range of different processes, where the integral part
of the process is the sulfonation of lignin with SO2 . The basic principle of the
procedure is that the lignocellulose is digested together with SO2 and MgO, CaO
or Na2 O and water at 140 °C, a pressure of 6 to 12 bar and a pH of 1 to 13 for 1.5 to
6 h. Acid, neutral and alkaline sulfite digestion are examples of this process [56].
The acid sulfite digestion occurs at a pH of 1 to 5, which corresponds to the state
of the art for chemical pulp production. Only with this digestion technology is the
extraction of all by-products economically viable [57]. Neutral sulfite digestion at
a pH of 6 to 7 is rarely applied and alkaline sulfite digestion at a pH of 8 to 13 is
still under development.
The products are pulp consisting of cellulose and hemicelluloses and the waste
liquor, which can either be used energetically or materially after recovery of the
digestion chemicals. Large quantities of lignosulfonates are fed into energy gen-
eration, which mainly supplies the process energy. The main application for the
purified lignosulfonates lies in their further processing to produce additives for ce-
ment, bitumen or for the polymer industry. Other constituents of the waste liquor
after chemical recovery include xylitol, furfural and acetic acid.
In principle, following the elaborate chemical recovery with associated high sul-
fur losses, the recovery of by-products is possible in this digestion process. There
is development potential for increasing the yield of by-products and black liquor
gasification. The technology readiness of the acidic magnesium bisulfite process

and the concomitant recovery of numerous by-products can be illustrated using the
example of the companies Lenzing AG and Borregaard, which produce chemical
pulp and various high-grade by-products in the form of a biorefinery on an indus-
trial scale [58, 59]. The technology of acid digestion has been established for over
one hundred years (TRL 9).
14.7.5 Digestion with Alcohols, Organosolv Process (Structure-

Preserving Process)
There are a number of processes for the digestion with alcohols, that use different
alcohols and are carried out at different temperatures, pressures and pH values.
These processes can be summarized under the term Organosolv process. The sol-
vents used are methanol (MeOH), ethanol (EtOH) or propanol, to which catalysts
such as H2 SO4 or NaOH are frequently added. The lignocellulose is in principle
digested in a reactor together with a solvent, water and a catalyst at 165 to 200 °C,
a pressure of 20 to 35 bar and a pH in the acid range of 2 to 4 or in the alkaline range
of 13 to 14 for 2 to 4 h. EtOH-water digestion, the Alcell method, the Organocell
method and the monoethanolamine (MEA) method are examples of this process.
The EtOH-water digestion (also known as the Alcell process with the aim of
pulp production) runs at 195 to 200 °C and 25 to 35 bar. There is usually no cata-
lyst used, but in exceptional cases 0.1 to 1% sulfuric acid is added. This process
was already implemented in the 1990s on a demonstration scale for Alcell. This ap-
proach is being pursued by the Fraunhofer-Gesellschaft as part of the biorefinery
development, and also by various other research institutions. There are two pilot
plants for Organosolv processes with EtOH/water: one plant with a capacity of one
metric ton of beechwood per day at Fraunhofer CBP in Leuna [60] (Fig. 14.18) and
one plant belonging to the company Lignol in Vancouver, Canada, with a capacity
of 350 metric tons per year [61].
The technology readiness of each is estimated to be at a level of 5 to 6. The
company LignoValue in the Netherlands has also planned a research project with
a pilot plant, although this has not yet been realized [62].
The Organocell method uses MeOH as a solvent and operates at 165 to 175 °C.
NaOH is also used as a catalyst, whereby a pH value of 13 to 14 is set. This process
was implemented for pulp production on an industrial scale in the 1990s by the
Bayerische Zellstoff GmbH in Kehlheim. The company had to declare bankruptcy
290 B. Kasal et al.
Fig. 14.18 Pilot plant at Fraunhofer CBP, Leuna
in 1993, however, due to implementation errors during the conversion from the
sulfite to the Organocell process and financial problems (TRL 8) [63].
The MEA process is carried out at 165 to 185 °C and works with mo-
noethanolamine as a solvent. It is therefore a special case of alkaline Organosolv
digestion and is currently still being studied (TRL 4–5).
The products that are formed in all processes are cellulose on the one hand
and the mother liquor on the other hand, from which the Organosolv lignin, hemi-
celluloses, furfural and acetic acid can be isolated after the chemicals have been
recovered. The advantages of these processes are the complete and straightforward
fractionation, highly purified and reactive lignin, easy solvent recovery and fewer
corrosion problems compared to digestion with organic acids. The disadvantages
are the relatively high pressures and the combustibility of the solvents used and the
concomitant safety requirements.
14.7.6 Digestion with Organic Acids (Structure-Preserving

Process)
There are also a great number of processes in the field of organic acid digestion pro-
cesses. In all of these, the lignocellulose-containing material is digested together
with formic acid or acetic acid, water and HCl or H2 O2 as catalysts, at 110 to
190 °C, a pressure of 1 to 13 bar and a pH of 1 up to 3 in a reaction time of 4 to 6 h.
Process examples include the Milox, Formacell, Acetosolv and Acetocell methods.
The products are cellulose and waste liquor, the latter consisting of xylose, furfural
and the corresponding solvents.
The advantages of these methods are that the production of chemical by-prod-
ucts is possible and that in some cases pressureless digestion can be carried out.
The disadvantages here are also high corrosion and the relevant explosion protec-
tion requirements.
The Milox process runs at 60 to 110 °C for 4 h and operates in two stages: the
first step runs with formic acid at atmospheric pressure and the second step, under
pressure, with peroxyformic acid, which is made from formic acid and hydrogen
peroxide [64]. This process is currently being developed further for annual plants,
in different variants on a pilot/demo scale by Chempolis in Finland. The technol-
ogy readiness level can thus be estimated to be 5 to 7.
In addition, there are three processes that use acetic acid as a solvent: the For-
macell, Acell, and Acetocell methods. The Formacell process has been brought
up to a technology readiness level of 5 to 6. The process runs at 150 to 180 °C
and works with acetic acid and 5 to 10% formic acid as a solvent. It is being de-
veloped further as an unpressurized variant by the company CIMV in France on
a pilot/demo scale for the use of 800 metric tons of straw per year (Fig. 14.19) and
is currently being transferred to industrial scale [65].
The Acetosolv digestion proceeds at 110 °C and ambient pressure or at 170–
190 °C and elevated pressure, and works with acetic acid as a solvent and hy-
drochloric acid as a catalyst. This process was developed in the 1980s, as was the
acetocell process, which proceeds at 190 °C with acetic acid as the solvent [64].
Although these processes have already been used on an industrial scale, the com-
plicated recovery of the chemicals used is problematic. For this reason, especially
with regard to the use of by-products in the pulp process, other digestion processes,
such as the Organosolv process, are preferred over the Milox and Formacell pro-
cesses [66].
292 B. Kasal et al.
Fig. 14.19 CIMV pilot plant for straw
14.7.7 Digestion with Ionic Liquids (Structure-Preserving

Process)
There are various methods for digestion with ionic liquids that differ according to
the ionic liquid that is used and the associated varying process parameters. Ligno-
cellulose is digested together with ionic liquids—e.g. EMIM (1-ethyl-3-methylim-
idazolium acetate) in the ratio of 10 : 1 or BMIM-Cl (1-butyl-3-methylimidazolium
chloride) [67] in the ratio of 9 : 1—and a variety of other salts [68] at 80 to 120 °C,
and a pressure between 0 and 60 bar for 0.5 to 6 h. The product is dissolved cellu-
lose, which is used as a starting material for further fractionation, derivatization or
saccharification. The hemicelluloses and lignin can thus be separated out relatively
cleanly, whether present in solution or still as solid components [67]. The advan-
tages of this digestion process are the almost complete solution of lignocellulose,
which facilitates new approaches to component separation and conversion. There
is high potential for development here in the digestion of the biomass for subse-
quent enzymatic conversion [69]. The disadvantages of the process are the cost,
which is still high and the recovery of the ionic liquids, which has not yet been
solved. Development potential in this area therefore lies above all in the recycling
of the solvents that are used. The technology of digestion with ionic liquids is cur-
rently at laboratory and pilot plant scale, which is why the technology readiness is
estimated to be at a level of 3 to 4.
14.7.8 Digestion by Calcium Oxide (Structure-Preserving

Process)
For pretreatment using lime, calcium oxide (CaO) or calcium hydroxide (slaked
lime, Ca(OH)2 ) is diluted with water and mixed with the lignocellulose in a tem-
pered reactor. The hemicelluloses are retained in the solid phase of the biomass,
while the acetyl groups and uronic acids are in large part removed. The process is
so mild that hardly any cleavage products are formed. A disadvantage is the recov-
ery of the lime, because it either has to be laboriously separated from the lignin
or, if the lignin is utilized energetically, it decreases the fuel value significantly
[70]. For lignocellulose with a high lignin content, it is necessary to add an oxidiz-
ing agent to achieve sufficient delignification, which turns the process into a “wet
oxidation” process [71] (Sect. 14.9).
14.7.9 Pretreatments for Subsequent Hydrolysis and

Utilization of the Sugar Fractions (Some Structure-
Degrading Processes)
When lignocellulose is to be hydrolyzed enzymatically or by means of acid, e.g. for

subsequent fermentation from cellulose to ethanol, it must be pretreated for the hy-
drolysis to be effective. In this way, accessibility can be increased for the enzymes
or the acid that are used. Pretreatment digestion methods can be of a physical
(grinding), physicochemical (steam explosion, hot water combined with acids),
chemical (acid, alkaline) or biological (fungi) nature, as the processes already men-
tioned [72].
Biological and physical pretreatment methods can however be excluded as the
sole pretreatment, at least when using wood. Fungal pretreatment is industrially
not feasible since it is too time consuming with a duration of several weeks, while
the use of a cleaver or mill for physical digestion without chemical pretreatment of
the wood is too energy intensive. The process of the steam explosion has become
established, as it is relatively energy efficient and can achieve a high yield of up to
95%. It is already being used in industrial practice [73].
294 B. Kasal et al.
Pretreatment with Ammonia

Pretreatment with ammonia in the AFEX (ammonia fiber/freeze explosion) process
is a modification of another method for pretreatment with subsequent enzymatic
hydrolysis. It operates at temperatures of 160 to 180 °C in a flow reactor with
aqueous ammonia in a concentration of 5 to 15%, with a retention time of about
15 min. Since almost all of the ammonia used can be recovered, the process is also
known as ARP (ammonia recycled percolation). The ammonia depolymerizes the
lignin and splits the existing lignin-carbohydrate bonds, without attacking the cel-
lulose. The advantages of the process are the small quantities of enzyme required
for the subsequent hydrolysis and relatively high conversion or digestion rates.
However, it is not suitable for coniferous woods and only to a limited extent for
deciduous woods. It was originally developed for agricultural raw materials and
herbaceous plants [34, 81]. The process is currently being transferred from the lab-
oratory to pilot scale in cooperation with universities and the technology company
MBI (TRL 5) [82].
Digestion with Mineral Acids

In the case of digestion with mineral acids with simultaneous saccharification, pos-
sibly following a previous digestion, the lignocellulose is converted to hydrolysis
lignin and a sugar mixture together with sulfuric, phosphoric or hydrochloric acid,
water and a catalyst e.g. hydrogen chloride. The digestion takes place at 140 to
170 °C and a pressure of 8 bar in 10 to 14 h [34, 81]. Variations of this digestion
process include the Bergius method, the Scholler method and the Verdia method
(until 03/2012 HCl Cleantech). The problem with all methods of this type is the
lignin separation or disposal, the acid recovery and the occurrence of corrosion on
the equipment, which increases with increasing concentration of the acids used.
Recent developments with concentrated hydrochloric acid have however been suc-
cessful and realized by companies on a demonstration scale (Weyland) and on an
industrial scale (Virdia, Bluefire).
The Verdia method (CASE process), for example, has been brought to an esti-
mated technology readiness level of 8 to 9. A first plant exists for this process with
a production capacity of 150,000 metric tons of sugar per year.
Bluefire operates a production facility in Japan and plans two more plants in
Lancaster, California and Fulton, Mississippi, USA, each with capacities of just
under 15 (cellulosic by-products) and 72 million liters (wood, etc.). The Bluefire
technology involves the hydrolysis of the purified and mechanically preshredded
biomass with sulfuric acid. The sugar-acid solution is then separated chromato-
graphically with ion exchange resins as the stationary phase and the acid is recycled
[85]. The technology readiness here is estimated to be 7 to 8.
Weyland operates a pilot plant in Bergen, Norway, which produces 200,000

liters of ethanol per year using concentrated acid in a total hydrolysis process.
A special feature of the process is a method for recovering up to 98% of the
acid used. There are also cooperations within Norway with ambitions to build
a bioethanol plant with a capacity of 25 to 30 million liters. The technology readi-
ness here is estimated to be 6 to 7.
The Bergius process is a forerunner of methods used today with mineral acids.
It was developed in the 1930s and runs at about 160 °C with highly concentrated
hydrochloric acid. The Scholler process runs at 170 °C, works with diluted sulfu-
ric acid and was implemented on an industrial scale in the USSR. Although these
technologies have already been used on an industrial scale in the past, they are no
longer economically viable today and do not meet current environmental require-
ments [85].
Pretreatment with Fungi or Lime

Pretreatment with fungi for subsequent enzymatic conversion is currently being
tested. There are promising approaches in which the use of energy (pressure and
heat), expensive technology (complex plant designs) and chemicals (especially
acid) can be reduced or eliminated [86]. Lime pretreatment can also be imple-
mented as a variation of a two-step process for subsequent enzymatic hydrolysis,
similarly to the previously described Organosolv processes and pretreatment with
organic acids.
In the enzymatic hydrolysis of the pretreated lignocellulose the latter is com-
bined with suitable microorganisms and water at a temperature of 40 to 60 °C, at
atmospheric pressure and a pH of 5 to 6 for 12 to 48 h to convert it to a carbohy-
drate fraction (mainly C6 sugars) and so-called hydrolysis lignin. The latter can
be filtered off and discharged, often together with the C5 sugars of the hemicellu-
loses, as well as extracts and ash components. No inhibiting degradation products
are formed during enzymatic hydrolysis and subsequent enzyme recovery in the
downstream process can make the process economical. An estimated Level 7 in
technology readiness was achieved at a first demonstration facility of the POET
and DSM joint venture in Emmetsburg, Iowa, USA, where agricultural residues
are fermented to ethanol [84] (Fig. 14.20). There is also a major commercial plant
being constructed by DuPont in Nevada, Iowa, USA, which will be the world’s
largest plant of this kind. It aims to produce just under a million metric tons of
ethanol per year, from straw by fermentation [85]. In the Kalundborg bioethanol
demonstration plant of Dong Energy and Inbicon in Denmark, 4 metric tons of
straw can be converted into ethanol, lignin pellets and C5 sugars per hour [86].
296 B. Kasal et al.
Fig. 14.20 POET-DSM demonstration facility, Emmetsburg, USA
Clariant has built a demonstration plant in Straubing, Germany for testing the
SunLiquid process for the production of ethanol from straw and put it into opera-
tion in 2011 with a production of 2000 metric tons per year. A special feature of
this is that C5 sugars can also be converted by fermentation [77]. The demo plants
of the companies Abengoa (no longer active), Babilafuente, Spain [52] and logen,
Birch Hills, Ottawa, Canada [51] (Sect. 14.9) also produce ethanol from straw and
other lignocellulosic biomass. Their processes have already been implemented in
existing commercial facilities or plants are currently under construction [78, 79].
Cellulosic ethanol has already been industrially produced for several years, mainly
through enzymes, and above all in the USA. Currently massive expansion is on-
going with the aim of building facilities with a total capacity of 60 million cubic
meters of ethanol from cellulose over the next ten years [80]. Although the pro-
cesses are being optimized, especially with regard to increasing the conversion
rate of the enzymes, the technology readiness can be rated at 7 to 9. On account
of sometimes very high subsidies, both in the USA and the EU, it is difficult to
ascertain the profitability of the processes. This is especially true with regard to the
marketability of the main product.
The sugar monomers obtained after the enzymatic hydrolysis of the sugar frac-
tion(s) can serve as a carbon source for fermentation processes. From the descrip-
tions of the digestion processes it can be seen that bioethanol was and in large part
still is the driver of these efforts worldwide. The more or less large amounts of by-
products formed in the digestion can usually only be removed with great effort and
thus its use as a fermentation raw material for many sensitive production organisms
is still in development. For each application verifications must be made as to how
tolerant the microorganism used may be or rather, how thoroughly the input sugar
fraction has to be purified.
The separate use of C5 and C6 sugar fractions, as is enabled by the Organosolv
process or a hydrothermolytic digestion, appears similarly lucrative. Conversion
to products with properties specifically associated with the xylan C5 structure of-
fers the best prospects for a high level value creation. Despite the generally high
availability of this raw material, however, it has been a niche area up to now. This
may also be due to the fact that these hemicelluloses are not yet available in suf-
ficient quantities for industrial applications. The high yield conversion to furans is
still a particular technical challenge, which has prevented the path to attractive and
interesting products of furan chemistry from being economically viable till now.
14.7.10 Utilization of the Lignin Fraction
A further fraction is formed during the pretreatment besides the sugar fractions:
lignin. Lignins are three-dimensionally linked networks of phenylpropane units
and represent the crosslinking component between the sugar polymers. Moreover,
lignins are hydrophobic and protect the more easily degradable carbohydrates from
microbial degradation. Depending on the pretreatment method used, the lignins
obtained differ greatly from each other, both qualitatively and structurally. Their
chemical and physical properties thus also vary.
Classic digestion processes such as the sulfate process or the sulfite process
are used primarily for pulp production and yield sulfur-containing lignins due to
the process chemicals used. High-quality material utilization, however, requires
especially those methods which yield sulfur-free lignins. The Organosolv process
and also the soda process or digestion variants that use organic acids are suitable
for this. Strongly acidic digestion conditions affect the lignin quality negatively
and lead to a considerable condensation of the lignin scaffold and thus virtually
preclude further material processing.
298 B. Kasal et al.
Some lignin types can be condensed into the resin with preservation of the poly-
meric structure as so-called liquid wood, by injection molding or extrusion either
thermoplastically [87] or by thermosetting. When used in adhesives for wood-
based materials, for example, the mechanical properties can even be improved
compared to the use of pure phenol [88]. In addition to their established use as
a dispersant, lignosulfonic acids (LSA) from the sulfite process were also used as
binders in wood-based materials in the past. The lack of reactivity of LSA’s makes
their use in adhesive systems tricky [89]. The Pedersen process, for example, did
not become established due to the long pressing and post-processing times. Phenol-
formaldehyde (PF) resins containing a proportion of kraft lignin can also produce
wood-based materials of DIN-compliant quality, although here too the reduced re-
activity compared to pure PF resins is problematic and must be compensated for by
the use of suitable hardener systems [90]. Other possible applications of lignin are
in polyurethane foams, epoxy resins or dispersants [91]. In addition, it was possi-
ble to show that lignin fibers may be spun from both Organosolv and Kraft lignin,
which can be further processed into carbon fibers [92]. A major market is foreseen
for the use of lignin-based carbon fibers [93].
Further structural degradation of the lignins obtained by digestion and isolation
processes leads to aromatic phenol-type building blocks. This broadens the appli-
cation possibilities and makes lignins usable even in more complex systems. Oxi-
dation, pyrolysis, hydrogenolysis, hydrolysis and enzymatic processes are known
depolymerization methods. Fundamental studies have been published on different
lignins or model substances, but only oxidative cleavage has been implemented as
a technical process, for the production of vanillin. The other approaches range from
being in the experimental stage up to pilot plant scale [94]. For example, studies
as part of the collaborative Lignoplast project have shown that aromatic synthe-
sis building blocks derived from various lignin types, prepared by base-catalyzed
hydrolysis, can be used in adhesive, coating, polyurethane and epoxy systems [95].
The process of base-catalyzed cleavage of lignin leads to the formation of
monomeric alkoxy phenols and catechols as well as to dimeric and oligomeric
alkyl-functionalized aromatic compounds [97] through hydrolysis of the aryl aryl
ethers and aryl methyl ethers of the lignin macromolecule, and is currently at
TRL 5 [97]. Targeted modification and further functionalization of the lignin cleav-
age products are required before they can be used in high-quality formulations such
as polyurethanes and adhesives. This can be achieved through biotechnological or
chemical conversion.
The particular challenge in the material processing of lignin lies in its inhomo-
geneity and widely varying chemical and physical properties, which depend on the
botanical origin of the starting material as well as the digestion and fractionation
conditions and the further refinement of the lignin fraction. It is therefore abso-
lutely essential for the entire process chain to be considered from both an economic
and ecological perspective for a comprehensive evaluation of every application. In
summary, lignins have huge market potential and are not only seen as an energy
carrier or niche product. However, industrial scale demonstration projects are re-
quired in order to explore and confirm the application potential.
It is evident from the reports that plant-based starting materials can be prepared
via fermentation or chemical processes either directly as a material, fractions, as
material components or, following digestion, completely new molecules (biopoly-
mers, solvents, etc.). The added value from renewable raw materials can be max-
imized through the procedural and product-oriented linkage of these processing
channels.
14.8 Pyrolysis to Oil (Non-Structure-Preserving Process)
Non-structure-preserving processes are an entirely different way of using plant-

based materials. Since the structural synthesis benefits provided by nature are not
exploited due to the harsh conditions used in these processes, such as pressures and
temperatures well above those of structure-preserving processes, a new synthesis
of the resulting molecules is necessary to produce the desired materials. These
methods are therefore mostly used for the synthesis of biofuels or basic chemicals.
Pyrolysis is a thermolytic process in which the lignocellulose is converted to
the products bio-oil, coal and various gases at high temperatures and under at-
mospheric pressure. The process can be carried out at different temperatures and
retention times, where the latter in particular significantly affects the product com-
position. In contrast to gasification, the process is carried out exclusively by intro-
duced heat under the exclusion of oxygen.
Reactors for the process are transported bed, circulating fluid bed, ablative, ro-
tating cone or vacuum reactors, which differ mainly in biomass input and heat
transfer [98]. The aim of pyrolysis is to produce a liquid intermediate (bio-oil),
which can be further processed using established refinery processes. The develop-
ment potential of this process lies in its decentralized implementation for increas-
ing the energy density and in the production of bio-oil as a “drop-in” raw material
for conventional refineries.
The main application is the conversion to the product pyrolysis oil (bio-oil).
In a “fast pyrolysis”, with a raw material retention time of about one second at
approx. 500 °C, about 75% is converted to oil. In an “intermediate pyrolysis” at
about 500 °C, about 50% is converted to oil. In a “slow pyrolysis” slightly lower
300 B. Kasal et al.
Fig. 14.21 BTG-BTL pilot plant
temperatures of about 400 °C are selected, with only about 30% of the raw material
being converted to oil and mainly gas being produced [99]. A disadvantage of these
processes is that bio-oils are thermolabile and prone to polymerization reactions,
which complicates their preparation and further processing.
The Dutch company, biomass technology group (BTG), delivered a fast pyrol-
ysis plant to Malaysia, which produces 1.2 t of pyrolysis oil per hour from two
metric tons of biomass. A larger plant with a capacity of 5 metric tons per hour
is planned in Hengelo, Netherlands [100] (Fig. 14.21). A fully integrated bio-oil
pilot plant was completed by the consortium Metso, VTT, Fortum and UPM in
Tampere, Finland in 2009 [101]. The process of fast pyrolysis is thus estimated to
be at a technology readiness level of 8 to 9. The company Pytec developed and
built a pilot plant with a capacity of 4 metric tons of oil per day in Hamburg, Ger-
many in 2005. The company discontinued operations in 2015. At the University of
Aarhus, Denmark, the HTL process (hydrothermal liquefaction) is currently being
scaled up to pilot scale. In this process, biomass of mixed origins can be converted
into pyrolysis oil [103].
14.9 Pyrolysis with Subsequent Gasification (Non-Structure-

