200

Assia Stefanova Gary Caldwell

Newcastle University Newcastle University

Ben Bridgens Rachel Armstrong

Newcastle University Newcastle University

Pichaya In-na
Newcastle University

Architectural
Laboratory Practice
for the Development
of Clay and Ceramic-
Based Photosynthetic
Biocomposites
TAD 4 : 2

This study outlines the development of

clay and ceramic-based living biocomposite
materials under minimal moisture environ-
ments. The biocomposites supported live
and metabolically active photosynthetic
microorganisms (microalgae). The work sets
out laboratory testing strategies to assess
the limiting conditions for life within living
materials to inform the conditions needed
for sustained operation. Combinations of
clay/ceramic-based substrate types, nutrient
loadings, amounts of moisture exposures,
and operation times were explored. As a
result, microalgae (Chlorella vulgaris) colonized
both clay and ceramic forms, showing
operational longevity of 100 days, even
with low nutrient exposure. Further itera-
tions of these sustainable living materials
may prove useful for local potential carbon
dioxide removal to improve air quality and
reduce the carbon footprint and operation
costs of mechanically ventilated spaces.
STEFANOVA ET AL.
v Opening Figure. Manually modeled porcelain biocomposite
prototype with living C. vulgaris; part of the Yggdrasil Exhibit, London
Design Festival, 2019.
Keywords: Living Materials, Photosynthesis, Biocomposites, Air
Pollution, Sustainable Living
Introduction
Living microorganisms have been employed in food fermenta-
tion and the production of medicines and are integrated within
a wide range of industries ranging from pharmaceuticals and
meat substitutes to low environmental impact alternatives for
everyday products.1 Areas of the construction industry are also
beginning to meet some of their needs through the integration of
living metabolic functions in a variety of ways, ranging from the
use of fungus mycelium to create loadbearing building bricks and
insulation to harnessing bacterial processes to produce materials
such as self-healing concrete.2 Another example is the develop-
ment of living biocomposite materials, incorporating a substrate
that provides a surface and structure in or on which living organ-
isms can live for the useful part of the material’s life cycle.
The clay/ceramic-based biocomposites described herein were
developed for the potential purposes of direct air carbon cap-
ture using photosynthesis. They comprise porous, hygroscopic
ceramic substrates that maintain a moist environment capable of
supporting photosynthetic microalgae—Chlorella vulgaris in this
instance, a species that can sequester an average of 6.24 g/L/d
of carbon dioxide (CO2).3 From a design perspective, hard sur-
faces that lend themselves to a variety of fabrication methods—
such as clay and ceramics (fired clay)—are of particular interest.4
Within modern, synthetic applications, microorganisms are often
combined with organic or inorganic substrates to increase their
resilience.5 The cells are attached to hard surfaces to increase
the cell density (i.e. cell number per unit space) relative to the cell
density that may typically be achieved when suspended within
liquid tank systems (the conventional method for growing micro-
algae). This study aimed to demonstrate the viability of the mate-
rial and to test the conditions to be met for the living material to
function correctly over sustained periods of operation.
To fulfil the organisms’ complex requirements, a new design
practice is emerging involving the use of laboratory protocols
to develop living materials.6 Designers and architects seek to
engage with laboratory practice and biological testing to estab-
lish key information for the development of interventions that
address pollution and energy consumption problems within
buildings. When designing for specialized species one particu-
larly needs to understand their vital requirements (such as tem-
perature, pH, light requirement, moisture levels, and nutrients) as
well as how to protect them from invading organisms (competi-
tors or pathogens).7
Algae Overview
There are in excess of 70,000 species of algae that have evolved
to survive and thrive within a wide variety of environments
where liquid water is available.8 Microalgae (as an umbrella term)
are either unicellular eukaryotes (organisms with cells containing
membrane-bound organelles) or simple unicellular prokaryotes
201
(organisms lacking membrane-bound organelles).9 Microalgae
are predominantly, although not exclusively, photosynthetic
and can fix between 10 and 50 times more CO2 from the air
per unit biomass than terrestrial plants depending on the spe-
cies.10 Cultivation of microalgae is typically conducted in open
ponds, lagoons, or raceways, which are unsuitable for the built
environment due to space requirements.11 Alternative cultiva-
tion approaches utilizing closed settings (photobioreactors) are
more amenable to integrate with buildings; however, the reliance
on liquid systems presents an engineering challenge that com-
plicates building integration.12 To address this issue, the study
proposes the adoption of living biocomposites as an alternative
cultivation method. The biocomposites minimize water use, in
essence enabling growth on damp solid surfaces that pose fewer
challenges when integrating into a building setting.
Carbon Capture Systems within the Built Environment
Design solutions for direct air carbon capture range from highly
engineered mechanical systems to biological approaches that uti-
lize photosynthetic species ranging from large terrestrial plants
to phytoplankton. Architects and designers such as Kengo Kuma
and Daan Roosegaarde have explored mechanical approaches
to carbon capture technology in the built environment through
PEER REVIEW / MAT TER
installations and prototypes.13 Roosegaarde’s smog cleaning vac-
uum cleaner tower is capable of cleaning more than 30,000 m3
(39,239 yd3) of air per hour and, through the use of electrolysis,
utilizes similar amounts of electricity as a water boiler.14
Engineered passive solutions include sorbent-based air collec-
tors, such as the filter material developed by Klaus Lackner that
has higher CO2 capture rates than trees.15 Absorption of CO2
also occurs from process treatment such as the production of
biochar and activated carbon that are used in their pure or modi-
fied state to remove CO2 from the air. However, these products
are derived from an energy intensive thermochemical conversion
of organic compounds such as timber waste from industry, sew-
age, and animal waste.16
Many of the chemical-based carbon capture systems provide
solutions that rely on removing carbon molecules from CO2,
whereas biological solutions employ naturally available meta-
bolic functions (starting with photosynthesis) that do not sim-
ply store carbon but convert it into biochemicals (including high
value products such as astaxanthin pigment and essential fatty
acids such as docosahexaenoic acid).17 There are many biological
solutions that are integrated into built environments—including
those that improve air quality—through urban farming or build-
ing interventions such as traditional green facades, double skin
facades, green roofs, and green walls.18 The reported cost esti-
mates vary depending on complexity.19
At the other end of the spectrum are biological solutions that
utilize photosynthetic microorganisms either as part of their
manufacturing processes or cultivation as part of the useful life
cycle of the system. Notable examples include the biomineraliza-
tion of building materials using cyanobacteria, bioreactor facades
such as the BIQ House by ARUP, and prototypes and art instal-
lations such as WaterLilly algae wall by Cesare Griffa and Living
Things by Jacob Douenias and Ethan Frier.20 It is estimated that
on average 1.8g of CO2 are removed from the air by every 1g of
 Architectural Laboratory Practice for the Development of Clay and Ceramic-Based Photosynthetic Biocomposites 202

