Preparation of chitosan
The purified chitin is subjected to a deacetylation process (Figure 2).
To this end, the CI is treated with a concentrated base, most often
NaOH,21 and then washed with water and dried. The
chitosan thus obtained is in the form of a white
powder.13
In the deacetylation process random acetyl groups are removed in favour
of the amino group, chitosan is a copolymer ofN– acetyl–D– glucosamine and D–glucosamine units. The
presence of the latter groups depends on the degree of deacetylation
(DD) that is used to characterise the CS. The more deacetylated the CS
is, the fewer N– acetyl–D– glucosamine groups. The value
of the DD of chitosan is in the range of
50–95%.17,28
Physicochemical properties
In its structure, chitosan contains functional groups: hydroxyl, amine
and acetyl. They have a significant impact on the chemical properties of
the polymer and the nature of interactions. The presence of hydrophobic
acetyl groups and hydrophilic amine and hydroxyl groups ensures the
amphiphilic nature of the polymer.13 The amine groups
that are responsible for the cationic nature of the polymer result in a
high surface tension of aqueous solutions of this
polysaccharide.29
CS is insoluble in water, in popular organic solvents (including
methanol, ethanol, isopropanol, toluene), however, it dissolves in
dilute aqueous acid solutions, e.g. in hydrochloric acid, acetic acid,
formic acid, citric acid.30–32 This property is due
to the presence of free amino groups that can protonate at a pH below
6.33 The amine groups become positively changed,
breaking the hydrogen bonds and allowing the polymer to dissolve. Above
this pH value, the –NH3+ groups
deprotonate, lose their charge, and the polymer solubility
decreases.16,34 The results of the solubility tests
are presented in Table 1.30
The solubility of chitosan also depends on the degree of deacetylation
(DD) and the molecular weight of the polysaccharide. The more amino
groups there are, i.e. the higher the deacetylation degree, the better
the chitosan solubility.23 The higher the molecular
weight of the polymer, the more internal hydrogen bonds there are and
the more difficult it is to dissolve.17 CS forms
water-soluble salts, including formates, acetates, hydrochlorides,
citrates.35 To characterise chitosan, its viscosity is
often used, which depends on various parameters such as concentration of
chitosan in the solution, temperature, degree of
deacetylation.36
The cationic nature of chitosan distinguishes it from other natural
polysaccharides. Due to its positive charge, chitosan has the ability to
form complexes with negatively charged compounds, e.g.
polylactide37, poly(glutamic
acid)38, DNA39,40,
collagen41, alginates42–44,
hyaluronic acid45–47, pectin48,49and many others.
The structure of chitosan, which is based on D– glucosamine andN– acetyl–D– glucosamine units, is similar to the
structure of naturally occurring glycosaminoglycans (GAG). GAG can build
the extracellular matrix and perform various functions in organisms,
e.g. they affect adhesion, growth factors and cell receptors, water
retention (influencing the resistance of cells to compression). It can
be expected that, thanks to its GAG-like structure, chitosan can also
interact with animal cells.7,34
Chitosan is readily used in tissue engineering because it is
characterised by features desired in biomaterials: biocompatibility,
non-toxicity, the ability to biodegrade into non-toxic compounds and
bioactivity.50–52 It also demonstrates
osteocompatibility (the presence of chitosan does not adversely affect
bone regeneration) and osteoconduction (the ability to connect with bone
tissue and stimulate its growth).13 CS also has
mucoadhesive properties, i.e. it promotes the adhesion of animal mucus
cells to the biomaterial. This property is due to the difference in
charges between positive chitosan and negative mucus, between which
electrostatic interactions are generated.53,54 CS is
also credited with the ability to inhibit the growth of neoplastic
cells55,56 and promote wound
healing.57–60 Chitosan also has antibacterial and
antifungal properties (against Gram-positive 61 and
Gram-negative bacteria62). These properties are
probably due to the interaction between the positively charged polymer
and negatively charged lipopolysaccharides or proteins in microbial
membranes.59 Interactions with the surface of the
bacterial cell reduce the permeability. The antibacterial properties of
chitosan may also result from the interaction with the DNA of the
bacterial cell, which results in inhibition of RNA
synthesis.50,59,63
Chitosan degradation
Chitosan is a natural polymer which decomposes under the influence of
both chemical and physical factors and enzymes. Chemical factors include
depolymerisation under the influence of strong acids, e.g. HCl,
HNO2, H3PO4,
CH3COOH,64–66 strong
oxidants67 or free radicals.68 The
products of such reactions are chitosan oligosaccharides with various,
low molecular weights.69 Chemical degradation is a
fast process, but expensive, poorly controlled (random cutting of the
polymer chain70) and highly polluting towards the
environment due to the chemical compounds
used.55,65,71
Physical methods of CS depolymerisation include, among others, γ
radiation, ultrasound. Contrary to chemical depolymerisation, mechanical
cuts occur more frequently inside the polymer,72 and
the obtained oligosaccharides are characterised by a narrower size
distribution.69 Gamma radiation can be used to
sterilise chitosan products. It was examined that under the influence of
gamma radiation the CS structure changes, there is a decrease in
molecular weight already at 15 kGy, which increases with radiation
dosage.73 Such changes may affect the properties of
the polymer and, consequently, the chitosan-containing medical device.
The use of ultrasound is a popular solution due to its low environmental
impact. Degradation of chitosan by ultrasound is based on mechanical
phenomena and depends on wave intensity, temperature, polymer
concentration, ionic forces.74 There are also studies
on CS depolymerisation using the cavitation
process.69,75,76
The use of enzymes for the depolymerisation of CS is associated with
carrying out the reaction in mild conditions and fairly good control of
the process. It is also an environmentally friendly
method.64,65 The enzyme specific chitosanase can be
used in the hydrolysis of β‑1,4‑glycosidic bonds, as well as
non-specific chitinases77, lysozym78or cellulases.71 The reduction of the CS molecular
weight under the influence of the protease enzyme changed the structure
of the oligomer, but did not affect the deacetylation
degree.79 Proteases can also be used for the
production of monomers (N– acetyl–D– glucosamine
units).80 The use of lipase, the enzyme present in the
human body, in CS depolymerisation leads to products with different
molecular sizes in the 30 – 50 kDa range, without major structural
changes to the polymer.81
Metabolism in living organisms
A biodegradable scaffold used in bone regeneration should degrade in the
body into known, non-toxic products that can be excreted from the body
or incorporated into the body’s natural metabolic cycles. The time of
scaffold degradation in the body depends on the place of implanting and
the patient (their age, chronic diseases), but on average in bone
regeneration it should take from 3 to 9 months.82Hydrolysis of glycosidic bonds in chitosan leads to the production of
non-toxic glucosamine oligosaccharides, which can be rapidly cleared
from the body in urine.83,84 Chitosan implants do not
elicit a long-lasting inflammatory or immune
response.52 The inflammation observed after the
introduction of chitosan scaffolds depends on the degree of
deacetylation of the polysaccharide. Chitosan with a lower DD induced a
stronger leukocyte response.85 The rate of degradation
of chitosan implemented in the form of microspheres, hydrogels or 3D
scaffolds varies. The onset of degradation of chitosan in the form of
microspheres was observed 4 weeks after introduction into the body of
rats, and after 12 weeks the microspheres disintegrated into small
fragments.86 For chitosan hydrogel, strong degradation
was observed after 4 weeks.87 In turn, the 3D porous
scaffold made of chitosan after 30 days in vivo was completely
biodegraded.88 Hydrolysis of bonds in chitosan takes
place in the human body under the influence of a non-specific enzyme
lysozyme.89 It has been noted that due to the cationic
nature of chitosan, it can interact with blood
cells. In the presence of
chitosan, blood tends to coagulate and form clots on the surface of
chitosan.90 After oral administration of a
polysaccharide in the organisms of rabbits, a specific response was
observed, i.e. an increase in lysozyme secretion by the organism, which
may prevent thrombosis.91 In vivo degradation
studies of chitosan-poly-L-lactide composites in rats showed that the
polysaccharide did not cause an unexpected, negative response, no blood
clots were detected after 1 week.92
Cell scaffolding
Tissue engineering is looking for solutions that could replace the need
for autographic or allographic transplants. One of the proposed
solutions is the use of scaffolds that mimic the basic roles of tissues
and enable their regeneration.93,94
Such scaffolding is usually a three-dimensional, porous structure made
of biomaterial. The main task of scaffolding is to support the
regenerating tissue by providing a rack on the surface of which cells
can settle.8 Scaffolds should imitate the tissue for
which they are used to regenerate, its structure and
properties.95 The implant must not cause a negative
immune response, it should be metabolised by the body, easy to shape and
sterilise, and it should be durable so that it can be stored in ex
vivo conditions.96
Scaffolds made of chitosan alone
Due to the properties of chitosan, such as non-toxicity,
biocompatibility, affinity to animal mucus and biodegradability, it is a
polymer very readily used in tissue engineering.4 It
supports cell adhesion and proliferation, and thus tissue regeneration.
It allows you to easily model the scaffolding to a specific shape and
create porous structures with open pores. These features make chitosan a
potential candidate for scaffolding material.97
The temperatures used during the production of 3D scaffolding
significantly affect, among others, the pore size or mechanical
properties related to the compressive modulus.98Chitosan with a higher degree of deacetylation was characterised by
higher mechanical resistance, slower degradation time and lower
absorbability of the scaffold.99 High DD (DD 88% and
95% tested) supports cell proliferation on the surface of the chitosan
scaffold.99 The porosity of the scaffold has a
significant influence on its mechanical properties. The more porous the
structure, the worse the mechanical properties. The decrease in the
porosity from 94.1% to 82.5% was related to an increase in Young’s
modulus to 5.2 kPa and 520 kPa, respectively, while the pore size
decreased. Cell adhesion was possible on both types of scaffolds, but
better conditions were provided by scaffolds with lower porosity, which
was associated with an increase in fibre density.100
Due to the properties of chitosan, it is a readily used biomaterial. In
vitro and in vivo studies are conducted on the use of chitosan scaffolds
in the treatment of damage to various tissues. The interactions and
influence of CS on cells are investigated, among others, spinal
cord52, bones, cartilage101,
tendons102, skin103.
The use of chitosan in the treatment of the spinal cord has been
studied. The implants were inserted into the meninges or directly into
the spinal cord for 6 or 12 months, respectively. The host cell response
(rat) was low, which indicates the inert nature of chitosan and the
possibility of using the material in long-term
therapies.52 In search of chitosan with the best
properties for the osteochondral treatment, porous chitosan scaffolds
were prepared. In vivo studies indicate that the cellular
response depended on the properties of the chitosan used, and chitosan
with a DD of 83%, a relatively low molecular weight of 11.49 KDa and a
high content of calcium, remaining due to the lack of demineralisation
during the preparation, turned out to be the best for applications.
After implant placement, its degradation and shaping of the subchondral
bone were observed.101
Active layers can be applied to the scaffolds to improve the interaction
at the biomaterial-tissue interface.104 The surface of
the chitosan scaffolds was mineralised to obtain apatite layers
imitating bone. After modification, the scaffolds are characterised by
greater rigidity and smaller pore sizes, yet sufficiently large to allow
unhindered migration of bone cells.105 The conductedin vitro studies indicate that the scaffold after apatite
modification supports the adhesion and proliferation of cells to a
greater extent than the unmodified version. Mineralised scaffolds are
better suited for use in bone tissue
engineering.106,107
The cultivation of chondrocyte cells is aided by the use of porous
chitosan microspheres (porosity ~90%, pore size 5 – 60
μm) with diameters of 180 – 280 μm108 and CS fibres
with diameters between 4 and 22
μm. The fibres maintained the
viability of chondrocytes (over 85%) and supported the formation of the
extracellular matrix (ECM). However, it was concluded that reducing the
size and the use of nanofibers could improve interactions with
chondrocytes.109
Chitosan as a scaffold surface modifier
Chitosan is often used as a coating material for implants or scaffolds.
Such surface modification significantly improves the surface properties
of an implant – its bioactivity, biocompatibility, corrosion
resistance, as well as, properties supporting bone regeneration, such as
osteoconductivity.110 Chitosan layers are stable, and
depending on the concentration of chitosan, its molecular weight, the
degree of deacetylation and application technique, they degrade at
different rates, even allowing for long-term
use.111,112
Surface modification significantly influences the mechanical properties
of the scaffolding. Three-dimensional, porous scaffolds made of
polycaprolactam (PCL) were covered with layers of chitosan at various
concentrations, from 1% to 3%. As the concentration of chitosan in the
layer increases, the compression strength and the compression modulus
factor increase, while scaffolding crystallinity decreases. The increase
in the first two is likely related to the increase in the amount of
amine groups that support PCL and improve the strength of the scaffold.