Preserving Process)
During gasification, lignocellulose is converted to synthesis gas or fuel gas and

slag or ash when combined with the conveying gas, steam or an oxidizing agent
(usually oxygen) at a temperature of 800 to 1800 °C and a pressure of 0 to 60 bar.
A distinction can be made between autothermal gasification, where the heat is
provided by partial oxidation of the starting material and allothermal gasification,
where the heat is provided by an externally supplied substance, such as steam or
gas.
Gasification reactors or processes include fluidized bed gasifiers, fixed bed gasi-
fiers and entrained flow gasifiers (Fig. 14.22). The aim of the process is to produce
a fuel gas which, after appropriate preparation, can either be fed into direct com-
bustion, or one of the versatile synthesis gases used in the chemical industry. The
gases can thus be processed into synthetic natural gas (BioSNG), Fischer-Tropsch
fuel (FT-diesel), hydrogen, methanol and dimethyl ether. The high raw material
flexibility is a great advantage of these processes. Some plants are already in op-
eration on a commercial industrial scale, although most plants are still unique and
are thus still seen as test facilities. The degree of technological readiness can thus
be estimated to be 8, although developmental potential exists, especially with re-
Fig. 14.22 Different types of gasifiers (moving bed gasifiers, fluidized bed gasifiers and
entrained flow gasifiers)
302 B. Kasal et al.
gard to increasing the plant efficiency and improving the efficiency of the reactor
architecture and/or biomass management.
The first commercial plant with a dual fluidized bed gasifier and subsequent
combustion of the gas for electricity and heat generation with a capacity of 2 met-
ric tons of biomass per hour (8 MW) was put into operation in Güssing, Austria in
2001 [104]. The two Finnish companies Stora Enso and Neste Oil commissioned
a plant in Varkaus, Finland within the scope of the NSE Biofuels project in 2009,
which produces 650 metric tons of FT diesel and generates 12 MW of electrical
power through combustion of the gas [105]. A plant, built by Choren in Freiberg,
Germany in 2010 with a capacity of 13,500 metric tons of lignocellulosic biomass
input for the production of gas and FT-diesel according to the CarboV process
[106] declared bankruptcy in 2011. The technology was further developed by the
Engineering Division of the Linde Group, which discontinued the development
after a few years. The company ECN commissioned a plant for the gasification
of 1800 metric tons of a variety of lignocelluloses in Petten, Netherlands in 2011
[107]. Enerkem built a plant in Westbury, Quebec, Canada in 2010 for the gasi-
fication and synthesis of alcohols from 12,000 metric tons of lignocellulose and
household waste per year, that produces 4000 metric tons of ethanol [108]. An
open pilot plant center for the gasification of up to 20 metric tons of a variety
of feedstocks per day was set up by the Southern Research Institute in Durham,
USA [109]. In 2008 the company Range Fuels in Soperton, Colorado, USA built
a plant for the gasification of wood and waste materials and subsequent synthe-
sis of 300,000 metric tons of ethanol and FT diesel per year, which however had
to be closed again in 2011 [110]. For the gasification of wood and subsequent
FT synthesis, Flambeau River Biofuels in Wisconsin, USA commissioned a plant
with a capacity of 51,000 metric tons per year in 2011 [111]. This company has
now ceased to operate. In 2009 the company Syngest planned the construction of
a plant to produce fuel and fertilizer from NH3 , mainly from corn, in a fluidized
bed reactor [112].
14.10 Fermentation (Non-Structure-Preserving Process)
In fermentation, the partially degraded lignocellulose together with cellulose and

hemicelluloses derived, for example, from the digestion of lignocellulose, are con-
verted, under usually mesophilic conditions and at atmospheric pressure, to biogas
and nutrient-rich liquid and solid fermentation residues. After purification, the gas
can then be introduced into the natural gas pipeline network. The basic principle is
anaerobic fermentation which, depending on the raw material, can be carried out as
a one- or two-stage wet fermentation or alternatively in a dry fermentation in a sin-

gle-stage or percolation process (container or horizontal plug flow process) [113].
Highly water-laden biomass, which cannot be directly used thermally, is especially
suitable as a substrate. Biomass such as wood and straw, which is rich in lignocel-
lulose, is difficult for microbes to access for degradation and therefore unsuitable
as a substrate for biogas production without prior digestion of the cellulose.
The production of biogas is widespread throughout the world. It is often located
decentralized for agricultural residues and individually adapted to raw materials,
size and climatic conditions. Germany alone, with over 9500 biogas plants, has an
installed electrical output producing more than 5000 MW [114].
The advantages of anaerobic fermentation are the short retention times of 1.5 to
2 weeks, the relatively small digestion chamber required, a high degree of degrada-
tion, a relatively high biogas yield and the multi-substrate capability of the process.
The main disadvantage is the higher cost of investment compared to conventional
systems. Development potential lies in scaling up the process for economically
viable plant operation and in pre-shredding.
Fig. 14.23 EtaMax process demonstration facility

304 B. Kasal et al.
The EtaMax plant, a high-load fermentation demonstration plant, which has

been in operation since the end of 2012 and has a throughput of 160 metric tons
per year, has achieved an estimated level 7 in technology readiness (Fig. 14.23).
It is part of the EtaMax research project towards a decentralized, sustainable and
resource-efficient overall concept for the use of waste biomass as fuel [115].

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Cognitive Biological Sensors
Learning from Nature for Nature
15
Albert Heuberger, Randolf Hanke, and Claudia Eckert
Summary
In biology, the targeted observation of the environment through organs of sight,

smell and touch is closely linked with the simultaneous steps of cognitive pro-
cessing of the data for acquisition of information and knowledge. In the dig-
itized economy, a variety of sensors are also used and networked with one
another to acquire information and make automated decisions. This chapter
addresses this duality between biology and technology from a variety of per-
spectives. Biological transformation has implications for technical systems, es-
pecially in the field of networked and cognitive sensor technology, where it has
an impact on the construction of novel sensors, on methods of establishing effi-
cient communication channels between them and not least on the objects being
observed themselves, which are increasingly plants as natural resources. This
chapter discusses research questions and recent findings in networked sensor
technology as part of the development of the Internet of Things, which can ul-
timately also be used to learn new things about biology as well as to advance
technology in this area.
A. Heuberger () R. Hanke

Fraunhofer Institute for Integrated Circuits IIS
Erlangen, Germany
C. Eckert
Fraunhofer Institute for Applied and Integrated Security AISEC
Garching, Germany

https://doi.org/10.1007/978-3-662-59659-3_15
312 A. Heuberger et al.
15.1 Introduction and Motivation
Biological transformation is a prime mover of technologically driven innovation.

This also applies to developments resulting from the advanced digitization of the
economy. A central building block of a digitized economy is modern, networked
sensor technology, which is increasingly forming the basis for the future Internet of
Things in the sense of networking the physical and virtual worlds. The principles
of biological transformation may be applied both to the development of sensors
and to the use, transmission and interpretation of the generated data volume. Not
least, also biological systems themselves are being monitored, evaluated and even
optimized by sensors as a part of economic cycles.
The following chapter covers, on the one hand, the bio-inspired perspec-
tive—that is, the question of constructing sensors based on biological models
and the value-added knowledge gained from their networking. However, it also
includes how biological systems can be better researched by means of modern
sensor technology. For both aspects, questions also arise about the utilization of
the data and information generated. This utilization is managed through the use of
intelligent algorithms. The latter are sometimes implemented directly in the sensor
and sometimes only at the point where the data from many sensors is combined.
In this context, sensor systems for the Internet of Things are to be developed
based on biological sensor systems in future. Biological, multimodal sensor sys-
tems operate on the basis of “sensors” for sight, hearing, smell or touch and are
inextricably linked to nervous systems for data processing.
This sensorium analyzes the large volumes of acquired data parallel to the data
acquisition. It modifies or adapts its “sensors” while simultaneously carrying out
the intrinsically given task. Intelligence of this nature thus leads to adaptive mea-
surement and reduces large volumes of data (big data) to quantities of important
data (relevant data), from which information and knowledge can subsequently be
generated. Leading on from this, in the future artificial intelligence will not only be
applied sequentially, following data acquisition for feature extraction; rather, sen-
sor systems will begin to “think” with the aid of the Cognitive Internet while the
data is being collected. In other words, they will analyze, plan, and draw conclu-
sions. Sensor-based artificial intelligence will in future ensure that from the start
only relevant data is captured, from which the required information and knowledge
can be extracted.
It will therefore be a future task to extract usable, relevant information as
quickly as possible from the exploding quantity of measured data coming from
the widest variety of sensor sources. This information must be selected simultane-
15 Cognitive Biological Sensors 313
ously from the data of any desired sensor system in order, for example, to adapt
the data acquisition and then use the intelligently extracted knowledge to derive
measures for concrete tasks, such as the control and optimization of the underlying
processes.
The rapid progress in the digital development of the world is already providing
a foretaste today of the possibilities of ubiquitous data acquisition, communication
and subsequent data analysis. For example, digitization in manufacturing (key-
word: Industry 4.0) is leading to the reorganization of production and logistics
processes, is increasing the efficiency of companies, facilitating individual prod-
uct design, enabling completely new business models and changing the daily work
routine of millions of employees.
It is expected that this development will progress step by step into all areas of
life. The digital development of the world requires the precise capture, processing
and interpretation of the environment—similar to human sensory perception—as
the basis for the next evolutionary level of networked intelligent systems. At the
same time, human perception needs to become more prominent. What is required
is a human-derived “Internet of Senses” that surrounds us. The assessment pro-
cesses that ultimately determine our human emotions, our choices and our behavior
are based on primary sensory processes and the related perception processes. An
improved understanding of these primary processes through technical-mechanical
developments can thus help to better understand or even predict human decision-
making processes, emotions and behavior (intuitive sensing) and to develop sys-
tems that are better equipped to deal with human decisions and reactions based on
this knowledge. Knowledge of this nature is a fundamental necessity, especially in
stress and crisis management. The Internet of Senses is intended to support people
in their daily routines. Furthermore, the aim is that in the future, intelligent, inter-
linked objects and processes shall be able to independently capture or model the
state and sensory perception of humans and to interact with them. It is even con-
ceivable that sensors will in future develop a kind of individual intuition similar to
people i.e. so not just cognitive but intuitive, anticipating sensors.
In summary, cognitive sensor systems of the Internet of Things will in future
learn, based on their high-dimensional data streams, to adapt to changing processes
(intelligent acquisition planning), monitor themselves in their own functioning and
network intelligently within a sensor network. With these capabilities and through
intelligent signal processing, cognitive sensor systems can generate precisely the
relevant information necessary for process optimization—from big data to relevant
data. Intelligent, self-learning algorithms can thus be used to formulate a new vi-
sion for the modern Internet. Based on its intelligent sensor systems, the Cognitive
Internet will in future become autonomous, yet remain transparent and compre-
hensible and will decide within a human-dictated framework what, when, where
and how it observes and records.
In the sections that follow, selected examples are used to show how biological
processes can serve as a model for technological processes, sensor technology and
sensor networks. Thereafter the current level of knowledge that has been reached in
terms of sensory perception and biological structures is presented, along with the
projects which are to apply this knowledge. In conclusion, a description is given
of how the Fraunhofer-Gesellschaft is meeting the increasing complexity of topics
above and beyond the field of biological transformation through a cluster of excel-
lence and how it is developing solutions for the Cognitive Internet for applications
in business and private life.
15.2 Learning from Biology
15.2.1 Insect Swarms as Multi-Modal Sensor Networks
Richard Schielein
For decades, the biological principles of swarms have been considered as candi-
dates for new technical sensor systems on account of their efficiency and robust-
ness. The underlying direct and indirect communication allows for task-specific
adaptation of the derived action, without depending on central nodes. In the follow-
ing, some terms from the field of swarm intelligence are briefly introduced. This
is done with the aim of demonstrating a particular pathway of biological transfor-
mation that has already yielded several useful artificial intelligence algorithms. In
addition to the already existing application of swarm intelligence in solving op-
timization problems, another application that can be considered is to transfer the
networking of the biological sensorium to technical sensor systems.
Ever since the mid-1990s, the concept of swarm intelligence can be found in-
creasingly in scientific literature [1]. The term refers to the observation that the
action or communication of individual members of a swarm can result in intel-
ligent behavior of the entire swarm. The “swarm” as an organizational form is
capable of actions that do not follow or only indirectly follow from the repertoire
of action or information content of individuals.
This phenomenon has been observed and studied in colonial insects such as
ants, bees or termites, but also in fish and bird swarms. This emergent property of
swarms can be described using the mathematical tools of self-organization. In this
way, the utilization of swarm intelligence has also been successful, for example
in the form of artificial intelligence algorithms. As early as the 1990’s, physicist
Hermann Haken demonstrated a connection to swarm intelligence with the princi-
ple of synergetics. The latter describes the phenomenon of self-organization with
mathematical precision [2].
The empirical observation of (natural) swarms shows that they behave re-
siliently and flexibly. This means that neither the loss of single components
(individuals) nor changing environmental conditions lead to system failure. Ro-
bustness and flexibility are therefore properties that are of paramount importance
where applicable to technological systems. Moreover, a biological transforma-
tion of swarm behavior is desirable under the working hypothesis that biological
processes are economically optimal.
When applying processes drawn from biology, as described here for swarm
intelligence, there is a special feature that should be pointed out explicitly. This
concerns their metaphorical character (This may apply to biological transformation
in general). In a highly interdisciplinary working environment, as found in applied
research and development, the advantage arises that biological metaphors facilitate
the communication and understanding of the developers.
Insect swarms can be considered self-organized systems. Self-organization phe-
nomena describe precisely the fact that higher structural order or global behavior
arises in complex systems. In the (full) description of the communicative behav-
ior of colonial insects, stigmergy—the indirect communication of individuals by
means of environmental modification—is important for understanding the self-
organizing system. Stigmergy is achieved, for example, via pheromone traces or
generally by means of any changes in the environment (e.g. degree of progress of
nest building).
Starting from the digitization of modern production systems, which contain an
large number of sensors (data generators), the question may be asked how the char-
acteristics of biological sensor systems can be adapted. The biological metaphor
of a “swarm of insects as a multimodal sensor network” may be considered and
this may be understood as a possible model for a system to monitor digitized pro-
duction (Fig. 15.1). The following working hypothesis can therefore be derived:
the design of biological sensor networks guarantees optimal and task-specific us-
age of the available sensorium. The resulting information is either processed “on
the spot” or communicated in an effective way. This means that the information,
which an individual gathers by means of its sensory organs, arrives exactly where
it is needed (one or more other individuals). The form of communication can be
direct or stigmergic.
Fig. 15.1 Future factories could take their communication structure from the “factories” of
nature. (Fraunhofer IIS)
The working hypothesis outlined here can be understood as a “biological data

economy”. To make such a data economy technologically usable, the rules of
communication in self-organized systems must be understood and transferred to
technical sensors. The fact that swarms do not require a central storage unit holds
potential. Task-Specific distribution of information across the agents of a net-
worked production facility generates robustness in the overall system.
However, when designing communications protocols derived from swarm com-
munication, there are also facts where the biological metaphor is not sufficient.
This is due to the low diversity of individuals in the biological swarm. Sensor
machines are found in greater variety. There is thus the additional challenge of
transferring the biological set of rules relating to communication in the self-orga-
nized system to the more diverse technological sensor network.
In summary, there is great technological and economic potential in transforming
the networking of the biological sensorium of swarms to technical sensor systems.
In doing so, the challenges lie in the exact understanding of such self-organized
systems and their communication methods, as well as the transfer to heterogeneous
technical sensor networks.
15.2.2 Beyond Human Vision—Multispectral Sensors and

Polarization
Michael Schöberl
The foundation for rapid development in imaging and processing was laid in 1975
with the first digital camera. At first, low spatial resolutions (100 × 100 pixels)
were still common and the further development and market penetration that digital
cameras would reach were not foreseeable.
In the years that followed, the goal was first to reach the performance of hu-
man vision and to develop cameras that could deliver a similarly convincing image
quality. Image qualities were gradually improved which enabled digital cameras
to be used in various industries. In the 1990s, Fraunhofer IIS made an important
contribution with the development of one of the first high-speed digital cameras,
which among other things led to the replacement of mechanical film cameras for
recording crash tests. At the turn of the millennium, projects were launched aiming
for a similar revolution in the field of digital cinema and the media industry. The
focus was on the best possible resolution, dynamic range and color reproduction.
Major further technical developments followed in the consumer market. As early
as 2003, more digital cameras were sold than analog consumer cameras and since
2006, the number of cameras in smartphones dominates. Intensive research and
development are being carried out in this segment, and advances will, with some
delay, also be usable beyond smartphones.
Digital imaging has reached a level of quality today, both in photography and
industrial inspection, that catches up with human perception. The cameras of today
have comparable abilities to those of our familiar visual apparatus.
What can the human visual apparatus actually accomplish? Despite the im-
pressive power of human vision, there are clear limits to human vision that are
surpassed in many places in nature. The usable spectral range of the human eye is
limited to 400 to 750 nm, whereas some animals are able to exploit an evolution-
ary advantage—perception in both the ultraviolet and in the infrared range. The
number of spectral bands (red, green and blue) is also exceeded in nature: up to
15 different receptors, for example, help a butterfly in its search for members of
its species and food. By contrast, nocturnal animals (active at night) have different
photoreceptors and their eyes have a different structure, so that they can see even
in much lower ambient light. Likewise, there are some animals that can use a nat-
urally occurring polarization of the ambient light to find water surfaces for laying
eggs or to orient themselves in flight [3].
It is possible to reproduce these properties using current imaging technology.