Table 1. Chemical composition, textural and aesthetic properties of the four studied clays. Chemical composition data provided by
Valentine Clays, Stoke-on-Trent, U.K.

Clay properties Porcelain White Fleck Crank (ES50) Crank Terracotta

(ES65)
Silicon dioxide (SiO2) % 68.96 70.28 59.01 53.99
Titanium dioxide (TiO2) % 0.36 1.3 1.24 0.97
Aluminium(I) oxide (Al2O) % 19.68 18.61 27.46 28.37
Iron oxide (Fe2O3) % 0.54 1.25 2.83 5.56
Phosphorus pentoxide (P2O5) % 0.04 0.06 0.06 0.08
Calcium oxide (CaO) % 0.26 0.27 0.43 0.53
Magnesium oxide (MgO) % 0.2 0.52 0.8 2.17
Potassium oxide (K 2O) % 4.95 1.84 1.96 1.45
Sodium oxide (Na2O) % 1.12 0.25 0.14 0.14
Density when fired at 1100˚C 3.94 g/cm3 4.37 g/cm3 4.58 g/cm3 4.52 g/cm3
Fired color White Off white-speckled Warm buff Medium-dark red

algae biomass, and while algae make up a mere 0.5% of the total
biomass on the planet they are responsible for over 50% of the
oxygen in the atmosphere.21 The method of cultivation proposed
here employs a high cell density cultivation technique onto clay
and ceramic hard surfaces capable of cultivating algae with min-
TAD 4 : 2
Germany). The assessments were conducted sequentially with
different aims to assess and evaluate the primary limiting factors
of the materials:
Ŏ Clay compatibility: This stage determined the ability of the
clay to sustain the algae; it served to assess whether ster-

imal water use compared to most traditional liquid suspension
cultivations hoping to be implemented in architectural settings.
Method
A crucial factor in the choice of material was the clay’s porous
structure that would directly impact the ability to retain and
deliver nutrient-enriched moisture to the algae cells. Four pot-
tery clays were chosen; Crank Terracotta (ES65), Crank (ES50),
Porcelain, and White Fleck (WF), which varied in texture
(ranging from a smooth homogenous consistency to a heavily
speckled texture), in chemical composition (Table 1), and had
various aesthetic properties (colors ranging from a white to a
deep red). The moisture retention of each ceramic from each
type of clay was tested using the procedure described in the
moisture level test.
Chlorella vulgaris (C. vulgaris) was chosen as the demonstra-
tion microalgae organism as its photosynthetic efficiency can
reach 20%, compared with 1% for typical terrestrial plants.22
This microalgae species is a promising candidate for the purpos-
es of potential CO2 fixation on clay as it is reasonably resilient
to stress from fluctuating environmental temperatures (the spe-
cies thrives in temperatures compatible with interior environ-
ments; 20-35°C; 68-95°F) and low moisture levels (can survive
full dehydration for periods of months). Many previous studies
have used C. vulgaris for CO2 capture in both suspension and
immobilized culture systems, and it is robustly tolerant of con-
tamination by other organisms.23
Tests were conducted to assess the effect of the clay sub-
strate and a set of potential limiting factors on the metabolic
function of C. vulgaris. Cellular health was assessed by determin-
ing in situ chlorophyll florescence using imaging pulse amplitude
modulated fluorometry (Imaging-PAM M-Series; Walz GmbH,
ilizing the clay prior to inoculation affected algae health
and to identify the most favourable pH for C. vulgaris
growth. Clay offers a malleable option that can change
its morphology and that can be recycled easily. The study
was conducted over a period of 14 days.
Ŏ Ceramic longevity: Following the initial compatibility
assessment, the clay was substituted with ceramic (fired
clay), offering a more robust base that opens a wider
range of applications, such as interior screens, ceiling tiles,
or lighting fixtures, that would not require future reshap-
ing. The test was conducted over 100 days to assess the
potential for long-term microalgae growth. Ceramic lends
itself to a different set of design solutions than clay; how-
ever, the study demonstrates that the microalgae used
have a similar response to both substrates.
Ŏ Moisture levels: The effect of moisture level on microalgae
growth was tested over a period of 12 days, establish-
ing minimal moisture content requirements to sustain the
algae on a hard ceramic substrate.
Ŏ Synthetic urine testing: This test was conducted to deter-
mine the effects of substituting enriched nutrient media
with a widely available and economical nutrient substitute
in the hope of assessing potential for wastewater treat-
ment. Ceramic was employed for this set of experiments
as it lends itself to a wider range of cleaning methods
likely to be necessary in an in vivo setting. The study was
conducted over a period of 14 days.
The tests undertaken in this study are summarized in Table 2
along with sample nomenclatures in Table 3.
Minitab 18 was used to conduct statistical analysis. None of
the data were normally distributed therefore the non-paramet-
ric Friedman test was employed.
STEFANOVA ET AL. 203

Table 2. Test parameters for the four groups of studies presented.

