Polysaccharide-Based Nanocomposites

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Polysaccharide-Based
Nanocomposites for Gene Delivery
and Tissue Engineering
Woodhead Publishing Series in Biomaterials

Polysaccharide-Based
Nanocomposites for Gene
Delivery and Tissue
Engineering

Edited by

Showkat Ahmad Bhawani

Faculty of Resource Science and Technology,
Universiti Malaysia Sarawak (UNIMAS), Kota
Samarahan, Sarawak, Malaysia
Zoheb Karim
MoRe Research AB, Sweden
Mohammad Jawaid
Biocomposite Technology Laboratory, Institute
of Tropical Forestry and Forest Products
(INTROP), University Putra Malaysia (UPM),
Serdang, Selangor, Malaysia

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Dedication

The editors are honored to dedicate this book to Dr. Kishwar Usmani
(Aunty of Dr. Mohammad Jawaid), who left this worldly life on January
14, 2021.
Allah SWT forgive her small and big mistakes and Give her a Higher
Place in Jannatul Firdous. Ameen.
Contributors

Aarzoo Department of Chemistry, School of Chemical and Life sciences, Jamia

Hamdard, New Delhi, India

Sadaf Afrin Department of Chemistry, Faculty of Sciences, Aligarh Muslim

University, Aligarh, India

Akil Ahmad School of Industrial Technology, Universiti Sains Malaysia, Penang,

Malaysia

Asrar Ahmad Center for Sickle Cell Diseases, College of Medicine, Howard
University, Washington, DC, United States

Khalid M. Alotaibi Department of Chemistry, College of Science, King Saud

University, Riyadh, Kingdom of Saudi Arabia

Mohammad Omaish Ansari Center of Nanotechnology, King Abdul Aziz

University, Jeddah, Saudi Arabia

Mohd Saquib Ansari Department of Biomedical Sciences, Shaheed Rajguru College

of Applied Sciences for Women, University of Delhi, New Delhi, India

M. Tarik Arafat Department of Biomedical Engineering, Bangladesh University of

Engineering and Technology (BUET), Dhaka, Bangladesh

Maya Asyikin Mohamad Arif Faculty of Resource Science and Technology,

University Malaysia Sarawak, Kota Samarahan, Sarawak, Malaysia

Mohd Razip Asaruddin Faculty of Resource Science and Technology, Universiti

Malaysia Sarawak (UNIMAS), Kota Samarahan, Sarawak, Malaysia

Mohammad Ashfaq BS Abdur Rahman Crescent Institute of Science and

Technology, Chennai, India

Erkan T€ urker Baran University of Health Sciences Turkey, Institute of Health

Sciences, Department of Biomaterials; University of Health Sciences Turkey,
Institute of Health Sciences, Department of Tissue Engineering, Istanbul, Turkey
xiv Contributors

Sunita Barik Department of Chemistry, Saraswati Degree Science College, Cuttack,

Odisha, India

Anant Narayan Bhatt Institute of Nuclear Medicine and Allied Sciences, Defence
Research and Development Organization, New Delhi, India

Showkat Ahmad Bhawani Faculty of Resource Science and Technology, Universiti

Malaysia Sarawak (UNIMAS), Kota Samarahan, Sarawak, Malaysia

Anuradha Biswal Department of Chemistry, Veer Surendra Sai University of

Technology, Sambalpur, Odisha, India

Afzal Hussain Department of Pharmacognosy, College of Pharmacy, King Saud

University, Riyadh, Saudi Arabia

Mohamad Nasir Mohamad Ibrahim School of Chemical Sciences, Universiti Sains

Malaysia, Penang, Malaysia

Iqbal I.M. Ismail Department of Chemistry, Faculty of Science, King Abdul Aziz
University, Jeddah, Saudi Arabia

Zoheb Karim MoRe Research AB, Sweden

Mohd Jahir Khan School of Biotechnology, Jawaharlal Nehru University,

New Delhi, India

Abhishek Kumar Department of Biomedical Sciences, Shaheed Rajguru College of

Applied Sciences for Women, University of Delhi; Institute of Nuclear Medicine and
Allied Sciences, Defence Research and Development Organization, New Delhi, India