Extensions of wavelengths, spectral resolution, sensitivity and polarization are pos-
sible and available in specialized cameras today. These effects are present in nature
and are already used among animals; although they still only occupy a niche in
imaging and analysis. One major reason lies in exceeding human possibilities. We
do not perceive these properties and therefore have no idea how our environment
would look in infrared or even multispectrally; the polarization of light is also un-
known to us in everyday life.
Fraunhofer IIS is working on the use of additional modalities such as polar-
ization and multispectral sensors for technical applications. These effects can be
exploited, for example, in the industrial inspection of manufactured products, but
also for natural objects e.g. in the food industry. Initially, extensive experiments
are the starting point for assessment of a research question. Laboratory tests can
determine whether the reliable and robust distinction of objects or the recognition
of defects in samples is possible. Camera laboratories have a range of systems, op-
tics, lighting and cameras for this purpose, with which a material sample may be
examined in a variety of ways.
When it comes to polarization, transmission effects (e.g. stress birefringence in
anisotropic materials such as glass or plastics [4]), reflection (in nanostructures,
carbon fibers [5], and thin-layer ellipsometry), and scattering by small particles are
common. Different wavelengths are used here (in visible light, near infrared and
short-wave infrared) to be able to penetrate objects. For example, silicon is only
transparent above 1000 nm and can then be examined for tension.
The use of selected wavelengths is also helpful in multispectral applications
without making use of the polarization property. The molecules of a material can
absorb light of specific wavelengths due to matching vibration frequency and thus
indicate the presence of specific substances. Point spectrometers are initially used
for this purpose, in order to then develop an industrial testing and sorting process
with the aid of multispectral cameras. Good results can be seen especially with
organic materials such as food, plants and tissue samples.
The laboratory evidence starts a process of industrial use of the effects. This
requires acquisition systems that can be implemented with sufficient speed and
industrial robustness. For example, when bottle glass is tested for proper thermal
stress treatment, the components such as the light source and optics must also be
able to deal with glass breakage. One aspect of integration into the customer’s
processes is often the handover to a supervising system integrator.
The development of imaging systems is currently on the threshold of expand-
ing the acquisition possibilities of human vision. More and more technical systems
are becoming available for this. Along with this, image analysis has to cope with
ever increasing volumes of data and hidden information in the high-dimensional

data sets. The challenges for the next few years lie in the further exploitation of the
effects beyond human vision and their use in industrial processes. Over and above
the intelligent processing of the sensor data, there is great potential for quality im-
provement and cost savings in linking the data to the entire IoT chain. This includes
the interference-free transmission of preprocessed and, if necessary, reduced data
to a secure data space and the evaluation of the data using artificial intelligence
methods (Sect. 15.4).
15.3 Learning for Biology
15.3.1 Digitization of Perception—Campus of the Senses
Jens-Uwe Garbas
The challenge in the digitization of human perception lies in the detection, de-
scription and transmission of the multiple mechanisms in sensory perception to the
machine world, i.e. the combination of physiological processes (e.g. smell and taste
while simultaneously perceiving a product by means of vision or hearing), stimulus
transfer and perception and interpretation in the brain. This requires a close inter-
play of the disciplines of pure physiological/psychological research, medical and
psychiatric research, taste physiology, multimodal sensing, reflex and facial ex-
pression analysis and analysis of body functions and behavior. Technical methods
and competencies include image and biosignal analysis, pattern recognition, ma-
chine learning, deep learning, big data analytics and the development of electronic
systems. There is a great need for the integration of chemical sensory perception
into the Internet of Senses, but it is also a particular challenge. The Campus of the
Senses, a newly formed joint project of Fraunhofer IIS, Fraunhofer IVV and the
Friedrich-Alexander-University, Erlangen-Nürnberg (FAU), will help to achieve
the required scientific and technological breakthrough here, by combining the nec-
essary interdisciplinary competencies and thereby also taking a leading position
in the international arena. Accelerating the digitization of chemical sensory per-
ception will thus complete the Internet of Senses on the campus, while giving
businesses access to a globally unique network of excellence, advanced knowledge
and improved access to technologies for innovation and new value propositions.
As outlined in Fig. 15.2, fields of action arise in at least three areas on the path
to the scientific, technical and economic success of the Internet of Senses:
Fundamental Research Applied Research Industry
Knowledge and method

development Realizaon
Percepon research, Applicaons, privacy,
psychology, neuroscience, regulaon, co-creaon,
medicine, biology, chemistry, workshops
AI methods
Technology
development
HW/SW development,
sensors, actuators,
databases, deep learning
Fig. 15.2 Fields of action towards creating the Internet of Senses. (Fraunhofer IIS/Jens-Uwe
Garbas)
Knowledge, method and competence development

Technology and prototype development
Realization.
These fields of action can only be tackled through an interdisciplinary, cross-do-

main and inter-institutional approach where joint infrastructure is built up. A cam-
pus is therefore being set up, which facilitates the cooperation of scientists from
Fraunhofer IVV, Fraunhofer IIS and the FAU.
There is an urgent need to catch up in the digitization particularly of the chem-
ical senses, which are extremely relevant for German companies—above all for
the food and beverage industry, the flavor and fragrance industry as well as manu-
facturers of bodycare and hygiene products, detergents and cleaning agents, car
manufacturers and the sports and leisure industry. The generic term “chemical
senses” encompasses perceptions evoked by volatile and non-volatile substances
in the form of odors and aromas, as well as taste sensations. Important knowledge
is lacking especially for the important food industry and the chemical industries
where the focus is on raw material and product manufacture for fragrances and fla-
vorings as well as consumer goods and articles of daily use. Basic human sensory
processes used in product perception as well as the effect of a product on humans

are not understood in detail and are only rudimentarily implemented in the applica-
tion. This is especially true in relation to human physiological chemosensors, i.e.
in the perception of smell, taste and trigeminal (burning, stinging) stimuli. There
is a lack of understanding of the basic scientific requirements, required technical
solutions and digital concepts. On the one hand, this understanding can facilitate or
even make chemical analysis and sensor technology possible in the first place, and
on the other hand it allows the detection, analysis and prediction of the (chemosen-
sory) perception and reactions triggered in humans based on suitable data sources.
Industry partners see an urgent need for action here. By closing this gap in science
and technology, the Campus of the Senses will be able to fulfill its mission to create
a digital representation of all human senses.
Primary goals of the Campus of the Senses are fundamental research into
chemosensory perception and its transfer to machine-based and digital concepts
as well as fundamental technology development for application in different fields.
The result is an interdisciplinary toolkit of methods for chemosensory stimula-
tion, for improved understanding of perception and the rapid and cost-effective
technical utilization of these findings (Fig. 15.3).
Another goal is to identify the basic challenges, opportunities and demands of
digitized sensory perception in the following areas of application: health, food and
agriculture, consumer goods, mobility and the environment, as well as work, en-
tertainment and leisure. Corporate partners help to define the specific development
requirements and initiate concrete product developments.
The Campus of the Senses will therefore produce new methods and technolo-
gies for the recording and interpretation of human senses and sensory perceptions,
especially in the field of chemical senses as well as for the evaluation of human
reactions to such sensory impressions. In this respect, the Campus of the Senses
offers tremendous opportunities both in the scientific field and in the implemen-
tation of the results. The greatest potential lies in multisensory integration. The
Campus of the Senses represents a holistic consideration of the origins, network-
ing and cross-influence of human sensory perceptions. The close interlocking of
pure and applied research and collaboration with industry result in practical tech-
nologies based on a strong scientific foundation.
About the Campus of the Senses Project

The machine recording and interpretation of human sensory perceptions, of the chemical
senses as well as visual, acoustic and haptic sensations, will play an important role in the
future. New microelectronic sensor and actuator solutions have to be developed for this pur-
pose, as well as new insights gained in perceptual research and brought together by the
algorithmic evaluation of machine learning. When individual sensory perceptions are linked
Fig. 15.3 The human sensory apparatus (center), methods toolkit (middle ring) and areas of
application (outer ring). (Fraunhofer IIS/Jens-Uwe Garbas)
for the purpose of a holistic sensation at a machine level too, based on the complex human
sensory system, the “Internet of Things” is augmented by the “human factor”.
Current cooperation partners:

Fraunhofer Institute for Integrated Circuits IIS | Fraunhofer Institute for Process Engineering
and Packaging IVV | Friedrich-Alexander-University Erlangen-Nuremberg FAU
Start: 12/01/2017
Duration: 6 years
Contact:
Bastian Dögl (bastian.doegl@iis.fraunhofer.de)
Dr. Tilman Sauerwald (tilman.sauerwald@ivv.fraunhofer.de)
15.3.2 Digitization of Self-Similar Biological Structures
Stefan Gerth and Joelle Claußen
The digitization of self-similar biological structures at Fraunhofer EZRT, a division

of Fraunhofer IIS, is an integral part of the establishment of sustainable, resource-
efficient plant cultivation systems. This is done through the phenotyping (quanti-
tative assessment of different characteristics) of plants in order to breed adapted
varieties [6]. The overall political objective is the sustainable use of existing bio-
logical resources in agriculture and all related sectors through innovative scientific
findings. The implementation of this requirement for sustainability is closely linked
to the “National Research Strategy Bioeconomy 2030” and the “Agenda 2030 for
Sustainable Development” of the United Nations.
Of the 17 development goals defined in Agenda 2030, the following goals are
especially driven by digital phenotyping:
Goal 2: “End hunger, achieve food security and improve nutrition and promote
sustainable agriculture”
Goal 8: “Promote ongoing, inclusive and sustainable economic growth, produc-
tive full employment and decent work conditions for all”
Goal 17: “Strengthen implementation means and revitalize the global partner-
ship for sustainable development”.
Breeding modern plant varieties adapted to climate change begins with the tar-
geted selection of parent plants. This is mostly accomplished by combining current
high-performance varieties with varieties or lines from regions suffering drought
stress or with known favorable properties relating to stress resistance. The resulting
offspring must then be tested over several years and in different environments for
their degree of adaptation, performance, processing quality and stress resistance.
A breeder of new grain varieties, for example, mostly still surveys these features
through visual assessment by individual experts followed, by laborious threshing of
individual lines with special combine harvesters. The subjective visual or destruc-
tive evaluation is not detailed enough when assessing complex features such as
biomass or yield potential, or it is impossible to assess features such as root growth

non-destructively. However, the non-destructive digitization of yield characteristics
and their application in the cultivation of stress-resistant plants is essential in order
to accelerate plant breeding in light of the global challenges of climate change.
Complex sensor systems, such as computed tomography, make it possible to
non-destructively visualize internal structures in 3D, such as root systems in the
surrounding soil [7]. The self-similarity of complex root systems means that clas-
sical image processing is only effective in rare cases [8].
The targeted use of cognitive sensor systems in phenotyping is a promising
approach for adequately addressing these challenges in the future. However, the
availability of annotated data sets is imperative for learning sensor systems. The
manual annotation of 3D CT datasets is often hardly possible. Also, due to the
complexity of the sensor system used, the public can only be included to a limited
extent. For this reason, a combination of simulation and “transfer learning” was
used at the Fraunhofer EZRT [9]. This chain is shown in Fig. 15.4 in simplified
form. The plant to be examined (a) has a complex root system. The underlying x-
ray physics can be realistically simulated by means of specialized software. Re-
alistic, virtual, yet completely annotated root systems can be created through the
simultaneous simulation of the surrounding substrate and the root structure typical
for the plant species (b). This approach makes it possible to generate input data for
Al-based sensor systems and to train them on plant-specific root systems. If real
measurements (c) are then carried out by means of the cognitive sensor systems
trained on this problem, visualization of the root system in question is possible
(d). Using classical image processing methods, digital features can now be calcu-
lated non-destructively based on real data, which can then be used directly for the
breeding of plant varieties with root systems that are regionally adapted.
Examples of plant characteristics of interest are root weight, root volume, root
angle and root length. In very dry growing areas, it can be beneficial for a plant
to develop a very deep and long root system in order to be able to utilize deep
groundwater reserves. The development of a deep root system, however, requires
resources that are then initially unavailable to the plant for the formation of fruits.
For this reason, it may make sense to rely on a very shallow, near-surface root
system which can use small amounts of water from dew formation or short rain
showers and thus make resources available for fruit formation much earlier on.
This example illustrates very clearly how strongly the regional peculiarities can in-
fluence underground root features, which are impossible or very difficult to achieve
through classical breeding properties.
Automated cognitive computed tomography systems are being developed at
Fraunhofer EZRT for this purpose, which are directly integrated into greenhouse
Fig. 15.4 Using the biological and self-similar root structures as a model (a), fully annotated
input data for the training of neural networks is generated through X-rays and substrate and
root simulation (b). Subsequent measurements of root systems in soil (c) can be virtually
separated from the soil by the previously trained neural network (d). Complex features may
be calculated based on binary data generated in this way and then used for the breeding of
adapted root systems. (Fraunhofer IIS)
conveyor belt systems. In this way, high-throughput measurements of root systems

and their subsequent statistical analysis have been made possible for the first time.
This example of cognitive sensor technology shows the great potential inherent
in the combination of intelligent sensors and AI methods for solving future chal-
lenges, especially in bioeconomy. The networking of intelligent sensors in a larger
context, for example across different test fields, requires the secure transmission
and storage of the data. A platform that can enable this is described in Sect. 15.4.
15.4 The Cognitive Internet
The biological systems described in the preceding sections, e.g. organisms with
swarm intelligence, are leading to developments of intelligently linked sensor sys-
tems in the technological transformation process, and are making new data-driven
value creation models and business models possible. This generates a major de-
mand for technology development in order to master the high degree of complexity.
The Fraunhofer-Gesellschaft is meeting the growing complexity of issues re-
lated to the Cognitive Internet of the future for industry with the Fraunhofer re-
search cluster “Cognitive Internet Technologies” (CIT). Fraunhofer is pursuing
new organizational paths using the format of a cluster of excellence, with the
goal of being able to offer a significantly broader range of expertise from a sin-
gle source. The research cluster is designed as an industrial collaboration platform
for the development of new key technologies. It efficiently accesses a wide range
of expertise from three CIT research centers:
IoT Communications: secure and reliable sensor data

Data Spaces: trustworthy data infrastructure and data sovereignty
Machine Learning: data, knowledge, simulation-driven learning for comprehen-
sible artificial intelligence.
Future industrial requirements will call for appropriate technology offerings,

including:
Secure, high-performance transmission technologies for networked and dis-

tributed sensors and systems
Optimized data generation through intelligent on-board implementation of ma-
chine learning algorithms
Use of expertise in informed machine learning
Secure data spaces as a basis for data-based business models.
The requirements listed above are to be clarified through the example of future
production systems. Current developments in the field of “Smart Factories” re-
veal a strong analogy with self-organizing biological systems. In conjunction with
the industrial Internet of Things, this approach opens up fundamentally new de-
sign possibilities for production processes e.g. processes that can be independently
controlled by the products to be manufactured. Likewise, this decentralized system
approach can achieve increased robustness against disturbance variables, analo-
gously to systems possessing swarm intelligence. Machine learning algorithms,
aided by the input of expert knowledge (Informed Machine Learning), will be used
to automatically optimize the overall production process in terms of continuous im-
provement and increased productivity. This opens up new ways of implementing
the principles of lean production with the corresponding economic benefits.
Technologically, self-organizing production has extremely high demands on
data rates, latency periods and the robustness of sensors and communication tech-
nology. Mobile products and machines, in contrast to conventional stationary pro-
duction lines, require appropriate wireless technologies and control systems. The
system complexity arising through the deployment and interaction of many dis-
tributed sensors will only remain manageable if intelligent sensor functions are
developed as well as sensor fusion with integrated AI functions. High volumes of
data must be processed with sovereignty and in a controllable manner in order to
be able to add value through new business models.
About the Fraunhofer Cluster of Excellence CIT

The Fraunhofer Cluster of Excellence “Cognitive Internet Technologies” (CIT) investigates
cognitive technologies for the industrial Internet. Researchers from 13 institutes and a range
of disciplines are developing key technologies along the value chain: from sensors, intelli-
gent learning processes in data processing up to the cloud. CIT empowers organizations to
future-proof their businesses by opening up market access with cognitive solutions and prod-
ucts. These highly intelligent solutions and products preserve the competitiveness of German
companies, strengthen their innovative power and safeguard digital sovereignty.
Through the special bundling of competencies in the CIT cluster, companies can be pro-
vided with customized cognitive solutions from the domain-specific expertise at every stage
of the transformation process.
Companies thus receive comprehensive support in managing complexity. This support
ranges from the analysis to the planning of a sustainable corporate strategy, to agile technol-
ogy development and testing in the CIT’s numerous innovation and application centers.
Current cooperation partners:

Fraunhofer Institute for Applied and Integrated Security AISEC | Intelligent Analysis and In-
formation Systems IAIS | Integrated Circuits IIS | Software and Systems Engineering ISST
| Applied Information Technology FIT | Telecommunications, Heinrich-Hertz-Institut HHI
| Experimental Software Engineering IESE | Material Flow and Logistics IML | Optronics,
System Technologies and Image Exploitation IOSB | Industrial Mathematics ITWM | Trans-
portation and Infrastructure Systems IVI | Machine Tools and Forming Technology IWU |
Algorithms and Scientific Computing SCAI
Start: 01/01/2018
Duration (1st approval phase): 3 years
Contact:
CIT Office
Dr. Uwe Wasmuth
uwe.wasmuth@cit.fraunhofer.de

1. Bonabeau E, Dorigo M, Théraulaz G, Theraulaz G (1999) Swarm intelligence: From nat-
ural to artificial systems (No. 1). Oxford University Press, Oxford
2. Haken H: Synergetik (1991) Die Lehre vom Zusammenwirken. Ullstein, Frankfurt, p
109 ff
3. Horváth G, Dezsö V (2004) Polarized light in animal vision: polarization patterns in na-
ture. Springer, Berlin
4. Ernst J (2015) Keine Scherben mehr – Polarisationskamera prüft Eigenspannung in
Glasprodukten. Qualität und Zuverlässigkeit (QZ) 5. www.qz-online.de/993378 Ac-
cessed: 22.09.2018
5. Schöberl M, Kasnakli K, Nowak A (2016) Measuring Beach Orientation in Carbon Fiber
Reinforced Plastics (CFRP) with Polarization. In: Proceedings of the World Conference
on Non-Destructive Testing (WCNDT), NDT.net, München, p 1–8
6. Fiorani F, Schurr U (2013) Future Scenarios for Plant Phenotyping. Annual review of
plant biology 64(1):267–291. https://doi.org/10.1146/annurev-arplant-050312-120137
7. Metzner R, Eggert A, Dusschoten D, Pflugfelder D, Gerth S, Schurr U, Uhlmann N,
Jahnke S (2015) Direct comparison of MRI and X-ray CT technologies for 3D imaging of
root systems in soil: Potential and challenges for root trait quantification. Plant methods
11:17. https://doi.org/10.1186/s13007-015-0060-z
8. Xu Z, Valdes C, Clarke J (2018) Existing and Potential Statistical and Computational
Approaches for the Analysis of 3D CT Images of Plant Roots. Agronomy 8(5):71. https://
doi.org/10.3390/agronomy8050071
9. Douarre C, Schielein R, Frindel C, Gerth S, Rousseau D (2018) Transfer Learning from
Synthetic Data Applied to Soil–Root Segmentation in X-Ray Tomography Images. Jour-
nal of Imaging 4(5):65. https://doi.org/10.3390/jimaging4050065
Prevention of Biofouling
Electrochemical and Anti-Adhesion Technologies
16
to Protect Ship Hulls and Membrane Modules
from Biofouling
Ralf B. Wehrspohn and Ulrike Hirsch
Summary
Biofouling is one of the key problems that many technical systems face. It
weighs down ships, clogs filtration modules for water treatment and causes
hygiene problems in clinical environments. At the Fraunhofer Institute for Mi-
crostructure of Materials and Systems IMWS, material science solutions have
been developed to prevent the adhesion of foulants. An electrically conductive
coating system applied to ships keeps the surfaces free of fouling effectively
for an extended time. Thin hydrophilic layers on the components of filtration
modules reduce the adsorption of microorganisms and can thus contribute to
increasing efficiency and energy saving.
Key Project Details: Antifouling Through Electrically Conductive Marine Paints

Objective: Development of polyurea-based electrically conductive paints to prevent biofoul-
ing on ship hulls
Cooperation partners: Schiffswerft Barth GmbH, FEW Chemicals GmbH, BLO Böhnstedt
GmbH and others
Research plan: Development of highly electrically conductive coatings for homogeneous
current density distribution, manufacture and characterization of electrochemically stable
coating layer systems, methods to prevent and destabilize biofilms, investigation of heavy
metal leaching from coating layer systems, testing of antifouling processes in seawater
Research budget: 1,340,000 C
Contact: Dr. Uwe Spohn
R. B. Wehrspohn () U. Hirsch

Fraunhofer Institute for Microstructure of Materials and Systems IMWS
Halle (Saale), Germany
https://doi.org/10.1007/978-3-662-59659-3_16
330 R. B. Wehrspohn and U. Hirsch
Key Project Details: Low-Fouling Membrane Modules for Reverse Osmosis

