Materials for tissue regeneration and wound healing management: A review

Rohan Ramesh Pawar

CHE 200
APRIL 28, 2023
Abstract
The biomedical field has made considerable achievement in developing materials in skin regeneration and
wound management areas during the previous few decades. Natural and synthetic based polymers are being
utilized for wound care and tissue regeneration due to their unique properties. Metal nanoparticles have also
been investigated for their applications in these areas. Due to limitations such as shortage of skin donor
sites, immune rejection and immunological responses, there is a need for alternate and appropriate skin
substitutes in dermal regeneration, therefore wound healing is essential. A significant amount of recent
research has also focused on integrating the various materials into skin/bone grafts, wound dressings, or
formulations (oral or topical) to combine the distinct properties and advantages that each material offers.
The ideal material would have the necessary mechanical and biological characteristics. The cellular
characteristics could possibly include not only speedier healing, but also avoiding infections and acting as a
skin graft for tissue regeneration, resulting in an overall recovery.

Fig 1-Graphical Abstract [1].

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Contents
1. Introduction.............................................................................................................................................. 2
1.1. Why alternate tissue regenerative and wound healing materials?...................................................... 2
1.2. Methods of preparation..................................................................................................................... 3
1.3. Hydrogel scaffold for tissue regeneration and wound healing .......................................................... 3
2. Applications of Chitosan in wound healing and tissue regeneration.......................................................... 4
3. Applications of Polyurethane in wound healing and tissue regeneration .................................................. 9
4. Applications of Silver in wound healing and tissue regeneration ........................................................... 11

5. Conclusions............................................................................................................................................. 13
6. Mini Proposal .......................................................................................................................................... 14
7. References............................................................................................................................................... 14

1. Introduction

1.1. Why alternate tissue regenerative and wound healing materials?

Several items and treatments including gauzes, lint, cloth, cotton, tulle, and so on, are already available in
the market. However, these materials do not give a perfect material that would combines the mechanical,
antiseptic, and healing promoter/tissue graft properties. Over the years, researchers have developed
materials, either natural or synthetic, that would combine many of these qualities [2]. Biopolymeric-based
technology employes biocompatible polymers with the ability to overcome the limitations of debridement,
autografting and allografting. They are important in wound healing and tissue regeneration. Skin
debridement is important because it aids in the process of re-epithelialization. The biopolymers d r e s s i n g
promote autolytic debridement, which directly improves the various biological properties such as keeping
the wound area moist, providing good oxygen supply, and water vapor permeability. They also avoid
maceration of tissue by removing the extreme exudates at the wound site, thus promoting the process of cell
proliferation, vascularization, and re-epithelialization [3].

Natural biopolymers are always advantageous and preferred over synthetic polymers due to their
biocompatibility, biodegradability, sustainability, hydrophilic nature, stability across the pH variations, and
low immunogenicity. They have extracellular matrix (ECM) mimicking properties at the time of dermal
regeneration [4]. Synthetic materials on the other hand, offer consistency/uniformity, customizable
mechanical support, and tunable porosity properties. An increased fluid uptake ability due to porosity aids
in cell proliferation, migration, differentiation, nutrition exchanges, and faster wound healing [5]. Figure
1 shows the different methods of wound healing. Different stages of wound healing such as hemostasis,
coagulation, inflammation, proliferation and migration, and remodeling is shown in figure 2.

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 Fig. 2. Schematic of wound healing [2]

1.2. Methods of preparation

There are several methods available for preparing these grafts or scaffolds for tissue regeneration and wound
healing. Solvent casting, particle leaching, electrospinning, and hydrogel creation are examples of these
techniques. Electrospinning is by far the most common technique because it aids in the fabrication of fibrous
scaffolds ranging from submicron to nano scales, resulting in high surface, high porosity, and better cell
adhesion properties, but it falls short of providing a three-dimensional (3D) environment for healing [6].
Hydrogels were created to provide a 3D environment that resembles the ECM of native tissues allowing for
quicker healing. Chemical and physical approaches can be used to cross-link hydrogels, with the physical
method involving either physical cross-linking of the polymer networks or physical interactions between the
polymer chains [7].

1.3. Hydrogel scaffold for tissue regeneration and wound healing

The epidermis, dermis, and subcutaneous layers make up the skin. It shields the body from damaging
external environmental assaults and serves as a physical support for the vascular and neurological systems.
Any skin injury destroys this protective layer, resulting in water loss, impaired sensitivity, thermoregulation,
and bacterial infection [8]. Wound healing is a complicated and dynamic process that is regulated by a
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variety of biomolecules and cells. Skin regeneration is influenced by both innate biological processes and
external stimuli. Hydrogels serve as temporary skin substitutes possessing specific properties to promote
these factors and hence promote wound healing. It also absorbs excess fluid to maintain a moist
microenvironment. Hydrogels can be customized and functionalized to include additional elements that can
increase wound healing properties. [9].

As shown in Fig. 3, the hybrid scaffold forms a network with the keratinocytes. This forms a
matrix that covers, shields, and heals the injury while keeping it wet, allowing re-epithelization of the
tissue [10].

Fig. 3. Schematic representation of wound healing and tissue regeneration using a hydrogel scaffold [1].