The decrease in crystallinity is a result of the formation of hydrogen
bonds between PCL and chitosan. Among the examined, the best osteoblast
differentiation was provided by the PCL scaffold covered with a 2.5%
chitosan layer.113
It is commonly known that the hydrophilicity/hydrophobicity of the
scaffold affects cell adhesion and proliferation. Polylactide (PLA) is a
polymer readily used in tissue engineering due to its biocompatibility
and bioresorbability. It is however, a hydrophobic polymer that does not
promote cell adhesion. In order to increase the hydrophilicity of the
polymer, the PLA surface can be modified by applying a layer of
chitosan.114 Modification of poly-L- lactide
surface with chitosan also improves mechanical properties of the
scaffold, as well as improves adhesion, differentiation, activity, and
morphology of chondrocyte cells115 and mouse bone
marrow stromal cells (mBMSCs) compared to the unmodified structure,
which accelerates the bone regeneration process.51,116The introduction of the PLLA scaffold with a CS layer into skull defects
of rats and the examination of bone changes after 12 weeks showed that
the modified scaffolds support greater growth of bone tissue with higher
density compared to the unmodified ones. This confirms the suspicions
that chitosan-coated polylactide scaffolds are sustainable for bone
regeneration,116 and modifying the scaffold surface
with chitosan significantly improves osteoblast adhesion and
proliferation.117
Chitosan surfaces exhibit antimicrobial properties, highly desirable in
biomedical applications. The antibacterial nature of CS was tested on
Gram-negative strains of E. coli 118,119 and
Gram positive Staphylococcus aureus .120 For
both groups, the use of chitosan layers reduced bacterial settling.
In addition to applying layers of chitosan itself, layers of mixtures of
chitosan and other compounds are also used to improve certain
properties. The use of a chitosan layer with ZnO particles on a titanium
implant increases the compatibility of such an implant and its corrosion
resistance, which results from the closure of pores on the implant
surface. The introduction of the oxide improves the antibacterial
properties of the layer compared to the layer made of chitosan
itself.119 Antibiotics can also be incorporated into
the chitosan layer. The compounds bind through weak intermolecular
interactions, which allows easy release of the antibiotic and,
consequently, the fight against bacteria.121
Complexes with other natural polymers
Due to the cationic nature of the polymer, CS has the ability to
spontaneously form stable polyelectrolyte complexes (PEC) with
negatively charged structures such as natural polymers. In solution,
permanent electrostatic interactions are formed between the cationic
amino group of chitosan and the negatively charged group of the
polyanion. Complexes are formed in stages, first the primary complex is
formed, with chains of polyions randomly arranged in space. Then the
chains arrange themselves in orderly structures, which is associated
with the formation of hydrogen bonds, van der Waalss forces and others.
Concurrently, aggregation into various structures may take place. The
wide range of properties, as well as the non-toxicity, biodegradability
and biocompatibility of PEC ensure a wide application of the complexes,
especially in tissue engineering.122,123 Examples of
PEC are shown in Table 2. PEC formation can be investigated, for
example, by FTIR (N–H, C=O bond shift study), DSC differential scanning
calorimetry (melting point shift) or by examining changes in zeta
potential.
The formation of complexes is influenced by the concentration of
polymers, the mass ratio of polyelectrolytes, the order of polymer
addition, the degree of CS deacetylation, the pH value of the solution,
presence of other ions and molecules in the solution. As the
concentration of polymers in the solution increases, the size of the
particles formed increases. In turn, the mass ratio of the polymers used
affects the size, charge and solubility of resulting
complexes.124,125 The molecular weight of chitosan and
the degree of deacetylation affect the size of the complex. The larger
the polymer, the thinner the packing of the compound and the larger the
size of the PEC.39 Other molecules or ions than the
polymers forming the complex may be present in the solution. They affect
the ease of formation of complexes, their stability and properties. An
example of such a compound is sodium lauryl sulphate surfactant, the use
of which increases the surface roughness and the thickness of the
chitosan/alginate (CS/Alg).125,126
The size of the nanoparticles of the chitosan CS/Pc with pectin complex
varied depending on the order of addition of polymers. At low polymer
concentrations, larger particles were formed when chitosan was added
first, the inverse was true at high concentrations. The size of the
complexes ranged from 460 nm (low polymer concentration) to 1110 nm
(high polymer concentration). The zeta potential depends on the mass
ratio of the polymers used, for complexes with different mass ratios it
was in the range between 40 and 60 mV, and the advantage of chitosan in
the complex increases the value of the potential, i.e. the stability of
the complex. CS/Pc nanocomplexes are characterised by low stability
compared to other polyelectrolyte complexes, they lose their stability
after 14 days, and they degrade after 30 days. The stability is
independent of particle size, but is dependent on the pH of the solution
(stable at pH 3.5 – 6.0).124
The introduction of a polyanion into a chitosan scaffold has many
advantages. The electrostatic interaction between the polymers increases
the stability of the scaffolds obtained.127Modification with alginate improves the mechanical properties of the
scaffold, the compressive modulus and yield strength after the
modification are, respectively, 8.16 MPa and 0.46 MPa, for the chitosan
only scaffold these values are about three times lower. Also, the
preparation of the scaffold from the complex is simpler, since the
environment is not necessarily acidic, it can just as well be alkaline
or neutral. The chitosan/alginate complex is non-toxic, has
anti-inflammatory effects,128 in many cases supports
adhesion, as well as osteoblast proliferation and
angiogenesis.129 However, some sources report the
CS/Alg surface as unfavourable for cell adhesion.43
Composites with chitosan
Often, implants made of purely natural or synthetic polymers do not have
the mechanical properties desired in tissue engineering, for instance,
they are not able to carry the required loads.13 To
change the properties of the scaffold, a new compound can be introduced
into the starting composition to obtain a composite. Examples of
complexes are presented in Table 3. The use of the composite influences
morphology, mechanical properties, porosity, swelling of the scaffolds
as well as biomineralisation and degradation processes. Chitosan is a
modifier commonly used in composites, a building material popular in the
animal world, composites of which support bone regeneration processes
such as adhesion and proliferation of bone cells, osteocalcin secretion
or biomineralisation.138–140 On the other hand,
nanoparticles are readily introduced into chitosan scaffolds, thanks to
which biomineralisation takes place more efficiently and the degradation
rate decreases.
The morphology of the scaffolds significantly depends on the
concentrations of the polymers used. For the poly-L-lactide + chitosan
(PLLA+CS) scaffold, the higher the concentration of chitosan in the
starting solution, the more chitosan was deposited on the pore surface,
making the pores surface more jagged compared to polylactide systems
alone, which is desirable due to an increase in the surface of the
implant.139
The use of the PLLA+CS composite significantly affects the rate of
scaffold degradation in comparison to the polylactide scaffold. This is
explained by the fact that when PLLA hydrolyses into lactic acid, it
lowers the pH of the environment, which causes the dissolution of the
CS. In this way, along with the depletion of chitosan, the pores of the
scaffold increase, which increases its surface contact with water. Also,
the hydrophilic properties of chitosan accelerate the diffusion of water
into the interior, making hydrolysis of the hydrophobic polylactide
faster.92
The use of composites has a significant influence on mechanical
properties. In order to strengthen the macrospheres made of chitosan,
montmorillonite (MMT) and/or hydroxyapatite (HAP) were introduced into
the system. Composite scaffoldings were characterised by much greater
compressive strength. The introduction of MMT and HAP also reduced the
swelling of scaffolds, which is probably related to the hydrophobicity
of the additives and the binding of hydrophilic amino and hydroxyl CS
groups by hydroxyapatite. Interactions with bone cells and cell
proliferation supported all systems.141
The chitosan/collagen polyelectrolyte complex (CS/Coll) was enriched
with MMT and modified organomontmorillonite (OMMT). The scaffolds made
of PEC had weakly connected large pores (about 1 mm). The introduction
of MMT and OMMT made it possible to obtain a structure with well
connected pores of smaller diameters (200 – 400 μm for CS/Coll+MMT and
50 – 200 μm for CS/Coll+OMMT), which are better suited for seeding
cells. Both composites were characterised by lower swelling ratio,
degradation rates and greater compressive strength, and they supported
biomineralisation better than the CS/Coll complex.142
The introduction of nanomaterials into scaffolds made of chitosan or its
complexes affects the properties of the structures. The use of a
chitosan-chitin composite with the addition of ZrO2nanoparticles (CS+CI+nZrO2) enables the production of
non-toxic scaffolds with a pore size of 150 – 200 μm (larger in the
CS+CI system) and improved bioactivity.143 Porous
scaffolds made of a combination of chitosan, gelatine and hydroxyapatite
(Cs+Ge+nHAP composite) with an average pore size of 100 – 180 μm
support the adhesion and development of bone cells. Hydroxyapatite
occurs naturally in human bones, while gelatine and chitosan mimic the
extracellular matrix (ECM) well. The introduction of nHAP significantly
affects the properties of the system, increases the susceptibility to
mineralisation of the scaffold, and as a result, significantly improves
cytocompatibility, reduces swelling and the rate of degradation.
Composite scaffolds are well suited for bone tissue
engineering.144 In the same way, the properties of the
Cs+Ge scaffolds change after the introduction of SiO2nanoparticles (Cs+Ge+nSiO2composite).145
Cellular response to chitosan materials
After a scaffold is implanted, the body reacts, which ultimately leads
to tissue reconstruction. The damage is immediately recognised and
triggers appropriate interactions with blood, clot formation, cellular
response, and eventually tissue reconstruction.157 A
thin layer of water forms on the scaffold surface, and proteins adhere
to it. The bone cells then recognise these proteins and settle on the
implant. Cell growth and integration enables tissue to regenerate on the
scaffold surface.158,159 Depending on the surface
chemistry and implant topography, cells may interact with the material
with different intensity.
Cell adhesion
The term “adhesion” describes two phenomena that occur during the
settlement of scaffolding. It concerns the short-term generation of
interactions between the cell membrane and the material (ionic forces,
van der Walls interactions) and long-term cell
attachment.159 Biomaterials may affect cell adhesion,
be osteoconductive or osteoinductive. Osteoconduction is the ability of
a material to connect to bone tissue, provide adequate support for cells
and influence the direction of the regeneration process. The
osteoconductive material also positively influences the processes of
cell proliferation and ECM formation. Osteoinduction involves triggering
a series of processes and reactions that lead to bone regeneration
through the use of biomolecular signalling devices. One such process is,
for example, cell differentiation.160–162
Implants are recognised by the body as foreign bodies and trigger an
immune response. The body’s response mechanism after implantation
(Figure 3) begins with the formation of a thin layer of water on the
surface of the biomaterial within a few nanoseconds. Then, adhesion
proteins are deposited, these are peptides with a specific structure,
containing the RGD group – the arginine-glycine-aspartic acid
combination.163,164 Proteins are deposited on the
implant surface within seconds to hours. Depending on the surface
properties of the biomaterial, proteins settle in different amounts,
densities and conformations.165 The cells of the
tissue then recognise the attached proteins, and cell-protein
interactions are formed, which last from a few minutes to several days.
Cell growth and integration, the final step, enables tissue formation on
the scaffold surface.157,158
In addition to the surface properties, the scaffold topography and
porosity have a significant impact on cell adhesion. The rough structure
increases the contact area between the implant and the surrounding
tissue and allows cells to settle. Osteoblast differentiation is
supported in particular by nanosized roughness.104,166The scaffold should be characterised by open porosity, which enables
cell migration and colonisation of pores. Open porosity supports the
process of vascularisation of the resulting tissue.160Cell adhesion is significantly affected by the surface energy of the
biomaterial. High surface energy enables very good wettability and
adhesion. Osteoblasts are more likely to settle, differentiate faster
and multiply on a surface with higher surface
energy.167–169
Interaction with groups in chitosan
The biomaterial can interact with the body’s cells in different ways.
Significant surface features that influence this are surface properties
such as hydrophilicity, the presence of functional groups or surface
charge.104,130
Functional groups on the surface of the biomaterial significantly affect
the adhesion of bone cells. The binding of osteoblasts to surfaces with
different functional groups was investigated. The hydroxyl–OH and
amino–NH2 groups present in chitosan supported the
adhesion and differentiation of stem cells into osteoblasts better than
the carboxyl –COOH and methyl–CH3groups.170 Stem cell subsidence was most strongly
supported by the amino group. Depending on the functional groups, the
shape of the cells may also change. Flattened cells were preferred for
the –NH2 group, and spherical cells for
–COOH.171 The surfaces with the amino group also
promoted and maintained the process of osteogenesis, both under normal
and osteogenesis-promoting conditions.172
The wettability of the biomaterial affects the adhesion of cells. Good
wettability facilitates the process of proteins settling on the surface
of the material and, as a result, cell adhesion. A greater number of
adhesive proteins, and a smaller number of hydrophobic proteins that do
not participate in the process, were deposited on the hydrophilic
surfaces.163,173
Proteins settle differently on the positively charged surface than on
the negatively charged one. Amino groups of the CS promote the
deposition of negatively charged proteins and cell membranes,
proliferation and differentiation, which results from electrostatic
interactions.159
Chitosan with a higher degree of deacetylation provides better
conditions for osteoblast subsidence, but to a lesser extent supports
the secretion of osteoprotegerin compared to CS with a lower degree of
deacetylation.161,174,175
Secreted cellular metabolites
There are several types of cells in bone: osteoblasts, osteocytes,
osteoclasts, and there are also bone lining cells.176Osteoblasts are mature, metabolically active bone-building cells that
are found at the surface of the bone. The task of osteoblasts is to
secrete compounds that make up the intercellular
substance.177,178 The most numerous cells in bone
tissue (over 90%179) are bone cells – osteocytes.