Objective: Innovative membrane spacers—chemical conditioning of spacer materials for re-
verse osmosis modules for water treatment
Cooperation partner: IAB Ionenaustauscher GmbH (LANXESS AG)
Research plan: Development of antifouling conditioning of membrane spacers for reverse
osmosis modules through the combination of plasma and wet coating to generate anti-adhe-
sive surface properties, application-oriented testing
Research budget: 470,000 C
Contact: Dr. Ulrike Hirsch
16.1 Introduction
Organisms are likely to settle on any surface that is damp and unsterile. This
settling is usually undesirable and leads to functional limitations of the material
or system. Such cases are referred to as biofouling. It occurs on practically all
technical surfaces in the marine sector, as well as in heat exchangers, in filtration
processes, on solar cells or medical devices.
Prior to the industrial revolution, the term biofouling almost exclusively re-
ferred to the accumulation of marine organisms to the hulls of ships. This leads
to increased flow resistance, increased energy and fuel consumption, a reduced
maximum speed, corrosion of the ship’s hull or even transport of harmful marine
organisms to other areas [8].
Written records on the treatment of ship keels date back to 500 BC [19]. It can
be assumed that the first attempts to prevent biofouling on boats and ships were
made even earlier than this. The early Phoenicians already used pitch to protect
their ships [22, 35]. The Greeks relied on tar and wax containing arsenic and sulfur
additives and from the 3rd century BC they clad their ships with lead, which was
attached with thick copper nails. Despite its strong tendency to corrode and espe-
cially the galvanic corrosive effect of lead on iron, for many centuries lead was
probably the most commonly used material for the protection of ship hulls [19].
The first truly successful and widely recognized antifouling surface was copper
cladding. The first authentic reports of this stem from the 18th century. In the long
term, however, this did not become established either, due to its highly corrosive
effect on the iron ships’ hulls. This problem could later be solved by dispersing
copper in the form of copper sulphate or copper oxide in electrically insulating
polymer matrices made of resins, tar or metal soaps [19]—the birth of antifouling
paints.
16 Prevention of Biofouling 331
Starting in the 1960s, the market for antifouling paints was revolutionized
through the development of petroleum-based plastics and, above all, the discovery
of tributyltin compounds (TBT) being highly effective against marine biofouling.
In the decades that followed, ship coatings with organotin compounds dominated
the market due to their almost universal antifouling effect, galvanic inertness and
optical transparency, with practically no competition [22]. However, after seri-
ous ecotoxic effects were proven, the use of the paints containing tributyltin was
banned throughout the EU in 2003 [10].
As negative environmental influences cannot be ruled out for any alternative, or-
ganic biocides [14, 31], there is an urgent need to develop biocide-free antifouling
systems—not only for ship hulls. Biofouling is also a major problem in filtration
processes such as reverse osmosis, through which seawater or brackish water may
be desalinated to produce drinking water. It leads to a heightened pressure drop, in-
creased energy expenditure, a reduced flow rate and therefore significantly reduces
the efficiency of the technology as well as the lifetime of the membrane modules
[3]. Combatting biofouling through the use of biocides is also not desirable here,
for reasons of food safety.
What is needed are antifouling systems that effectively prevent the adhesion of
fouling organisms by modifying the surface properties or by influencing the natu-
ral adhesion mechanisms. In doing so, the strategy for a material-based antifouling
modification depends on the application and the specific goal. Thus, it may be nec-
essary for some applications to suppress primary adhesion, i.e. the formation of
the conditioning film through the adsorption of organic and/or mineral foulants.
For other applications, however, only the adsorption of microorganisms or the for-
mation of a differentiated biofilm needs to be suppressed in order to operate the
relevant technical system without disruptions.
16.2 Formation and Control of Biofouling
Although the term “fouling” describes all interfering organic and inorganic de-
posits on technical surfaces, it is mostly the biological deposits (i.e. biofouling)
that are particularly problematic. It is very beneficial for microorganisms to live in
a stable biofilm network, because nutrients can be efficiently stored and exchanged
within the biofilm [11] and biofilms demonstrate a greatly increased tolerance to
antimicrobial reagents [7].
The colonization of surfaces is often described as a linear, successive process,
since the organisms accumulate essentially in order of size and thus form a multi-
layered composite [5, 32]. However, biofouling is an extremely dynamic process
Fig. 16.1 Phases of biofilm formation. (Fraunhofer IMWS, according to [12])
and the composition of the biofilm that is formed significantly depends on the
substrate, the geographical location, the season and the “survival of the fittest” [6].
In the filtration of saline water, e.g. by reverse osmosis, the water is already pre-
filtered, so here the “microfouling” mostly occurs through microorganisms such as
bacteria or unicellular algae. For marine applications, e.g. on ship hulls, “macro-
fouling” by mussels, barnacles, plants and multicellular algae is added to this. The
macroorganisms require an already formed microbiofilm of proteins, polysaccha-
rides and microorganisms in order to accumulate. Preventing the development of
this is thus also a promising strategy.
The initial biofilm formation takes place in three phases [5] (Fig. 16.1).
Induction Phase
Fouling on surfaces overflowed by water begins with the formation of a lam-
inar boundary layer in which the flow velocity in the direction of the surface
drops completely. The microorganisms arrive at this boundary layer via convection.
A conditioning film of various organic substances such as proteins, polysaccha-
rides and humic acids begins to form, which builds a foundation for the adhesion
of the microorganisms. This is initially still reversible, but becomes irreversible
after a short time due to the increasing interactions between cell surfaces and the
substrate.
Accumulation or Growth Phase

This phase comprises the proliferation of microorganisms, the production of ex-
tracellular polymeric substances (EPS) and the development of a differentiated
biofilm. The bacteria react to the changed environmental conditions by adapting
their metabolism and adopt a biofilm-specific phenotype. The EPS produced en-
sure the physical cohesion and adhesion of the biofilm on the surface and act as
a medium for the exchange of nutrients and signals. The composition of the EPS
2 < pH < 12
a
Fig. 16.2 Antifouling concepts: a Chemical antifouling through toxic agents; b Steric an-
tifouling using the example of a hydrophilic polymer coating; c Electrochemical antifouling
through applying a voltage to conductive surfaces. (Fraunhofer IMWS)
matrix thereby strongly depends on the microorganisms involved and the environ-
mental conditions.
Plateau or Subsistence Phase

From a certain thickness of the biofilm, an equilibrium is established between
growth and redetachment. Shear forces acting on the biofilm can cause large por-
tions of the biofilm to be detached (so-called sloughing). Some bacteria near the
surface of the film also actively leave the biofilm network to colonize new surfaces
as swarming cells.
The material-based suppression of biofilm formation—i.e. the preparation of
the surface to prevent colonization by microorganisms—can be achieved by differ-
ent strategies. Besides equipping the surface with biocidal agents, which are not
desirable for the targeted applications in the marine sector and in drinking water
treatment, the physicochemical or even electrochemical properties of the material
surface can be adjusted. It is important to understand the surface parameters that
influence biofouling, such as the interfacial energy (hydrophilicity or hydropho-
bicity), surface roughness and surface charge.
The following material-based antifouling concepts arise as a result (Fig. 16.2).
Chemical Antifouling
Incorporation of or coating with inorganic or organic toxic materials (Fig. 16.2a).
As a rule, a very good antifouling effect is achieved in this way and it is highly
effective against harmful organisms. However, the concerns and risks with regard
to environmental protection, food safety and the long-term effects on other organ-
isms mean that non-biocidal technologies are increasingly being used. Coatings
and paints for ships containing copper or organic biocides will in future rely on
the lowest possible emissions of toxic agents, so that these only act through direct
contact. The long-term perspective is that marine biocides should increasingly be
replaced with anti-adhesive technologies such as nanostructured and/or self-regen-
erating polymer coatings [13].
Steric Antifouling
Modification or coating of the surfaces to be protected through anti-adhesive prop-
erties. This includes the deposition of highly hydrophobic or highly hydrophilic
layers as well as the adjustment of certain surface structures (e.g. lotus effect)
(Fig. 16.2b). For applications in an aqueous medium, of the variants mentioned,
the deposition of a thin, hydrophilic or hygroscopic polymer layer is particularly
useful. This is able to absorb water and thus form a hydrogel, the surface tension
of which is very similar to that of the surrounding water [4]. As a result of this and
owing to the steric effects of the polymer chains, adhesion of molecules such as
proteins and polysaccharides as well as microorganisms is thermodynamically un-
favorable [1, 20]. Another effect that contributes to steric repulsion is the osmotic
pressure between the overlapping polymer layers [34]. If the polymer film con-
tains zwitterionic head groups, the neutral surface charge additionally suppresses
electrostatic interactions that could lead to attraction and adhesion. Since the an-
tifouling effect is based on a nonspecific suppression of the interactions required
for adhesion, it is universally effective, provided that the surface is in contact with
an aqueous medium.
Electrochemical Antifouling
Application of a conductive coating, which serves as an electrode, and electrolysis
of the seawater by applying a voltage between two electrodes (Fig. 16.2c). As
a result of the oxidation of water taking place at the anode and the chloride ions
contained in the sea water, as well as the reduction of water and dissolved oxygen
occurring at the cathode, the pH is either locally reduced (oxidation at the anode)
or increased (reduction at the cathode). The pH is lowered even further due to
the formation of hypochlorous acid (HClO). The variable pH conditions between
pH 2 and pH 12 and the resulting reactive species make colonization of the surface
'HYHORSPHQWSURJUHVV
5JQTVVGTOVGUV .QPIVGTOVGUV (KGNFVGUV

NCDQTCVQT[ NCDQTCVQT[ CRRNKECVKQPUKVG
• Small sample size or • Small sample size • Sample dimensions and
material model • Application of optimized application site correspond
• Variation of modification or modification or coating to the final product
coating parameters parameters (prototype)
• Testing of many samples in • Testing of a few samples • Scale up of modification
a short time under static with adapted test duration tools or outsourced
conditions under dynamic conditions manufacture
• Evaluation of the overall
Goal: Parameter optimization Goal: Application-oriented system
simulation
Goal: Marketability and
product launch
Fig. 16.3 Evaluation methods for modified antifouling materials according to development
progress. (Fraunhofer IMWS)
very unattractive to bacteria [27, 29]. In addition, the electric fields themselves also
have an antifouling effect [33]. The combination and controllability of the different
effects make this antifouling system universally effective too. The prerequisites are
a conductive surface (intrinsic or due to a coating) and the installation or operation
of a voltage source and its control.
The evaluation of the antifouling or antimicrobial performance of surface-modi-
fied materials is carried out in line with the progress in development on a laboratory
or application scale (Fig. 16.3).
Short-term microbiological tests are an excellent tool to quickly evaluate the
antifouling effect of surfaces, to compare different coatings or coating parameters
and to optimize the modification process. The method allows for the rapid test-
ing of a larger number of samples. As a rule, these samples are small sections of
the larger product, as is the case for films or membranes. Products with complex
geometries, for example implants or nets, are only suitable for material analysis
to a limited extent. Material models can also be used here, whereby the material
is processed into suitable sample forms. The incubation is usually discontinuous,
i.e. with nutrients supplied once off at the beginning of the culture period and is
conducted with defined model bacteria. After a maximum of 24 h, the biomass that
has grown on the samples is quantified; either directly by microscopy or indirectly

by peeling off, streaking and counting the microorganisms.
For the application-related testing of selected samples that have been modified
with optimized parameters, continuous long-term tests are available that simulate
field conditions under laboratory conditions. As a rule, a suitable test bench is
required for this, which allows for operation-related and dynamic adjustment of
environmental parameters such as pressure, temperature, irradiation, surrounding
flow or nutrient supply. Untreated tap water or seawater can be used, for example,
instead of defined cultures of model bacteria. The test is carried out over a period of
weeks or months. For this reason and because the tests are conducted significantly
differently, the results can differ from those of the standard, short-term microbio-
logical tests. The evaluation can in turn be done either visually or indirectly, for
example by determining the pressure drop in the system, which allows conclusions
to be drawn about the biofilm formation.
If the long-term tests are also satisfactory with regard to the antifouling effect of
the samples, field tests can be carried out at the application site. As a rule, not just
a sample section is tested here, but ideally a prototype with dimensions comparable
to those of the end product. Manufacturing the prototype may involve scaling up
the modification tools or require outsourced production. The aim of these tests,
which can be designed entirely differently depending on the product, is to test the
marketability of the product and, above all, its market introduction.
The criteria according to which an antifouling or antibacterial modification or
a corresponding product is considered to be effective can vary greatly depending on
the application. Thus, a percentage reduction in biofilm growth within a given time
can be set as a criterion. An increase in the length of cleaning intervals, reduced
energy expenditure and the prolonged life of components can also represent deci-
sive criteria. As a rule, not just individual components benefit from the reduction
in biofouling, but entire systems and processes.
Other important evaluation criteria for products modified to have an antifouling
effect are, in addition to their effectiveness in preventing fouling, their stability
and sustainability, versatility of application and approval under current guidelines
and laws. Research into related markets and trends is therefore required before
designing and introducing an appropriate new product.
16.3 Market Development
Biofouling is a problem that affects a variety of systems and thus also impacts
different markets. There is high demand for antifouling paints not only in the ma-
rine sector, but also in the oil and gas sector, as well as in photovoltaics and the
construction industry. The global market for antifouling paints exceeded US$5.68
billion in 2016 and is projected to reach US$9.21 billion in 2021 [21, 37]. The
compound annual growth rate (CAGR) for the period from 2016 to 2021 is indi-
cated to be over 8.4% [21, 37]. On account of the increasingly strict regulations
with regard to environmental protection and the fact that they are constantly under-
going further technical development, biocide-free products and formulations using
natural raw materials will experience significant growth and are expected to domi-
nate the market in future. Special emphasis will be placed on making the solutions
sustainable and as flexible as possible in their application [13]. The highest demand
worldwide comes from the Asia Pacific region [30].
In contrast to commercially available antifouling paints, which represent a stan-
dalone market, a very large proportion of antifouling technologies can be seen
as a feature of other products, which can significantly improve devices and sys-
tems from the fields of desalination, biomedical engineering and energy production
and make them more efficient and sustainable. Demand for energy-efficient tech-
nologies is particularly high in the desalination segment, as rapidly increasing
industrialization and the growing world population are in contrast with the wor-
rying decline in and increasing pollution of natural freshwater resources [18]. The
water desalination market had a volume of US$13.31 billion in 2016 and is pro-
jected to reach US$26.81 billion in 2025 [18]. The average annual growth rate
indicated is over 8% on account of increasing water scarcity in many regions of
the world, [15, 23]. The biggest growth engine is the demand for drinking water in
the Middle East and Africa [18] and in the Asia-Pacific region [23, 24]. Thereby,
the market is clearly dominated by the market segment of reverse osmosis, which
amounted to more than 58% in 2016 [15], with a strong upward trend.
The greatest challenge of membrane-based reverse osmosis is the high energy
demand to set the required high pressure of 15 to 80 bar depending on the salinity
of the water, in order to overcome the natural osmosis process. Energy expenditure
accounts for about half of the total cost of the process in the case of seawater
desalination [9]. As a result, there is great demand for energy savings through
process optimization and prevention of the most serious problem in membrane
separation processes: biofouling.
In the following sections, results are presented from projects conducted by the
Fraunhofer Institute for Microstructure of Materials and Systems IMWS, in which
innovative antifouling technologies in the form of electrically conductive marine
paints and non-stick coatings for components of reverse osmosis modules are being
developed.
16.4 Example: Antifouling Through Electrically Conductive

Marine Paints
Electrically conductive and seawater-resistant marine paints are a promising al-

ternative to the widespread antifouling coatings with toxic activity. Such systems
have only been applied in technical operations in a few experiments up to now, but
they function without biocides, act against both micro and macroorganisms and
can be controlled in an intelligent manner via the electrolysis current.
The antifouling mechanism is based on the electrolysis of seawater, which is
initiated when a voltage is applied between two electrodes. The electrical current
that flows in this process is a consequence of the oxidation of the water and chlo-
ride ions contained in the seawater at the anode, as well as the reduction of water
and dissolved oxygen taking place at the cathode (Eqs. 16.1 to 16.4). The anodic
oxidation of water results in a surplus of protons and lowers the pH, while the pro-
tons reduced at the cathode cause the pH in the local environment of the electrode
to rise (Fig. 16.4). Chlorine then reacts with water as a consequence of the oxida-
tion to form hypochlorous acid, which causes a further decrease in pH at the anode
(Eq. 16.5).
2HC C 2e • 2H2 (16.1)
O2 C 2H2 O C 4e • 4OH (16.2)
C
2H2 O • 4H C O2 C 4e (16.3)
2Cl • Cl2 C 2e (16.4)
Cl2 C H2 O • OCl C Cl C 2HC (16.5)
For maritime electrochemical antifouling, the electrodes required for the elec-
trochemical reactions must be insulated from each other as large area segments
and isolated or mounted on the underwater surface, for example on ship hulls
(Fig. 16.4). The disinfecting action of such a system is based primarily on the
pH-stress that is generated [29] as well as on the hypochlorous acid and hypochlo-
rite ions that are formed [27]. This combination produces a universal antifouling
Fig. 16.4 Schematic representation of the sequential subdivision of a ship’s hull into cathode
and anode areas. (Fraunhofer IMWS)
effect, which acts against marine bacteria, algae, bryozoa and barnacles as well
as mussels. The effect is also permanently retained in waters of lower and higher
salinity such as brackish water and the Baltic Sea.
Although the electrochemical antifouling effect has long been known, its ap-
plication has not yet been successfully established. The most important reason for
this is that the conductive paints required were not available in sufficient quanti-
ties and could not be applied with the usual application methods for marine paints.
The objective was therefore set at the Fraunhofer Institute for Microstructure of
Materials and Systems IMWS to develop sufficiently electrochemically stable and
conductive outer coatings. The paint layers must also have a low surface roughness
and the interfacial energy should be in the range of 20 to 30 mN/m. It was crucial
for the functionality and long-term stability of the electrochemical antifouling sys-
tem to obtain as homogeneous a current density as possible over large areas. In
addition, the coating was to be implemented using coating methods suitable for
dockyard applications such as spray coating or roll coating.
The path to achieving this has led to the development of multi-layer paint
systems with electrical conductivities that are coordinated in such a way that ho-
mogeneous current density distributions can be achieved. The first step demanded
by the long operating times required for technical operation, is the choice and pro-
duction of paint systems that are stable in sea water i.e. that are stable with respect
to hydrolysis and only swell minimally. The electrical conductivity of such paint
systems must be optimally and stably adjustable by addition of electrically con-
ductive and corrosion-resistant particles.
More than 200 coatings were developed within the scope of an intensive
paint screening and their mechanical and chemical stability with regard to tensile
strength, elongation, abrasion resistance and water permeability were determined
using selected examples. Three polymer coatings based on acylate, epoxide and
polyurea were selected as pertinent for the electrochemical prevention of fouling.
The acylate paint is inexpensive and easy to handle. It is therefore very well suited
as an inner coating. The outer coating is exposed to waves and the electrochemical
splitting of water, which is why a very durable polyurea coating is applied here.
However, because of its fast reaction times of less than ten minutes, it must be
processed using high-pressure spraying equipment. The positive qualities of the
epoxy paint that was developed make it applicable in all relevant areas, both as an
inner and an outer coating.
In order to adjust the conductivity over large areas, metal particles (silver-coated
copper, nickel, iron and glass particles) were dispersed within the already defined
polymer matrices. To develop an outer coating that is stable in the long-term, but
less electrically conductive, carbon-based additives such as graphite flakes, carbon
fiber fragments and carbon black were dispersed, since they are subject to less
anodic dissolution compared to the metals used, even at high voltages.
In this way, an outer coating that is stable in seawater could be developed in
combination with a highly conductive inner coating. The latter has a specific con-
ductivity of more than 400 Scm1 for a layer thickness of 50 to 200 µm. The outer
coating then only has to achieve a specific electrical conductivity of at least 0.02
Scm1 with a layer thickness of 400 to 1000 µm. The newly developed coating
system, including the insulating primer, can be applied to marine steel by spray
coating. The composition of the multi-layer coating system is shown in Fig. 16.5.
The most promising coating systems were tested under stable continuous opera-
tion for more than 16 months in calm, Baltic Sea water in the Kühlungsborn harbor
where a very high risk of fouling is present. In this process, different electrolytic
current-time regimes with respect to antifouling efficacy and destabilization of al-
ready existing biofilms were applied to test areas on a pilot scale (25 cm × 65 cm)
in two field tests and examined for the duration of the test period. It was shown
that with a current density of 0.2 mAcm2 the marine growth could be completely
suppressed over the entire surface.
Fig. 16.5 Schematic struc-

ture of a multi-layer coating
system for electrochemi-
cal antifouling, consisting
of substrate (1a—original
surface, 1b—roughened
surface), electrically insu-
lating primer (2), highly
conductive inner coating,
(3) seawater-stable outer
coating (4). (Fraunhofer
IMWS)
A current-time progression with switching times in the minute range proved to

be the most effective against fouling.
The best result was shown by the polyurea outer coating, where the current-
carrying test plates could be kept almost completely free of fouling over the entire
test period of 16 months. However, identical test plates, which were exposed to the
fouling conditions without current applied, demonstrated growth of mussels and
barnacles several centimeters thick (Fig. 16.6).
Fig. 16.6 Progression of biofouling on steel plates with an intermittent current-carrying

polymer composite coating layer (left) and two currentless reference coating layers (right).
a) start of the experiment; b) after 35 days; c) after 237 days; d) after 514 days. (Fraunhofer
IMWS)
The overgrowth after 16 months seen in Fig. 16.6 (left) comes from the un-
coated back of the steel plates and does not adhere to the paint surface. This
underlines the need for good antifouling coatings and shows that the antifouling
effect of the current-carrying test plates is based on the electrochemical splitting
of sea water. The voltage curves recorded during the field test show that an equi-
librium state sets in after two weeks. Thereafter electrochemical antifouling takes
place at relatively constant voltages and stable conditions.
If the current intervals and current densities are adjusted to the actual ship op-
eration, this can be extrapolated to a continuous operation time of five years and
more. In so doing, the electrochemical antifouling system would be able to com-
pete with conventional concepts already established on the market. A successful
experiment on a motorboat has demonstrated the operability of the layered coat-
ing, electrode-based antifouling system. The operational stability and economic
viability of the electrochemical antifouling system must be further improved in or-
der to be able to equip even larger ships. The paint application in particular must
be optimized before commercialization.
16.5 Example: Low-Fouling Membrane Modules for Reverse

