Natural polymers in TE

applications
 Introduction
• Natural polymers are essential in so many life processes.
• First to be used as scaffold materials for tissue regeneration, in tissue engineering
applications because they are either components of, or have properties similar to, the
natural ECM.
• Natural polymers -renewable resources- plants, animals and microorganisms- widely
distributed in nature.
• Exhibit a large diversity of unique and complex structures, different physiological
functions- offer a variety of potential applications in the field of tissue engineering
due to their various properties - pseudoplastic behavior, gelation ability, water
binding capacity, biodegradability, among many others.
 Introduction cont..
• In addition, they possess many functional groups (amino, carboxylic and hydroxyl groups)
available for chemical (hydrolysis, oxidation, reduction, esterification, etherification,
cross-linking reactions, etc.) and enzymatic modification and/or conjugation with other
molecules, which allows a variety of products with tailorable chemistries and properties
to be obtained.
• The creation of hybrid materials– by means of combining the advantages of different
natural polymers– may constitute a useful approach to mimicking the natural
environment of the ECM and to obtain scaffolding materials with superior mechanical and
biological properties.
 Introduction cont..
• Living organisms are able to synthesize a vast variety of polymers which can be divided into
eight major classes according to their chemical structure:
(1) polysaccharides,
(2) proteins and other polyamides,
(3) polyoxoesters (polyhydroxyalkanoic acids),
(4) polythioesters,
(5) polyanhydrides (polyphosphate),
(6) polyisoprenoids,
(7) lignin, and
(8) nucleic acids.
 1. Polysaccharides
• Polysaccharides, also known as glycans, consist of monosaccharides (aldoses or ketoses)
linked together by O-glycosidic linkages.
• Each monosaccharide is classified according to number of carbons in monosaccharide
chain (usually 3 to 9).
• Polysaccharides can be classified as homopolysaccharides or heteropolysaccharides if
they consist of one type or more than one type of monosaccharide.
• Because glycosidic linkages can be made to any of the hydroxyl groups of a
monosaccharide, polysaccharides form linear as well as branched polymers.
• Differences in the monosaccharide composition, linkage types and patterns, chain shapes,
and molecular weight, dictates their physical properties, including solubility, flow behavior,
gelling potential, and/or surface & interfacial properties.
• In the living organisms, polysaccharides perform a range of biological functions, such as
maintenance & structural integrity (cellulose, chitin), energy reserve storage (starch,
glycogen) & biological protection and adhesion (gum exudates, extracellular microbial
polysaccharides).
Alginate:
• Alginate is a biological material derived from sea algae, composed of linear block
copolymers of 1-4 linked β-D-mannuronic acid (M) and α-L-guluronic acid (G) -
Divalent ions form cross-links by binding the guluronic residues, inducing a sol–gel
transition in the material.
• Alginate can be ionically cross-linked by the addition of divalent cations (like Ca2+) in
aqueous solution.
• The gelation and cross-linking of the polymers are mainly achieved by the exchange of
sodium ions from the guluronic acids with the divalent cations, and the stacking of
these guluronic groups to form the characteristic egg-box structure- and they impart
viscoelastic solid behavior to the material.
• The properties of alginate derive from this behavior, and include: a relatively inert
aqueous environment within the matrix; a high gel porosity which allows for high
diffusion rates of macromolecules; the ability to control this porosity with simple
coating procedures.
Alginate:
• Alginate has also been widely studied for engineering liver tissue.
• The bioartificial liver- assist device substitutes for liver tissue engineering requires a
suitable ECM for hepatocyte culture because hepatocytes are anchorage- dependent cells,
highly sensitive to ECM milieu for maintenance of their viability and differentiated
functions.
• A potential approach to facilitate the performance of implanted hepatocytes is to enable
their aggregation and re-expression of their differentiated function prior to implantation
and alginate has been shown to allow hepatocyte culture and function.

Egg-box structure
Dextran:
• Dextran is a bacterial-derived polysaccharide, consisting essentially of a-1,6 linked D-
glucopyranose residues with a few percent of a-1,2-, a-1,3-, or a- 1,4-linked side
chains synthesized from sucrose by Leuconostoc mesenteroides streptococcus.
• Dextran hydrogels can be created by either physical or chemical cross-linking, taking
advantage of the hydroxyl groups present on the a-1,6-linked D-glucose residues.
• Dextran particles have been widely used as separation matrices, such as Sephadex, as
cell microcarriers, such as Cytodex, and as drug-delivery vehicles.
• There has been considerable interest on dextran scaffolds for tissue engineering
applications.
• The possibility of having an injectable, in-situ -gellifying material than can serve as a
filler and template for the regeneration/ repair of tissues such as cartilage is a very
attractive one.
• Alginate and dextran have shown and continue to show excellent properties for this
purpose.
• Because of their biocompatibility, abundance and low prices, they have been widely
used in the food industry as thickeners and emulsifying agents.
Chitosan:
• Chitosan is a semi-crystalline polymer and the degree of crystallinity is a function of the
degree of deacetylation.
• Crystallinity is maximum for both chitin (i.e. 0% deacetylated) and fully deacetylated
(i.e.100%) chitosan.