Vannessa Lawai Faculty of Resource Science and Technology, Universiti Malaysia

Sarawak, Sarawak, Malaysia

Zainab Ngaini Faculty of Resource Science and Technology, Universiti Malaysia

Sarawak, Sarawak, Malaysia

Mehvish Nisar Faculty of Resource Science and Technology, Universiti Malaysia

Sarawak (UNIMAS), Kota Samarahan, Sarawak, Malaysia

Mohammad Oves Center of Excellence in Environmental Studies, King Abdulaziz

University, Jeddah, Saudi Arabia

Md. Wahidur Rahman Department of Biomedical Engineering, Bangladesh

University of Engineering and Technology (BUET), Dhaka, Bangladesh
Contributors xv

Mohd Rashid School of Chemical Sciences, Universiti Sains Malaysia, Penang,

Malaysia

Chinmayi Rath Department of Chemistry, Saraswati Degree Science College,

Cuttack, Odisha, India

Mohd Ahmar Rauf Use-Inspired Biomaterials & Integrated Nano Delivery

(U-BiND) Systems Laboratory, Department of Pharmaceutical Sciences, Eugene
Applebaum College of Pharmacy and Health Sciences, Wayne State University,
Detroit, MI, United States

M. Samim Department of Chemistry, School of Chemical and Life sciences, Jamia

Hamdard, New Delhi, India

Rahul Kumar Sharma Department of Chemistry, Government Shyam Sundar

Agrawal PG College, Sihora, Madhya Pradesh, India

Laishram Rajendra Kumar Singh Dr. B.R. Ambedkar Center for Biomedical
Research, University of Delhi, New Delhi, India

Sarat K. Swain Department of Chemistry, Veer Surendra Sai University of

Technology, Sambalpur, Odisha, India

Aydin Tahmasebifar University of Health Sciences Turkey, Institute of Health

Sciences, Department of Biomaterials; University of Health Sciences Turkey,
Institute of Health Sciences, Department of Tissue Engineering, Istanbul, Turkey

Abu Tariq Department of Chemistry, Aligarh Muslim University, Aligarh, Uttar

Pradesh, India

Mohiuddin Khan Warsi Department of Biochemistry, College of Science,

University of Jeddah, Jeddah, Saudi Arabia

Asim Ali Yaqoob School of Chemical Sciences, Universiti Sains Malaysia, Penang,
Malaysia

Bengi Yilmaz University of Health Sciences Turkey, Institute of Health Sciences,

Department of Biomaterials; University of Health Sciences Turkey, Institute of Health
Sciences, Department of Tissue Engineering, Istanbul, Turkey
Editors’ biographies