Osmosis
The project presented here contributes to increasing the efficiency of reverse os-
mosis, which is applied in the desalination of water. Antifouling coatings are being
developed for components of these modules in close cooperation with IAB Ione-
naustauscher GmbH (LANXESS AG).
In addition to the actual separation membranes, the membrane spacers in the
flow channels are especially affected by biofilm formation (Fig. 16.7). The modifi-
cation or coating of the membrane spacers instead of the membranes offers certain
advantages: a membrane spacer made of, for example, polyethylene/polypropylene
is less sensitive to the conditions of coating or modification than the highly sensi-
tive membrane, and the separation properties of the membrane and the module are
also not directly influenced.
There are several approaches to reducing biofouling in reverse osmosis modules
that make use of the properties of the membrane spacer. State of the art modules
have particularly thick membrane spacers compared to the standard. As a result, the
affinity for particle fouling in particular can be reduced. Such membrane modules
are currently being developed and sold by Lanxess, Hydranautics/Nitto Denko,
Toray and DOW, among others. Equipping feedspacers with biocides is another
Fig. 16.7 Diagram of a membrane module. Biofouling on the membrane spacers and mem-
branes causes an increasing loss of pressure and the flow rate drops. (Fraunhofer IMWS)
option. Spacers equipped with triclosan are available, for example, from Hydranau-
tics/Nitto Denko [28]. Their effect is however controversial [2]. Promising results
were also obtained with zinc oxide or silver particles [25, 26]. However, metallic
particles have limited effectiveness, since the release of metallic cations decreases
over time. Metal toxicity therefore does not make much sense in the long term
[17]. In addition, dead adherent biomass also forms a nutrient bed for microor-
ganisms. The integration of toxic components is moreover questionable in as far
as contamination of the permeate—which subsequently may be prepared for use,
among other things, as drinking water—due to tiny imperfections in the filtration
membrane cannot be entirely ruled out.
In contrast to this, the thin, hydrophilic, neutrally charged polymer coatings
developed in the project presented here have a steric antifouling effect (Sect. 16.2).
The coatings are applied to commercially available membrane spacers made
of a blend of high-density polyethylene (HDPE, 80%) and polypropylene (PP,
20%) and, as a reference, to extruded films of 100% HDPE, 100% PP and HDPE
(80%)/PP (20%). The surfaces are first treated by means of dielectric barrier dis-
charge under atmospheric conditions, thereby producing functional groups such
as hydroxyl or peroxide groups. These functional groups generated by plasma
chemical reactions form the points of attachment for the subsequent grafting of the
hydrophilic, neutrally charged polymer, poly-[2-(methacryloyloxy)ethyl]dimethyl-
(3-sulfopropyl)ammonium hydroxide (poly-SBMA), that is expected to have an
antifouling effect. In order to achieve optimal coating, process parameters such
as temperature, monomer concentration or treatment time are systematically var-
a b
Fig. 16.8 Atomic force micrographs of the surfaces of an uncoated membrane spacer (a)
and a poly-SBMA coated membrane spacer (b). (Fraunhofer IMWS)
ied during dip coating and the effect on the adhesion behavior of the poly-SMBA
layer is systematically investigated as well as the antifouling activity.
Innovative, plasma-based approaches to layer deposition are also being pursued
in addition to conventional dip coating processes. The deposition of poly-SBMA is
therefore also carried out via plasma-assisted aerosol deposition as well as through
the process of plasma polymerization at low pressure.
Diverse material science methods such as X-ray photoelectron spectroscopy
(XPS), atomic force microscopy (AFM), quartz crystal microbalances (QCM) and
contact angle measurements are used to test the layer formation and for character-
ization of the poly-SBMA. Fig. 16.8 shows the surface morphology of membrane
spacers before and after poly-SBMA coating (atomic force microscopy). The un-
treated and uncoated surface is primarily characterized by lamellar structures typ-
ical of HDPE and PP. Spherulite-like polymer structures can be seen after coating
with poly-SBMA, in addition to smooth, amorphous layer regions.
The surface energies of the samples, which are determined by means of con-
tact angle measurements, increase up to about 55 mN/m in the presence of poly-
SBMA (uncoated: ca. 32 mN/m, water: ca. 72 mN/m [16]). The assumption is
made that the surface energy of the poly-SBMA layers will increase further in
a wet or swollen state. A swelling factor of about 1.3 to 1.7 was determined for
the poly-SBMA layers using quartz crystal microbalancing. The swelling factor
is calculated from the ratio of the swollen mass to the dry mass of the layers and
a b
Fig. 16.9 a biofilms on uncoated and poly-SBMA-coated spacer materials. The scale of all
images is identical. b degree of biofilm coverage determined by gray scale analysis (Fraun-
hofer IMWS)
depends inter alia on the crosslinking density of the coating, which can be varied
depending on deposition conditions and the use of initiators.
The antifouling properties of the modified samples are investigated both in dis-
continuous short-term tests (batch) as well as in a continuously operated test bench
with subsequent quantification of the biomass found on the samples.
The discontinuous short-term tests of the coated materials provide a quick as-
sessment of the antifouling effect. For purposes of this testing, the samples are
incubated with Pseudomonas species in a culture medium with a composition
based on the site of application. The biofilms that have formed are subsequently vi-
sualized using confocal laser scanning microscopy (CLSM). In Fig. 16.9 the degree
of biofilm coverage is shown on the reference films (HDPE, PP and HDPE/PP) and
after poly-SBMA coating. The SBMA coating significantly reduces biofilm forma-
tion: by more than 80% for the HDPE/PP blend and HDPE and over 75% for PP.
A continuous test bench was set up in order to test the antifouling effect of the
poly-SBMA-modified membrane spacers in an application-related way (membrane
fouling simulator MFS, Fig. 16.10a). This is specially designed to allow suitable
test water to bathe reverse osmosis membranes and membrane spacers in long-term
operation in a test cell (marked in red). This takes place under continuous moni-
toring of the pressure, pressure drop, temperature and flow rate. Biofilm formation
can be accelerated by adding nutrients. The most important parameter at this ap-
plication-oriented measuring station is the loss of pressure due to the deposition of
biomass on the spacers or membranes in the test cell, which increases in the course
of operation (Fig. 16.10b).
a b
Fig. 16.10 a Membrane fouling simulator (MFS), the test cell is marked. b variation in
pressure drop in the test cell as a function of the membrane spacer used. (Fraunhofer IMWS)
An important prerequisite for the industrial application of the technology and

a convincing improvement of the reverse osmosis process is adequate stability of
the antifouling coatings, which, however, strongly depends on the properties of the
initial material, i.e. the surface roughness as well as the cleanliness of the mem-
brane spacers. There is still a need for optimization here, especially with regard to
the thickness and the adjustment of the cross-linking between the coatings. While
a layer thickness of 20 nm is theoretically sufficient to suppress the physical-chem-
ical interactions necessary for the adhesion of microorganisms [36], a substantially
higher layer thickness may be required in practice in order to compensate for
roughness of the substrate.
At the time of writing, the project was still in progress. For the remainder of
the project, the focus will be on transferring the coating technologies to a continu-
ous roll coating system, in cooperation with the industrial partner, with the aim of
incorporating them into the membrane modules’ manufacturing process. The suc-
cessful equipment of membrane spacers with antifouling properties is expected to
significantly reduce the pressure drop in the membrane module, which will allow
lower pressures to be applied, thereby significantly reducing energy consumption.
The modules also need to be cleaned less frequently and therefore have a signifi-
cantly longer service life. The desalination process thus becomes more efficient
overall and at the same time less expensive.
16.6 Outlook
Non-toxic, alternative antifouling solutions are becoming increasingly important.

Over and above their environmental friendliness and food compatibility, the most
important aspects of their development and application are their sustainability and
energy efficiency.
The antifouling coating technologies presented in this chapter are designed for
future industrial application. The electrochemical antifouling paint system for ship
hulls is extremely effective, but requires continuous operation, especially during
rest periods of ships in the harbor, in narrow channels or in case of low speed
(a few knots). In order to optimize the coating system even further, it could in
future be combined with or extended to include fouling-release surfaces, so that
adhering substances can be removed relatively easily. The long-term stability of
the coatings must also be improved.
The spacer materials modified to have an antifouling effect in membrane mod-
ules, which lead to demonstrably less biofouling could reduce maintenance cycles,
minimize material damage and excessive material wear, and enable significantly
more energy-efficient and cost-effective operation of the membrane modules over-
all. Since the antiadhesive mode of action of the coatings that have been developed
is non-specific, it can also be transferred to other materials that are in contact with
an aqueous medium. However, here too the stability of the coatings must be opti-
mized.
The coating methods are potentially also applicable to other polymer-based ma-
terials, for example in medical technology. Other markets with a high demand for
antifouling solutions are, for example, offshore technologies such as wind power
plants, food technology and the medical and hospital sectors.

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Urban Agriculture
The Future of Agriculture – Local, High-Quality
17
and Value-Adding
Eckhard Weidner, Görge Deerberg, and Volkmar Keuter
Summary
At the Fraunhofer Institute for Environmental, Safety and Energy Technology

UMSICHT research into new forms of agriculture has been ongoing for several
years. At UMSICHT, the concepts of “Indoor Farming”, “Urban Agriculture”
and “Smart Farming” encompass not only the development of technology build-
ing blocks for illumination new plant breeding systems, new materials and
non-destructive analysis or nutrient recovery, but also include the adaptation of
plants to cultivation without soil. This chapter presents examples of approaches
that can shape a future, transformation-oriented agricultural economy.
Key Project Details

Project title: inFARMING®
Goal: Development of technology building blocks for the efficient operation of building
integrated agricultural systems
Cooperation partners: Fraunhofer, commercial enterprises, higher education institutions and
universities
Research plan, funding volume: Continuous development of technology building blocks
since 2011
Contact: Volkmar Keuter (Fraunhofer UMSICHT)
E. Weidner () G. Deerberg V. Keuter

Fraunhofer Institute for Environmental, Safety and Energy Technology UMSICHT
Oberhausen, Germany

https://doi.org/10.1007/978-3-662-59659-3_17
352 E. Weidner et al.
17.1 Introduction
Our lives are changing rapidly. Germany will face considerable challenges in the
coming decades: global population growth, demographic change, energy transi-
tion, digitization, urbanization, an increase in single households and more. At the
same time, and as a result of this, there is a growing need for high-quality, fresh
active ingredients and medicinal products, which are often still obtained from open
cultivation and wild plants today, sometimes under precarious conditions. The need
to ensure that the population is supplied with high-quality, fresh foodstuffs and
active ingredients while at the same time reducing environmental pollution and
conserving resources puts additional pressure on agricultural production.
Most experts agree that the “green revolution” in agriculture is gradually reach-
ing its limits. This, in general, refers to the development of modern high-perfor-
mance and/or high-yield agricultural crops started in the 1960s and successfully
spread into developing countries. In this way, the yields of global agricultural sys-
tems could be dramatically increased. However, the consequences, for example for
the environment and also for the producers, are not only seen as positive. Water
pollution and over-fertilization as well as pesticide and herbicide residues found
in agricultural products are increasingly causing problems. Several current studies
suggest that the challenges faced by developed countries will not be maximizing
yields, but rather reducing environmental impact while maintaining similar yields
and increasing quality. Security of supply can therefore only be achieved through
sustainable agricultural production. Various approaches are in discussion, ranging
from integrated solutions close to the consumer, the use of intelligent nutrient man-
agement solutions, to pure organic farming or agricultural communities.
17.2 New Approaches to Knowledge-Based Urban

Agriculture
More and more findings from research into plant physiology show that in addition
to the central parameters addressed today, such as the availability of water, nutri-
ents and light, many other factors can be used to influence the growth of plants and
control and optimize the formation of nutritionally important constituents or active
ingredients.
In this way, stress factors such as a lack of water can support the accumulation
of certain active ingredients in plants. Biomass accumulation or active ingredi-
ent concentration may be significantly increased through controlled illumination,
17 Urban Agriculture 353
Fig. 17.1 Comparison of the concentrations of secondary plant constituents by example of

rutin in lovage exposed to illumination with and without an infrared component. (IR compo-
nent) as a function of the illumination (PPFD: photosynthetically active photon flux density).
Results from the project LEDs-grow-quality® . (Fraunhofer UMSICHT)
which is precisely selected with regard to light color and intensity intervals (see
Sect. 17.5 and Fig. 17.1).
Since such strategies are not feasible in the open field, excellent synergies
are possible here with closed urban production systems (Controlled Environment
Agriculture, CEA) such as the inFarming® approach, provided that the neces-
sary knowledge of the principles of action and corresponding technology building
blocks is available.
17.3 Local Production Sites
By 2050, it is expected that 66% of the world’s population will be living in cities.
Recent findings suggest that urban and peri-urban agricultural-based economies
will play a key role in providing a stable food supply for the city of the future [1,
2, 3]. These forms of agricultural-based economies are part of a so-called “City
Region Food System” (CRFS), a planning approach of the Food and Agriculture
Organization of the United Nations (FAO), which is seen as a core issue in the
implementation of Agenda 2030 and the New Urban Agenda (NUA) [9]. The pro-
duction of higher value-adding active ingredients and medicinal plants are to be
integrated here, so that the growing demand for these can also be satisfied.
A key element of the potential sustainability of CRFS is the production of prod-
ucts in the place where they are consumed—in the city. Traditional peri-urban
agriculture in metropolitan areas is faced with difficult conditions in view of in-
creasingly limited availability of cultivation area and legal restrictions. The role
played by the local supply of food and active ingredients is nevertheless becoming
increasingly important due to global trends, such as the growing world population
and increasing soil sealing. A sustainable supply of agricultural products will need
to be ensured through systemic approaches in the future.
So-called “Controlled Environment Agriculture” (CEA) processes, also known
as indoor farms, combined with soil-free cultivation methods such as hydroponics,
make resource-efficient and space-saving plant cultivation possible in the place
where it is needed. Hydroponic culture systems allow for precise control of the
water and nutrient supply of horticultural crops. The use of inert substrates pre-
vents sorption of dissolved ions, and by recycling the nutrient solution uncontrolled
movement of substances into the substrate is avoided and water is saved. The
first systems of this kind are already integrating agricultural production into ur-
ban areas, some using vertical approaches and some also partially integrated into
buildings [4, 5].
In Oberhausen a unique research and demonstration center for building-inte-
grated agriculture is currently being created (Fig. 17.2). Oberhausener Gebäudem-
anagement GmbH (OGM) is following the plans of Berlin-based architects, Kuehn
Malvezzi, to build the “Altmarktgarten” at the Altmarkt in Oberhausen. This is
based on the inFARMING® concept developed by Fraunhofer UMSICHT, which
essentially links production systems and building infrastructures. For example,
gray and rainwater streams are supposed to be redirected and used as a water
source for local crop production, and CO2 from the job center in the building be-
low will later be used to fuel plant growth. The production area is complemented
by a research area, with a total area of more than 1000 m2 . It will be possible
to research future urban agriculture systems in the “Altmarktgarten” and develop
technical systems within a metropolitan context. The concept was so convinc-
ing that the German Federal Ministry for the Environment, Nature Conservation,
Building and Nuclear Safety (now: Federal Ministry of the Interior, Building and
Community, BMI) decided to promote the project as one of the lighthouse projects
of the federal program “National Projects of Urban Development” in 2015. The
“Altmarktgarten” is in operation since September 2019.
Fig. 17.2 The “Altmarktgarten” project in Oberhausen. (Kuehn Malvezzi, Visualization:

Reindeer Renderings, Jonas Marx)
Plant production systems which can be integrated spatially into intensive sys-
tems directly where they are needed, irrespective of location and type of plant, can
also be of great interest to other sectors of the economy. Wherever there is year-
round need for consistently high plant quality, regardless of the environmental con-
ditions, CEA processes will be applied more and more in the future.
17.4 Circular Farming
Agriculture consumes around 104 million metric tons of nitrogen, 46 million met-
ric tons of phosphate (P2 O5 ) and 33 million metric tons of potassium (K2 O) world-
wide every year [8]. Due to the rapidly growing world population’s need for fresh
food and other agricultural products an average annual increase of 2 to 3% per
component is currently expected.
Many scientists assume that so-called “peak phosphorus” will be reached in
about 20 years. Other sources say that this point of maximum phosphorus pro-
duction will be reached later, but still within this century [6]. Europe is not self-
sufficient in its supply of phosphorus and nitrogen, but relies on imports from
Fig. 17.3 Process diagram of the demonstration plant ROOF WATER-FARM (MBR: Mem-
brane bioreactor, MBBR: moving bed biofilm reactor). (Fraunhofer UMSICHT)
Morocco, China, the USA and other countries, which are in turn influenced by fluc-
tuations in the global fertilizer and energy market [7]. Although the main source
of synthetic nitrogen fertilizer—elemental nitrogen from the atmosphere—is virtu-
ally inexhaustible, the energy and hydrogen for fixing elemental nitrogen through
the chemical synthesis of ammonia are obtained from natural gas or coal. The ni-
trogen fertilizer industry is considered the main customer and exporter of natural
gas in Europe [7].
At the national and European level (AbfKlärV, DüMV), foundations are being
laid for the use of greater proportions of phosphorus from secondary sources of
raw materials such as waste water in the agricultural sector. The spreading of waste
water sludge on agricultural land will, however, be phased out.
Fresh vegetables and fresh fish directly from our city rooftops—that is the
vision of ROOF WATER-FARM. The project, funded by the German Federal Min-
istry of Education and Research (BMBF), has put decentralized wastewater treat-
ment and its use in buildings and residential areas to the test. Instead of transport-
ing domestic waste water from the shower, washing machine, bathtub or toilet to
sewage treatment plants, they can also be recycled directly on site.
Fig. 17.4 Comparison of yields using conventional NPK fertilizer and the “gold water” fer-
tilizer from the ROOF WATER-FARM. (Fraunhofer UMSICHT)
Two independent test series were conducted, one with toilet waste water (also
called black water) and one with waste water from the shower, washing machine
and bath (gray water), where the waste water was treated to such an extent that
high-quality cucumbers and lettuce could be harvested in the ROOF WATER-
FARM (Fig. 17.3).
All concepts and processes developed for the ROOF WATER-FARM project
were tested in a building complex at the pilot site in Berlin-Kreuzberg. This is also
where the black water liquid fertilizer plant developed by Fraunhofer UMSICHT
is located, which produces liquid plant fertilizers from black water with a high
nutrient content (nitrogen, phosphate and potassium). Since its commissioning, the
plant has reliably treated the waste water of 50 residents to produce liquid fertilizer,
also known as gold water due to its yellow color. This in turn is used for vegetable
production in a greenhouse, which is also located in Berlin-Kreuzberg, although
in this initial pilot phase it is still located on the ground instead of on the roof. In
addition, gray water generated by 250 people (all domestic waste water excluding
toilet waste water) is treated and used for the irrigation of plants.
The yield of lettuce heads in the ROOF WATER-FARM could even be increased
slightly when self-produced gold water was used, without affecting the appearance
or taste (Fig. 17.4).
The positive experiences from the ROOF WATER-FARM project are currently
flowing into further project developments e.g. to develop so-called NEWtrient®
centers. In the future these centers could treat resource flows that include all nutri-
ents in cities e.g. also biodegradable waste. The overarching goal of the vision is
to establish urban circular agricultural production, within the framework of a cir-
cular urban economy, as an innovative future sector of the bio-based economy in
Germany.
17.5 Digital Horticulture
In the future, digitization will permeate every aspect of life and change all areas
of life and economic activity such as value-creation processes, science, consumer
behavior and social and cultural interactions. German agriculture and its upstream
and downstream economic sectors are at a high level of productivity and automa-
tion. Here, too, digitization is progressing. The keywords “precision farming” or
“smart farming” stand for the optimization of complex processes with the aid of
IT solutions in traditional open field cultivation. Similar to the forecasts for Indus-
try 4.0 or the Internet of Things, components in agriculture and horticulture will
also communicate with each other more and more in the future.
“Digital horticulture” is a new term that refers to the use of a range of digital
technologies (including plant health measurement devices) used in various horti-
cultural applications such as phenotyping and precision horticulture (often referred
to as site-specific cultivation [9]).
The data that is collected can provide information, for example, on nutrient re-
quirements, plant damage and the necessary irrigation intervals. This information
can be cohesively evaluated, especially in CEA and it may be used purposefully
for optimized production in combination with appropriate actuators. In this way,
the quantity of biomass and the active ingredient content can be deliberately in-
creased. To this end, however, deeper insights into plant-physiological effects and
the further development of suitable measurement and evaluation methods are still
required today.
Plant growth is critically influenced by light. However, for photosynthesis, re-
ferred to as PAR (photosynthetically active radiation), the entire spectrum of sun-
light is not needed, but primarily the range of blue light (about 420 to 480 nm) and
red light (about 630 to 790 nm), (Fig. 17.5). Professional horticulture still predom-
Fig. 17.5 Comparison of the spectra of natural sunlight in summer with high-intensity
discharge lamps (SON-T), a specific LED module (10-channel module), and the photosyn-
thetically active radiation (PAR) required by plants. Measurements from LED-grow-quality®
project. (Fraunhofer UMSICHT)
inantly uses high intensity discharge lamps today. New, high-performance LEDs
offer tremendous potential in plant cultivation, as it is possible for only the re-
quired spectrum to be emitted. Suitable LEDs can be selected depending on the
illumination scenarios required and the optimum illumination spectrum set up for
the plant varieties.
The aim of the LEDs-grow-quality® project at Fraunhofer UMSICHT was to
develop a needs-based plant illumination system that automatically adjusts accord-
ing to the plant’s light requirements based on the content and concentration of
secondary plant ingredients, thus providing optimal growth conditions as well as
ideal flavor and active ingredient concentrations. It was demonstrated that both the
concentration of the secondary plant ingredient, rutin (substance class: flavonoid)
and the plant biomass obtained could be doubled depending on the illumination
parameters applied. The project results are particularly relevant for professional
horticulture. Their relevance relates to the directed adjustment of the concentra-
Fig. 17.6 Basil under artificial LED lighting in the project LEDs-grow-quality® . (Fraun-
hofer UMSICHT)
tions of flavoring agents and active ingredients in the cultivation of vegetables,

fruits, herbs (Fig. 17.6) and medicinal plants.
17.6 Outlook
Biological and technological modes of action will in future be integrated into plant
production systems even more. The biological transformation approach in produc-
tion and the interaction between vastly different value chains to create circular
systems will shape the future agricultural economy.