They arise from transformed osteoblasts trapped in the bone matrix, in
the bone cavities. Osteocytes form a communication network of bone
cells, thanks to which they regulate bone tissue homeostasis. Signals
are sent via cleft junctions, chemical secretion, or direct dendrite
association.180–182 The bone lining cells are
osteoblasts that have not undergone apoptosis or developed into
osteocytes. They perform functions related to bone
remodelling.176,183 The last group of cells are
multinucleated osteoclasts, whose task is to maintain the calcium
balance in the tissue and to secrete compounds that break down the
bone.184–187
The living cells of the body conduct metabolic processes to produce the
energy necessary for life from carbohydrates, proteins and fats. For
this, the energy carrier adenosine-5′-triphosphate (ATP), produced in
various processes, is most often used. Glucose is the main source of
energy in mesenchymal stem cells, which is converted into pyruvic acid
and ATP in the process of glycolysis. Osteoblasts obtain ATP through
glycolysis and the Krebs cycle, with predominance of the former process.
The Krebs cycle is used mainly in the period of higher energy demand,
during bone formation. In addition to glucose, osteoblasts obtain ATP
from the transformation of amino acids (in particular glutamine) and
fats. Osteocyte metabolism is not a well understood process. Osteoclasts
generate energy in the processes of glycolysis and the Krebs
cycle.188
The extracellular matrix is mostly produced by osteoblasts. Osteocytes
contribute to a lesser extent to the production of matrix, which is due
to their structure – mature osteocytes do not have many organelles
responsible for secretion.182 ECM consists mainly of
collagen (90%) and non-collagen proteins (10%). The vast majority of
collagen is type I collagen, however, type III and V are also found in
the ECM. The non-collagenous organic part consists of proteoglycans,
osteocalcin (also known as bone gamma-carboxyglutamic acid-containing
protein (BGLAP)), glycoproteins and
Small Integrin-Binding LIgand
N-linked Glycoproteins (SIBLINGs).189–191Proteoglycans are proteins with which saccharide glycosaminoglycans
(GAGs) are linked. They are mainly secreted by osteoblasts, e.g.
biglikan192 or decorin.193Osteocalcin is secreted mainly by mature osteoblasts, but also by
osteocytes.194 SIBLINGs are small hydrophilic proteins
that contain the same set of amino acids (Arg-Gly-Asp) found in the bone
matrix. These compounds are mainly produced by osteocytes (Matrix
Extracellular Phosphoglycoprotein195), but to a lesser
extent also by osteoblasts (osteopontin, Dentin Matrix
Protein-1196,197).198
Osteoclasts are capable of secreting H+–adenosine
triphosphate responsible for the dissolution and cathepsin K protease
responsible for dissolving the demineralised organic
part.199,200
Chitosan has a positive effect on the metabolic behaviour of cells. In
vitro studies of MCF-7 cells on chitosan scaffolds were performed. The
metabolism of cells in such a system is very similar to that in tissue,
which proves the positive effect of the chitosan matrix on the vital
processes of cells.201 Similarly, for dental pulp
stromal cells, CS has a positive effect on metabolism and
proliferation.202
CONCLUSION
Chitosan, a chitin derivative, is a unique polysaccharide. It is
obtained in the process of deacetylation of chitin, most often derived
from the shells of sea crustaceans. It is a readily used compound in
tissue engineering due to its similarity to the glycosaminoglycans
present in the body, biocompatibility, bioactivity, non-toxicity,
antibacterial properties and non-toxic degradation products that may
occur under the influence of human enzymes. Amine groups of chitosan
improve the surface properties and provide better adhesion of bone
cells. CS can be used in the production of cell scaffolds for bone
regeneration as a direct substrate or component of a composite. The
cationic nature of the polysaccharide also enables the formation of
polyelectrolyte complexes with other natural polymers. It can also be
used for surface modification to improve the properties of the scaffold.
AUTHOR INFORMATION
Corresponding Author
*Email:agnieszka.gajadhur@pw.edu.pl
Author Contributions
The manuscript was written through contributions of all authors. All
authors have given approval to the final version of the manuscript.
Funding Sources
This scientific research was financed from the National Centre for
Research and Development as a research project “Lider 11”
(LIDER/4/0010/L-11/19/NCBR/2020) titled “Porous, biodegradable implants
for the regeneration of spongy bone”.
Notes
The authors declare no competing financial interest. Thanks to Karolina
Kalbarczyk for help with graphics.
ABBREVIATIONS
Alg, alginate;
ATP, adenosine triphosphate;
BCNs, bacterial cellulose nanocrystals;
BGLAP, bone gamma-carboxyglutamic acid-containing protein;
CHAP, carbonated hydroxyapatite;
CI, chitin;
Coll, collagen;
CS, chitosan;
CSA, chondroitin sulfate;
DSC, differential scanning calorimetry;
ECM, extracellular matrix;
FTIR, fourier transform infrared spectroscopy;
GAGs, glycosaminoglycans;
Ge, gelatin;
HA, hyaluronic acid
HAP, hydroxyapatite;
MMT, montmorillonite;
nBG, nanobioglass;
nHAP, nano hydroxyapatite;
nPP, nano-pearl powder;
nSiO2, nano-silica;
OMMT, organomontmorillonite;
Pc, pectin;
PCL, polycaprolactone
PCS, phosphorylated chitosan;
PEC, polyelectrolyte complex;
PGS, polyglycerol sebacate;
PLLA, poly-L-lactide;
PVA, poly(vinyl alcohol);
SIBLINGs, Small Integrin-Binding LIgand N-linked Glycoproteins;
REFERENCES
(1) Alonzo, M.; Alvarez Primo, F.; Anil Kumar, S.; Mudloff, J. A.;
Dominguez, E.; Fregoso, G.; Ortiz, N.; Weiss, W. M.; Joddar, B. Bone
Tissue Engineering Techniques, Advances, and Scaffolds for Treatment of
Bone Defects. Curr. Opin. Biomed. Eng. 2021 , 17(August 2020), 100248. https://doi.org/10.1016/j.cobme.2020.100248.
(2) Kanczler, J. M.; Wells, J. A.; Gibbs, D. M. R.; Marshall, K. M.;
Tang, D. K. O.; Oreffo, R. O. C. Bone Tissue Engineering and Bone
Regeneration ; INC, 2020.
https://doi.org/10.1016/B978-0-12-818422-6.00052-6.
(3) Schemitsch, E. H. Size Matters: Defining Critical in Bone Defect
Size! J. Orthop. Trauma 2017 , 31 (10), S20–S22.
https://doi.org/10.1097/BOT.0000000000000978.
(4) Kozusko, S. D.; Riccio, C.; Goulart, M.; Bumgardner, J.; Jing, X.
L.; Konofaos, P. Chitosan as a Bone Scaffold Biomaterial. J.
Craniofac. Surg. 2018 , 29 (7), 1788–1793.
https://doi.org/10.1097/SCS.0000000000004909.
(5) Giannoudis, P. V.; Dinopoulos, H.; Tsiridis, E. Bone Substitutes: An
Update. Injury 2005 , 36 Suppl 3 , 20–27.
https://doi.org/10.1016/j.injury.2005.07.029.
(6) Pape, H. C.; Evans, A.; Kobbe, P. Autologous Bone Graft: Properties
and Techniques. J. Orthop. Trauma 2010 , 24(SUPPL. 1), 36–40. https://doi.org/10.1097/BOT.0b013e3181cec4a1.
(7) Preethi Soundarya, S.; Haritha Menon, A.; Viji Chandran, S.;
Selvamurugan, N. Bone Tissue Engineering: Scaffold Preparation Using
Chitosan and Other Biomaterials with Different Design and Fabrication
Techniques. Int. J. Biol. Macromol. 2018 , 119 ,
1228–1239. https://doi.org/10.1016/j.ijbiomac.2018.08.056.
(8) Deb, P.; Deoghare, A. B.; Borah, A.; Barua, E.; Das Lala, S.
Scaffold Development Using Biomaterials: A Review. Mater. Today
Proc. 2018 , 5 (5), 12909–12919.
https://doi.org/10.1016/j.matpr.2018.02.276.
(9) Hudecki, A.; Kiryczyński, G.; Łos, M. J. Biomaterials, Definition,
Overview. Stem Cells Biomater. Regen. Med. 2018 , No. ii,
85–98. https://doi.org/10.1016/B978-0-12-812258-7.00007-1.
(10) Budnicka, M.; Szymaniak, M.; Kołbuk, D.; Ruśkowski, P.;
Gadomska-Gajadhur, A. Biomineralization of Poly-l-Lactide Spongy Bone
Scaffolds Obtained by Freeze-Extraction Method. J. Biomed. Mater.
Res. - Part B Appl. Biomater. 2020 , 108 (3), 868–879.
https://doi.org/10.1002/jbm.b.34441.
(11) Navarro, M.; Michiardi, A.; Castaño, O.; Planell, J. A.
Biomaterials in Orthopaedics. J. R. Soc. Interface 2008 ,5 (27), 1137–1158. https://doi.org/10.1098/rsif.2008.0151.
(12) Larry L, H.; Julia M, P. Third-Generation Biomedical Materials.Science (80-. ). 2002 , 295 (February),
1014–1017.
(13) Islam, M. M.; Shahruzzaman, M.; Biswas, S.; Nurus Sakib, M.;
Rashid, T. U. Chitosan Based Bioactive Materials in Tissue Engineering
Applications-A Review. Bioact. Mater. 2020 , 5(1), 164–183. https://doi.org/10.1016/j.bioactmat.2020.01.012.
(14) Lalzawmliana, V.; Anand, A.; Mukherjee, P.; Chaudhuri, S.; Kundu,
B.; Nandi, S. K.; Thakur, N. L. Marine Organisms as a Source of Natural
Matrix for Bone Tissue Engineering. Ceram. Int. 2019 ,45 (2), 1469–1481.
https://doi.org/10.1016/j.ceramint.2018.10.108.
(15) Bastiaens, L.; Soetemans, L.; D’Hondt, E.; Elst, K. Sources of
Chitin and Chitosan and Their Isolation. Chitin and Chitosan2019 , 1–34. https://doi.org/10.1002/9781119450467.ch1.
(16) Zargar, V.; Asghari, M.; Dashti, A. A Review on Chitin and Chitosan
Polymers: Structure, Chemistry, Solubility, Derivatives, and
Applications. ChemBioEng Rev. 2015 , 2 (3),
204–226. https://doi.org/10.1002/cben.201400025.
(17) Wang, W.; Xue, C.; Mao, X. Chitosan: Structural Modification,
Biological Activity and Application. Int. J. Biol. Macromol.2020 , 164 , 4532–4546.
https://doi.org/10.1016/j.ijbiomac.2020.09.042.
(18) Yadav, M.; Goswami, P.; Paritosh, K.; Kumar, M.; Pareek, N.;
Vivekanand, V. Seafood Waste: A Source for Preparation of Commercially
Employable Chitin/Chitosan Materials. Bioresour. Bioprocess.2019 , 6 (1). https://doi.org/10.1186/s40643-019-0243-y.
(19) Bakshi, P. S.; Selvakumar, D.; Kadirvelu, K.; Kumar, N. S. Chitosan
as an Environment Friendly Biomaterial – a Review on Recent
Modifications and Applications. Int. J. Biol. Macromol.2020 , 150 , 1072–1083.
https://doi.org/10.1016/j.ijbiomac.2019.10.113.
(20) Pochanavanich, P.; Suntornsuk, W. Fungal Chitosan Production and
Its Characterization. Lett. Appl. Microbiol. 2002 ,35 (1), 17–21. https://doi.org/10.1046/j.1472-765X.2002.01118.x.
(21) Kou, S. (Gabriel); Peters, L. M.; Mucalo, M. R. Chitosan: A Review
of Sources and Preparation Methods. Int. J. Biol. Macromol.2021 , 169 , 85–94.
https://doi.org/10.1016/j.ijbiomac.2020.12.005.
(22) Younes, I.; Rinaudo, M. Chitin and Chitosan Preparation from Marine
Sources. Structure, Properties and Applications. Mar. Drugs2015 , 13 (3), 1133–1174.
https://doi.org/10.3390/md13031133.