Dr. Showkat Ahmad Bhawani is presently working as an Associate Professor at the

Department of Chemistry, Faculty of Resource Science and Technology, UNIMAS
Malaysia. He received his Ph.D. in Applied Analytical Chemistry from Aligarh
Muslim University, Aligarh, India. He is working on the synthesis of molecular
imprinting polymers for the removal/extraction of dyes, fungicides, and various nat-
ural products from environmental and biological samples. In addition to this, he is
working on the development of new test methods and determining standard conditions
for analysis (separation, isolation, and determination) of various analytes from envi-
ronmental and biological samples. He is involved in the analysis of samples such as
surfactants, amino acids, drugs, vitamins, sugars, and metal ions. He has edited two
books and has written 12 book chapters, and he has published more than 40 papers in
various journals. He is a lifetime member of the Asian Polymer Association and an
editorial board member of several journals.
Dr. Zoheb Karim worked as Senior Researcher at MoRe Research Ornskoldsvik
AB. His current research is process-property-application correlation understanding of
nanocellulose (crystals and fibers)-based assembled dimensional structures. Fabrica-
tions of functionally and structurally sound dimensional architecture for a predefined
application using a best appropriate route and starting nanocellulose (required intrinsic
properties) is another goal of his research. Recently, he patented a mechanism of metal
ions removal by nanocellulose-based functionalized membranes with tailored pore-size
which form metal ions-cluster on membranes and help to remove metal ions in high
capacity (Patent application no. 1657849-9, AWAOATENT AB). Dr. Karim carried
out his postdoctoral research at KTH Royal Institute of Technology Stockholm,
Sweden, and before that he worked as a researcher at Luleå University of Technology,
Sweden. His research has been supported by some well-known funding agencies such as
the European Commission (NanoSelect under the 7th Framework Programme), Kempe
Foundation, Mistra Foundation (MisraTerraClean), R&D Council of Processum, etc. He
has given various plenary and invited talks related to his research interest in various
reputed institutes including the University of Oxford, Imperial College London, Indian
Institute of Technology Delhi, University of Guelph, EMPA-Swiss Federal Laboratory
of Material Sciences and Technology, etc. He attended various international and
national conferences/symposiums/workshops/invited lecturers around the world to pre-
sent his research works. He is a member of the International Association of Advanced
Materials, the Technical Association of Pulp and Paper Industry (TAPPI), and Sveriges
Ingenj€orer. Moreover, he organized two workshops at Lueå University of Technology
Luleå in 2013 and 2014. He was a member of the International Committee of Bioma-
terials for Tomorrow, B4T (2018) organized by Mahatma Gandhi University, India.
xviii Editors’ biographies

At the same time, he was also a member of an international advisory committee of

SafeBioPack (2018) organized by the Institute of Tropical Forestry and Forest Products
(INTROP), University Putra Malaysia. Currently, he is also serving as an external
expert/consultant of an Egypt-based company, inTEXive® (intexive.com). He has pub-
lished about 31 peer-reviewed research articles of international repute and 10 book
chapters with well-known publishers such as Elsevier, Springer Nature, CRC Press,
TAPPI Press, and World Scientific. He also won the best paper award in 2012, awarded
by Aufau Periodicals Publication USA.
Dr. Mohammad Jawaid is currently working as a Senior Fellow (Professor) at
Biocomposite Technology Laboratory, Institute of Tropical Forestry and Forest
Products (INTROP), University Putra Malaysia (UPM), Serdang, Selangor, Malaysia,
and also has been a Visiting Professor at the Department of Chemical Engineering,
College of Engineering, King Saud University, Riyadh, Saudi Arabia since June
2013. He has more than 20 years of experience in teaching, research, and industries.
His area of research interests includes hybrid composites, lignocellulosic reinforced/
filled polymer composites, and advanced materials: graphene/nanoclay/fire retar-
dants, modification and treatment of lignocellulosic fibers and solid wood, biopoly-
mers and biopolymers for packaging applications, nanocomposites and nanocellulose
fibers, and polymer blends. So far, he has published 40 books, 65 book chapters, more
than 350 peer-reviewed international journal papers, and several published review
papers in “Top 25 Hottest Articles” in Science Direct during 2013–19. He also
obtained two patents and six copyrights. He has Scopus H-index-55 (total citation
14171) and in Google scholar H-index-66 (total citation 20242). He is founding Series
Editor of the Composite Science and Technology Book Series from Springer Nature,
also Series Editor of Springer Proceedings in Materials, Springer Nature, and an
International Advisory board member of the Springer Series on Polymer and Compos-
ite Materials. He worked as a guest editor on special issues of SN Applied Science,
Frontiers in Sustainable Food Systems, Current Organic Synthesis and Current Ana-
lytical Chemistry, International Journal of Polymer Science, and IOP Conference
Proceeding. He is also an Editorial Board Member of the Journal of Polymers and
the Environment, Journal of Plastics Technology, Applied Science and Engineering
Progress Journal, Journal of Asian Science, Technology and Innovation, and Recent
Innovations in Chemical Engineering. In addition, he is a reviewer of several high-
impact international peer-reviewed journals of Elsevier, Springer, Wiley, Saga,
ACS, RSC, Frontiers, etc. Presently, he is supervising 12 Ph.D. students (five as Chair-
man, and seven as Member) and six Master’s students (one as Chairman, and five as
Member) in the fields of hybrid composites, green composites, nanocomposites, nat-
ural fiber-reinforced composites, nanocellulose, etc. Twenty-six Ph.D. and 13 Mas-
ter’s students graduated under his supervision in 2014–20. He has received several
research grants at university, national, and international levels on polymer composites
of around 3 million Malaysian ringgits (USD 700,000). He also delivered plenary and
invited talks in international conferences related to composites in India, Turkey,
Malaysia, Thailand, the United Kingdom, France, Saudi Arabia, Egypt, and China.
Besides that, he is a member of technical committees of several national and interna-
tional conferences on composites and material science. Recently, Dr. Mohammad
Editors’ biographies xix