Parameters Experimental Tests

Clay Compatibility Ceramic Longevity Moisture Levels Synthetic Urine

Substrate Type (x3 samples) Clay: Porcelain, Ceramic: Porcelain, Ceramic: Porcelain, Ceramic: Porcelain,
White Fleck (WF), White Fleck (WF), White Fleck (WF), White Fleck (WF),
Crank ES50, Crank Crank ES50, Crank Crank ES50, Crank Crank ES50, Crank
ES65 ES65 ES65 ES65
Algae Slurry Amount 20µl 20µl 20µl 20µl
Nutrient Type BG11 BG11 BG11 Synthetic Urine
(SU)
Nutrient Amount 0.5ml 0.5ml 1ml, 0.5ml, 0.25ml 0.5ml
and 0.125ml
Nutrient Dilution full, 1/2, 1/4, 1/8 full, 1/2, 1/4, 1/8 Full 1:25, 1:50, 1:100,
and 1/10 and 1/10 1:200
Suspension Type (x3 Samples) Substrate+BG11 Substrate+BG11 Substrate+BG11 Substrate + UV
(as per dilutions) (as per dilutions) Sterilized Synthetic
Urine (UV SU)

Table 3. Samples nomenclature.

PEER REVIEW / MAT TER

Sample Name Porcelain White Fleck Crank ES50 Crank ES65 Suspension Synthetic Urine UV Light Sterilized
Synthetic Urine
Abbreviation P WF ES50 ES65 S SU UV SU

Clay Compatibility Test
The clay compatibility test was conducted according to the
presented protocol. One litre of 14-day-old liquid algae cul-
ture (optical density of 0.243 ± 0.006, equivalent to 4.56 × 10 8
cells/ml) that had been grown in full strength BG-11 medium
(full strength BG-11 contains 1.5  g/L NaNO3, 0.036 g/L
CaCl2 · 2H2O, 0.075 g/L MgSO4 · 7H2O, 0.04 g/L K 2HPO4,
and 0.02 g/L Na2CO3) was placed in 50 ml Falcon tubes and
centrifuged at 1620 RCF (Relative Centrifugal Force) for 10
minutes to produce a wet dense algae slurry.24 The culture
was checked under a microscope for any contamination before
centrifugation to prevent other species from influencing the
results.
The clays were prepared as 7 mm cubes, weighing approxi-
mately 1.6 g (± 0.15). The experiment was split into four sets
of primary treatments: 1) not autoclaved/ not pH balanced, 2)
not autoclaved/ pH balanced, 3) autoclaved/ not pH balanced,
and 4) autoclaved and pH balanced. Autoclaving is a process of
sterilization by applying high temperature (125°C; 257°F for one
hour) and pressure (0.2 MPa; 30psi) within a pressure chamber.
The pH of ES65 and Porcelain was adjusted to pH 7 using 0.2 ml
of 0.1 M acetic acid solution per 0.84 g of clay, while White
Fleck used 0.25 ml of 0.2 M ammonium hydroxide solution per
0.7 g to stabilize as its initial pH was acidic. ES50 did not require
balancing as it had a pH of 7 which is neutral and compatible
with C. vulgaris. In addition to pH, the clays also differ in their
composition, with ES50 and ES65 containing higher amounts of
aggregate (added granular material).
Cubes were placed into individual wells (3 ml capacity) with-
in 24-well tissue culture plates. For the biological treatments
(i.e. treatments with added algae cells), 0.5 ml of media and 20
µl of wet algae slurry were added. Each sample was mirrored
by a non-biological negative control (clay plus media but no
added algae cells). For the biological positive controls, suspen-
sion controls were also run in parallel consisting of only 0.5 ml
nutrient media with the same amount of algae cells (Figure 1).
The four clay types were tested at the same time, alongside
five enriched nutrient conditions: full strength BG-11 medium,
and 1/2, 1/4, 1/8 and 1/10 strengths. All treatments were in
triplicate. The well plates were sealed with a plastic lid to limit
contamination and evaporation. The samples were incubated
in a constant temperature and humidity-controlled room (tem-
perature 19 ±2°C; 66 ±4°F; relative humidity 50%) under a
16:8 hours light:dark photoperiod from mixed cool and warm
fluorescent LED tubes with a mean light intensity of 2,500
lux (≈ 30.5 µmol/m/s).
The lids were removed during PAM imaging analysis as the
plastic interfered with the fluorescence signal. The PAM read-
ings were taken in two day intervals for 14 days.
Ceramic Longevity Test
The ceramic (fired clay) longevity test followed the same pro-
tocol as the initial clay compatibility test; however, the ceramic
samples were not autoclaved or pH balanced. All four types of
clay (Porcelain, White Fleck, ES50, ES65) in the previous test
were fired at 1100°C (2012°F) and left unglazed to maintain
 Architectural Laboratory Practice for the Development of Clay and Ceramic-Based Photosynthetic Biocomposites
TAD 4 : 2
r Figure 1. Example images of the 24-well plate compatability
assays. Biological assays (i.e. with algae) are presented in the first
three columns, non-biological controls (no algae) are presented
in the second three columns. A) Different clays with full strength
BG-11 medium; B) Nutrient dilution (½, ¼, ⅛, and ⅟10) treatments with
Porcelain; and C) BG-11 suspension controls with a range of nutrient
dilutions (½, ¼, ⅛, and ⅟10).
204
their porosities. The average weight of the ceramic samples
was around 1.45 g (± 0.15 g). The ceramic samples were tested
using five BG-11 concentrations (full, 1/2, 1/4, 1/8, and 1/10
strengths). The test was conducted for 100 days with readings
taken every two days to assess the longevity of the materials
and the cell viability. During the test, 0.1 ml of autoclaved deion-
ized water (around 7 pH) was reintroduced every 14 days to
compensate for evaporation occurring within the well plates.
There were no additional nutrients after the start of the test
as the test aimed to establish at what point nutrients would be
depleted and cell development would be affected.
Determination of Moisture Levels
The ceramic moisture test followed the same preparation and
incubation protocols as the clay compatibility test, with the
objective to establish a minimal level of moisture necessary to
sustain cell vitality. The clay, as per the ceramic longevity testing,
was fired at 1100°C (2012°F) and left unglazed to maintain its
porosity. Before the moisture test, the moisture retention capa-
bilities of the ceramics were determined by calculating change
in mass before and after immersion in sterile deionized water. A
cubic ceramic sample was submerged in 2 ml for five minutes.
After the immersion, the samples were removed and quickly
blotted with absorbent paper to remove excess surface water
before weighing. The ceramic samples were then tested using
four BG-11 volumes (1 ml, 0.5 ml, 0.25 ml, and 0.125 ml) and the
average weight of the samples was 1.45 g (± 0.15 g). The test
was conducted for 12 days with readings taken every two days.
Synthetic Urine Assay
In the final assessment stage, the performance of the microal-
gae was exposed to four different synthetic urine dilutions and a
UV sterilized control (sterilized through exposure to UV light for
1.5 hours, wavelength between 185-280 nm). The purpose of
this experiment was to study the effects of different synthetic
urine dilutions on the ceramic biocomposites. The experiment
followed the same protocol and incubation conditions as the clay
compatibility test. The synthetic urine was diluted using sterile
deionized water to be 1:25, 1:50, 1:100, and 1:200 of synthetic
urine to water. These dilutions were tested for each 1.45 g (±
0.15 g) ceramic sample (Porcelain, White Fleck, ES50 and ES65).
Results
Clay Compatibility Test: Effect of Different Clay Treatments and
Preparation Conditions
The four different treatments offered various degrees of mate-
rial adjustment. The non-sterilized clays without pH adjustment
(Treatment 1) are described here as they required the least
preparation and energy input, making them the most acces-
sible option out of the four for future building applications.
Despite the treatment offering the least favorable conditions,
the cells on the living biocomposite samples had a significantly
higher maximum fluorescence yield in all cases (n = 60, d.f. =
3, ChiSq = 20.84, P = <0.001) except the ES65 treatment
with 1/2-strength BG-11 media. During the first two days,
fluorescence yield decreased, remaining low in all suspension
 STEFANOVA ET AL. 205