(23) Kumari, S.; Kishor, R. Chitin and Chitosan: Origin,
Properties, and Applications ; INC, 2020.
https://doi.org/10.1016/b978-0-12-817970-3.00001-8.
(24) Younes, I.; Hajji, S.; Frachet, V.; Rinaudo, M.; Jellouli, K.;
Nasri, M. Chitin Extraction from Shrimp Shell Using Enzymatic Treatment.
Antitumor, Antioxidant and Antimicrobial Activities of Chitosan.Int. J. Biol. Macromol. 2014 , 69 , 489–498.
https://doi.org/10.1016/j.ijbiomac.2014.06.013.
(25) Arbia, W.; Arbia, L.; Adour, L.; Amrane, A. Chitin Extraction from
Crustacean Shells Using Biological Methods – A Review. Food
Technol. Biotechnol 2013 , 51 (1), 12–25.
(26) Valdez-Peña, A. U.; Espinoza-Perez, J. D.; Sandoval-Fabian, G. C.;
Balagurusamy, N.; Hernandez-Rivera, A.; de-la-Garza-Rodriguez, I. M.;
Contreras-Esquivel, J. C. Screening of Industrial Enzymes for
Deproteinization of Shrimp Head for Chitin Recovery. Food Sci.
Biotechnol. 2010 , 19 (2), 553–557.
https://doi.org/10.1007/s10068-010-0077-z.
(27) Doan, C. T.; Tran, T. N.; Nguyen, V. B.; Vo, T. P. K.; Nguyen, A.
D.; Wang, S. L. Chitin Extraction from Shrimp Waste by Liquid
Fermentation Using an Alkaline Protease-Producing Strain, Brevibacillus
Parabrevis. Int. J. Biol. Macromol. 2019 , 131 ,
706–715. https://doi.org/10.1016/j.ijbiomac.2019.03.117.
(28) Aravamudhan, A.; Ramos, D. M.; Nada, A. A.; Kumbar, S. G.Natural Polymers: Polysaccharides and Their Derivatives for
Biomedical Applications ; Elsevier Inc., 2014.
https://doi.org/10.1016/B978-0-12-396983-5.00004-1.
(29) Xu, H.; Yang, Y. 3D Electrospun Fibrous Structures from
Biopolymers. ACS Symp. Ser. 2014 , 1175 , 103–126.
https://doi.org/10.1021/bk-2014-1175.ch007.
(30) Tsai, H.-S.; Wang, Y.-Z.; Lin, J.-J.; Lien, W.-F. Preparation and
Properties of Sulfopropyl Chitosan Derivatives with Various Sulfonation
Degree. Wiley Intersci. 2009 . https://doi.org/DOI
10.1002/app.31689.
(31) Grant, J.; Allen, C. Chitosan as a Biomaterial for Preparation of
Depot-Based Delivery Systems. ACS Symp. Ser. 2006 ,934 , 201–205. https://doi.org/10.1021/bk-2006-0934.ch010.
(32) Lu, S.; Song, X.; Cao, D.; Chen, Y.; Yao, K. Preparation of
Water-Soluble Chitosan. J. Appl. Polym. Sci. 2004 ,91 (6), 3497–3503. https://doi.org/10.1002/app.13537.
(33) Leedy, M. R.; Martin, H. J.; Norowski, P. A.; Jennings, J. A.;
Haggard, W. O.; Bumgardner, J. D. Use of Chitosan as a Bioactive Implant
Coating for Bone-Implant Applications. Adv. Polym. Sci.2011 , 244 (1), 129–166.
https://doi.org/10.1007/12_2011_115.
(34) Chicatun, F.; Griffanti, G.; McKee, M. D.; Nazhat, S. N.Collagen/Chitosan Composite Scaffolds for Bone and Cartilage
Tissue Engineering , Second Edi.; Elsevier Ltd., 2017.
https://doi.org/10.1016/b978-0-08-100752-5.00008-1.
(35) Kumar, M. N. V. R.; Muzzarelli, R. A. A.; Muzzarelli, C.; Sashiwa,
H.; Domb, A. J. Chitosan Chemistry and Pharmaceutical Perspectives.Chem. Rev. 2004 , 104 (12), 6017–6084.
https://doi.org/10.1021/cr030441b.
(36) Raquel Madureira, A.; Sarmento, B.; Pintado, M. Current State of
the Potential Use of Chitosan as Pharmaceutical Excipient. Handb.
Polym. Pharm. Technol. 2015 , 3 , 275–297.
https://doi.org/10.1002/9781119041450.ch9.
(37) Wan, Y.; Fang, Y.; Wu, H.; Cao, X. Porous Polylactide/Chitosan
Scaffolds for Tissue Engineering. J. Biomed. Mater. Res. - Part A2007 , 80 (4), 776–789.
https://doi.org/10.1002/jbm.a.31025.
(38) Antunes, J. C.; Pereira, C. L.; Molinos, M.; Ferreira-Da-Silva, F.;
Dessi, M.; Gloria, A.; Ambrosio, L.; Gonca̧lves, R. M.; Barbosa, M. A.
Layer-by-Layer Self-Assembly of Chitosan and Poly(γ-Glutamic Acid) into
Polyelectrolyte Complexes. Biomacromolecules 2011 ,12 (12), 4183–4195. https://doi.org/10.1021/bm2008235.
(39) Strand, S. P.; Danielsen, S.; Christensen, B. E.; Vårum, K. M.
Influence of Chitosan Structure on the Formation and Stability of
DNA-Chitosan Polyelectrolyte Complexes. Biomacromolecules2005 , 6 (6), 3357–3366.
https://doi.org/10.1021/bm0503726.
(40) Maurstad, G.; Danielsen, S.; Stokke, B. T. The Influence of Charge
Density of Chitosan in the Compaction of the Polyanions DNA and Xanthan.Biomacromolecules 2007 , 8 (4), 1124–1130.
https://doi.org/10.1021/bm0610119.
(41) Kaczmarek, B.; Sionkowska, A.; Gołyńska, M.; Polkowska, I.;
Szponder, T.; Nehrbass, D.; Osyczka, A. M. In Vivo Study on Scaffolds
Based on Chitosan, Collagen, and Hyaluronic Acid with Hydroxyapatite.Int. J. Biol. Macromol. 2018 , 118 , 938–944.
https://doi.org/10.1016/j.ijbiomac.2018.06.175.
(42) Wang, J. Z.; Huang, X. B.; Xiao, J.; Yu, W. T.; Wang, W.; Xie, W.
Y.; Zhang, Y.; Ma, X. J. Hydro-Spinning: A Novel Technology for Making
Alginate/Chitosan Fibrous Scaffold. J. Biomed. Mater. Res. - Part
A 2010 , 93 (3), 910–919.
https://doi.org/10.1002/jbm.a.32590.
(43) Phoeung, T.; Spanedda, M. V.; Roger, E.; Heurtault, B.; Fournel,
S.; Reisch, A.; Mutschler, A.; Perrin-Schmitt, F.; Hemmerlé, J.; Collin,
D.; et al. Alginate/Chitosan Compact Polyelectrolyte Complexes: A Cell
and Bacterial Repellent Material. Chem. Mater. 2017 ,29 (24), 10418–10425.
https://doi.org/10.1021/acs.chemmater.7b03863.
(44) Park, J. W.; Park, K. H.; Seo, S. Natural Polyelectrolyte
Complex-Based PH-Dependent Delivery Carriers Using Alginate and
Chitosan. J. Appl. Polym. Sci. 2019 , 136 (43),
1–7. https://doi.org/10.1002/app.48143.
(45) Lallana, E.; Rios De La Rosa, J. M.; Tirella, A.; Pelliccia, M.;
Gennari, A.; Stratford, I. J.; Puri, S.; Ashford, M.; Tirelli, N.
Chitosan/Hyaluronic Acid Nanoparticles: Rational Design Revisited for
RNA Delivery. Mol. Pharm. 2017 , 14 (7),
2422–2436. https://doi.org/10.1021/acs.molpharmaceut.7b00320.
(46) Oyarzun-Ampuero, F. A.; Brea, J.; Loza, M. I.; Torres, D.; Alonso,
M. J. Chitosan-Hyaluronic Acid Nanoparticles Loaded with Heparin for the
Treatment of Asthma. Int. J. Pharm. 2009 , 381(2), 122–129. https://doi.org/10.1016/j.ijpharm.2009.04.009.
(47) Gennari, A.; de la Rosa, J. M. R.; Hohn, E.; Pelliccia, M.;
Lallana, E.; Donno, R.; Tirella, A.; Tirelli, N. The Different Ways to
Chitosan/Hyaluronic Acid Nanoparticles: Templated vs Direct
Complexation. Influence of Particle Preparation on Morphology, Cell
Uptake and Silencing Efficiency. Beilstein J. Nanotechnol.2019 , 10 , 2594–2608.
https://doi.org/10.3762/bjnano.10.250.
(48) Pandey, S.; Mishra, A.; Raval, P.; Patel, H.; Gupta, A.; Shah, D.
Chitosan-Pectin Polyelectrolyte Complex as a Carrier for Colon Targeted
Drug Delivery. J. Young Pharm. 2013 , 5 (4),
160–166. https://doi.org/10.1016/j.jyp.2013.11.002.
(49) Bombaldi de Souza, F. C.; Bombaldi de Souza, R. F.; Drouin, B.;
Mantovani, D.; Moraes, Â. M. Comparative Study on Complexes Formed by
Chitosan and Different Polyanions: Potential of Chitosan-Pectin
Biomaterials as Scaffolds in Tissue Engineering. Int. J. Biol.
Macromol. 2019 , 132 , 178–189.
https://doi.org/10.1016/j.ijbiomac.2019.03.187.
(50) Kumar, M.; Brar, A.; Vivekanand, V.; Pareek, N. Possibilities
and Perspectives of Chitosan Scaffolds and Composites for Tissue
Engineering ; Elsevier Inc., 2019.
https://doi.org/10.1016/B978-0-12-816909-4.00007-5.
(51) Li, M.; Mondrinos, M. J.; Chen, X.; Gandhi, M. R.; Ko, F. K.;
Lelkes, P. I. Fabrication and Characterization of PLLA–Chitosan Hybrid
Scaffolds with Improved Cell Compatibility. J. Biomed. Mater. Res.
Part A 2006 , 79 (4), 963–973.
https://doi.org/10.1002/jbm.a.
(52) Kim, H.; Tator, C. H.; Shoichet, M. S. Chitosan Implants in the Rat
Spinal Cord: Biocompatibility and Biodegradation. J. Biomed.
Mater. Res. - Part A 2011 , 97 A (4), 395–404.
https://doi.org/10.1002/jbm.a.33070.
(53) Luo, K.; Yin, J.; Khutoryanskaya, O. V.; Khutoryanskiy, V. V.
Mucoadhesive and Elastic Films Based on Blends of Chitosan and
Hydroxyethylcellulose. Macromol. Biosci. 2008 , 8(2), 184–192. https://doi.org/10.1002/mabi.200700185.
(54) Wu, Q. X.; Lin, D. Q.; Yao, S. J. Design of Chitosan and Its Water
Soluble Derivatives-Based Drug Carriers with Polyelectrolyte Complexes.Mar. Drugs 2014 , 12 (12), 6236–6253.
https://doi.org/10.3390/md12126236.
(55) Yuan, X.; Zheng, J.; Jiao, S.; Cheng, G.; Feng, C.; Du, Y.; Liu, H.
A Review on the Preparation of Chitosan Oligosaccharides and Application
to Human Health, Animal Husbandry and Agricultural Production.Carbohydr. Polym. 2019 , 220 (February), 60–70.
https://doi.org/10.1016/j.carbpol.2019.05.050.
(56) Shi, G. N.; Zhang, C. N.; Xu, R.; Niu, J. F.; Song, H. J.; Zhang,
X. Y.; Wang, W. W.; Wang, Y. M.; Li, C.; Wei, X. Q.; et al. Enhanced
Antitumor Immunity by Targeting Dendritic Cells with Tumor Cell
Lysate-Loaded Chitosan Nanoparticles Vaccine. Biomaterials2017 , 113 , 191–202.
https://doi.org/10.1016/j.biomaterials.2016.10.047.
(57) Abd El-Hack, M. E.; El-Saadony, M. T.; Shafi, M. E.; Zabermawi, N.
M.; Arif, M.; Batiha, G. E.; Khafaga, A. F.; Abd El-Hakim, Y. M.;
Al-Sagheer, A. A. Antimicrobial and Antioxidant Properties of Chitosan
and Its Derivatives and Their Applications: A Review. Int. J.
Biol. Macromol. 2020 , 164 , 2726–2744.
https://doi.org/10.1016/j.ijbiomac.2020.08.153.