Jawaid received the Excellent Academic Award in the Category of International

Grant-Universiti Putra Malaysia—2018 and also the Excellent Academic Staff Award
in Category of industry High Impact Network during Community and Industry Net-
work (ICAN 2019). Beside that he also won Gold Medal during JINM Showcase
under Community and Industry Network Category at Universiti Putra Malaysia,
Malaysia. He also received the Publons Peer Review Awards 2017, and 2018 (Mate-
rials Science), Certified Sentinel of Science Award Recipient—2016 (Materials
Science) and 2019 (Materials Science and Cross Field). He is the winner of the
Newton-Ungku Omar Coordination Fund: UK-Malaysia Research and Innovation
Bridges Competition 2015. Recently he was recognized with a Fellow and Chartered
Scientist Award from the Institute of Materials, Minerals and Mining (IOM), UK. He
is also a lifetime member of the Asian Polymer Association, and the Malaysian
Society for Engineering and Technology. He has professional membership of the
American Chemical Society (ACS) and the Society of Plastics Engineers (SPE), USA.
Preface

This book provides solid, quantitative descriptions and reliable guidelines, reflecting
the maturation and demand of the field and the development of new polysaccharide
nanocomposites. It focuses on the different types of polysaccharide nanocomposites
of cellulose, chitosan, alginate, etc. for gene delivery and tissue engineering. The book
also covers polysaccharide hydrogels for tissue engineering and polysaccharide mag-
netic nanocomposites for gene delivery and highlights the most exciting applications
in the field of gene delivery and tissue engineering. This book will be of interest to
researchers working in the fields of material science, biomaterials, regenerative med-
icines, drug delivery, tissue engineering, polymer science/chemistry, and chemical
engineering, and in the polymer industry. It will be useful for scientists working on
polysaccharide nanocomposites for gene delivery and tissue engineering. The book
will be very helpful for students in the development of new polysaccharide
nanocomposites as well as graduates in polymer technology, biomedical science,
and biotechnology.
The two introductory chapters cover basic information about polysaccharides and
nanocomposites, to provide a foundational understanding. The second section of this
book covers chitosan and its derivatives-based dimensional frameworks as carriers for
gene delivery, alginate- and hyaluronic-based hydrogels for tissue engineering,
heparin- and cellulose-based nanocomposites for and dextran, pullulan, gellan,
xanthan, and xanthun gum-based nanocomposites for tissue engineering applications.
The last section describes polysaccharide-based 3D bioprinter inks for tissue
engineering, polysaccharide-based nanocomposites for gene delivery, chitosan- and
starch-based nanocomposites for gene delivery, and hyaluronic acid magnetic
nanocomposites for gene delivery.
Finally, we assure the readers that the information provided in this book can serve
as a very important tool for anyone wishing to select/design polysaccharide-based
nanocomposites to fulfil the requirements of gene delivery and tissue engineering
applications. We are grateful to all the authors who contributed chapters to this book
and who helped to turn our thoughts into reality. Lastly, we are grateful to the Elsevier
team for their continuous support at every stage to make it possible to publish on time.
Showkat Ahmad Bhawani
Zoheb Karim
Mohammad Jawaid
Polysaccharides
Sadaf Afrina and Zoheb Karimb
a
Department of Chemistry, Faculty of Sciences, Aligarh Muslim University,
1
Aligarh, India, bMoRe Research AB, Sweden