0.8
v Figure 2. Maximum fluorescence yield
of Chlorella vulgaris grown on clays from
MAXIMUM FLUORESCENCE YIELD, FM'

0.7 Treatment 1 (non-sterilized clays without

pH adjustment) with diluted BG-11 medium
0.6 (½, ¼, ⅛, and ⅒). P = Porcelain (mean StDev
= 0.097 ± 0.054), WF = White Fleck (mean
0.5 StDev = 0.076 ± 0.026), ES50 = Crank ES50
(mean StDev = 0.072 ± 0.043), ES65 = Crank
Terracotta ES65 (mean StDev = 0.029 ±
0.4 0.007) and S = Suspension control (mean
StDev = 0.041 ± 0.010).
0.3

0.2

0.1

0.0
Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14

TIME (DAYS)

P BG11 P 1/2BG11 P 1/4BG11

P 1/8BG11 P 1/10BG11 S BG11
S 1/2BG11 S 1/4BG11 S 1/8BG11
S 1/10BG11 WF 1/2BG11 ES50 1/2BG11
ES65 1/2BG11

Day 0 Day 14 v Figure 3. Photographs (top) and PAM

PEER REVIEW / MAT TER

P P fluorescence images (bottom) of the four clay
types on days 0 and 14 in Treatment 1 with
full-strength BG-11 medium. A visible increase
in fluorescence and a wider distribution of
cells was evident for all clay types including
WF WF the higher BG11 dilutions tested, except for
ES65 where warmer colors were observed
at day 14. Cooler colors signify higher
fluorescence levels. P = Porcelain, WF = White
Fleck, ES50 = Crank ES50, and ES65 = Crank
Terracotta ES65.
ES50 ES50