(58) Duan, C.; Meng, X.; Meng, J.; Khan, M. I. H.; Dai, L.; Khan, A.;
An, X.; Zhang, J.; Huq, T.; Ni, Y. Chitosan as A Preservative for Fruits
and Vegetables: A Review on Chemistry and Antimicrobial Properties.Chitosan as A Preserv. Fruits Veg. A Rev. Chem. Antimicrob. Prop.2019 , 4 (1), 11–21.
https://doi.org/10.21967/jbb.v4i1.189.
(59) Kucharska, M.; Sikora, M.; Brzoza-Malczewska, K.; Owczarek, M.
Antimicrobial Properties of Chitin and Chitosan. Chitin and
Chitosan 2019 , 169–187.
https://doi.org/10.1002/9781119450467.ch7.
(60) Kim, J.-S.; Shin, D.-H. Inhibitory Effect on Streptococcus Mutans
and Mechanical Properties of the Chitosan Containing Composite Resin .Restor. Dent. Endod. 2013 , 38 (1), 36.
https://doi.org/10.5395/rde.2013.38.1.36.
(61) Raafat, D.; Von Bargen, K.; Haas, A.; Sahl, H. G. Insights into the
Mode of Action of Chitosan as an Antibacterial Compound. Appl.
Environ. Microbiol. 2008 , 74 (12), 3764–3773.
https://doi.org/10.1128/AEM.00453-08.
(62) Helander, I. M.; Nurmiaho-Lassila, E. L.; Ahvenainen, R.; Rhoades,
J.; Roller, S. Chitosan Disrupts the Barrier Properties of the Outer
Membrane of Gram-Negative Bacteria. Int. J. Food Microbiol.2001 , 71 (2–3), 235–244.
https://doi.org/10.1016/S0168-1605(01)00609-2.
(63) Ahmad, S. I.; Ahmad, R.; Khan, M. S.; Kant, R.; Shahid, S.; Gautam,
L.; Hasan, G. M.; Hassan, M. I. Chitin and Its Derivatives: Structural
Properties and Biomedical Applications. Int. J. Biol. Macromol.2020 , 164 , 526–539.
https://doi.org/10.1016/j.ijbiomac.2020.07.098.
(64) Harish Prashanth, K. V.; Tharanathan, R. N. Chitin/Chitosan:
Modifications and Their Unlimited Application Potential-an Overview.Trends Food Sci. Technol. 2007 , 18 (3), 117–131.
https://doi.org/10.1016/j.tifs.2006.10.022.
(65) Je, J. Y.; Kim, S. K. Chitooligosaccharides as Potential
Nutraceuticals. Production and Bioactivities , 1st ed.; Elsevier Inc.,
2012; Vol. 65. https://doi.org/10.1016/B978-0-12-416003-3.00021-4.
(66) Qandil, A. M.; Marji, T. J.; Al-Taani, B. M.; Khaled, A. H.;
Badwan, A. A. Depolymerization of HMW into a Predicted LMW Chitosan and
Determination of the Degree of Deacetylation to Guarantee Its Quality
for Research Use. J. Excipients Food Chem. 2018 ,9 (2), 51–63.
(67) Ma, Z.; Wang, W.; Wu, Y.; He, Y.; Wu, T. Oxidative Degradation of
Chitosan to the Low Molecular Water-Soluble Chitosan over
Peroxotungstate as Chemical Scissors. PLoS One 2014 ,9 (6), 3–9. https://doi.org/10.1371/journal.pone.0100743.
(68) Hsu, S.-C.; Don, T.-M.; Chiu, W.-Y. Free Radical Degradation of
Chitosan with Potassium Persulfate. Polym. Degrad. Stab.2002 , 75 (1), 73–83.
https://doi.org/10.1016/S0141-3910(01)00205-1.
(69) Yan, J.; Ai, S.; Yang, F.; Zhang, K.; Huang, Y. Study on Mechanism
of Chitosan Degradation with Hydrodynamic Cavitation. Ultrason.
Sonochem. 2020 , 64 (October 2019), 105046.
https://doi.org/10.1016/j.ultsonch.2020.105046.
(70) Tsao, C. T.; Chang, C. H.; Lin, Y. Y.; Wu, M. F.; Han, J. L.;
Hsieh, K. H. Kinetic Study of Acid Depolymerization of Chitosan and
Effects of Low Molecular Weight Chitosan on Erythrocyte Rouleaux
Formation. Carbohydr. Res. 2011 , 346 (1),
94–102. https://doi.org/10.1016/j.carres.2010.10.010.
(71) Pandit, A.; Indurkar, A.; Deshpande, C.; Jain, R.; Dandekar, P. A
Systematic Review of Physical Techniques for Chitosan Degradation.Carbohydr. Polym. Technol. Appl. 2021 , 2(January), 100033. https://doi.org/10.1016/j.carpta.2021.100033.
(72) Lim, L.; Khor, E.; Koo, O. γ Irradiation of Chitosan. J.
Biomed. Mater. Res. 1998 , 43 (3), 282–290.
https://doi.org/10.1002/(sici)1097-4636(199823)43:3<282::aid-jbm9>3.3.co;2-x.
(73) San Juan, A.; Montembault, A.; Gillet, D.; Say, J. P.; Rouif, S.;
Bouet, T.; Royaud, I.; David, L. Degradation of Chitosan-Based Materials
after Different Sterilization Treatments. IOP Conf. Ser. Mater.
Sci. Eng. 2012 , 31 , 2–7.
https://doi.org/10.1088/1757-899X/31/1/012007.
(74) Wu, T.; Zivanovic, S.; Hayes, D. G.; Weiss, J. Efficient Reduction
of Chitosan Molecular Weight by High-Intensity Ultrasound : Underlying
Mechanism. 2008 , No. I, 5112–5119.
https://doi.org/10.1021/jf073136q.
(75) Yan, J.; Xu, J.; Ai, S.; Zhang, K.; Yang, F.; Huang, Y. Degradation
of Chitosan with Self-Resonating Cavitation. Arab. J. Chem.2020 , 13 (6), 5776–5787.
https://doi.org/10.1016/j.arabjc.2020.04.015.
(76) Chen, R. H.; Huang, J. R.; Tsai, M. L.; Tseng, L. Z.; Hsu, C. H.
Differences in Degradation Kinetics for Sonolysis, Microfluidization and
Shearing Treatments of Chitosan. Polym. Int. 2011 ,60 (6), 897–902. https://doi.org/10.1002/pi.3055.
(77) Sørbotten, A.; Horn, S. J.; Eijsink, V. G. H.; Vårum, K. M.
Degradation of Chitosans with Chitinase B from Serratia Marcescens:
Production of Chito-Oligosaccharides and Insight into Enzyme
Processivity. FEBS J. 2005 , 272 (2), 538–549.
https://doi.org/10.1111/j.1742-4658.2004.04495.x.
(78) Nawrotek, K.; Tylman, M.; Adamus-Włodarczyk, A.; Rudnicka, K.;
Gatkowska, J.; Wieczorek, M.; Wach, R. Influence of Chitosan Average
Molecular Weight on Degradation and Stability of Electrodeposited
Conduits. Carbohydr. Polym. 2020 , 244 (December
2019), 116484. https://doi.org/10.1016/j.carbpol.2020.116484.
(79) Li, J.; Du, Y. M.; Liang, H. B.; Yao, P. J.; Wei, Y. A. Effect of
Immobilized Neutral Protease on the Preparation and Physicochemical
Properties of Low Molecular Weight Chitosan and Chito-Oligomers.J. Appl. Polym. Sci. 2006 , 102 (5), 4185–4193.
https://doi.org/10.1002/app.24555.
(80) Kumar, A. B. V.; Gowda, L. R.; Tharanathan, R. N. Non-Specific
Depolymerization of Chitosan by Pronase and Characterization of the
Resultant Products. Eur. J. Biochem. 2004 , 271(4), 713–723. https://doi.org/10.1111/j.1432-1033.2003.03975.x.
(81) Shin, S. S.; Lee, Y. C.; Chan, L. The Degradation of Chitosan with
the Aid of Lipase from Rhizopus Japonicus for the Production of Soluble
Chitosan. J. Food Biochem. 2001 , 25 (4),
307–321. https://doi.org/10.1111/j.1745-4514.2001.tb00742.x.
(82) Budnicka, M.; Gadomska-Gajadhur, A.; Ruśkowski, P. Wytwarzanie
Polimerowych Substytutów Kości. Tworzywa Sztuczne w Przemyśle2018 , 43 (1), 56–61.
(83) Onishi, H.; Machida, Y. Biodegradation and Distribution of
Water-Soluble Chitosan in Mice. Biomaterials 1999 ,20 (2), 175–182. https://doi.org/10.1016/S0142-9612(98)00159-8.
(84) Szymańska, E.; Winnicka, K. Stability of Chitosan - A Challenge for
Pharmaceutical and Biomedical Applications. Mar. Drugs2015 , 13 (4), 1819–1846.
https://doi.org/10.3390/md13041819.
(85) Barbosa, J. N.; Amaral, I. F.; Águas, A. P.; Barbosa, M. A.
Evaluation of the Effect of the Degree of Acetylation on the
Inflammatory Response to 3D Porous Chitosan Scaffolds. J. Biomed.
Mater. Res. - Part A 2010 , 93 (1), 20–28.
https://doi.org/10.1002/jbm.a.32499.
(86) Mi, F. L.; Tan, Y. C.; Liang, H. F.; Sung, H. W. In Vivo
Biocompatibility and Degradability of a Novel Injectable-Chitosan-Based
Implant. Biomaterials 2002 , 23 (1), 181–191.
https://doi.org/10.1016/S0142-9612(01)00094-1.
(87) Moura, M. J.; Brochado, J.; Gil, M. H.; Figueiredo, M. M. In Situ
Forming Chitosan Hydrogels: Preliminary Evaluation of the in Vivo
Inflammatory Response. Mater. Sci. Eng. C 2017 ,75 , 279–285. https://doi.org/10.1016/j.msec.2017.02.050.
(88) Qasim, S. B.; Husain, S.; Huang, Y.; Pogorielov, M.; Deineka, V.;
Lyndin, M.; Rawlinson, A.; Rehman, I. U. In-Vitro and in-Vivo
Degradation Studies of Freeze Gelated Porous Chitosan Composite
Scaffolds for Tissue Engineering Applications. Polym. Degrad.
Stab. 2017 , 136 , 31–38.
https://doi.org/10.1016/j.polymdegradstab.2016.11.018.
(89) Halim, A. S.; Keong, L. C.; Zainol, I.; Rashid, A. H. A.
Biocompatibility and Biodegradation of Chitosan and Derivatives.Chitosan-Based Syst. Biopharm. Deliv. Target. Polym. Ther.2012 , 57–73. https://doi.org/10.1002/9781119962977.ch4.
(90) Hirano, S.; Noishiki, Y.; Kinugawa, J.; Higashijima, H.; Hayashi,
T. Chitin and Chitosan for Use as a Novel Biomedical Material. InAdvances in Biomedical Polymers ; Gebelein, C. G., Ed.; Springer
US: Boston, MA, 1987; pp 285–297.
https://doi.org/10.1007/978-1-4613-1829-3_26.
(91) Hirano, S.; Iwata, M.; Yamanaka, K.; Tanaka, H.; Toda, T.; Inui, H.
Enhancement of Serum Lysozyme Activity by Injecting a Mixture of
Chitosan Oligosaccharides Intravenously in Rabbits. Agric. Biol.
Chem. 1991 , 55 (10), 2623–2625.
https://doi.org/10.1080/00021369.1991.10871007.
(92) Guo, Z.; Bo, D.; He, Y.; Luo, X.; Li, H. Degradation Properties of
Chitosan Microspheres/Poly(L-Lactic Acid) Composite in Vitro and in
Vivo. Carbohydr. Polym. 2018 , 193 (January),
1–8. https://doi.org/10.1016/j.carbpol.2018.03.067.
(93) Chan, B. P.; Leong, K. W. Scaffolding in Tissue Engineering:
General Approaches and Tissue-Specific Considerations. Eur. Spine
J. 2008 , 17 (SUPPL. 4).
https://doi.org/10.1007/s00586-008-0745-3.
(94) Salgado, A. J.; Oliveira, J. M.; Martins, A.; Teixeira, F. G.;
Silva, N. A.; Neves, N. M.; Sousa, N.; Reis, R. L. Tissue
Engineering and Regenerative Medicine: Past, Present, and Future ; 2013;
Vol. 108. https://doi.org/10.1016/B978-0-12-410499-0.00001-0.
(95) O’Brien, F. J. Biomaterials & Scaffolds for Tissue Engineering.Mater. Today 2011 , 14 (3), 88–95.
https://doi.org/10.1016/S1369-7021(11)70058-X.
(96) Middleton, J. C.; Tipton, A. J. Synthetic Biodegradable Polymers as
Orthopedic Devices. Biomaterials 2000 , 21 (23),
2335–2346. https://doi.org/10.1016/S0142-9612(00)00101-0.