1.1 Introduction
Natural polysaccharides are generally constituted of simple carbohydrates with long
chain molecules attached via glycosidic linkages. Carbohydrates are made up of alde-
hydes or ketones, and have multiples hydroxyl groups. They make up the majority of
organic matter and play a crucial role in human life. Carbohydrates serve as energy
stores, fuel, and various metabolic intermediates. In parallel, various sugars like ribose
and deoxyribose form part of the structural framework of RNA and DNA. Polysac-
charides are also present in the cell walls of bacteria and plants, and cellulose is
the main constituent of plant cell walls [1, 2].
The most authentic classification performed for polysaccharides is based on degree
of polymerization (DP). Polysaccharides may reach a DP of 105, whereas, by defini-
tion, the maximum DP for oligosaccharides is 10. Hence, by convention, compounds
having a DP of 11 or more are designated polysaccharides. A monosaccharide is a
monomer of saccharides, and a disaccharide is two conjugated monomers [1, 3].
Many studies are available related to the general introduction of polysaccharides,
their chemical structures, chemical compositions, methods of isolation, types, etc.
[4–7]. Hence, in this chapter, a very interesting aspect is explained: the nano-form of
these polysaccharides, which is an area of emerging interest in scientific studies. Very
few studies in the literature have been summarized related to nano-polysaccharides
characterization, the isolation process, and their properties and applications. Conse-
quently, this chapter explains the collective efforts and provides a summary of
nano-polysaccharide understanding and their utilization in various applications.

1.2 Natural polysaccharide nanomaterials

Polysaccharides are molecules composed of monomer sugar units linked by glyosidic
bonds; this bond is responsible for the attachment of monomers and forms polymers.
They are the most abundant natural polymer on Earth [8, 9]. The amount of polysac-
charide produced by plants per year exceeds by several orders of magnitude the syn-
thetic production by the chemical industry [10]. Natural polysaccharides (Table 1.1)
are synthesized to fulfill many different functions, such as energy storage in plants
(i.e., starch), structural support of vegetal cells (i.e., cellulose), and gelling agents for-
ming the intercellular matrix and containing several ions such as sodium, calcium, and

Polysaccharide-Based Nanocomposites for Gene Delivery and Tissue Engineering

https://doi.org/10.1016/B978-0-12-821230-1.00010-4
Copyright © 2021 Elsevier Inc. All rights reserved.
2 Polysaccharide-Based Nanocomposites for Gene Delivery and Tissue Engineering

Table 1.1 Natural polysaccharides classified according to their origin.

Origin Polysaccharide

Plants Starch, cellulose, glucomannan, pectin, hemicellulose, gums, mucilage

Algae Agar, galactans, alginate, carrageenans
Animals Chitin, chitosan, hyaluronic acid, glycosaminoglycans, cellulose
Bacteria Dextran, levan, polygalactosamine, gellan, xanthan, cellulose
Fungi Elsinan, chitin, chitosan, pullulan, yeast glucans

Reproduced with permission from Torres FG, Troncoso OP, Anissa P, Francesca G, Giuseppe B. Natural polysaccharide
nanomaterials: an overview of their immunological properties. Int J Mol Sci 2019;20:5092–9.

magnesium (i.e., alginate in the brown algae). Some of them, such as cellulose, starch,
chitin, and polysaccharides from seaweeds, are commercially important in several
markets, ranging from paper production to food industry products. We will restrict
our description to those polysaccharides and their interactions with the immune sys-
tem. In this chapter, we shall discuss some of them in detail.

1.3 Polysaccharides in nano-form

It is well-known that controlling the size of polysaccharides is the biggest challenge in
current scientific study. At the time of writing, there is no universally accepted size
range that particles must fall within to be classified as microparticles or nanoparticles.
However, many researchers classify particles smaller than 1 μm as nanoparticles and
particles larger than 100 mm as macroparticles [11–23]. Nanoparticles are then a sub-
micronic colloidal system, ranging in size from a few nanometers to 1 μm, and made
of artificial or natural polymers [24]. Nanoscale typically has a greater surface area per
mass. The nanometric size effect has led to intensive research in the area of nano-sized
particles. The high surface area of nano-size particles opens various routes for novel
interactive behavior with polymers/particles. Hence, this chapter will also discuss the
novel interactive behaviors of nano-sized polysaccharides with other species. Various
origins of polysaccharides are given in Table 1.1, thus in this chapter, the nano-forms
of possible listed polysaccharides will be explained in detail [25].