ES65 ES65

Day 0 Day 14

P P

WF WF

ES50 ES50

ES65 ES65
 Architectural Laboratory Practice for the Development of Clay and Ceramic-Based Photosynthetic Biocomposites
0.7
MAXIMUM FLUORESCENCE YIELD, FM'
0.6
0.5
0.4
0.3
0.2
0.1
0.0
Day 0 Day 10 Day 20 Day 30 Day 40 Day 50 Day 60 Day 70 Day 80 Day 90 Day 100
TIME (DAYS)
P BG11 P 1/2 BG11
S BG11 S 1/2 BG11
controls and in the Crank Terracotta ES65. The White Fleck
with 1/2-strength BG-11 media maintained a steady fluores-
TAD 4 : 2
cence signal throughout the experiment. The Porcelain treat-
ments exhibited some variation in chlorophyll fluorescence,
although the 1/10 strength BG-11 treatment had a consis-
tently increasing fluorescence yield and was the only treatment
with a net increase in fluorescence over the 14-day period
(Figures 2 and 3). This could be due to the cells adjusting to
their environment, in which the cells may disperse along the
substrate surface and migrate into areas with higher nutrient
concentrations. In all cases, smoother clays (Porcelain and White
Fleck) outperformed clays with higher amounts of aggregates
(ES50 and ES65).
Longevity of Ceramics
The living biocomposite materials survived throughout the 100-
day period of the longevity test, with the Porcelain ceramic
proving to be the best performing substrate, although there
were no significant differences between treatments (n = 48, d.f.
= 3, ChiSq = 4.40, P = 0.221). The fluorescence yield remained
broadly consistent for all samples, with a clear decline evident
after day 90 (Figure 4). Fluorescence values peaked between
days 40 and 50, indicating at which point nutrients should be
added so that nutrient availability does not become a limiting
factor for cell survival. In contrast with the clay treatment, the
suspension control outperformed the ceramic treatments, albeit
with a small improvement. The impact of nutrient dilution was
also more evident in the ceramic treatment, with more dilute
media supporting weaker fluorescence.
Moisture Levels
The moisture retention of the ceramics without the addition
of algae was significantly different (n = 48, d.f. = 3, ChiSq =
10.50, P = 0.015), with 15.19% for Porcelain, 11.94% for ES50,
206
v Figure 4. Maximum
fluorescence yield of Chlorella
vulgaris grown on ceramics
with full strength and diluted
BG-11 medium (½, ¼, and ⅛)
over a 100-day period. Data
for Porcelain (P) (mean StDev
= 0.067 ± 0.009) displayed
alongside the suspension
controls (S) (mean StDev =
0.043 ± 0.014). Porcelain
samples outperformed other
clay type samples with ES50
(mean StDev = 0.086 ± 0.037)
and ES65 (mean StDev =
0.082 ± 0.023) presenting the
least satisfactory results, but
following a similar growth and
decline pattern.
P 1/4 BG11 P 1/8 BG11
S 1/4 BG11 S 1/8 BG11
4.56% for ES65, and 2.61% for WF. It is important to determine
whether these differences in retention capacity could affect cell
viability within a biocomposite, and in so doing, setting tolerance
levels within an in vivo setting as the response to evaporation,
combined with nutrient availability, would dictate any mainte-
nance schedule. Low moisture content had a negative effect,
with treatments containing less than 0.25 ml showing evidence
of stress (low fluorescence) by six to eight days (Figure 5);
although there may have been scope for recovery by the addi-
tion of water. In contrast, the Porcelain treatments with either
0.5 or 1 ml displayed a progressive increase in fluorescence over
the 12-day period, albeit lower than the suspension controls in
all instances.
Synthetic Urine Assay
The test investigated the impact of diluted synthetic urine on
cell vitality in the living materials over 14 days. Notable chlo-
rophyll fluorescence was observed even with 1:100 and 1:200
dilutions, indicating that within an in vivo scenario a living sur-
face could be sustained with minimal amounts of nutrients from
urine. The UV treated samples presented a consistently higher
level of chlorophyll fluorescence than the biocomposite samples
(n = 48, d.f. = 3, ChiSq = 32.50, P = <0.001; Figure 6).
Discussion
The data serves to inform a process for testing a wider range of
material substrates and photosynthetic organisms that may be
used within design practices, and builds upon existing research
of living photosynthetic biocomposites.25 The clay compatibility
test showed that Porcelain and White Fleck clays and ceramics
are better suited for cultivating C. vulgaris compared with higher
aggregate substrates (ES50 and ES65), which may also influence
performance through variations in chemical composition, and
porosity for capillary absorption and moisture retention. Using
STEFANOVA ET AL. 207

v Figure 5. Maximum
fluorescence yield of Chlorella
vulgaris grown on Porcelain
(mean StDev = 0.057 ± 0.017)
0.9 ceramic with decreasing
MAXIMUM FLUORESCENCE YIELD, FM'

volumes of full-strength
0.8 BG-11 media over a 12-day
period. Suspension controls
(mean StDev = 0.159 ± 0.035)
0.7 represented in broken lines are
shown for comparison.
0.6

0.5

0.4

0.3

0.2

0.1

0.0
Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12

TIME (DAYS)
P 1ml P 0.5ml P 0.25ml P 0.125ml

PEER REVIEW / MAT TER

BG11 1ml BG11 0.5ml BG11 0.25ml BG11 0.125ml
v Figure 6. Maximum
fluorescence yield of Chlorella
vulgaris grown on Porcelain
0.8 (ceramic) with four dilutions
of synthetic urine (1:25, 1:50,
MAXIMUM FLUORESCENCE YIELD, FM'

1:100, and 1:200) (mean StDev

0.7 = 0.085 ± 0.017). UV sterilized
artificial urine (mean StDev
0.6 = 0.143 ± 0.093) treatment
controls are shown for
comparison in dotted lines.
0.5