(97) Cruz, D. M. G.; Gomes, M.; Reis, R. L.; Moratal, D.;
Salmerón-Sánchez, M.; Ribelles, J. L. G.; Mano, J. F. Differentiation of
Mesenchymal Stem Cells in Chitosan Scaffolds with Double Micro and
Macroporosity. J. Biomed. Mater. Res. - Part A 2010 ,95 (4), 1182–1193. https://doi.org/10.1002/jbm.a.32906.
(98) Reys, L. L.; Silva, S. S.; Pirraco, R. P.; Marques, A. P.; Mano, J.
F.; Silva, T. H.; Reis, R. L. Influence of Freezing Temperature and
Deacetylation Degree on the Performance of Freeze-Dried Chitosan
Scaffolds towards Cartilage Tissue Engineering. Eur. Polym. J.2017 , 95 (July), 232–240.
https://doi.org/10.1016/j.eurpolymj.2017.08.017.
(99) Thein-Han, W. W.; Kitiyanant, Y. Chitosan Scaffolds for in Vitro
Buffalo Embryonic Stem-like Cell Culture: An Approach to Tissue
Engineering. J. Biomed. Mater. Res. - Part B Appl. Biomater.2007 , 80 (1), 92–101.
https://doi.org/10.1002/jbm.b.30573.
(100) Xu, Y.; Xia, D.; Han, J.; Yuan, S.; Lin, H.; Zhao, C. Design and
Fabrication of Porous Chitosan Scaffolds with Tunable Structures and
Mechanical Properties. Carbohydr. Polym. 2017 ,177 (March), 210–216.
https://doi.org/10.1016/j.carbpol.2017.08.069.
(101) Abarrategi, A.; Lópiz-Morales, Y.; Ramos, V.; Civantos, A.;
López-Duŕn, L.; Marco, F.; López-Lacomba, J. L. Chitosan Scaffolds for
Osteochondral Tissue Regeneration. J. Biomed. Mater. Res. - Part
A 2010 , 95 (4), 1132–1141.
https://doi.org/10.1002/jbm.a.32912.
(102) Chen, E.; Yang, L.; Ye, C.; Zhang, W.; Ran, J.; Xue, D.; Wang, Z.;
Pan, Z.; Hu, Q. An Asymmetric Chitosan Scaffold for Tendon Tissue
Engineering: In Vitro and in Vivo Evaluation with Rat Tendon
Stem/Progenitor Cells. Acta Biomater. 2018 , 73 ,
377–387. https://doi.org/10.1016/j.actbio.2018.04.027.
(103) Intini, C.; Elviri, L.; Cabral, J.; Mros, S.; Bergonzi, C.;
Bianchera, A.; Flammini, L.; Govoni, P.; Barocelli, E.; Bettini, R.; et
al. 3D-Printed Chitosan-Based Scaffolds: An in Vitro Study of Human Skin
Cell Growth and an in-Vivo Wound Healing Evaluation in Experimental
Diabetes in Rats. Carbohydr. Polym. 2018 , 199(April), 593–602. https://doi.org/10.1016/j.carbpol.2018.07.057.
(104) Budnicka, M.; Szymaniak, M.; Gadomska-Gajadhur, A. A. Metody
Modyfikacji Powierzchni Implantów Polimerowych Do Regeneracji Tkanki
Kostnej. 2018 , No. September, 79–96.
(105) Dash, M.; Samal, S. K.; Douglas, T. E. L.; Schaubroeck, D.;
Leeuwenburgh, S. C.; Van Der Voort, P.; Declercq, H. A.; Dubruel, P.
Enzymatically Biomineralized Chitosan Scaffolds for Tissue-Engineering
Applications. J. Tissue Eng. Regen. Med. 2017 , 11(5), 1500–1513. https://doi.org/10.1002/term.2048.
(106) Aday, S.; Gümüşderelioğlu, M. Bone-like Apatite-Coated Chitosan
Scaffolds: Characterization and Osteoblastic Activity. Polym.
Compos. 2009 , 16 (2), NA-NA.
https://doi.org/10.1002/pc.20927.
(107) Manjubala, I.; Ponomarev, I.; Wilke, I.; Jandt, K. D. Growth of
Osteoblast-like Cells on Biomimetic Apatite-Coated Chitosan Scaffolds.J. Biomed. Mater. Res. - Part B Appl. Biomater. 2008 ,84 (1), 7–16. https://doi.org/10.1002/jbm.b.30838.
(108) Lu, G.; Zhu, L.; Kong, L.; Zhang, L.; Gong, Y.; Zhao, N.; Zhang,
X. Porous Chitosan Microcarriers for Large Scale Cultivation of Cells
for Tissue Engineering: Fabrication and Evaluation. Tsinghua Sci.
Technol. 2006 , 11 (4), 427–432.
https://doi.org/10.1016/S1007-0214(06)70212-7.
(109) Ragetly, G. R.; Slavik, G. J.; Cunningham, B. T.; Schaeffer, D.
J.; Griffon, D. J. Cartilage Tissue Engineering on Fibrous Chitosan
Scaffolds Produced by a Replica Molding Technique. J. Biomed.
Mater. Res. - Part A 2010 , 93 (1), 46–55.
https://doi.org/10.1002/jbm.a.32514.
(110) Di, A.; Sittinger, M.; Risbud, M. V. Chitosan : A Versatile
Biopolymer for Orthopaedic Tissue-Engineering. 2005 , 26 ,
5983–5990. https://doi.org/10.1016/j.biomaterials.2005.03.016.
(111) Kumari, S.; Tiyyagura, H. R.; Pottathara, Y. B.; Sadasivuni, K.
K.; Ponnamma, D.; Douglas, T. E. L.; Skirtach, A. G.; Mohan, M. K.
Surface Functionalization of Chitosan as a Coating Material for
Orthopaedic Applications: A Comprehensive Review. Carbohydr.
Polym. 2021 , 255 (November 2020), 117487.
https://doi.org/10.1016/j.carbpol.2020.117487.
(112) Gallyamov, M. O.; Chaschin, I. S.; Bulat, M. V.; Bakuleva, N. P.;
Badun, G. A.; Chernysheva, M. G.; Kiselyova, O. I.; Khokhlov, A. R.
Chitosan Coatings with Enhanced Biostability in Vivo. J. Biomed.
Mater. Res. - Part B Appl. Biomater. 2018 , 106 (1),
270–277. https://doi.org/10.1002/jbm.b.33852.
(113) Poddar, D.; Jain, P.; Rawat, S.; Mohanty, S. Influence of Varying
Concentrations of Chitosan Coating on the Pore Wall of Polycaprolactone
Based Porous Scaffolds for Tissue Engineering Application.Carbohydr. Polym. 2021 , 259 , 117501.
https://doi.org/10.1016/j.carbpol.2020.117501.
(114) Zhu, H.; Ji, A.; Shen, J. Surface Engineering of Poly (DL-Lactic
Acid) by Entrapment of Biomacromolecules. Macromol. Rapid Commun.2002 , 23 (14), 819–823.
https://doi.org/10.1002/1521-3927(20021001)23:14<819::AID-MARC819>3.0.CO;2-9.
(115) Ma, Z.; Gao, C.; Gong, Y.; Ji, J.; Shen, J. Immobilization of
Natural Macromolecules on Poly-L-Lactic Acid Membrane Surface in Order
to Improve Its Cytocompatibility. J. Biomed. Mater. Res.2002 , 63 (6), 838–847.
https://doi.org/10.1002/jbm.10470.
(116) Chen, S.; Zhao, X.; Du, C. Macroporous Poly (L-Lactic
Acid)/Chitosan Nanofibrous Scaffolds through Cloud Point Thermally
Induced Phase Separation for Enhanced Bone Regeneration. Eur.
Polym. J. 2018 , 109 (381), 303–316.
https://doi.org/10.1016/j.eurpolymj.2018.10.003.
(117) He, J.; Hu, X.; Cao, J.; Zhang, Y.; Xiao, J.; Peng, li J.; Chen,
D.; Xiong, C.; Zhang, L. Chitosan-Coated Hydroxyapatite and Drug-Loaded
Polytrimethylene Carbonate/Polylactic Acid Scaffold for Enhancing Bone
Regeneration. Carbohydr. Polym. 2021 , 253(September 2020), 117198. https://doi.org/10.1016/j.carbpol.2020.117198.
(118) Al-Nabulsi, A.; Osaili, T.; Sawalha, A.; Olaimat, A. N.; Albiss,
B. A.; Mehyar, G.; Ayyash, M.; Holley, R. Antimicrobial Activity of
Chitosan Coating Containing ZnO Nanoparticles against E. Coli O157:H7 on
the Surface of White Brined Cheese. Int. J. Food Microbiol.2020 , 334 (July), 108838.
https://doi.org/10.1016/j.ijfoodmicro.2020.108838.
(119) Lin, M. H.; Wang, Y. H.; Kuo, C. H.; Ou, S. F.; Huang, P. Z.;
Song, T. Y.; Chen, Y. C.; Chen, S. T.; Wu, C. H.; Hsueh, Y. H.; et al.
Hybrid ZnO/Chitosan Antimicrobial Coatings with Enhanced Mechanical and
Bioactive Properties for Titanium Implants. Carbohydr. Polym.2021 , 257 (August 2020), 117639.
https://doi.org/10.1016/j.carbpol.2021.117639.
(120) Foss, B. L.; Ghimire, N.; Tang, R.; Sun, Y.; Deng, Y. Bacteria and
Osteoblast Adhesion to Chitosan Immobilized Titanium Surface: A Race for
the Surface. Colloids Surfaces B Biointerfaces 2015 ,134 , 370–376. https://doi.org/10.1016/j.colsurfb.2015.07.014.
(121) Zarghami, V.; Ghorbani, M.; Pooshang, K. Melittin Antimicrobial
Peptide Thin Layer on Bone Implant Chitosan-Antibiotic Coatings and
Their Bactericidal Properties. Mater. Chem. Phys. 2021 ,263 (December 2020), 124432.
https://doi.org/10.1016/j.matchemphys.2021.124432.
(122) Wu, D.; Zhu, L.; Li, Y.; Zhang, X.; Xu, S.; Yang, G.; Delair, T.
Chitosan-Based Colloidal Polyelectrolyte Complexes for Drug Delivery :
A Review. Carbohydr. Polym. 2020 , 238 (December
2019). https://doi.org/10.1016/j.carbpol.2020.116126.
(123) Kulkarni, A. D.; Vanjari, Y. H.; Sancheti, K. H.; Patel, H. M.;
Belgamwar, V. S.; Surana, S. J.; Pardeshi, C. V. Polyelectrolyte
Complexes: Mechanisms, Critical Experimental Aspects, and Applications.Artif. Cells, Nanomedicine Biotechnol. 2016 , 44(7), 1615–1625. https://doi.org/10.3109/21691401.2015.1129624.
(124) Birch, N. P.; Schiffman, J. D. Characterization of Self-Assembled
Polyelectrolyte Complex Nanoparticles Formed from Chitosan and Pectin.Langmuir 2014 , 30 (12), 3441–3447.
https://doi.org/10.1021/la500491c.
(125) de Vasconcelos, C. L.; Bezerril, P. M.; dos Santos, D. E. S.;
Dantas, T. N. C.; Pereira, M. R.; Fonseca, J. L. C. Effect of Molecular
Weight and Ionic Strength on the Formation of Polyelectrolyte Complexes
Based on Poly(Methacrylic Acid) and Chitosan. Biomacromolecules2006 , 7 (4), 1245–1252.
https://doi.org/10.1021/bm050963w.
(126) Kaygusuz, H.; Micciulla, S.; Erim, F. B.; von Klitzing, R. Effect
of Anionic Surfactant on Alginate-Chitosan Polyelectrolyte Multilayer
Thickness. J. Polym. Sci. Part B Polym. Phys. 2017 ,55 (23), 1798–1803. https://doi.org/10.1002/polb.24429.
(127) Park, H.; Choi, B.; Hu, J.; Lee, M. Injectable Chitosan Hyaluronic
Acid Hydrogels for Cartilage Tissue Engineering. Acta Biomater.2013 , 9 (1), 4779–4786.
https://doi.org/10.1016/j.actbio.2012.08.033.
(128) Hardy, A.; Seguin, C.; Brion, A.; Lavalle, P.; Schaaf, P.;
Fournel, S.; Bourel-Bonnet, L.; Frisch, B.; De Giorgi, M.
β-Cyclodextrin-Functionalized Chitosan/Alginate Compact Polyelectrolyte
Complexes (CoPECs) as Functional Biomaterials with Anti-Inflammatory
Properties. ACS Appl. Mater. Interfaces 2018 , 10(35), 29347–29356. https://doi.org/10.1021/acsami.8b09733.
(129) Li, Z.; Ramay, H. R.; Hauch, K. D.; Xiao, D.; Zhang, M.