1.4 Nano-starch
Starch is a polymer of carbohydrates consisting of numerous glucose units joined by
glycoside bonds. Starch nanoparticles (St-NPs) have attracted much attention due to
their unique properties that are different significantly their bulk material [26]. In a
study, St-NPs were extracted from starch granules by means of physical or chemical
treatments. Utilization of chemical treatments for preparation of St-NPs has received
more attention than physical treatments [25]. Isolation of St-NPs has been reported in
various scientific studies [27–29] and the fabricated nanoparticles are in the shape of
5–7 nm thick, 20–40 nm long, and 15–30 nm. In a study, St-NPs were synthesized
Polysaccharides 3

with native starch using a solvent displacement method modified with an aqueous
alkaline medium as a solvent and ethanol as the organic phase [30], as shown in
Fig. 1.1.
Surface modifications of St-NPs have also been performed using various chemis-
tries, in a study, surface of waxy maize starch isolated using H2SO4 hydrolysis was
performed using two different reagents, namely, alkenyl succinic anhydride and phe-
nyl isocyanate and various sophisticated techniques are employed for the character-
ization of modified St-NPs [31]. Furthermore, St-NPs with high specific area and
surface hydroxyl groups have good potential for surface chemical modification to
introduce any desired surface functionality. The accessible hydroxyl group content
on the surface of St-NPs was reported to be around 14% of the total hydroxyl group
in polymer structure, which is around 0.0025 mol/g of St-NPs [31, 32]. The main
reported surface modification is grafting different reagents on the surface of the par-
ticles [33, 34] to prepare new material for different applications, especially in blends
with polymers.

1.5 Nanocelluloses
Our group has been working with various kinds of functionalized nanocelluloses for
many years. As discussed and reported in various scientific studies, the nano-form of
cellulose exists mainly as crystalline cellulose (CNC) and cellulose nanofibers (CNFs)
[22–24]. In our study, we have also used an integrated route of bioethanol for the iso-
lation of crystalline cellulose having highly negative surface ζ-potential compared to
sludge-based crystalline cellulose in acidic pH (pH 5.0) (Fig. 1.2) [14].
Surface modification of nanocellulose is performed using various chemistries, and
very detailed report is published early [35]. In our lab, a green approach using enzyme
hexokinase was studied for the introduction of phosphate (PO4 2 + ) groups on the sur-
face of cellulosic fibers [21, 22]. Furthermore, in situ TEMPO functionalization of

Fig. 1.1 Transmission electron microscopy (TEM) of fabricated St-NPs: (A) the histogram
represents the size distribution of fabricated nanoparticles; (B) using dynamic light
scattering (DLS).
4 Polysaccharide-Based Nanocomposites for Gene Delivery and Tissue Engineering

Fig. 1.2 Isolation routes of cellulose in nano-forms, crystalline cellulose (CNC), cellulose
nanofibers (CNFs), and highly functional crystalline cellulose called bioethanol CNC (CNCBE)
are reported in this chapter. Available functional groups on the surface of isolated celluloses are
denoted on the right-hand side.
Reproduced with permission from Mathew AP, Oksman K, Karim Z, Liu P, Khan SA, Naseri
N. Process scale up and characterization of wood cellulose nanocrystals hydrolysed using
bioethanol pilot plant. Ind Crop Prod 2014;58:212–19.

Fig. 1.3 In situ TEMPO functionalization of crystalline cellulose isolated using a bioethanol
integrated route. High surface charge density and more negative ζ-potential indicate highly
effective reactions.