0.4

0.3

0.2

0.1

0.0
Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14

TIME (DAYS)
SU 1:25 SU 1:50 SU 1:100 SU 1:200
UV SU 1:25 UV SU 1:50 UV SU 1:100 UV SU 1:200

PAM as the method of evaluation provided an overall indica-
tion of the development of the living materials compared with
suspension controls. PAM (which is a non-sacrificial sampling
method) works by exciting chlorophyll molecules in photosys-
tem II; however, algae cells closer to the biofilm surface will yield
a stronger signal due to masking of cells deeper in the biofilm.
Consequently, the PAM method could underestimate the per-
formance of the most densely populated biocomposites. To
explore the response of cells within deeper layers of the biofilm
would require sacrificial sampling that would have risked desta-
bilizing the biocomposite structure. The study has also dem-
onstrated that biocomposites can be cultivated with synthetic
urine instead of optimized laboratory growth media (BG-11 in
this instance). From a cost and chemicals access standpoint,
this is an important consideration, as alternate nutrient sources
(such as human urine or indeed other wastewater streams)
are economically more attractive than optimized media which
tend to contain expensive and difficult to prepare components.
 Architectural Laboratory Practice for the Development of Clay and Ceramic-Based Photosynthetic Biocomposites
r Figure 7. Manually modeled porcelain biocomposite prototype (fired
at 1100°C, dimensions: 25 cm x 25 cm x 15 cm) with living C. vulgaris;
part of the Yggdrasil Exhibit, London Design Festival, 2019. (Credit:
Assia Stefanova)
Further, a supply of human urine (or a greywater source) should
be easily accessible within an appropriately designed building.26
However, contamination is likely to be a factor that may affect
the health of the algae as suggested by the UV-treated samples.
The presented results partially assess the biocomposite
behavior and further investigations are required to examine
TAD 4 : 2
in-depth effects of an uncontrolled environment on the algae
cells. This will enable the fabrication of working prototypes that
would give a better understanding of how the living material
could be affected by human-made environments. The current
assessment is limited to the level of algae chlorophyll fluores-
cence, inferring a healthy photosynthetic function and potential
CO2 capture for improving air quality in interior spaces. Further
tests are needed to quantify the exact amount of CO2 fixation
in order to assess the efficiency of the living materials. In addi-
tion, factors such as temperature fluctuations, sudden evapo-
ration on the surface, maintenance of the materials, and the
effects of contamination if incubated in an open container will
be integrated into the development of a working scaled-up pro-
totype. The longevity and moisture level tests provide prelimi-
nary information ahead of a wider investigation into materials
development. The design of moisture and nutrient distribution
systems would provide a next step in the development of the
living materials, enabling scaling and cost-effective integration.
A nutrient distribution system must take into account the algae’s
tendency to migrate towards (and hence wash off) the direction
of higher moisture levels as observed in the experiments.
This initial fabrication of living materials in this study forms
an essential part of future exploration. This could be achieved
through traditional manual fabrication techniques (Figure 7 and
8)27 or 3D fabrication practices that utilize clay, enabling the cre-
ation of multifaceted structures with large surface areas provid-
ing enhanced space for a greater number of cells to live on.28
These surfaces are likely to manifest as unit-based solutions
that can be easily dismantled and maintained within tempera-
ture-controlled environments such as partition wall construc-
tion, surface tiles, and ceiling treatments.
208
r Figure 8. Manually modeled porcelain biocomposite prototype (fired
at 1100°C, diameter 12 cm, height 2 cm) with living C. vulgaris; part
of the Yggdrasil Exhibit, London Design Festival, 2019. (Credit: Assia
Stefanova)
Conclusion
The study has demonstrated that C. vulgaris microalgae can
be cultivated on a range of ceramic substrates, performing
particularly well with Porcelain-based clay and ceramics. The
durability of the biocomposites and their nutrient require-
ments translated to a period in excess of three months for
uninterrupted material use. The living materials were suf-
ficiently resilient to withstand fluctuations in moisture and
nutrient levels, thereby enabling the findings to be useful for
informing bio-design practice.
The study offers a quantitative method for species-task
selection, e.g. enhanced air quality via potential CO2 fixation
or wastewater remediation. Further development is necessary
to enable the photosynthetic biocomposites to function in an
uncontrolled environment which would require prototype
development and evaluation.
The study demonstrates a way of engaging with laboratory
practice for the purposes of developing photosynthetic liv-
ing materials. The work offers a particular example that could
be modified to enable wider testing of substrates and organ-
isms. Engagement with scientific methods and interdisciplin-
ary collaboration is necessary to foster a better understanding
of living systems if designers are to actively participate in the
development of such materials. Therefore, laboratory testing
and development of protocols become necessary steps with-
in the design process that would offer solutions to architec-
tural problems, where building-scale constraints become an
integral part of formulating methodologies on a fundamental
level. The study outlined a methodological approach to the
development of living materials where the next steps would
require the design of life-support systems for these species
to be sustained in ways that do not place great burdens upon
inhabitants.
Acknowledgments
This research was funded by Research England as part of the
Hub for Biotechnology in the Built Environment.
STEFANOVA ET AL.
Notes
1. R.C Ray and V.K. Joshi, “Fermented Foods: Past, Present and
Future,” in Microorganisms and Fermantation of Traditional
Foods, ed. R.C. Ray and M. Didier (Boca Raton: CRC Press,
2015), 1–36; W. Fox, Toward a Transpersonal Ecology:
Developing New Foundations for Environmentalism (Albany,
NY: State University of New York Press, 1995); V. Joshi and
S. Kumar, “Meat Analogues: Plant Based Alternatives to