Chitosan-Alginate Hybrid Scaffolds for Bone Tissue Engineering.Biomaterials 2005 , 26 (18), 3919–3928.
https://doi.org/10.1016/j.biomaterials.2004.09.062.
(130) Wang, C.; Huang, W.; Zhou, Y.; He, L.; He, Z.; Chen, Z.; He, X.;
Tian, S.; Liao, J.; Lu, B.; et al. 3D Printing of Bone Tissue
Engineering Scaffolds. Bioact. Mater. 2020 , 5(1), 82–91. https://doi.org/10.1016/j.bioactmat.2020.01.004.
(131) Liao, I. C.; Wan, A. C. A.; Yim, E. K. F.; Leong, K. W. Controlled
Release from Fibers of Polyelectrolyte Complexes. J. Control.
Release 2005 , 104 (2), 347–358.
https://doi.org/10.1016/j.jconrel.2005.02.013.
(132) Patil, T.; Saha, S.; Biswas, A. Preparation and Characterization
of HAp Coated Chitosan-Alginate PEC Porous Scaffold for Bone Tissue
Engineering. Macromol. Symp. 2017 , 376 (1), 1–9.
https://doi.org/10.1002/masy.201600205.
(133) Unagolla, J. M.; Alahmadi, T. E.; Jayasuriya, A. C. Chitosan
Microparticles Based Polyelectrolyte Complex Scaffolds for Bone Tissue
Engineering in Vitro and Effect of Calcium Phosphate. Carbohydr.
Polym. 2018 , 199 (April), 426–436.
https://doi.org/10.1016/j.carbpol.2018.07.044.
(134) Hu, Y.; Chen, J.; Fan, T.; Zhang, Y.; Zhao, Y.; Shi, X.; Zhang, Q.
Biomimetic Mineralized Hierarchical Hybrid Scaffolds Based on in Situ
Synthesis of Nano-Hydroxyapatite/Chitosan/Chondroitin Sulfate/Hyaluronic
Acid for Bone Tissue Engineering. Colloids Surfaces B
Biointerfaces 2017 , 157 , 93–100.
https://doi.org/10.1016/j.colsurfb.2017.05.059.
(135) Sellgren, K. L.; Ma, T. Perfusion Conditioning of
Hydroxyapatite-Chitosan-Gelatin Scaffolds for Bone Tissue Regeneration
from Human Mesenchymal Stem Cells. J. Tissue Eng. Regen. Med.2012 , 6 (1), 49–59. https://doi.org/10.1002/term.396.
(136) Coimbra, P.; Ferreira, P.; de Sousa, H. C.; Batista, P.;
Rodrigues, M. A.; Correia, I. J.; Gil, M. H. Preparation and Chemical
and Biological Characterization of a Pectin/Chitosan Polyelectrolyte
Complex Scaffold for Possible Bone Tissue Engineering Applications.Int. J. Biol. Macromol. 2011 , 48 (1), 112–118.
https://doi.org/10.1016/j.ijbiomac.2010.10.006.
(137) Li, Q. L.; Chen, Z. Q.; Darvell, B. W.; Liu, L. K.; Jiang, H. B.;
Zen, Q.; Peng, Q.; Ou, G. M. Chitosan-Phosphorylated Chitosan
Polyelectrolyte Complex Hydrogel as an Osteoblast Carrier. J.
Biomed. Mater. Res. - Part B Appl. Biomater. 2007 , 82(2), 481–486. https://doi.org/10.1002/jbm.b.30753.
(138) Kara, A.; Tamburaci, S.; Tihminlioglu, F.; Havitcioglu, H.
Bioactive Fish Scale Incorporated Chitosan Biocomposite Scaffolds for
Bone Tissue Engineering. Int. J. Biol. Macromol. 2019 ,130 , 266–279. https://doi.org/10.1016/j.ijbiomac.2019.02.067.
(139) Mano, J. F.; Hungerford, G.; Gómez Ribelles, J. L. Bioactive
Poly(L-Lactic Acid)-Chitosan Hybrid Scaffolds. Mater. Sci. Eng. C2008 , 28 (8), 1356–1365.
https://doi.org/10.1016/j.msec.2008.03.005.
(140) Rajan Unnithan, A.; Ramachandra Kurup Sasikala, A.; Park, C. H.;
Kim, C. S. A Unique Scaffold for Bone Tissue Engineering: An Osteogenic
Combination of Graphene Oxide–Hyaluronic Acid–Chitosan with
Simvastatin. J. Ind. Eng. Chem. 2017 , 46 ,
182–191. https://doi.org/10.1016/j.jiec.2016.10.029.
(141) Vyas, V.; Kaur, T.; Thirugnanam, A. Chitosan Composite Three
Dimensional Macrospheric Scaffolds for Bone Tissue Engineering.Int. J. Biol. Macromol. 2017 , 104 , 1946–1954.
https://doi.org/10.1016/j.ijbiomac.2017.04.055.
(142) Cao, X.; Wang, J.; Liu, M.; Chen, Y.; Cao, Y.; Yu, X.
Chitosan-Collagen/Organomontmorillonite Scaffold for Bone Tissue
Engineering. Front. Mater. Sci. 2015 , 9 (4),
405–412. https://doi.org/10.1007/s11706-015-0317-5.
(143) Jayakumar, R.; Ramachandran, R.; Sudheesh Kumar, P. T.; Divyarani,
V. V.; Srinivasan, S.; Chennazhi, K. P.; Tamura, H.; Nair, S. V.
Fabrication of Chitin-Chitosan/Nano ZrO2 Composite Scaffolds for Tissue
Engineering Applications. Int. J. Biol. Macromol. 2011 ,49 (3), 274–280. https://doi.org/10.1016/j.ijbiomac.2011.04.020.
(144) Dan, Y.; Liu, O.; Liu, Y.; Zhang, Y. Y.; Li, S.; Feng, X. bo;
Shao, Z. wu; Yang, C.; Yang, S. H.; Hong, J. bo. Development of Novel
Biocomposite Scaffold of Chitosan-Gelatin/Nanohydroxyapatite for
Potential Bone Tissue Engineering Applications. Nanoscale Res.
Lett. 2016 , 11 (1), 1–6.
https://doi.org/10.1186/s11671-016-1669-1.
(145) Kavya, K. C.; Jayakumar, R.; Nair, S.; Chennazhi, K. P.
Fabrication and Characterization of Chitosan/Gelatin/NSiO2 Composite
Scaffold for Bone Tissue Engineering. Int. J. Biol. Macromol.2013 , 59 , 255–263.
https://doi.org/10.1016/j.ijbiomac.2013.04.023.
(146) Beşkardeş, I. G.; Hayden, R. S.; Glettig, D. L.; Kaplan, D. L.;
Gümüşderelioğlu, M. Bone Tissue Engineering with Scaffold-Supported
Perfusion Co-Cultures of Human Stem Cell-Derived Osteoblasts and Cell
Line-Derived Osteoclasts. Process Biochem. 2017 ,59 , 303–311. https://doi.org/10.1016/j.procbio.2016.05.008.
(147) Yan, H.; Chen, X.; Feng, M.; Shi, Z.; Zhang, D.; Lin, Q.
Layer-by-Layer Assembly of 3D Alginate-Chitosan-Gelatin Composite
Scaffold Incorporating Bacterial Cellulose Nanocrystals for Bone Tissue
Engineering. Mater. Lett. 2017 , 209 , 492–496.
https://doi.org/10.1016/j.matlet.2017.08.093.
(148) Thuaksuban, N.; Nuntanaranont, T.; Suttapreyasri, S.; Pattanachot,
W.; Sutin, K.; Cheung, L. K. Biomechanical Properties of Novel
Biodegradable Poly ε-Caprolactone-Chitosan Scaffolds. J. Investig.
Clin. Dent. 2013 , 4 (1), 26–33.
https://doi.org/10.1111/j.2041-1626.2012.00131.x.
(149) Bhowmick, A.; Jana, P.; Pramanik, N.; Mitra, T.; Banerjee, S. L.;
Gnanamani, A.; Das, M.; Kundu, P. P. Multifunctional Zirconium Oxide
Doped Chitosan Based Hybrid Nanocomposites as Bone Tissue Engineering
Materials. Carbohydr. Polym. 2016 , 151 , 879–888.
https://doi.org/10.1016/j.carbpol.2016.06.034.
(150) Januariyasa, I. K.; Ana, I. D.; Yusuf, Y. Nanofibrous Poly(Vinyl
Alcohol)/Chitosan Contained Carbonated Hydroxyapatite Nanoparticles
Scaffold for Bone Tissue Engineering. Mater. Sci. Eng. C2020 , 107 , 110347.
https://doi.org/10.1016/j.msec.2019.110347.
(151) Shakir, M.; Jolly, R.; Khan, M. S.; Rauf, A.; Kazmi, S.
Nano-Hydroxyapatite/β-CD/Chitosan Nanocomposite for Potential
Applications in Bone Tissue Engineering. Int. J. Biol. Macromol.2016 , 93 , 276–289.
https://doi.org/10.1016/j.ijbiomac.2016.08.046.
(152) Li, X.; Xu, P.; Cheng, Y.; Zhang, W.; Zheng, B.; Wang, Q.
Nano-Pearl Powder/Chitosan-Hyaluronic Acid Porous Composite Scaffold and
Preliminary Study of Its Osteogenesis Mechanism. Mater. Sci. Eng.
C 2020 , 111 (January).
https://doi.org/10.1016/j.msec.2020.110749.
(153) Saravani, S.; Ebrahimian-Hosseinabadi, M.; Mohebbi-Kalhori, D.
Polyglycerol Sebacate/Chitosan/Gelatin Nano-Composite Scaffolds for
Engineering Neural Construct. Mater. Chem. Phys. 2019 ,222 , 147–151. https://doi.org/10.1016/j.matchemphys.2018.10.010.
(154) Akmammedov, R.; Huysal, M.; Isik, S.; Senel, M. Preparation and
Characterization of Novel Chitosan/Zeolite Scaffolds for Bone Tissue
Engineering Applications. Int. J. Polym. Mater. Polym. Biomater.2018 , 67 (2), 110–118.
https://doi.org/10.1080/00914037.2017.1309539.
(155) Maji, K.; Dasgupta, S.; Pramanik, K.; Bissoyi, A. Preparation and
Evaluation of Gelatin-Chitosan-Nanobioglass 3D Porous Scaffold for Bone
Tissue Engineering. Int. J. Biomater. 2016 , 2016 .
https://doi.org/10.1155/2016/9825659.
(156) Khan, S.; Garg, M.; Chockalingam, S.; Gopinath, P.; Kundu, P. P.TiO2 Doped Chitosan/Poly (Vinyl Alcohol) Nanocomposite Film with
Enhanced Mechanical Properties for Application in Bone Tissue
Regeneration ; Elsevier B.V, 2020; Vol. 143.
https://doi.org/10.1016/j.ijbiomac.2019.11.246.
(157) Bosco, R.; Van Den Beucken, J. V.; Leeuwenburgh, S.; Jansen, J.
Surface Engineering for Bone Implants: A Trend from Passive to Active
Surfaces. Coatings 2012 , 2 (3), 95–119.
https://doi.org/10.3390/coatings2030095.
(158) Paital, S. R.; Dahotre, N. B. Calcium Phosphate Coatings for
Bio-Implant Applications: Materials, Performance Factors, and
Methodologies. Mater. Sci. Eng. R Reports 2009 ,66 (1–3), 1–70. https://doi.org/10.1016/j.mser.2009.05.001.
(159) Anselme, K. Osteoblast Adhesion on Biomaterials.Biomaterials 2000 , 21 (7), 667–681.
https://doi.org/10.1016/S0142-9612(99)00242-2.
(160) Gadomska-Gajadhur, A.; Łojek, K.; Szymaniak, M.; Gadomska, A.
Materiały Porowate Do Regeneracji Tkanki Chrzęstnej i Kostnej.Wyr. Med. 2018 , No. August, 51–58.
(161) Cao, S.; Zhao, Y.; Hu, Y.; Zou, L.; Chen, J. New Perspectives:
In-Situ Tissue Engineering for Bone Repair Scaffold. Compos. Part
B Eng. 2020 , 202 (September), 108445.
https://doi.org/10.1016/j.compositesb.2020.108445.
(162) Habibovic, P.; de Groot, K. Osteoinductive
Biomaterials—Properties and Relevance in Bone Repair. J. Tissue
Eng. Regen. Med. 2007 , 1 (1), 25–32.
https://doi.org/10.1002/term.5.
(163) Nowacka, M. Biomateriały Stosowane W Inżynierii Komórkowej I
Medycynie Regeneracyjnej Biomaterials for Tissue Engineering and
Regenerative Medicine. Wiadomości Chem. 2012 , 66 ,
9–10.