CNCBE-based dimensional structures was also reported, where mild acid-based intro-
duction of carboxylic groups (COO
) was performed to produce highly active and
effective dimensional frameworks [11]. Fig. 1.3 represents the possible catalytic
reaction for the in situ TEMPO oxidation of cellulose-based dimensional structures.
Polysaccharides 5

The functionalization of produced frameworks indicates the catalytic transformation

of hydroxyl groups to carboxylic groups after mild acid reactions. Furthermore, more
negative ζ-potential (
32 to
65 mV) and high density of carboxylic groups
(36–321 mmol/kg) was reported after functionalization [11].

1.6 Nano-alginate
Alginate is a naturally occurring anionic polymer typically obtained from brown sea-
weed. It has been extensively investigated and used for many biomedical applications
due to its biocompatibility, low toxicity, relatively low cost, and mild gelation by addi-
tion of divalent cations such as Ca2+ [36]. Alginate gels are also promising for cell
transplantation in tissue engineering, which aims to provide man-made tissue and
organ replacements to patients who suffer the loss or failure of an organ or tissue
[37]. In this approach, hydrogels are used to deliver cells to the desired site, provide
a space for new tissue formation, and control the structure and function of the
engineered tissue [38].
Alginate-based nanoparticles have been widely used in various applications [39,
40]. Two promising applications discussed in the literature are targeted drug delivery
and tissue engineering, and due to their nontoxic nature, high surface area of
nanoparticles (high loads of drugs and other active molecules), and effective biocom-
patibility, alginates in nano-forms are an effective candidate in these research fields
[41–44]. In a study, a novel procedure for the fabrication of alginate nanoparticles in
the aqueous phase of water-in-oil (w/o) nanoemulsion was developed and
reported [45].
Moreover, the hybrid of alginate nanoparticles with metals nanoparticles/metals is
also reported in the literature. In a study, stable nanoparticles of alginate were fabri-
cated in mild acid conditions, and stability was further increased after crosslinking
with zinc [46]. Fig. 1.4 provides a complete step-by-step production of alginate-zinc
nanoparticles.

1.7 Nano-pectin
Pectin is a natural polysaccharide structurally composed of large amounts of poly-D-
galacturonic acid bonded via α-1,4-glycosidic linkage. According to its degree of
methyl esterification, pectin can be classified as high methoxyl pectin or low methoxyl
pectin, which yields some differences in its properties [47, 48]. Pectin has commonly
been used as a gelling agent, a thickening agent, and a colloidal stabilizer in the food
industry [49]. Its applications in the pharmaceutical industry have been increased in
the last decade [50–52].
Nanoparticles of pectin are prepared using mechanical homogenization for disso-
lution enhancement of itraconazole [53] and further used for drug loading. It has been
concluded that nanoemulsions were achieved when chloroform was used as an
internal phase. The obtained emulsion was freeze-dried to obtain solid porous
Fig. 1.4 Step-by-step fabrication of alginate-zinc nanoparticles. Highly stable alginate nanoparticles were prepared using zinc-based cross-linking.
Reproduced with permission from Pistone S, Qoragllu G, Smistad M, Hiorth. Formulation an preparation of stable cross-linked alginate-zinc
nanoparticles in the presence of a monovalent salt. Soft Matter 2015;11:5765–74.
Polysaccharides 7

nanoparticles, and further various sophisticated analytical techniques were applied for
characterizations. Scanning electron microscopy and optical images of produced
nanoparticles are shown in Fig. 1.5. Furthermore, in a study, citrus-derived pectin
nanoparticles were produced via the ionotropic gelation method, using magnesium
(Mg2+) as the divalent cross-linker [54].

1.8 Nano-chitin and chitosan

The most abundant natural polymer available in the world is cellulose [22], with chitin
and chitosan in second position [9]. The sources used for the isolation of chitin or
chitosan are marine crustaceans, shrimp, and crabs. A study reported that about
1000 tons per year (70% of total) of these polymers comes from marine species.
Chitosan is mainly produced by deacetylation or removing of acetyl groups from