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urban and rural areas for sustainable and resilient development. Deutsche Gesellschaft für
Internationale Zusammenarbeit, Eschborn
2. Eigenbrod C, Gruda N (2015) Urban vegetable for food security in cities. A review. Agron
Sustain Dev 35(2):483–498
3. Jennings S et al (2015) Food in an urbanised world. The role of city region food systems
in resilience and sustainable development. Urban Agriculture Magazine 29: 5–7
4. Despommier DD (2011) The vertical farm: Feeding the world in the 21st century. Picador,
New York
5. Aerofarms: The world’s largest Vertical Farm. http://aerofarms.com/ Accessed:
16.02.2018
6. Childers DL et al (2011) Sustainability Challenges of Phosphorus and Food: Solutions
from Closing the Human Phosphorus Cycle. BioScience 61(2):117–124
7. Malingreau J-P, Eva H, Maggio A (2012) NPK: Will there be enough plant nutrients to
feed a world of 9 billion in 2050? Publications Office, Luxembourg
8. Industrieverband Agrar (2017) Wichtige Zahlen 2016–2017: Düngemittel, Produktion,
Markt, Landwirtschaft
9. Colaco AF, Molin JP, Rosell-Polo JR, Escolà A (2018) Application of light detection and
ranging and ultrasonic sensors to high-throughput phenotyping and precision horticulture:
current status and challenges. Hortic Res 5: 35
Digital Villages
How Digital Ecosystems Are Structured and What
18
They Can Do
Mario Trapp and Steffen Hess
Summary
The Fraunhofer Institute for Experimental Software Engineering IESE demon-

strates, in its “Digital Villages” project, how digitization is opening up new
opportunities for rural regions. The project began in summer 2015 with the aim
of examining the challenges of modern life in rural areas in terms of digitization.
Since then, concepts and solutions have emerged that reveal the possibilities
inherent in taking a holistic view of the topic of digitization, in the sense of
a digital ecosystem. The project, funded by the Rhineland-Palatinate Ministry
of the Interior and for Sport, Fraunhofer IESE and the Rhineland-Palatinate De-
velopment Agency, is considered a pioneer for many other initiatives that have
since emerged in Germany. Their mutual aim is to put digital services in rural
areas to the test and make them sustainable.
18.1 Introduction
It does not seem so long ago that software systems were almost exclusively devel-
oped as stand-alone products. The focus was on the individual product, from word
processing through to complex ERP systems. The interaction of software products
from different manufacturers remains a challenge to this day. Particularly in the
M. Trapp ()
Fraunhofer Institute for Embedded Systems and Communication Technologies ESK
München, Germany
S. Hess
Fraunhofer Institute for Experimental Software Engineering IESE
Kaiserslautern, Germany

https://doi.org/10.1007/978-3-662-59659-3_18
364 M. Trapp and S. Hess
case of professional products, it is even a significant part of the business model to

generate a substantial proportion of turnover through adaptations and integrating
other software systems. The effort and costs associated with connecting systems
and data from other providers when introducing a large ERP system are signifi-
cant.
In the past, all were firmly convinced of the established business models and the
focus was on strategic sales of products and related services. Part of this strategy
was also the rigorous protection of in-house software and its interfaces in order
to be able to make money from selling extensions and adaptations and to keep
competitors out of the market.
These rules of success applied until the “Big Four”—Google, Apple, Facebook
and Amazon, GAFA for short—heralded the age of the platform economy, chang-
ing the very foundations of the market. When Amazon began to open its online
marketplace also for products from other providers, German companies in par-
ticular shook their heads in disbelief. What reputable bookseller would open all
its doors to the competition? However, Amazon was concerned with turning its
marketplace into an open platform that would attract many more buyers with the
widest possible range of offerings than a single online store could ever do. Ama-
zon was convinced that this would ultimately generate significantly more sales.
And ultimately, as a platform operator, it would also make money on every other
sale through charging commission and profit from a multiplier effect. Today we
know how this initially ridiculed strategy has worked out.
This example shows that today it is no longer about the development of individ-
ual software systems and services, but rather about the development and operation
of digital ecosystems based on platforms. Even though the idea of a platform econ-
omy goes back many decades, it was only through digitization that the necessary
technology was found to be able to establish it across the board on a large scale. As
the Amazon example shows, it is however not enough to consider digital ecosys-
tems only from a technological point of view. Rather, all aspects—from business
models to corporate culture—need to do justice to this new way of thinking in
order to succeed.
The success of the GAFA companies is no longer based on focusing on and pro-
tecting individual products, but rather on creating open digital ecosystems, which
a very wide range of companies and customer groups can participate in. In a dig-
ital ecosystem such as this, as in a biological ecosystem, there are a multitude of
different participants each with their own specific interests. Some participants of
the ecosystem are cutthroat competitors. And yet, precisely through the interaction
of the individual participants, through the competition, but also through the com-
plementarity of strengths and offers, disruptive business models have emerged that
18 Digital Villages 365
offer customers real added value and open up completely new markets for compa-
nies. In this process, the participants are not only software systems and services,
but also people who participate in the ecosystem with different roles and different
objectives.
In contrast to a biological ecosystem, digital ecosystems are created; the cre-
ator of the ecosystem determines the rules and benefits from all transactions in the
ecosystem as the operator of the underlying platform. Besides being a new way
of thinking and incorporating alternative business models, a digital ecosystem is
also based on another development compared to a monolithic software system.
The focus is on coordinating the interaction of individual products and services.
In contrast to a single system, in a digital ecosystem there is no way of deter-
mining and controlling all aspects of the system. New participants are constantly
being added while existing ones leave the system. The strategies of the individual
participants change continuously. And the interaction of the individual products
and services creates emergent effects that are extremely hard to predict. Analogies
are therefore drawn to social associations as known in biology. There are no fixed
structures, but rather “behavioral rules” and common “languages” and “values”
that all participants must adhere to, and their compliance is monitored. Those who
do not adhere to these are sanctioned or even excluded. At the same time, each
participant can take on an individual role with an individual behavioral repertoire
within the framework of these basic rules.
Both the behavior of the individual participants and the rules of the ecosys-
tem can and should be continuously adapted and further developed in the process.
This requires completely new architectural principles. There are no longer any rigid
structures, but rather flexible platforms that enable open collaboration between ser-
vices and provide basic services. These platforms are highly adaptive in order to
meet the continuous change. They continuously monitor developments and adapt
accordingly. This results in the merging of the previously separate phases of de-
velopment and operation, which is reflected in the new concept of DevOps (coined
from development and operations). Many systems already adapt dynamically and
autonomously based on biological concepts—these are referred to as self-adaptive
systems. This applies both to the platform and to the individual participants. To
do this, the software has to become self-aware. Although this is far removed from
the human concept of consciousness, it is a special form of artificial intelligence
that mimics biological intelligence. The systems do not only blindly perform the
programmed or trained behaviors, they are aware of their role and their overriding
tasks and goals and can therefore optimize themselves.
There are many example applications for such platforms—from online mar-
ketplaces to the Smart City. One particular application that is within the focus of
Fraunhofer IESE is Smart Rural Areas. Because only around a third of the German
population live in urban areas, digital ecosystems and the underlying platforms can
generate economic and, above all, social added value, especially for rural regions.
This chapter will therefore explain the concepts of digital ecosystem platforms
using this fascinating application as an example. In doing so, we not only focus on
the technical foundations, but also draw on the experience gained from the actual
operation of the platforms in real rural pilot regions. Services, people and systems
already feed into this in a digital ecosystem, thereby creating added value for rural
areas, which would not be possible to develop by means of individual systems.
18.2 Smart Ecosystems as the Basis of Digital Ecosystems
Before we turn to the concrete application example, however, let us first go deeper
into the fundamental concepts of digital ecosystems. As already mentioned, plat-
forms are an essential building block for the implementation of digital ecosystems.
The “Big Four” lead the way. A successful platform forms the technical basis for
the digital business model and is therefore an integral part of digital transforma-
tion. If the idea of the platforms is expanded holistically, the concept of smart
ecosystems emerges as the technological basis for digital ecosystems.
18.2.1 The Platform Strategy—More than the Sum of All Parts
A platform strategy is different from a product strategy: the platform strategy

requires an external ecosystem to generate comprehensive product or service in-
novations and synergies between these options and the platform that is enabling
them. If successful, the result can have greater potential for innovation and growth
than a single company with its product-oriented strategy.
Value creation takes place through digital partners and communities of users
who use the platform and contribute to it. The platform concept is not new in and
of itself. It has existed in various forms since the 1960s. It is receiving a lot of
attention now, however, because of the success of companies like Facebook and
LinkedIn in the area of social networking, Amazon and eBay in retail, or Uber in
transportation.
18.2.2 From Platforms to Smart Ecosystems
Platforms are however only one aspect of digital ecosystems. Ecosystems also need
to be considered holistically in terms of technology in order for them to be to
successfully implemented. For example, the future vision of a “digital society” as-
sumes that all operational, private and social activities and services will in future
be consistently supported by software-based systems without media disruption,
and that the integration of the most varied applications will enable the creation of
further, entirely new synergies. This trend, which can be observed across almost all
domains, requires complete integration of all systems into intelligent ecosystems.
Driven by a common goal, these ecosystems offer cross-company customized so-
lutions.
This idea extends far beyond the idea of platforms. These are then referred to as
“Smart Ecosystems”, which break up the previously isolated solutions for control-
ling business processes and technical processes and allow them to converge in an
integrated overall solution. Good examples of this trend can be found everywhere,
whether they are Smart Energy, Smart Health, Smart Mobility, Smart Farming or
Smart Production—also known as Industry 4.0.
The holistic view of digitization in terms of a “Smart Ecosystem” is also
a model for the platform developed in the project “Digital Villages”, which is the
focus of this chapter. The term Smart Ecosystem has its roots in ancient Greek,
where “oikos” stands for “house” and “systema” for the “assembled” or the “in-
terconnected”. In economics, the term “ecosystem” refers to the community of all
stakeholders within an industry. The behavior of biologically complex systems is
mimicked in the digital smart ecosystem in order to build up an overall system that
is dynamically adaptable.
Fig. 18.1 shows the underlying technical reference model for a smart ecosys-
tem that illustrates the holistic approach. Anchored at the core of the model are the
fundamental definition of the business model, the technology applied and clarifi-
cation of the legal framework. These constitute the DNA of a Smart Ecosystem,
so to speak. The organizational layer deals with structuring the content and with
strategic aspects. In the next layer, partners providing ecosystem services and the
community of users form further key operational building blocks for the sustain-
able operation and success of an ecosystem. The analysis of competitors and other
stakeholders in a smart ecosystem is therefore linked with activities that need to be
carried out continuously [2].
Fig. 18.1 The Smart Ecosystems reference model. (Fraunhofer IESE)
This basic reference model can be applied to the design of smart ecosystems in
all application areas and provides a conceptual roadmap for their systematic design
that takes many stumbling blocks into account early on in the development.
The following section shows an example of how this reference model was ap-
plied in the project “Digital Villages in rural areas”. Major challenges include both
the integration of public services with services that focus on economic aspects and
also the cooperation of partners from the public sector (e.g. municipalities or dis-
tricts) with private companies on a single platform.
18.3 The Smart Ecosystem of Digital Villages
The term Smart City is currently on everyone’s lips—relevant solutions are already
being applied in practice in many places. However, Germany is a country in which
about two thirds of the population call rural areas their home. The structure of
these regions with villages and towns of typically less than 100,000 inhabitants
make them less suitable for the classic Smart City approaches.
Digital Villages [9], or more generally the idea of Smart Rural Areas, are there-
fore an important complement to Smart Cities and address a large proportion of
the population through their focus on rural areas. Today they are already increas-
ing the attractiveness of many regions by enabling the use of digital services on
a single platform. The first step towards a digital ecosystem of the “rural area” has
thus been taken.
The development of such digital ecosystems brings numerous challenges with
it, especially non-technical challenges. In this way, the whole initiative should be
understood as an innovation process that can bring changes in the form of technical
systems, but also in the form of organizational processes and lifestyles. There must
therefore be a fundamental willingness to question the status quo and also to permit
profound changes. These may, for the most part, relate to organizational processes,
such as the work practices and processes of public authorities or the business mod-
els of regional providers and require the involvement of citizens. This also means
that one has to be prepared to take a certain amount of risk [7].
18.4 Success Factors for a Smart Rural Area
A successful Smart Rural Area—that is, a rural area characterized by intelligent

digitization—can emerge if the central variable, namely the local population, is
included in the process of change right from the start. In doing so, it is all about
creating a culture of cooperation and thereby recognizing people’s most urgent
needs. These needs should then be met, where appropriate, with the aid of digital
services. Of course, it should also be mentioned at this point that digitization is
no panacea: particularly in rural areas, successful approaches are characterized
by a combination of social and digital innovation. Digitization can only ever act
as an amplifier for changes and innovations that take place in the analog world.
A project like the Digital Villages project, which seems to have a technical focus
at first glance, is actually chiefly about social innovation [1].
However, the involvement of the local population is not the only success fac-
tor. The collaborative development of digital solutions by citizens, municipalities,
business and science is very promising. It creates a culture of innovation that takes
local needs into account, involves local stakeholders, and increases the acceptance
of digital solutions.
Local authorities or regions should also pay particular attention to their own
communication, which should take place using a variety of channels and address
target groups appropriately. The use of social media, for example, is indispensable
nowadays as a large proportion of the target group in a rural region is active there.
Organizational steps are also definitive success factors, for example the establish-
ment of a CDO (Chief Digital Officer) or a comparable role. This person oversees
the digitization of a region at a political level and is equipped with decision-making
power and provided with a budget.
Once the appropriate framework conditions have been created, the question
arises as to which services should be offered or developed in order to set up a Smart
Rural Area. Frequently, idea generation begins by studying approaches that have
already been successfully tested or by referring to a neighboring region. In doing
so, it is quite legitimate to copy existing good ideas and then ideally to adapt them
to the local context. However, real innovation and added value for the region can
only truly be achieved by combining different ideas to create something new. This
is the point at which digital ecosystems and platforms come into play: it is possible
to combine different services to create new added value. Once developed, services
can also be used in different regions. This can save costs and allow the regions to
learn from each other in operation.
18.5 Smart Rural Areas—More than a Theoretical Construct
In order to be able to make a contribution to the future viability of regions through

digitization, the Digital Villages platform was developed in the Smart Rural Areas
research program. This enables municipalities in Germany to leverage the poten-
tial of digitization in the area of public services and thereby tackle the challenges
of developing their region. However, the platform is far more than a theoretical
construct or an academic prototype. It was developed together with citizens, was
piloted in test regions and is now available to municipalities throughout Germany.
The Digital Villages project was launched already in 2015 in the Rhineland-
Palatinate pilot municipalities of Betzdorf-Gebhardshain, Eisenberg and Göllheim
with the aim of jointly developing and piloting the platform. Approximately 26,000
inhabitants live in the association of municipalities Betzdorf-Gebhardshain with an
area of 73.5 square kilometers. The city of Betzdorf with its district Dauersberg
and sixteen local municipalities are included in it. The association of municipal-
ities is located in the north of Rhineland-Palatinate in the border area between
Siegerland and Westerwald and belongs to the district of Altenkirchen. The associ-
ation of municipalities Eisenberg and Göllheim, about a seven-minute drive apart,
belong to the Donnersbergkreis rural district and is located between Kaiserslautern
and Worms. About 13,000 people live in three municipal areas in the Eisenberg
association of municipalities. The Göllheim association of municipalities with its
13 small municipalities, in which between 140 and just under 3800 inhabitants live,
accounts for approximately 12,000 people in total.
The digital services created in this project have also been available to other mu-
nicipalities for their cost-efficient use since the beginning of 2018. The platform
has thus been made transferable to other regions and a sustainable proposal is sup-
posed to be created. Participating Digital Villages can thus benefit directly from
the further development of the platform and new services. Fraunhofer IESE has
taken on a pioneering role with the Digital Villages. The concept is reflected in
many funding programs today.
18.6 A Unified Platform with Flexibly Usable Services
One key success factor for a rural Digital Ecosystem is the use of a common digital
services platform in rural areas. The flexible use of the services available on the
platform allows the rural region individual development based on the given starting
conditions. Thus, each region can take its “own DNA” into account in structuring
the digital services, while at the same time using substantial parts of the existing
ecosystem in order to create high-quality services faster and more cost-effectively
than would be possible with individual development.
The Digital Villages project is currently focusing on the areas of communica-
tion and local supply as the central building blocks of the platform. Communication
especially between citizens, but also the communication between citizens and mu-
nicipal administration is made possible in an innovative way through the DorfNews
(VillageNews) and DorfFunk (VillageRadio) services of the Digital Villages plat-
form (Fig. 18.2). In this way not only is information about regional events spread
quickly and easily, but citizens are also enabled to actively participate in shaping
the future of their region.
DorfNews is a local online news portal. The aim is to deliver local news to
citizens quickly. Event information, current news, the local weather and opening
Fig. 18.2 The services of the Digital Village Platform. (Fraunhofer IESE)
hours e.g. for the citizens office can be accessed. In doing so, citizens, clubs and
companies are involved. The news is edited by the local administration.
DorfFunk, the village radio, acts as the communication center of the region,
where citizens can offer their help, put in requests or just chat informally with each
other. The latest news from the DorfNews is also broadcast, so residents always
remain up to date. Thanks to DorfFunk, there is a direct line between municipality
and citizens—in future it will be possible to send suggestions or complaints that
automatically go to the right municipal employees with a single wireless transmis-
sion. The testing of chatbots is also envisaged, which would be used to answer
incoming questions automatically outside of the administrative office hours.
In combination, DorfNews and DorfFunk offer the opportunity to raise com-
munication between the municipality and citizens to a whole new level and to im-
plement innovative mechanisms. For example, push messages sent to citizens’ cell
phones can implement daily updates, making use of direct communication chan-
nels. Digital contributions are also conceivable through solutions such as these, if
there is a willingness to also replace the internal administrative processes.
Fig. 18.3 Scene from a Living Lab workshop. (Fraunhofer IESE)
A sustainable local supply is made possible by using the BestellBar (play of

words: order bar/orderable). An online marketplace that can be flexibly configured
may be used as is reasonable for the region. Especially in combination with the
LieferBar (play of words: delivery bar/deliverable), a flexible delivery service, of-
fers particularly innovative added value and the opportunity to achieve more in the
community.
The BestellBar is a brand-new kind of online marketplace. It combines the ad-
vantages of online shopping with those of shopping at a local retailer. Retailers in
the region present and sell their products online. If you do not wish to collect your
goods in person, packages can be delivered via the LieferBar by volunteers or pro-
fessional couriers. In this way you can support well-known dealers in your region,
whose products you know and appreciate while at the same time conveniently fill-
ing your shopping cart from home, and having it delivered to you. The LieferBar
is the community delivery service. Here, citizens can see which packages from the
local online shop, BestellBar, are still waiting for delivery and deliver them to their
neighbors. The resident receives bonus points in the form of DigiTalers for deliver-
ing packages, although these are only a part of their motivation to participate. The
voluntary deliveries not only give you the feeling that you have done something
good, but you get to know lots of new people from the community.
Further digital solutions are being created in the Living Labs together with the
citizens of the association of municipalities, broadly following the project motto
“From the countryside for the countryside” (Fig. 18.3). For example, in the area of
mobility an app is currently being worked on that can network current short notice
mobility offers and requests and support established concepts such as the citizens’
bus. In addition, the possibilities for autonomous rural transport is gradually being
tested. To begin with, the project is dealing with the needs of citizens in rural areas
for interaction with autonomous vehicles and how these can be integrated into
everyday life.
In addition to mobility in rural areas, the platform will in future be expanded to
include building blocks in the areas of digitally supported working environments,
medicine and nursing care.
18.7 The “Rural Areas” Smart Ecosystem
In addition to the applications already used by the citizens, recommendations for

action in the form of a conceptual model for the Smart Rural Areas ecosystem have
emerged from the “Digital Villages” project. In this model there are five levels of
digitization that need to be addressed in the context of the formation of a Smart
Rural Area: society, digital services, technical platform, infrastructure, and the or-
ganizational ecosystem (Fig. 18.4) [6].
Level 1: Society as the Engine of the Smart Rural Area

For the successful implementation of a digital ecosystem, it is necessary to in-
volve key groups of stakeholders in the design right from the start. The residents
in particular should be involved so that the structuring of the services within the
ecosystem addresses their needs. The interaction between local economy, admin-
istration and residents is especially relevant when the services that are developed
are public services.
Level 2: Digital Services Raise the Potential of the Smart Rural Area
Today, digitization offers many opportunities to solve problems or provide support.
In the development of digital services in rural areas the focus is often on public ser-
vices. However, it is crucial to see digital services not as a panacea, but rather as
a tool for structuring domains of application such as local supply, communications,
mobility, administration, work, school, medicine, and nursing care. In the current
Fig. 18.4 The “Digital Villages” Smart Ecosystem. (Fraunhofer IESE)
wave of digitization, it is important not to lose sight of the holistic, strategic design
of the various application domains. It is therefore essential to integrate a sophisti-
cated digitization strategy (level 5).
Level 3: The Technical Platform as a Link Between Services and