• It is a linear polysaccharide of (1-4) linked d-glucosamine and N-acetyl-d–glucosamine
residues derived from chitin, a high molecular weight, the second most abundant natural
biopolymer commonly found in arthropod exoskeletons such as shells of marine
crustaceans and cell walls of fungi.
• Chitosan has been proven to be biologically renewable, biodegradable, bioadhesive &
biocompatible used in wound dressing and healing and various tissue engineering
applications.
• The cationic nature of chitosan is primarily responsible for electrostatic interactions
with anionic GAGS, proteoglycans and other negatively charged molecules.
• This property is of great interest because a large number of cytokines/ growth factors are
linked to GAG, and a scaffold incorporating a chitosan GAG complex may retain and
concentrate growth factors secreted by colonizing cells.
Chitosan-based scaffolds for different tissue engineering applications
Cellulose:
• Cellulose is the main component of plant cell walls.
• It also constitutes the most abundant, renewable polymer resource available today, existing
mainly as lignocellulosic material in forests, with wood being the most important source.
• Primary structure of linear polymer consists of up to 15,000 D-glucose residues linked by
(1-4)-glycosidic bond.
• The fully equatorial conformation of β -linked glucopyranose residues stabilizes the chain
structure, minimizing its flexibility.
• It is the ability of these chains to hydrogen-bond together into fibres (microfibrils) that
gives cellulose its unique properties of mechanical strength and chemical stability, leading
also to insoluble materials with small degradability in-vivo.

Structure of cellulose
Cellulose:
• The biodegradability of cellulose is considered to be limited, if it occurs at all, because of
the absence of hydrolases that attack the (1, 4) linkage.
• Cellulose and its derivatives have been employed with success as biomaterials - an
adequate source for tissue engineering applications - In orthopaedic applications, it has
been shown that cellulose sponges could support bone tissue ingrowths, suggesting that it
could be used in bone tissue engineering
• Bacterial cellulose (BC) is a biotechnological method for producing pure nanofibrilar
cellulose structures that have high mechanical strength, high water content and high
crystallinity.
• BC is excreted extracellularly by the Acetobacter xylinum & the pellicle formed has
been used in TE related applications.
Starch:
• Starch is the dominant carbohydrate reserve material of higher plants- leaf
chloroplasts, amyoplasts of storage organs such as seeds and tubers.
• Most of the starch utilized world-wide comes from a relatively small number of crops-
corn, potato, wheat, tapioca, rice, sorghum, sweet potato, arrowroot, sago, and mung
beans.
• One of the cheapest biopolymers, totally biodegradable into carbon dioxide and
water, abundantly available.
• Native starch is composed of granules of variable sizes and shapes depending on the
source of the starch.
• Chemically starch is a polysaccharide consisting only of homoglucan units.
Starch:
• One of the most important properties of native starch is its semi-crystallinity -
depending on the source & moisture content, the degree of crystallinity ranges
between 15% to 50%- crystallinity is due to amylose & amylopectin, but mostly
depends on amylopectin.
• To make a thermoplastic starch (TPS) that can be processed by conventional
processing techniques - extrusion or injection moulding, it is necessary to disrupt the
granule and melt the partially crystalline nature of starch in the granule by adding
plasticizers - water & polyols such as glycerol and glycol.
• A wide range of starch based scaffolds have been developed exhibiting different
properties and porous architectures.
Use of starch based polymers in tissue engineering research
Hyaluronan:
• Hyaluronan(HA), is a natural and highly hydrophilic polysaccharide, - a key
constituent of native ECM & tissues.
• Found naturally in the ECM of skin, cockscomb, cartilage, vitreous humor, and other
body tissue and plays a role in the movement and proliferation of cells.
• HA belongs to the family of glycosaminoglycans and is synthesized as a large, negatively
charged and linear polysaccharide of varying chain length (2 - 25 μm ) composed of
repeating disaccharide units.
• Interactions between HA with other ECM macromolecules and chondrocytes, its
hydrodynamic characteristics, especially its viscosity and ability to retain water are critical
for the maintenance of both cartilage homeostasis & biomechanical integrity.
• The characteristics of HA are, to a great extent, responsible for the regulation of the
porosity and malleability of these matrices.
• HA is cleaved by enzymes called hyaluronidases, showing that the cells of the host or the
ones present in the engineered tissue, may regulate the local clearance of the material,
while the new tissue is being formed.
• HA is biocompatible and has a greater bacteriostatic effect when compared with other
matrices namely, collagen type I, poly(lactide-co-glycolide) (PLGA) & hydroxyapatite.
Hyaluronan and its derivatives in tissue engineering
2. Proteins
• Proteins are the most abundant organic molecules within the cell extracellular and
intracellular medium, where they ensure multiple biological functions such as transport,
regulation of pathways, protection against foreign molecules, structural properties,
protein storage, as well as being the catalyst for a great diversity of reactions, acting as
biocatalysts.
Collagen:
• Collagen is the most abundant protein in mammalian tissues (cornea, blood vessels, skin,
cartilage, bone, tendon and ligament) and is the main component of ECM.