(164) Cavalcanti-Adam, E. A.; Aydin, D.; Hirschfeld-Warneken, V. C.;
Spatz, J. P. Cell Adhesion and Response to Synthetic Nanopatterned
Environments by Steering Receptor Clustering and Spatial Location.HFSP J. 2008 , 2 (5), 276–285.
https://doi.org/10.2976/1.2976662.
(165) Roach, P.; Eglin, D.; Rohde, K.; Perry, C. C. Modern Biomaterials:
A Review - Bulk Properties and Implications of Surface Modifications.J. Mater. Sci. Mater. Med. 2007 , 18 (7),
1263–1277. https://doi.org/10.1007/s10856-006-0064-3.
(166) Xiao, D.; Guo, T.; Yang, F.; Feng, G.; Shi, F.; Li, J.; Wang, D.;
Duan, K.; Weng, J. In Situ Formation of Nanostructured Calcium Phosphate
Coatings on Porous Hydroxyapatite Scaffolds Using a Hydrothermal Method
and the Effect on Mesenchymal Stem Cell Behavior. Your Article Is
Registered as a Regular Item and Is Being Processed for Inclusi.Ceram. Int. 2017 , 43 (1), 1588–1596.
https://doi.org/10.1016/j.ceramint.2016.10.023.
(167) Tian, Y.; Liu, H.; Sheldon, B. W.; Webster, T. J.; Yang, S.; Yang,
H.; Yang, L. Surface Energy-Mediated Fibronectin Adsorption and
Osteoblast Responses on Nanostructured Diamond. J. Mater. Sci.
Technol. 2019 , 35 (5), 817–823.
https://doi.org/10.1016/j.jmst.2018.11.009.
(168) Comelles, J.; Estévez, M.; Martínez, E.; Samitier, J. The Role of
Surface Energy of Technical Polymers in Serum Protein Adsorption and
MG-63 Cells Adhesion. Nanomedicine Nanotechnology, Biol. Med.2010 , 6 (1), 44–51.
https://doi.org/10.1016/j.nano.2009.05.006.
(169) Yang, L.; Li, Y.; Sheldon, B. W.; Webster, T. J. Altering Surface
Energy of Nanocrystalline Diamond to Control Osteoblast Responses.J. Mater. Chem. 2012 , 22 (1), 205–214.
https://doi.org/10.1039/c1jm13593g.
(170) Keselowsky, B. G.; Collard, D. M.; García, A. J. Integrin Binding
Specificity Regulates Biomaterial Surface Chemistry Effects on Cell
Differentiation. Proc. Natl. Acad. Sci. U. S. A. 2005 ,102 (17), 5953–5957. https://doi.org/10.1073/pnas.0407356102.
(171) Curran, J. M.; Chen, R.; Hunt, J. A. Controlling the Phenotype and
Function of Mesenchymal Stem Cells in Vitro by Adhesion to
Silane-Modified Clean Glass Surfaces. Biomaterials 2005 ,26 (34), 7057–7067.
https://doi.org/10.1016/j.biomaterials.2005.05.008.
(172) Curran, J. M.; Chen, R.; Hunt, J. A. The Guidance of Human
Mesenchymal Stem Cell Differentiation in Vitro by Controlled
Modifications to the Cell Substrate. Biomaterials 2006 ,27 (27), 4783–4793.
https://doi.org/10.1016/j.biomaterials.2006.05.001.
(173) Sandra, S.; Brygida, Z.; Łagan, S. Porównanie Zwiżalności Oraz
Swobodnej Energii Powierzchniowej Biomateriałów i Tkanki Kostnej.Aktualne Problemy Biomechaniki . 2013.
(174) Govindasamy, K.; Dahlan, N. A.; Janarthanan, P.; Goh, K. L.; Chai,
S. P.; Pasbakhsh, P. Electrospun Chitosan/Polyethylene-Oxide
(PEO)/Halloysites (HAL) Membranes for Bone Regeneration Applications.Appl. Clay Sci. 2020 , 190 (March), 105601.
https://doi.org/10.1016/j.clay.2020.105601.
(175) Sukul, M.; Sahariah, P.; Lauzon, H. L.; Borges, J.; Másson, M.;
Mano, J. F.; Haugen, H. J.; Reseland, J. E. In Vitro Biological Response
of Human Osteoblasts in 3D Chitosan Sponges with Controlled Degree of
Deacetylation and Molecular Weight. Carbohydr. Polym.2021 , 254 (November 2020).
https://doi.org/10.1016/j.carbpol.2020.117434.
(176) Florencio-Silva, R.; Sasso, G. R. da S.; Sasso-Cerri, E.; Simões,
M. J.; Cerri, P. S. Biology of Bone Tissue: Structure, Function, and
Factors That Influence Bone Cells. Biomed Res. Int.2015 , 2015 (6), 1–17.
https://doi.org/10.1155/2015/421746.
(177) Ducy, P.; Schinke, T.; Karsenty, G. The Osteoblast: A
Sophisticated Fibroblast under Central Surveillance. Science (80-.
). 2000 , 289 (5484), 1501–1504.
https://doi.org/10.1126/science.289.5484.1501.
(178) Dirckx, N.; Moorer, M. C.; Clemens, T. L.; Riddle, R. C. The Role
of Osteoblasts in Energy Homeostasis. Nat. Rev. Endocrinol.2019 , 15 (11), 651–665.
https://doi.org/10.1038/s41574-019-0246-y.
(179) Datta, H. K.; Ng, W. F.; Walker, J. A.; Tuck, S. P.; Varanasi, S.
S. The Cell Biology of Bone Metabolism. J. Clin. Pathol.2008 , 61 (5), 577–587.
https://doi.org/10.1136/jcp.2007.048868.
(180) Aarden, E. M.; Burger, E. H.; Nijweide, P. J. Function of
Osteocytes in Bone. [Review] [75 Refs]. J. Cell. Biochem.1994 , 55 (3), 287–299.
(181) Bonewald, L. F. Cell–Cell and Cell–Matrix Interactions in Bone.
In Handbook of Cell Signaling ; Elsevier, 2010; Vol. 1, pp
2647–2662. https://doi.org/10.1016/B978-0-12-374145-5.00313-2.
(182) Klein-Nulend, J.; Bonewald, L. F. The Osteocyte ; Elsevier
Inc., 2019. https://doi.org/10.1016/B978-0-12-814841-9.00006-3.
(183) Brown, J. L.; Kumbar, S. G.; Laurencin, C. T. Bone Tissue
Engineering , Third Edit.; Elsevier, 2013.
https://doi.org/10.1016/B978-0-08-087780-8.00113-3.
(184) Kąkol, P. T. Biologia Kompedium , 1st ed.; Świat Książki:
Warszawa, 2007.
(185) Kalfas, I. H. Principles of Bone Healing. Neurosurgical
focus . 2001, pp 10–13. https://doi.org/10.3171/foc.2001.10.4.2.
(186) Morgan, E. F.; Barnes, G. L.; Einhorn, T. A. The Bone Organ
System. Form and Function , Fourth Edi.; Elsevier, 2013.
https://doi.org/10.1016/B978-0-12-415853-5.00001-7.
(187) Jang, J. H.; Castano, O.; Kim, H. W. Electrospun Materials as
Potential Platforms for Bone Tissue Engineering. Adv. Drug Deliv.
Rev. 2009 , 61 (12), 1065–1083.
https://doi.org/10.1016/j.addr.2009.07.008.
(188) Yang, J.; Ueharu, H.; Mishina, Y. Energy Metabolism: A Newly
Emerging Target of BMP Signaling in Bone Homeostasis. Bone2020 , 138 (June), 115467.
https://doi.org/10.1016/j.bone.2020.115467.
(189) Lin, X.; Patil, S.; Gao, Y. G.; Qian, A. The Bone Extracellular
Matrix in Bone Formation and Regeneration. Front. Pharmacol.2020 , 11 (May), 1–15.
https://doi.org/10.3389/fphar.2020.00757.
(190) Johansen, J. S.; Williamson, M. K.; Rice, J. S.; Price, P. A.
Identification of Proteins Secreted by Human Osteoblastic Cells in
Culture. J. Bone Miner. Res. 1992 , 7 (5),
501–512. https://doi.org/10.1002/jbmr.5650070506.
(191) Gentili, C.; Cancedda, R. Cartilage and Bone Extracellular Matrix.Curr. Pharm. Des. 2009 , 15 (12), 1334–1348.
https://doi.org/10.2174/138161209787846739.
(192) Hua, R.; Ni, Q.; Eliason, T. D.; Han, Y.; Gu, S.; Nicolella, D.
P.; Wang, X.; Jiang, J. X. Biglycan and Chondroitin Sulfate Play Pivotal
Roles in Bone Toughness via Retaining Bound Water in Bone Mineral
Matrix. Matrix Biol. 2020 , 94 , 95–109.
https://doi.org/10.1016/j.matbio.2020.09.002.
(193) Li, X.; Pennisi, A.; Yaccoby, S. Role of Decorin in the
Antimyeloma Effects of Osteoblasts. Blood 2008 ,112 (1), 159–168. https://doi.org/10.1182/blood-2007-11-124164.
(194) Hosseini, S.; Naderi-Manesh, H.; Vali, H.; Baghaban Eslaminejad,
M.; Azam Sayahpour, F.; Sheibani, S.; Faghihi, S. Contribution of
Osteocalcin-Mimetic Peptide Enhances Osteogenic Activity and
Extracellular Matrix Mineralization of Human Osteoblast-like Cells.Colloids Surfaces B Biointerfaces 2019 , 173(October 2018), 662–671.
https://doi.org/10.1016/j.colsurfb.2018.10.035.
(195) Siggelkow, H.; Schmidt, E.; Hennies, B.; Hüfner, M. Evidence of
Downregulation of Matrix Extracellular Phosphoglycoprotein during
Terminal Differentiation in Human Osteoblasts. Bone2004 , 35 (2), 570–576.
https://doi.org/10.1016/j.bone.2004.03.033.
(196) Singh, A.; Gill, G.; Kaur, H.; Amhmed, M.; Jakhu, H. Role of
Osteopontin in Bone Remodeling and Orthodontic Tooth Movement: A Review.Prog. Orthod. 2018 , 19 (1).
https://doi.org/10.1186/s40510-018-0216-2.
(197) Saito, K.; Nakatomi, M.; Ohshima, H. Dentin Matrix Protein 1
Compensates for Lack of Osteopontin in Regulating Odontoblastlike Cell
Differentiation after Tooth Injury in Mice. J. Endod.2020 , 46 (1), 89–96.
https://doi.org/10.1016/j.joen.2019.10.002.
(198) Bellahcène, A.; Castronovo, V.; Ogbureke, K. U. E.; Fisher, L. W.;
Fedarko, N. S. Small Integrin-Binding Ligand N-Linked Glycoproteins
(SIBLINGs): Multifunctional Proteins in Cancer. Nat. Rev. Cancer2008 , 8 (3), 212–226. https://doi.org/10.1038/nrc2345.
(199) Teitelbaum, S. L. Bone Resorption by Osteoclasts. Science
(80-. ). 2000 , 289 (5484), 1504–1508.
https://doi.org/10.1126/science.289.5484.1504.
(200) Bossard, M. J.; Tomaszek, T. A.; Thompson, S. K.; Amegadzie, B.
Y.; Hanning, C. R.; Jones, C.; Kurdyla, J. T.; McNulty, D. E.; Drake, F.
H.; Gowen, M.; et al. Proteolytic Activity of Human Osteoclast Cathepsin
K: Expression, Purification, Activation, and Substrate Identification.J. Biol. Chem. 1996 , 271 (21), 12517–12524.
https://doi.org/10.1074/jbc.271.21.12517.
(201) Dhiman, H. K.; Ray, A. R.; Panda, A. K. Three-Dimensional Chitosan
Scaffold-Based MCF-7 Cell Culture for the Determination of the
Cytotoxicity of Tamoxifen. Biomaterials 2005 , 26(9), 979–986. https://doi.org/10.1016/j.biomaterials.2004.04.012.
(202) Amir, L. R.; Suniarti, D. F.; Utami, S.; Abbas, B. Chitosan as a
Potential Osteogenic Factor Compared with Dexamethasone in Cultured
Macaque Dental Pulp Stromal Cells. Cell Tissue Res.2014 , 358 (2), 407–415.
https://doi.org/10.1007/s00441-014-1938-1.
Figure LegendsFigure 1. Chitin (a) and chitosan (b).
Figure 2. Production of chitosan.
Figure 3. Cell adhesion to biomaterial. After biomaterial implementation
thin water layer is formed on the surface, followed by protein adhesion.
Cells recognise proteins and adhere to the biomaterial, allowing the
tissue to regenerate.Table LegendsTable 1. Chitosan solubility.
Table 2. Examples of polyelectrolite complexes with chitosan in tissue
engineering
Table 3. Examples of chitosan composites in bone tissue engineering