Infrastructure
Smart Rural Areas can gain a significant qualitative advantage over other regions
if they succeed in operating their services on a single platform. These platforms,
which intelligently integrate services from different application domains, therefore
often represent the actual technical innovation. Frequently these are highly com-
plex software systems that are produced under tremendous time pressure as they
are often taken into account too late in the development of services. Well-designed
platform architecture based on industrial standards makes all the difference. The
underlying platform is what makes it possible to transfer the concept to other re-
gions. The shared use of the digital foundation thus also allows for global resource
saving, which makes many sustainable business or operating models possible for
the first time.
Fig. 18.5 The Living Lab concept. (Fraunhofer IESE)
Level 4: Infrastructure as a Basic Requirement

The technical platform is thus also the level that connects digital services with
the underlying infrastructure in the sense of network connection—in short, the
Internet of Things. We see well-developed infrastructure as an enabler for Smart
Rural Areas and, as a result, also for the digital innovation of a region. However,
the relevance of the infrastructure goes well beyond network coverage and will
become even more important in the context of new trends such as Smart Home,
Smart Energy or Smart Mobility.
Level 5: The Organizational Ecosystem Connects the Levels

Before we can speak of a Smart Rural Area, the fifth level must be addressed—the
organizational ecosystem. In light of this, it is important to continue to develop
the structures needed for successful implementation in rural areas. In the context
of “Digital Villages”, for example, a transferable concept for the development of
Living Labs or Real-World Laboratories has emerged [3, 4]. Living Labs refer to
a user-centered open innovation ecosystem in which representatives of different
groups of people can network with each other in an open and flexible environment
(Fig. 18.5). In realistic situations, joint contributions can be made continuously to
the development of innovative pilot applications through the use of software engi-
neering user-centered methods on an ongoing basis. Over and above this, a network
of partners should be systematically created to help rural areas draw up a roadmap
for digitization.
18.8 Outlook
This work forms a foundation for the establishment of digital ecosystems in more
rural regions. If we look to the future, digital ecosystems offer huge potential that
is far from being exhausted. For example, the Bertelsmann study on future mobil-
ity carried out by Fraunhofer IESE reveals fascinating possibilities that can arise
through digital platforms. Within this, an example of a major trend in future digital
ecosystems is also revealed. While today’s ecosystems tend to focus on a single
industry such as mobility, in the future people and their needs will become the
central focus. If we consider the need for “health” as an example, ecosystems that
have been separate so far need to be interconnected—for example, the Smart Home
for health monitoring and as a switchboard of everyday life, telemedicine for pro-
fessional health care, the doctors’ administration software for appointments, the
mobility platform for transport to the doctor and the billing platforms of insurers
[7].
This need, like most other needs, shows that people do not live within industrial
sectors and their ecosystems. Expanding on current trends, therefore, the indi-
vidual person with their unique needs will become the connecting element and
central point of all services. Customers expect affordable services that are intu-
itively and individually tailored to meet their personal needs. Current markets that
are still highly supplier-oriented and industry-specific will be replaced by highly
individualized, user-oriented and cross-industry markets. This will present not only
entirely new challenges both technically and from the perspective of business mod-
els, but also completely new possibilities. This transition is reflected in many other
trends—from autonomous driving to Industry 4.0.
In such a scenario, even greater momentum will develop. Continually chang-
ing service providers from a wide range of sectors will need to cooperate within
an ecosystem. Suppliers and customers are involved in partnerships much more.
To this end, processes, some of which are still manual today, will be increas-
ingly automated by intelligent algorithms so that ecosystems can develop more
autonomously and manage themselves. The individual person will take on a com-
pletely different, much more central role. This applies especially to the user experi-
ence as well. While users today still have to adapt extensively to technical systems
such as smartphones, web pages, etc., in the future technical systems are supposed
to be unobtrusive, concealed and adapt highly intuitively to the everyday life of
the user. This trend can already be seen in initial approaches in the form of voice
assistants or chatbots as alternatives to complex user interfaces.
Digital ecosystems have already changed established rules of the game of big
tech companies in many places, opening up entirely new opportunities and mar-
kets. And yet we are only at the beginning of exploiting their potential. To harness
this potential, established companies in particular will have to rethink the role of
their customers and competitors, their business models and their technology devel-
opment at all levels in terms of digital ecosystems.

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Alternatives to Growth
Climax Economy Modelled on Ecology
19
Christoph Schäfers, Kristina Bette, Florian Herrmann, and
Georg Nawroth
Summary
In any consideration of biological transformation, it is worthwhile to include

a perspective on how populations manage the habitats in which they live, as
described by population ecology. Strategies for growth and capacity can be un-
derstood in this context as extreme ways of responding to the fundamental ques-
tion of how to make limited resources of energy and materials available to the
next generation. In ecological succession research, the term climax designates
a final state, which represents the hypothetical end stage of the developmental
succession in plant, animal and soil communities. This is only achieved under
stable environmental conditions and resource availability. Current globalization
trends require that resources must also be considered globally. When we con-
sider the human population and its demands, we are heading towards a climax
situation with regard to our planet’s carrying capacity. This steady-state situ-
ation will require a paradigm shift in the way we think about economy, if we
want to avoid or at least attenuate the cyclical collapses observable in nature in
rapidly growing populations. The global system as a whole exists under quasi-
constant conditions and at the limits of its capacity it requires a circular econ-
omy, which grows in stability by networking as many niche-adapted economic
entities as possible.
C. Schäfers () K. Bette

Aachen, Germany
F. Herrmann G. Nawroth
Fraunhofer Institute for Industrial Engineering IAO
Stuttgart, Germany

https://doi.org/10.1007/978-3-662-59659-3_19
380 C. Schäfers et al.
The present text describes the concept of climax economy, which can serve
as a model for optimal resource utilization. Its principles may also be transferred
to the current developments and challenges of the modern economy—such as
increasing digitization and customization. Intelligent value networks that are
based on diversity and occupy all available niches are prerequisites for a climax
economy. These are illustrated using examples in agriculture and agroforestry
as well as new developments in the automotive industry.
19.1 Introduction: Limits of Growth, Limits of Growth

Economy?
Economic growth is the basis of our economic thinking, regardless of whether

we are neo-liberal or Marxist. All growth requires resources. As long as resource
bottlenecks could be overcome by continuous expansion of economic areas, the
growth paradigm was never questioned. Once a global scale was reached, the
fundamental limitations of planetary resources became obvious and have been ad-
dressed since the 1970s. The limits of growth [10] were also highlighted by the
US government [2]. Although economics has used analogies with evolution in the
so called evolutionary economics [12] to explain the competitive advantages of
companies through their individual appliance of their resources, as well as the im-
pact of diverse market participants on the innovative activity within a market, no
rethinking has taken place in economic policy in relation to the finite nature of
global resources. Instead, a number of paths have been taken to continue following
the growth paradigm, even with increasingly limited resources:
1. Reliance on the development of new resources, be it further deposits of raw

materials on earth, new methods of exploiting raw materials, energy sources or
even extraterrestrial raw material storage
2. Reliance on the possibility of “green growth” or “sustainable growth”, with
ever greater inclusion of renewable resources, renewable energy and a circular
economy, which is reflected in the formulation of the UN Sustainable Develop-
ment Goal 8: “Promote sustained, inclusive and sustainable economic growth,
full and productive employment and decent work for all.”
3. Increased decoupling of financial and real economies with the possibility of
creating virtual assets whose growth does not require real resources.
All three options avoid dealing with the finite nature of real resources as the ab-
solute upper limit of our economic activity. They shift the possibility of an upper
19 Alternatives to Growth 381
limit into a distant future that is not yet relevant to our economic models, whether
through the isolated consideration of individual resources outside of systemic rela-
tionships (1.) or through the idealistic notion of there being immense potential for
transforming quantitative growth into qualitative growth (2.). Or they ignore the
repercussions of financial capitalist growth on the real economy, because without
real values, financial bubbles arise and when they burst it is clear that a virtual
world cannot replace the real one (3).
In any consideration of biological transformation, it is worth including a per-
spective on how populations manage the habitats in which they live, as described
by population ecology1 . This assumes that each species is evolutionarily adapted
with regard to its specific needs for particular resources within a habitat. The term
resource as it is used here is broad: light, oxygen, water, available nutrients (as
salts in the soil or in the form of prey organisms), space, structures and connec-
tivity (for nest building, overnight shelter, overwintering). The habitat is managed
by the production and survival of offspring. A habitat has a carrying capacity for
each species, which is determined by the availability of specific limiting resources.
Resource availability depends on current consumption due to the intensity of man-
agement, which is determined by the densities and demands of the interacting
populations within a habitat. The capacity for each species is therefore a dynamic
variable. Mobility and migration play a crucial role in the settlement and manage-
ment of new habitats.
19.2 The Population Ecology Perspective: Growth Strategy

Versus Capacity Strategy
The growth of natural populations is determined by the growth rate r (as the dif-
ference between birth and death rates) and the specific habitat capacity K (logistic
population growth [22, 24]). Both variables are population-specific and subject to
selection. During phases of rapid expansion when sufficient resources are avail-
able e.g. in spring or after disasters, an r-selection that favors a growth strategy
sets in. Populations with rapid and excessive reproduction are at an advantage.
Small, short-lived organisms are predestined for an r- or growth strategy. Their
opportunistic strategy of surplus production of less-developed offspring with high
mortality rates results in unstable, frequently oscillating populations whose regula-
tion is not dependent on density [13]. Individual competitive ability is not essential.
1
Populations: communities of mutually reproducing individuals; spatially definable subset
of individuals of a single species.
Instead, migration is a phenomenon that is often observed, as the capacity limit is

inevitably exceeded by the exponential growth, leading to a sharp decline in or
collapse of the population.
By contrast, in habitats that are continuously densely populated due to constant
conditions, K-selection results in an adaptation or capacity strategy being favored:
highly competitive populations and population densities that optimally exploit re-
sources prevail. In the broader systematic framework, Southwood [20] identifies
K-strategists by their relatively large body size, longevity, reduced mortality and
renewal, high competitiveness, and greater investment in their offspring. Differen-
tiated behaviors requiring communication, such as brood care, rearing their young
or territorial behavior are typical. Their population density corresponds to a state
of equilibrium dependent on the habitat capacity and their mating behavior corre-
sponds to the equilibrium density.
r- and K-strategies can thus be understood as extreme ways of responding to
the fundamental question of how to make limited resources of energy and materi-
als available to the next generation. According to Remmert [15], character traits of
population dynamic strategies are never realized in their extremes, but can be clas-
sified within an r/K continuum, whereby different characteristics of populations
may well be selected in different directions. Here, allocations to the r- and K-strat-
egy are to be understood fundamentally in comparison with one another and can
be carried out within the various taxonomic levels as well as within a population.
The two strategies offer different possibilities for dealing with threats. The
reproductive performance of r-strategists is largely independent of density. A neg-
ative influence on the reproduction or survival rate is not compensated for indi-
vidually, but has a direct effect on the intra- and interspecific competition and
leads to population collapses [16, 17]. K-strategists, on the other hand, have ad-
vanced individual compensation abilities and reaction to disturbances in the intra-
and interspecific network, so their populations are comparatively stable. However,
exercising compensation requires additional resources, which reduces habitat ca-
pacity.
19.2.1 Economic Development from a Resource Perspective
Humans evolved in a variable environment (savannah, rainy and dry seasons) and
from there settled both in habitats characterized by constant conditions with a va-
riety of resources (e.g. tropical rainforests) as well as in those that feature signifi-
cantly more extreme fluctuations in environmental conditions and resources, such
as steppes, semi-deserts or temperate climates with a seasonally limited vegeta-
tion period. This was possible because their evolving cognitive abilities allowed
humans to adapt their needs and thanks to their mobility they could exploit new
habitats. Humans expanded their natural abilities and improved their use of re-
sources through the development of culture and technology: toolmaking and raw
material extraction, livestock breeding and domestication, agriculture, preservation
and storage processes. The storage needed for overwintering expanded the oppor-
tunity for trade development, made humans increasingly independent of seasons
and climates and led to their global proliferation and resource exploitation.
In the temperate climates of the northern hemisphere, especially in Europe, r-
selection predominated (growth strategy). This first developed the necessity and
then also the technical and ethical conditions for expansion and claiming of exter-
nal resources. Thus, economic relationships were always also classified according
to religion or ideology, which led to Christian colonialism, to Calvinism with its
basis for mercantilism and capitalism and also to Marxism. At the same time, there
was increased decoupling of population density and resource demand. Human cul-
tural development increases individual demands on the quality of food, clothing
and standards of living as well as on the living space itself, on consumer goods
and experiences and therefore also on resources (global footprint). In this way
the goals for human population development (slowdown in growth, reducing birth
rate towards capacity strategy) and economic development (postulate of perpetual
growth = density- and resource-independent growth strategy) diverge.
In cases of resource surplus, for example following wars or epidemics, a growth
strategy of this type leads to predictable successes and stabilizes the system. If the
carrying capacity is reached by using up the limiting resource, the predictability
ends. A growth strategy will then destabilize the system and inevitably lead to
collapse.
19.2.2 Characteristics of Climax Ecosystems
A Model for Optimal Resource Utilization

In ecological succession research2 , the term climax designates a final state, which
represents the hypothetical end stage of the developmental succession in plant,
animal and soil communities. This is only achieved under stable environmental
conditions and resource availability. The example of a tropical rainforest illustrates
2
Succession: sequential changes over time of the characteristic compositions of a living
community (plants, animals, fungi) with full manifestation from the initial state (new habitat
to be colonized) up to climax community.
this clearly. Here there is a constant environment over long periods of time, which
ensures a stable situation with regard to resources and enables optimal adaptation
of all species to this environment and to their cooperative existence. As a result,
highly diverse ecological niches emerge with a high degree of specialization and
distinctive interactions between different species (commensalism3 , symbioses4).
Since nutrients are reassimilated immediately after their release, a small-scale
short-circuited circular economy arises, where all resources are utilized close to
optimally. The dense occupation of the space and networking of the nutrient cycles
across the different levels of the rainforest result in fewer nutrients reaching the
soil. Tropical soils are very poor in nutrients. A scenario such as this, of highly
networked communities of highly competitive populations with low reproductive
rates is ideal for K-strategists.
Local Disasters as Refuges for the Growth Strategy

According to the mosaic cycle concept [15], a climax ecosystem is made up of
a rough pattern of local early succession stages that were created as new habitats
after local disasters such as floods or forest fires and reclaimed by the structures
and life contexts of the adjacent climax ecosystem in the medium term. R-strate-
gists survive in these small, temporary partial habitats through their great ability
to multiply and distribute themselves. As distribution or resettlement is part of the
growth strategy, r-strategists can jump from one disaster site to the next to stay
alive. After succession has taken place, the annidated capacity strategists of the
climax state prevail again and again. The relationship between growth and capac-
ity strategies is dynamic in the peripheral areas of the climax ecosystem: changes
in environmental conditions can shift balances and alter networks.
Buffer for Optimal Resource Utilization of Dynamic Ecosystems

Climax ecosystems are sensitive to major destruction, since their recovery requires
long-term stable environmental conditions to set in, which takes a long time. At
the same time, the complexity of their species networks makes them resilient to
temporary challenges. This can be illustrated by the example of migratory birds
from temperate climates: as K-strategists, migratory birds use the seasonal food
surpluses in temperate climates for rearing their young, but overwinter in tropical
3
Commensalism: a form of interaction between individuals of different species, which is
positive for the members of one species and neutral for the other.
4
Symbiosis: a form of interaction between individuals of different species, which is positive
for the members of both species.
climax ecosystems in low-food periods. The local populations adapt the time when
they rear their own young to that of the migratory birds.
19.3 Transfer to the Economy: Climax Economy

Characteristics Such as Niches, Networking, Material
Cycles
When we consider the human population and its demands, we are heading towards
a climax situation with regard to our planet’s carrying capacity. This steady-state
situation will require a paradigm shift in the way we think about economy, if we
wish to avoid or at least attenuate the cyclical collapses observable in nature in
rapidly growing populations. This requires reinforcing such characteristics as in-
creased investment in fewer offspring, the longevity and flexibility of life concepts,
the optimal exploitation of habitat capacity through the occupation of all available
niches and networking the lives of as many participants as possible. A medium-
sized economic structure enables the small-scale, regionally adapted utilization of
resources, annidation and networking.
Innovation Pressure, Niches and Networking

Where a system poses new challenges, there is pressure to meet them with new de-
velopments (innovation pressure). If enough resources are available, specifications
arise from new demands that are met by economic growth. Above all, technical
innovations are in demand. By contrast, in a system that is generally constant and
where resources are limited, greater demands are placed on efficiency and resource
management. Over and above technical innovations, the need also arises for socio-
economic innovation, which may involve the inclusion of former, more resource-
efficient economies and lifestyles. Niches are thus no longer creative spaces for
disruptive technical innovations protected by resource surpluses. In fact, innova-
tions that have the potential to change sociotechnical regimes and shape economic
landscapes in the long term can now only be developed in a resource-friendly man-
ner and serve to optimize the distribution of resources. In this sense, niches should
be seen as socio-economic (cultural) and ecological (natural space) specialization
spaces that need to be preserved. The goal is no longer to isolate niches from dom-
inant, regime-forming currents, but rather to network them intelligently. A climax
economy uses and supports both socio-economic and natural spatial diversity and
requires political intermediaries. Besides technological innovation, it must rely on
a primacy of social interactions, more so than a growth economy.
Growth as an Interim Strategy and Material Cycles

Current globalization trends require that resources must also be considered glob-
ally. The global system as a whole exists under quasi-constant conditions and—like
a climax ecosystem—requires a circular economy at the limits of its capacity that
gains stability by networking as many niche-adapted economic units as possible.
Growth does not need to be abandoned as a driver of innovation here: analogous to
the mosaic theory in climax ecosystems, growth strategists enjoy temporary suc-
cesses after disasters. Growth economics is thus possible as an interim strategy
when coordinated with the climax economy. The capacity strategists are always
more competitive after the reintroduction of constant conditions and resilient net-
works. In this process, political coordination is required to determine resource
utilization priorities and upper usage limits that preclude overuse. As with biolog-
ical K-strategists, it is only possible to secure capacity for subsequent generations
through the successful implementation of intelligent structures.
Diversification Instead of Standardization

The diversification of demands and the possibilities to achieve them are already re-
flected in global economic trends: personalized medicine takes people’s individual
conditions into account and personalized production addresses individual wishes.
Both are made possible by digital networking and processing of large amounts of
data, which leads to individually differentiated control of diagnostic and therapeu-
tic procedures or production processes. In agriculture, large seed companies are
now also working with niche plants and their regional requirements. Plant protec-
tion agent companies are moving their products and services away from the pure
sale of pesticides towards customized crop protection advice. Large agricultural
machinery manufacturers are also increasingly developing smaller and more spe-
cific solutions. The era of planning global technological and economic solutions is
being replaced by that of planning according to a biological model: diverse, small-
scale and interactive.
19.4 Agriculture and Forestry as Pioneers of the Climax