Fig. 1.5 Optical images (left-hand column and middle column) and SEM images
(right-hand column) of particles prepared from caprylic/capric triglyceride (CCT)-based and
chloroform-based nanoemulsion using (A) high methoxy pectin, (B) amidated LM pectin
with degree of esterification 29, and (C) low methoxy pectin (degree of esterification 38).
Reproduced with permission from Burapapadh K, Takeuchi H, Sriamornsak, P. Development
of pectin nanoparticles through mechanical homogenization for dissolution enhancement
of itraconazole. Asian J Pharm Sci 2016;11:365–75.
8 Polysaccharide-Based Nanocomposites for Gene Delivery and Tissue Engineering

the chitin by alkali treatment [55]. However, deacetylation of chitin is almost never
complete, and chitosan or deacetylated chitin still contains acetamide groups to some
extent. Unlike cellulose, chitin and chitosan contain 5%–8% N2, which in chitin is in
the form of acetylated amine groups and in chitosan is in the form of primary aliphatic
amine groups; this makes chitin and chitosan suitable for typical reaction of amines
[56]. However, chitosan is chemically more active than chitin due to the presence of
primary and secondary hydroxyl groups on each deacetylated unit (Fig. 1.6). These
reactive groups are readily subject to chemical modification to alter the mechanical
and physical properties of chitosan.
The synthesis of chitosan nanoparticles in the presence of sodium tripolyphosphate
(TPP) as a reducing agent was discussed [57]. In another study, eco-friendly spherical
chitin nanoparticles were synthesized using oxidative Oxone. This versatile-triple salt
was effective in the cleavage of glycosidic bonds and etching out individualization of
elementary fibrils to form chitin nanoparticles and selective oxidation occur at C6
hydroxyl of glucose molecule submit to form carboxylic acid [58]. In a very interest-
ing study, the size of chitosan-TPP nano and microparticles has been controlled using
various molar ratios of chitosan and TPP. Furthermore, the degree of acetylation (DA)
is reported to be the second most influential parameter of tuning the size of produced
nano/micro particles [59]. The sizes of produced nanoparticles are shown in Fig. 1.7.
In the same study, in which very interesting results were discussed, the authors
explored the influence of chitosan concentrations on the hydrodynamic radii of fab-
ricated nanoparticles, as shown in Fig. 1.8. A high concentration of chitosan is respon-
sible for the greater size of produced chitosan nanoparticles.

Fig. 1.6 Structure relationship between chitin and chitosan.

Reproduced with permission from Islam S, Bhuiyan MAR, Islam MN. Chitin and chitosan;
structure, properties and applications in biomedical engineering. J Polym Environ
2017;25:854–66.
Polysaccharides 9

Fig. 1.7 TEM micrographs of chitosan-TPP particles prepared from chitosan samples at
varying DA and concentration with a NH2/PO4 molar ratio of 1.5 for DA 20% and 50%, and
with a NH2/PO4 molar ratio of 1 for DA 35%: (A) DA 20% at 0.5 mg/mL chitosan
concentration; (B) DA 20% at 2.5 mg/mL chitosan concentration; (C) DA 20% at 5 mg/mL
chitosan concentration; (D) DA 35% at 0.5 mg/mL chitosan concentration; (E) DA 35% at
2.5 mg/mL chitosan concentration; (F) DA 35% at 5 mg/mL chitosan concentration; (G) DA
50% at 0.5 mg/mL chitosan concentration; (H) DA 50% at 2.5 mg/mL chitosan concentration;
(I) DA 50% at 5 mg/mL chitosan concentration. Scale bars (A, D, G) ¼ 200 nm; (B, E, H) ¼
500 nm; (C, F, I) ¼ 1 μm.
Reproduced with permission from Sreekumar S, Goycoolea FM, Moerschbacher BM, Rivera-
Rodriguez GR. Parameters influencing the size of chitosan-TPP nano and microparticles. Sci
Rep 2018;8:4695.

Physical modifications of these polymers can be achieved by blending, which

involves physical mixing of two or more polymers. It is one of the oldest and easiest
modification approaches. Some common hydrophilic polymers that can be blended
with chitosan to achieve oral drug delivery are poly (vinyl alcohol) (PVA), poly (vinyl
pyrrolidone) (PVP), and poly (ethyl oxide) (PEO). Blending of chitosan and PVA
improves the mechanical (tensile strength) and barrier properties (water vapor
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