Meat Products - A Review,” International Journal of Food
and Fermentation Technology 5, no. 2 (2015): 107-119; G.
Holt, D. Mcintyre, D. Flagg, E. Bayer, J. Wanjura, and M.
Pelletier, “Fungal Mycelium and Cotton Plant Materials in the
Manufacture of Biodegradable Molded Packaging Material:
Evaluation Study of Select Blends of Cotton Byproducts,”
Journal of Biobased Materials and Bioenergy 6, no. 4 (2012):
431–439.
2. A. Stefanova, T.H. Arnardottir, D. Ozkan, and S. Lee,
“Approach to Biologically Made Materials and Advanced
Fabrication Practices,” in International Conference on Emerging
Technologies In Architectural Design (ICETAD2019), ed. M. Asefi
and M. Gorgolewski (Toronto: Ryerson University, 2019),
193–200; A. Frearson, “Tower of ‘Grown’ Bio-Bricks by The
Living Opens at MoMA PS1,” Dezeen, July 1, 2014, https://
www.dezeen.com/2014/07/01/tower-of-grown-bio-bricks-
by-the-living-opens-at-moma-ps1-gallery; Y. Xing, M. Brewer,
H. El-Gharabawy, G. Griffith, and P. Jones, “Growing and
Testing Mycelium Bricks as Building Insulation Materials,” IOP
Conference Series: Earth and Environmental Science 121, no.
2 (2018); N. De Belie and J. Wang, “Bacteria-Based Repair
and Self-Healing of Concrete,” Journal of Sustainable Cement-
Based Materials 5, no. 1 (2016): 35–56.
3. A. Ghorbani, H.R. Rahimpour, Y. Ghasemi, S. Zoughi,
and M.R. Rahimpour, “A Review of Carbon Capture and
Sequestration in Iran: Microalgal Biofixation Potential in
Iran,” Renewable and Sustainable Energy Reviews 35 (2014):
73–100.
4. E.W. Smith and G.S. Austin, Adobe, Pressed-Earth, and
Rammed-Earth Industries in New Mexico (Revised Edition),
Bulletin 159, New Mexico Bureau of Mines and Mineral
Resources (1996).
5. I. Holzmeister, M. Schamel, J. Groll, U. Gbureck, and E.
Vorndran, “Artificial Inorganic Biohybrids: The Functional
Combination of Microorganisms and Cells with Inorganic
Materials,” Acta Biomaterialia 74, (2018): 17–35.
6. A. Umar, “The Screening, Fabrication and Production
of Microalgae Biocomposites for Carbon Capture and
Utilisation,” Thesis, Newcastle University, 2018.
7. C. Reynolds. Ecology of Phytoplankton (Cambridge:
Cambridge University Press, 2006).
8. M.D. Guiry, “How Many Species of Algae Are There?” Journal
of Phycology 48, no. 5 (2012): 1057–1063.
9. Reynolds, Ecology of Phytoplankton.
10. W.Y. Cheah, P. Loke Show, J-S. Chang, T.C. Ling, and J.C.
Juan, “Biosequestration of Atmospheric CO2 and Flue Gas-
Containing CO2 by Microalgae,” Bioresource Technology 184
(2015): 190–201.
11. H. Wolkers, M.J. Barbosa, D.M.M. Kleinegris, R. Bosma,
R.H. Wijffels, and P.F.H. Harmsen, “Microalgae: The Green
Gold of the Future?: Large-Scale Sustainable Cultivation of
Microalgae for the Production of Bulk Commodities,” report,
Wageningen University & Research (2011).
12. K. Lofgren, “World’s First Algae-Powered Building by
209
Splitterwerk Architects Opens in Germany,” Inhabitat, March
5, 2013, https://inhabitat.com/splitterwerk-architects-
design-worlds-first-algae-powered-building-for-germany/
algae-powered-house-biofacade-splitterwerk-arup-colt-
international-scc-green-power-building.
13. B. Hobson, “Dassault Systèmes Presents Pollution-
Absorbing Architecture by Kengo Kuma and Daan
Roosegaarde,” Dezeen, June 8, 2018, https://www.dezeen.
com/2018/06/08/video-interview-dassault-systemes-
pollution-architecture-kengo-kuma-daan-roosegaarde-
movie.
14. L. Garfield and C. Thompson, “The World’s First ‘Smog
Vacuum Cleaner’ Can Suck Up Air Pollution and Turn It
into Jewellery,” The Independent, January 29, 2018, https://
www.independent.co.uk/environment/smog-vacuum-
cleaner-air-pollution-daan-roosegaarde-netherlands-china-
poland-a8183236.html.
15. K.S. Lackner, “Capture of Carbon Dioxide from Ambient
Air,” The European Physical Journal Special Topics 176 (2009):
93–106.
16. P.D. Dissanayake, S. You, A.D. Igalavithana, Y. Xia, A.
Bhatnagar, S. Gupta, H.W. Kua, S. Kim, J-H. Kwon, D.C.W.
Tsang, and Y.S. Ok, “Biochar-Based Adsorbents for
Carbon Dioxide Capture: A Critical Review,” Renewable
and Sustainable Energy Reviews 119 (2020): 109582; A.U.
Rajapaksha, Y.S. Ok, A. El-Naggar, H. Kim, F. Song, S. Kang,
PEER REVIEW / MAT TER
and Y.F. Tsang, “Dissolved Organic Matter Characterization
of Biochars Produced from Different Feedstock Materials,”
Journal of Environmental Management 233, (2019): 393–399;
T.M. Melo, M. Bottlinger, E. Schulz, W.M. Leandro, A.M. de
Aguiar Filho, H. Wang, Y.S. Ok, and J. Rinklebe, “Plant and
Soil Responses to Hydrothermally Converted Sewage Sludge
(Sewchar),” Chemosphere 206 (2018): 338–348; X. He, Z.
Liu, W. Niu, L. Yang, T. Zhou, D. Qin, Z. Niu, and Q. Yuan,
“Effects of Pyrolysis Temperature on the Physicochemical
Properties of Gas and Biochar Obtained from Pyrolysis of
Crop Residues,” Energy 143 (2018): 746–756; C. Sophia and
E.C. Lima, “Removal of Emerging Contaminants from the
Environment by Adsorption,” Ecotoxicology and Environmental
Safety 150 (2018): 1–17.
17. E.H. Murchie and T. Lawson, “Chlorophyll Fluorescence
Analysis: A Guide to Good Practice and Understanding Some
New Applications,” Journal of Experimental Botany 64 (2013):
3983–3998; A. Ilavarasi, D. Mubarakali, R. Praveenkumar, E.
Baldev, and N. Thajuddin, “Optimization of Various Growth
Media to Freshwater Microalgae for Biomass Production,”
Biotechnology 10 (2011): 540–545.
18. G. Pérez, J. Coma, I. Martorell, and L.F. Cabeza, “Vertical
Greenery Systems (VGS) for Energy Saving in Buildings: A
Review,” Renewable and Sustainable Energy Reviews 39 (2014):
139–165; M. Manso and J. Castro-Gomes, “Green Wall
Systems: A Review of Their Characteristics,” Renewable and
Sustainable Energy Reviews 41 (2015): 863–871.
19. T.A.M. Pugh, R.A. Mackenzie, D.J. Whyatt, and C.N. Hewitt,
“Effectiveness of Green Infrastructure for Improvement of
Air Quality in Urban Street Canyons,” Environmental Science
and Technology 46, no. 14 (2012): 7692–7699; K. Perini, M.
Ottelé, E.M. Haas, and R. Raiteri, “Greening the Building
Envelope, Façade Greening and Living Wall Systems,” Open
Journal of Ecology 1, no. 1 (2011): 1–8.
20. Lofgren, “World’s First Algae-Powered”; C.M. Heveran,
S.L. Williams, J. Qui, J. Artier, M.H. Hubler, S.M. Cook,
J.C. Cameron, and W. Srubar III, “Biomineralization and
Successive Regeneration of Engineered Living Building
Materials,” Matter 2, no. 2 (2020): 481–494; P. Bogias, “Algae
Textile: A Lightweight Photobioreactor for Urban Buildings,”
 Architectural Laboratory Practice for the Development of Clay and Ceramic-Based Photosynthetic Biocomposites 210