Economy
The primary economic sector, i.e. the extraction of resources, is particularly strik-
ing when it comes to the orientation of the economy towards resources due to its
own evident dependence on them. The agriculture and forestry sector is an espe-
cially vivid example, where human needs for food, housing and energy translate
directly into the consumption of the resources of land, soil and water through the
use of arable land, building land and wood for construction and firewood. The
concept of sustainability formulated at the beginning of the 18th century by Carl
von Carlowitz [3] described the need to adjust wood consumption to the amount
of wood being regrown. The urgent implications of this idea were initially dimin-
ished by the increasing use of fossil resources as wood combustion became less
economically important. However, the fundamental idea derived from forestry has
now, 300 years on, become established as the basis for future-oriented economic
activity. Thus, it stands to reason that the decisive impulses for a paradigm shift
from the economy of growth to a climax economy come from agriculture:
The global carrying capacity is limited by the availability of water and food.
The strictest technical limits are set with regard to the availability/usability of
agricultural land. Soil erosion caused by deforestation, open pit mining, surface
sealing and fragmentation or salinization due to irrigation are examples of the
reasons for land shortage through overuse.
Recreation and leisure activities as well as nature conservation measures com-
pete for space just as much as measures for energy production.
In agriculture, not only the production of food and animal feedstuff compete
with each other, but these also compete with the production of biomass for
energy generation or industrial raw materials as well as the production of fibers,
active ingredients or value-added compounds under a modern bioeconomy.
Global warming through overuse of fossil resources puts agriculture under par-
ticular pressure due to climate change.
The steady increase in agricultural production through the increased use of fer-
tilizers, pesticides and high-performance seeds, as well as the adaptation of the
agricultural landscape to the use of large machinery leads to an increase in
monocultures and a decline in agrobiodiversity, which in turn leads to a de-
crease in agricultural value added per area in the medium to long term.
The intensive livestock farming of industrial meat production requires soya im-
ports and generates manure, which is increasingly polluting our ground and
surface water.
The agricultural production of biomass, active ingredients and value-added

compounds or industrial raw materials is therefore only justifiable if it is sustain-
able both within the global system of the Earth and under regional conditions. It
must be ensured that:
all ecosystem services remain protected, which is only possible with great di-
versity and interaction,
the competition for land for food production and raw material extraction is min-
imized,
the use of agricultural products is as complete as possible (coupling, cascading,
waste processing and recycling of nutrients back into agricultural production
systems and soils),
the socio-economic conditions and impacts are taken into account (access to
resources, distributive justice, regional value creation, participation).
Optimization of Land Use Through Diversity of Use and Development

of Intelligent Value Networks
The perception of the global habitat as an overall system of finite resources with
regional differences and dynamics requires land use to be optimized to specific
regional conditions, maximum diversity of use within small-scale cultivation con-
cepts and strong social interactions to coordinate these uses. Regional crops and
crop rotation represent adaptation to the respective soils and ecology of the region.
Diversification can thus be understood as decentralized value creation based on
a networked system of differentiated land use and supported by a self-modifying
data and communication infrastructure that promotes medium-sized businesses as
well as agroecology.
A small-scale assessment of potential cultivation areas using geo-referenced
data analysis, based on the specific plant’s needs and the regional economic situ-
ation, may in fact allow for regional value creation that is optimized in terms of
resource use, environmental impact, and the use of infrastructure. Plants that devi-
ate from the main crops in terms of their location requirements can also grow on
marginal land that is poorly suited for efficiency-optimized agriculture. In addition,
they can support the urgently-needed expansion of agrobiodiversity, which would
reduce the spread of pests and diseases in diversified, composite landscapes. The
expansion of the spectrum of cultivated species and varieties, can improve ecosys-
tem services in the close crop rotations of intensive central European agriculture
(provisioning services, cultural services, regulating and supporting services) [9].
The formation of functioning niche markets and development of new, spe-
cialized value chains in an agricultural economy characterized by diversity and
regionality requires close networking between the different players at all stages of
the value chain and between different value chains. The aim is to bring together
suppliers of the widest variety of agricultural products and associated cultivation
systems and customers i.e. the various processing technologies for raw material
production and/or waste processing. The close networking of highly specialized
companies in the processing industry with farms responsive to their needs increases
the regional value creation as well as the resilience of this expanded economy
to crises. At the same time, societal goals can be incorporated into the optimiza-
tion process, allowing various ecosystem services to be taken into account. Trade-
offs, where benefits in certain sections of a value chain correspond to damage to
national or global sustainability goals (greenhouse gas balances, scarcity of ele-
mentary resources), can be systematically addressed as the trade-off is visualized
quantitatively.
The most important development goal is thus the networking of agricultural
operations and companies within a value chain as well as the networking be-
tween different value chains via data provision, data analysis and mathematical
algorithms for optimization. Fraunhofer’s organizational structures can make a va-
riety of contributions here: The Fraunhofer ICT Group can evaluate and provide
data on the landscape (soil quality, water, microclimate, topography, infrastruc-
ture, position to commodity and sales market), market development (needs, sales
opportunities) and management (seed quality and price, operating costs, customer
management). In addition, the Fraunhofer Water Alliance (agricultural production)
and Food Chain Management Alliance (processing and management of agricultural
products), the Fraunhofer Groups for Life Sciences (active ingredients), Materials
(renewable raw materials) and Production (processing methods) should also be
mentioned. In the field of Precision Farming, a number of Fraunhofer institutes
have already been cooperating with leading manufacturers of agricultural machin-
ery for many years. The Fraunhofer Group for Innovation Research can provide
support for systemic approaches, forward thinking strategies and ongoing critical
evaluation of the paths taken and their secondary effects.
Fraunhofer would be an ideal mediator between state environmental land use
planning and private medium-sized businesses. The decentralized structure of
Fraunhofer institutes, akin to medium-sized companies, can contribute to the de-
velopment of a wide range of value chains from agricultural production to the end
product. Examples of such value chains are “high quality food”, “energy produc-
tion from agricultural residues”, “bulk chemicals from renewable raw materials”
or “active ingredients from the field”. This requires close cooperation with external
specialists, in particular from the fields of crop production, agroecology and socio-
economics. The concept of “active ingredients from the field” was developed dur-
ing a workshop held in Schmallenberg on March 17–18, 2016 as a product of such
cooperation and as an example for the development of value chains, which could
form part of a networked niche economy. This workshop was hosted by Fraun-
hofer IME and IVV together with representatives of JKI, ZALF, IRI THESys (HU
Berlin), FH Südwestfalen, TLL Jena and IfA (RLP AgroScience).
Agroforestry Project Example: AUFWERTEN

Modern agroforestry systems are an example of land use systems that combine
different cultivation systems or usage forms. In principle, agroforestry, i.e. the com-
bination of crops and trees on an area of land, is a long-established land use form,
as woody plants have always been an important component of agricultural land
use—for example as windbreak strips or along the edges of water. However, in the
course of the industrialization of agriculture, agroforestry was displaced by mono-
cultures, intensive land management and increasing land pressure. As a result, the
remaining agricultural trees and shrubs were protected and prohibited from being
used, which meant they became unprofitable landscape features for farmers. In the
context of agroforestry systems, usable trees and shrubs with associated livestock
alternating with arable crops on the same area of land, constitute multifunctional
land usage. In this way, diversification of agricultural products is achieved while
simultaneously increasing productivity if sustainably managed in terms of climate,
groundwater, soil and species protection.
From a societal point of view, agroforestry systems also contribute to improving
the landscape and thus also to recreation quality. Modern agroforestry systems,
which are laid out in alternating strips of trees or shrubs and arable crops, can
be efficiently managed with modern agricultural machinery and power technology.
For example, the development of easy-to-transport machines for the cultivation and
harvesting of agroforestry systems is being promoted, such as the mower chipper
developed by the Leibniz Institute for Agricultural Engineering in Potsdam [14].
Agroforestry systems are therefore also of interest to operations that only have
small strips of short-rotation coppices in the agroforestry system.
While agroforestry has been discussed and successfully promoted and sup-
ported by politicians in France for years, such cultivation systems have rarely been
put into practice in Germany—chiefly because of legal hurdles, the lack of regional
value creation and processing chains, economic uncertainties, technical equipment
and deficiencies in advisory services and, to some extent, the lack of a culture of
innovation towards shaping the agriculture of tomorrow [19, 21]. The successful
establishment of agroforestry requires knowledge about agroforestry to be dissem-
inated in the population and within agriculture, so that business models may be
created with awareness of sustainability and particularly of the value of the arable
crops, the products of the woody plants or livestock kept in an agroforestry system.
In order to make the advantages of agroforestry feasible for modern agriculture,
the Innovation Group AUFWERTEN (Agroforestry Environmental Services For
Increasing Value and Energy [25]), for example, provides practical agroforestry
land use concepts. Topics include nature conservation and environmental protec-
tion, agriculture, landscape design, technology, logistics, processing, marketing,

cost-effectiveness, societal and legal control tools, as well as consultation with and
participation in equivalent fields of investigation.
In the context of the climax economy, agroforestry creates agro-ecological and
economic niches. The cultivation of agricultural trees and shrubs for the production
of quality wood, trunk wood, wood fuel or fruit requires longer-term economic
planning and reduced use of pesticides or fertilizers. For example, for short rotation
strips, the cuttings of the trees or shrubs are planted once and provide a regular
yield every four to seven years if they are harvested, for example, as wood chips. In
so doing, only the trunks of the trees are removed, so that the roots can sprout again
for the next harvest. Additional land usage is possible if cattle, pigs or chicken are
kept on the strips of trees and shrubs, providing natural soil fertilization as well as
the possibility of mushroom cultivation in addition to the harvesting of the woody
plants [11, 23].
The strips of trees and shrubs increase biodiversity by creating longer-term
habitats for associated plants, insects, birds, reptiles and small mammals. By con-
trast, the cultivation of arable crops represents the r-strategy aspect of an agro-
forestry system in terms of the mosaic theory. For arable crops there are signifi-
cantly shorter planting and harvesting cycles, although the effort required for plant-
ing, cultivation and harvesting is much greater than for the strips of woody plants.
In contrast to woody crops, the selection of arable crops can be adapted at short
notice to local economic and climatic constraints, which is why the woody strips
of agroforestry systems planted for decades of growth still allow for flexible man-
agement.
One goal of modern agroforestry that is in keeping with the climax economy is
to optimize the interactions between woody and arable crops for maximum yet sus-
tainable productivity. The trees and shrubs provide shade for the crops, increase the
water retention capacity of the soil and through their deep roots draw the ground-
water from depths that would not be accessible to the arable crops. In addition, crop
yields are significantly increased by limiting soil erosion, and crop failure due to
storms is significantly reduced. The trees and shrubs act as a windbreak and their
roots stabilize the ground, especially in the winter when the field is lying fallow.
Thus, even in drier or more sandy areas which are strongly affected by erosion,
equal or even greater yields of arable crops can be harvested on the same field,
even though the arable area has been reduced in favor of the strips of trees and
shrubs [6].
The value chains based on agroforestry systems are an important aspect in the
context of the climax economy. New ways of removing the harvest from the strips
of woody plants—whether in the form of wood chips, fruit or livestock that are
kept in the strips—also need to be established as well as technical process chains,

e.g. from the chipping of the harvested wood to its use in a wood chip power plant
or from harvesting nuts and transporting them to the oil mill through to sales of the
nut oil. This includes the search for new customers, such as by selling arable crops
under a higher quality agroforestry label (e.g. through direct marketing in a farm
shop or through a digital sales platform) or by expanding the farmers’ product
portfolio. The automotive industry in Germany, which is currently facing massive
changes due to the digitization, individualization and electrification of mobility [7,
8], can serve as an example of how new value networks can be established in newly
emerging economic sectors.
19.5 Value Networks for a Climax Economy
The speed of development and exponentially increasing complexity are presenting

companies with new challenges, especially SMEs, which can only operate with
strictly limited resources. On the one hand, planning is almost impossible, and on
the other, traditional modes of cooperation can no longer keep pace with the rate of
development. Thus, in practice, companies are finding it difficult to follow ground-
breaking trends. In the context of the climax economy this means that companies
are creating the conditions for maximizing value creation potential through new
forms of cooperation, not only internally but also externally.
The implementation of new technologies in the automotive industry is opening
up the generation of a variety of new products and services. In this way, attrac-
tive fields of innovation are emerging such as artificial intelligence for achieving
autonomous driving functions, digital networking solutions (e.g. to increase the
information level during driving in the event of accidents or traffic jams) or elec-
tric drive concepts for a wide range of applications. Many of these technologies
are being introduced to the automotive industry by “lateral entrants” or “industry
newcomers”, such as Google, Apple, or Tesla, who are putting immense pressure
on classic original equipment manufacturers (OEMs). Over and above new tech-
nologies, however, innovative business models are also being established [5]. For
example, car sharing solutions, the first approaches to driverless transport services
and intermodal transport services are leading to significantly altered user behav-
ior. This affects mobility itself as well as the requirements for mobility systems
including their services.
More and more new manufacturers, suppliers and (digital) service providers
are pouring into existing markets on account of convergent technology develop-
ment (merging of previously independent technology fields). In this way a value
network is created between existing and new players, which, in addition to the de-
velopment and production of physical goods, is generating more and more data-
driven business models. New ways of cooperation are needed to exploit the poten-
tial of the data generated by products, production or users for services.
The stakeholders in the value networks are thus characterized in particular by
the fact that they no longer only develop, research and test in-house, but increas-
ingly use open structures both within and outside of the value network and combine
the respective best characteristics of the companies. These develop from being
controllers of the respective value chain which they dominate to becoming equal
stakeholders in a value network. Its members occupy special fields of competence
(niches), are distributed globally and networked through digital technologies. This
allows them to collaborate locally in multidisciplinary teams across companies to
innovate and maximize added value, as for example within the framework of the re-
search factory ARENA2036 [26]. As customers are substantially involved in these
value networks, targeted and reactive product development is guaranteed.
The integration of startups into the network extends the niche diversity and in-
creases synergy effects as well as motivation, commitment and therefore also the
competitive advantage of the entire value network. New formats such as innova-
tion hubs, co-creation and on-campus concepts are taking on great importance [1,
4]. While services and developments defined in classic automotive value chains
were implemented with the aim of maximizing the utilization of business capaci-
ties, with customer requirements not being the first priority [18], the goal of open
value networks is for them to develop an individual dynamic. This guarantees max-
imum added value with the most suitable partners and reacts quickly and flexibly
to changing conditions.
In the context of the climax economy, innovative start-ups and small businesses
often take on the role of niche occupants in such value networks, with large com-
panies playing the role of networkers. The great versatility of small companies
and startups also has the advantage that new niches can be filled quickly. As a re-
sult, many large companies are founding startups to strategically occupy emerging
niches. The goal of this kind of cooperation within value networks is that the broad
win-win situation and diversification as a solution for increasing complexity will
increase the motivation of all network stakeholders and their commitment to this
form of stable cooperation in order to jointly stand up to innovation pressure. In
a climax economy, the innovation landscape and therefore the economy should
reach a steady state for the network, in line with all dynamic changes in individual
expertise.
These open, dynamic value networks require traditional companies to change
their way of thinking. Lately, you could end up sitting at the same development
table with a competitor or one of their partners, and must constantly reckon with
the network taking on a stakeholder who could replace your own technology or
service with something else. Agile working methods allow bilateral cooperations
within the value network or a team to be terminated quickly and built-up anew. The
goal is for this network to develop its own internal dynamics, so that according to
the climax economy, maximum value creation is reached in cooperation with the
most appropriate value creation partners and furthermore that it reacts quickly to
changing conditions.
Innovation pressure is mounting for all participating companies in the direction
of further specialization (development of niches) and networking competence. This
undermines the security of the market power of old structures, but constantly offers
new possibilities for diversification and thus new opportunities. Digital technolo-
gies are the main drivers for this development. Virtual platforms enable different
stakeholders to collaborate from different continents (crowd engineering), by us-
ing the necessary expertise and technical equipment to formulate and process new
tasks. Former organizational models are too rigid and more of a hindrance for these
structural constraints. New, agile forms of organization are therefore being applied,
such as Scrum, for example, which offers a lot of leeway both for the methodol-
ogy and the individual elaboration of each task. On the technical side, prototypes
can be developed and tested quickly and independently of location based on digital
models and using generative manufacturing processes. The merging of the digital
world and the real world is creating new possibilities for services, so that data pro-
cessing is taking on immense significance as a secondary market. The question of
who owns the data and who is allowed to use it, must however be addressed, which
opens up further service opportunities for consulting and data security companies.
19.6 Outlook
Fraunhofer currently has the opportunity to play a decisive role in developing the
economy of the 21st century in terms of biological transformation. This does not
only apply to production processes, but also to the structure of business units and
value chains, as the agricultural example has shown. Agriculture is particularly
challenged by the finite nature of resources, climate change and conflicts of in-
terest between competing land uses, not least because of the growing importance
of the bioeconomy. Fraunhofer commands the technologies, the concepts and an
understanding of the system needed to facilitate a turnaround in agriculture, which
means a departure from the philosophy of growth to the optimal use of resources in
a diversified and networked climax economy. With its relatively small-scale struc-
Fig. 19.1 Climax economy in agriculture: levels of diversification and niche formation
ture and heterogeneous agriculture and with an economy based on medium-sized

businesses, Germany can act as a driver for innovations in networked bioeconomic
value chains.
A central element for this development is the Landscape Data Space (LDS)
a space that provides controlled access to and connection of landscape-related dig-
ital data that can be used for a variety of services (Fig. 19.1). The LDS is being
developed as a verticalization of the Industrial Data Space (IDS) and provides
interfaces for accessing sensor networks and public data. Developments in field
technology (sensors, controls, image analysis, mechanical engineering/robotics)
are required to achieve automated small-scale and efficient field processing with
reduced and more targeted use of fertilizers and pesticides. At the same time, the
public sector is beginning to make existing landscape-related data available. This
data can be used for compliance with stipulations, landscape planning and opera-
tional planning and networking, so that land users will be able to make use of it
autonomously. In various areas of agricultural development, new added value is
emerging that uses the LDS and the sensor networks that feed into it. Cultivation
can thus be diversified by the production of active ingredients and value-added
compounds from niche plants.
Smaller-scale cultivation of a variety of plants with suitable crop rotations has
a positive effect on the biodiversity of insects, birds and small mammals, reduces
monoculture-enhanced infestation rates, reduces pesticide use and contributes to

soil conservation. Cascading usage makes traditionally cultivated plants econom-
ically attractive again if processes and facilities are developed for the primary re-
fining of high-grade target substances as well as the secondary refining of residues
for on-farm operations. Optimal land areas for cultivation are identified through
the LDS. Fraunhofer research activities into vertical or contained farming and in
plant biotechnology are also valuable additions.
In terms of circular economy in animal production, the production of insect pro-
tein from slurry, liquid manure and silage effluent has been made technically and
legally possible on a scale suitable for agricultural enterprises. As a result, slurry
storage and transport as well as groundwater pollution are reduced and the use of
imported soya as an animal feed additive is replaced. With the aid of the LDS,
soil nutrient sensor signals are processed and optimal operating sizes and loca-
tions determined for alliances. The design and fulfillment of legal requirements for
crop protection management is another example of LDS services based on public
data and field sensor data e.g. the spatiotemporal coordination of spraying events
smoothens peak loads in bodies of water.
A shift in the direction of the climax economy thus has the potential to solve
many global environmental issues and address current economic challenges.
A growth economy, on the other hand, would only be possible under climax con-
ditions if the latter were temporarily suspended—be it through the development of
new (e.g. extraterrestrial) resources that may be used compatibly with the system,
or through a drastic reduction in competition, i.e. by reducing population density.
The former is an illusion that bursts in the face of increasing knowledge of system
relations and the divergence of the universe, the latter merely cynical. There is
therefore no alternative to the transformation to a climax economy, which relies
more than ever on human innovation.

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26. https://www.arena2036.de/de/ Accessed: 22.09.2018

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Fraunhofer-Forschungsfokus

Reimund Neugebauer Ed.

ISBN 978-3-662-59658-6 ISBN 978-3-662-59659-3 (eBook)

Printed on acid-free paper

1 From Contrast to Convergence . . . . . . . . . . . . . . . . . . . . . . 1

3 Biomimetics Research for Medical Engineering . . . . . . . .... 19

3.2 Main Strands of Biomimetics . . . . . . . . . . . . . . . . . . .. 21

4 Innovative Food Products . . . . . . . . . . . . . . . . . . . . . . . . . . 37

5 Technical Homes for Human Cells . . . . . . . . . . . . . . . . . . . . 63

5.2 Requirements from a Biological Aspect and Modeling

6 Phenotyping and Genotyping of Plants . . . . . . . . . . . . . . . . . 91

7 Cells as Sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105

7.5.2 Analysis of Cytomechanical Changes

8 Biopolymers – Function Carriers in Materials Research . . . . . . 129

9 Biogenic Plastic Additives . . . . . . . . . . . . . . . . . . . . . . . . . . 161

10 Organisms as Producers . . . . . . . . . . . . . . . . . . . . . . . . . . . 179

10.2 Plant Stem Cells for the Cosmetic Industry . . . . . . . . . . . 184

11 Biologized Robotics and Biomechatronics . . . . . . . . . . . . . . . 199

13 Insect Biotechnology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 247

14 The Resource Principle . . . . . . . . . . . . . . . . . . . . . . . . . . . 261

14.7 An Overview of the Digestion Processes . . . . . . . . . . . . . 283

15 Cognitive Biological Sensors . . . . . . . . . . . . . . . . . . . . . . . . 311

16 Prevention of Biofouling . . . . . . . . . . . . . . . . . . . . . . . . . . . 329

17 Urban Agriculture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 351

18 Digital Villages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 363

19 Alternatives to Growth . . . . . . . . . . . . . . . . . . . . . . . . . . . 379

Reimund Neugebauer and Martin Thum

1.1 Origins: Technology Helps in the Fight for Survival

© Springer-Verlag GmbH Germany, part of Springer Nature 2020 1

1.2 A New Approach: Nature and Technology to Cooperate

1.3 Latest: Biological and Digital Transformation Cross-

complexity or even imitating it by means of technology. The current acceleration

1.4 The Goal: Efficient and Sustainable Value Creation

The adoption of biological models, principles and processes rapidly results in

1.5 Biological Transformation at Fraunhofer and Outlook

The Fraunhofer-Gesellschaft has recognized the developmental technological po-

mentally friendly production of natural rubber from dandelions was discussed in

The application of biological principles has resulted in profound changes in sec-

2.1 The Meaning of Biological Transformation

The term “biological transformation” refers to the increasing use of materials,

P. Dieckhoff () S. Hippmann R. Klingner M. Leis

© Springer-Verlag GmbH Germany, part of Springer Nature 2020 9

2.2 Nature as Inspiration for Innovations

Using nature as a source of innovation has a long tradition in Germany. The

technology”, although in the USA in particular, “bionics” is largely understood as

2.3 The Contribution to Global Challenges and

Currently humanity uses, on average, an equivalent of “1.7 Earths” for resource

2.4 Concepts and Examples from Fraunhofer Research

tory function of a nose. Here Fraunhofer EMFT is developing miniaturized hybrid

Biological systems are characterized in particular by unique abilities such as adap-

Sources and Bibliography

Thomas Bauernhansl and Oliver Schwarz

T. Bauernhansl () O. Schwarz

© Springer-Verlag GmbH Germany, part of Springer Nature 2020 19

Biomimetics can be understood as a creative technology. Although many people

3.1.1 Biomimetics as Creative Technology

3.1.2 Biomimetics as a Discipline with Many Sub-Disciplines

in the biomimetics community resulted in a series of VDI guidelines1 (Associa-