• The individual triple helices are arranged to form fibrils which are of high tensile strength
and can be further assembled and cross-linked (collagen fibrils are stabilized in the
ECM by the enzyme lysyl oxidase).
• In tissues that have to resist shear, tensile, or pressure forces, such as tendons, bone,
cartilage, and skin, collagen is arranged in fibrils, with a characteristic 67 nm axial
periodicity, which provides the tensile strength.
2. Proteins cont..
• Only collagen types I, II, III, V, and XI self-assemble into fibrils.
• The fibrils are composed of collagen molecules, which consist of a triple helix of
approximately 300 nm in length and 1.5 nm in diameter.
• 1970s and 1980s - academics and commercial researchers began to use collagen as a
biomaterial in a variety of connective tissue applications because of its excellent
biocompatibility, low antigenicity, high biodegradability and good haemostatic and cell-
binding properties.
• These enzymes are secreted by neutrophils during the foreign body reaction, allowing the
collagen degradation to be controlled by the cells present at the implantation site.
• Collagen is naturally degraded by matrix metalloproteinases, specifically collagenase and
serine proteases.
Application of collagen scaffolds in tissue engineering
research.
Elastin:
• Elastin is another of the key structural proteins found in
the ECM of connective tissues that need to stretch and
retract following mechanical loading and release.
• It is found predominantly in the walls of arteries, lungs,
intestines and skin, as well as other elastic tissues.
• However, unlike type I collagen, elastin has found little
use as a biomaterial, due to two main reasons:
(i) elastin preparations have a strong tendency to calcify
upon implantation, and
(ii) the purification of elastin is complex.
• Recombinant protein technologies have allowed the
synthesis of well-defined elastin-derived polypeptides.
• Elastin-like polypeptides (ELPs) are artificial
polypeptides with unique properties that make them
attractive as biomaterial for tissue-engineering.
Application of elastin based scaffolds in tissue engineering
research
Soybean:
• Soybeans can be processed into three kinds of protein-rich products: soy flour, soy
concentrate and soy isolate.
• Soy protein, the major component of the soybean (30 - 45%) is readily available from
renewable resources, is economically competitive, and presents good water resistance as
well as storage stability.
• About 90 - 95% of the soy is storage protein, with two subunits, namely 35% conglycinin
(7S) and 52% glycinin.
• The combination of its properties with a similarity to tissue constituents and a reduced
susceptibility to thermal degradation, makes soy an ideal template for use in
biodegradable polymer for biomedical applications.
• Membranes, microparticles and thermoplastics-based soy materials have been developed
for tissue regeneration.
• Biodegradable soy plastics have been developed by melt-based methods such as
extrusion and injection moulding.
Silk fibroin:
• Silk fibroin is a highly insoluble fibrous protein produced by domestic silk worms
(Bombyx mori) containing up to 90% of the amino acids glycine, alanine, and serine
leading to antiparallel- pleated sheet formation in the fibers.
• Fibroin is a structural protein of silk fibers and sericins are the water-soluble glue-like
proteins that bind the fibroin fibers together.
• High purity silk fibroin fiber can be obtained easily from degummed silk (boiling-off) -
partial or complete removal of the sericin- removal of the sericin coating before use
removes the thrombogenic and inflammatory response of silk fibroin.
Silk fibroin:
• Bombyx mori silk fibroin can be dissolved with neutral salt solutions such as lithium
bromide (LiBr), lithium thiocyanate (LiSCN), hexafluroisopropryl alcohol (HFIP) and
calcium nitrate-methanol [Ca(NO 3 ) 2 -MeOH].
• Their mixtures are dialyzed to get pure fibroin solution, which can be used to prepare silk
fibroin membranes, fiber, hydrogel, scaffolds.
• Traditionally, silk fibroin has been used for decades as suture material.
• Applications of silk fibroin are related to its permeability to oxygen & water, cell
adhesion & growth characteristics, slow degradability, low inflammatory response, and
high tensile strength with flexibility.
• Porous three-dimensional scaffolds with silk fibroin have been obtained using various
processing techniques- salt leaching, electro-spinning, freeze-drying & gas-foaming
processing.
Application of soy and silk based materials in tissue engineering
studies
Polyhydroxyalkanoates:
• Polyhydroxyalkanoates (PHAs) are naturally occurring biodegradable polymers.
• PHAs are synthesized and stored as water-insoluble inclusions in the cytoplasm of several
bacteria and used as carbon and energy reserve materials.
• The first PHA to be identified was poly(3-hydroxybutyric acid) (P[HB]).
• This homopolymer is the most abundant bacteria synthesized polyester and its 3-
hydroxybutyrate (HB) monomer was thought to be the unique PHA constituent in
bacteria.
• The introduction of other units in the PHA chain (besides 3HB) has a significant effect
on mechanical behavior of the polyester.
• The homopolymer of PHB is a brittle material, while the increase in 3-
hydroxyvalerate(HV) content turns the HB-co-HV copolymer more ductile.
• The wide range of mechanical properties coupled with the biodegradable &
biocompatible behaviors of PHAs makes them potential biomedical candidates including
drug delivery and tissue engineering.
Thank you