UWSpace, University of Waterloo, 2014; P. Peruccio and M.

Vrenna, “Design and Microalgae: Sustainable Systems for
Cities,” Agathón | International Journal of Architecture, Art and
Design 6 (2019): 2019, 218–227.
21. Wolkers et al., “Microalgae”; G.M. Elrayies, “Microalgae:
Prospects for Greener Future Buildings,” Renewable and
Sustainable Energy Reviews 81 (2017): 1175–1191.
22. M. Adamczyk, J. Lasek, and A. Skawińska, “CO2
Biofixation and Growth Kinetics of Chlorella vulgaris
and Nannochloropsis gaditana,” Applied Biochemistry and
Biotechnology 179 (2016): 1248–1261.
23. A. Umar, P. In-na, A. Wallace, M. Flickinger, G. Caldwell,
and J. Lee, “Loofah-Based Microalgae and Cyanobacteria
Biocomposites for Intensifying Carbon Dioxide Capture,”
SSRN Electronic Journal (2019), doi:10.2139/ssrn.3489079.
24. R.Y. Stanier, R. Kunisawa, M. Mandel, and G. Cohen-Bazire,
“Purification and Properties of Unicellular Blue-Green Algae
(order Chroococcales),” Bacteriology Reviews 35, no. 2 (1971):
171–205.
25. Umar, “The Screening, Fabrication and Production.”
26. Ilavarasi et al., “Optimization of Various Growth”; S. Jaatinen,
A-M. Lakaniemi, and J. Rintala, “Use of Diluted Urine for
Cultivation of Chlorella vulgaris,” Environmental Technology
37, no. 9 (2016): 1159–1170; S. Månsson, “Cultivation of
Chlorella vulgaris in Nutrient Solution from Greenhouse
Tomato Production,” Swedish University of Agricultural
Sciences, 2012; T. Fazal, A. Mushtaq, F. Rehman, A.U.
Khan, N. Rashid, W. Farooq, M.S. Ur Rehman, and J. Xu,
TAD 4 : 2
Pichaya In-na is a Chemical Engineering PhD Researcher in the
School of Engineering at Newcastle University. Her research
interest is in the development of living algal biocomposites for
carbon capture applications. She has recently been involved
in organizing the Postgraduate Research Conference 2020 at
Newcastle University and was highly commended as the best
host of a live industrial speaker session.
Gary Caldwell is a Senior Lecturer in Applied Marine Biology at
Newcastle University. He works at the interface of biology and
engineering to develop novel solutions to societal challenges.
He is an authority on low water demand algae technologies
and has developed biocomposites suited to a range of
environmental applications.
Rachel Armstrong is Professor of Experimental Architecture
at the School of Architecture, Planning and Landscape,
Newcastle University. Her work is characterized by design
thinking as a fusion element for interdisciplinary expertise
and pioneers in the field of “living” architecture that materially
and technologically directly engages the potency of life within
spatial agendas.

“Bioremediation of Textile Wastewater and Successive

Biodiesel Production Using Microalgae,” Renewable and
Sustainable Energy Reviews 82, no. 3 (2018): 3107–3126.
27. A. Stefanova, T. Arnardottir, D. Ozkan, and S. Lee, “Biodesign
Here Now | Yggdrasil,” London Design Festival: Open Cell
(2019), accessed September 25, 2019, https://www.opencell.
bio/ldf/yggdrasil.
28. R. van Broekhoven and O. van Herpt, “Solid Vibrations,”
Olivier van Herpt, accessed April 24, 2019, http://
oliviervanherpt.com/solid-vibrations; B. Gürsoy, “From
Control to Uncertainty in 3D Printing with Clay,” in
Computing for a Better Tomorrow - Proceedings of the 36th
eCAADe Conference - Volume 2, eds. A. Kepczynska-Walczak
and S. Bialkowski, Lodz University of Technology, 2018,
21–30.

Assia Stefanova is an ARB registered architect and a creative

practice PhD Researcher at Newcastle University. Her work
focuses on the development of biological material alternatives
and their integration into the urban realm. She is the architect
for the Hub for Biotechnology in the Built Environment (HBBE)
and a Resident Artist at the Wellcome Centre for Mitochondrial
Research, Newcastle University.

Ben Bridgens is a Senior Lecturer in Architectural Technology

in the School of Architecture, Planning and Landscape
at Newcastle University. He is a member of the Hub for
Biotechnology in the Built Environment, which aims to create
a new generation of Living Buildings which respond to their
environment, are grown using living engineered materials,
metabolize their own waste, and modulate their microbiome.