This literature review compares three types of materials that are commonly used in tissue
regeneration and wound healing: Natural polymers such as chitosan, synthetic polymers such as
polyurethane, and metal nanoparticles made of silver. Comparison table 1 outlines the applications of these
three materials in the wound healing and tissue regeneration area along with their salient features. Natural
polymers are the most used materials, due to their ease of availability and biocompatibility with the body.
The advent of polymers in biological treatment led to the invention of polyesters and polyurethanes, that
are now being widely used in wound healing and tissue regeneration. Metals such as gold and silver, while
known for their antibacterial properties, were not used for wound healing and tissue regeneration until the last
decade. Silver nanoparticles are the most used, either alone or in combination with other materials to
combine the antiseptic properties along with their wound healing and tissue regeneration properties.

2. Application of Chitosan in wound healing and tissue regeneration

Kim Mirae et al. [11] examined the use of hydrogels in wound healing and adhesives. The tissue adhesive
properties of the hydrogels were examined using in vivo and ex vivo models as tissue sealants. The wound
recovery process was observed for five days. A rat incision model was utilized to examine the in vivo
biocompatibility and skin wound closure capabilities of Cs-PEG hydrogel as tissue sealants.
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 Three different hydrogels encapsulated with epidermal growth factors (EGF) were tested: PEG 750 (0.7) g-
Cs, PEG2k (0.7) g-Cs and PEG 2k (0.7) g-Cs. Control groups included untreated, sutured and Beriplast- P
treated conditions. The untreated wound gap was big and did not close naturally in the absence of any sealant.
For suture, the gap was closed, and considerable wound healing occurred by day 3.

Fig. 4. (a) Cs-PEG hydrogel was applied to a rat skin wound model via syringe to close the wound gap. (b) OCT imaging
was used to visualize the detailed skin tissue around the wound area.

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Fig. 4. (c) After applying tissue sealants, the wound closure and subsequent healing process was monitored over time
for up to 5 days. (c-1) untreated, (c-2) suture, (c-3) Beriplast-P, (c-4) PEG750-g-Cs hydrogel, (c-5) PEG2K-g-Cs
hydrogel, (c-6) PEG@K-g-Cs encapsulated with EGF. the wound area.

Beriplast-P’s hydrogel tissue sealant closed the wound gap and healed the wound by day 5. PEG 750 (0.7) g-
Cs caused slower wound healing and less inflammation compared with Beriplast-P. Because of its increased
moisture retention property, PEG 2k (0.7) g-Cs cured the wound and significantly reduced inflammation.
PEG 2k-g-Cs (EFG) expedited the wound process due to presence of EGF and formed less fibrotic tissue.
As shown in Fig. 4 (c), untreated and Beriplast-P treated tissues showed the highest degree of fibrosis. PEG
750 (0.7)-g-Cs and PEG 2k -g-Cs showed less fibrosis. EGF encapsulated PEG 2k-g-Cs treated tissue showed
the least amount of fibrosis. These findings show that the Cs-PEG hydrogel is an effective tissue sealant
when combined with a controlled release of a therapeutic drug.

The findings also show that the presence of PEG in the hydrogels in the form of grafting or crosslinker
synergistically improved the antimicrobial activity due to hydrophobicity and anti-fouling properties of
PEG. Rheological analysis of the gelation kinetics revealed that all hydrogels with a few exceptions formed
quickly in a minute. The adhesive properties of Cs pEG hydrogels were generally in line with the mechanical
properties and were like those of a commercial hydrogel-based sealant.

Sundaram N et al. [12] studied the effects of addition of inorganic materials like propanoic acid (PA) and
calcium (Ca) to increase hemostatic potential of calcium (Cs) based hydrogel. In vivo hemostatic evaluation
of composite hydrogel with commercial hemostatic agents like Fibrin and Flosaeal was observed. Rat liver
hemorrhage models that depict oozing bleeding and femoral artery hemorrhage were used. The time taken by
hydrogel to stop bleeding completely was noted. A smooth injectable composite hydrogel was created by
consistently mixing 2% Cs hydrogel with 0.25% PA and 0.25% Ca. The injectability of the generated was
visually inspected, revealing a smooth, uninterrupted flow of hydrogel. Rheological studies on 2% Cs and
2% Cs-0.25% PA-0.25% Ca hydrogels showed high elastic modulus value indicating its gel like property.
The prepared hydrogel flowed only under external pressure. The concentration of Cs hydrogel, PA, and Ca
to form Cs-PA-Ca composite hydrogel was optimized using an in vitro blood clotting experiment. The
hydrogels with 2% Cs and 3% Cs concentrations had the shortest clotting times.

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Fig. 5. In vivo evaluation of Cs-PA-Ca hydrogel in liver hemorrhage model, (a) exposed rat liver, (b) injury created
with 5mm biopsy punch, (c) initiation of bleeding, (d) Cs-PA-Ca composite hydrogel application, (e) stable clot
formation, (f) time for hemostasis and (g)blood loss during the procedure.

Fig. 6. In vivo evaluation of cs-Pa-Ca hydrogel in femoral artery hemorrhage model. (a) femoral artery was exposed,
(b) hemorrhage caused by puncturing artery, (c) initiation of bleeding

Different concentrations of Ca (0.0625%, 0.125%, 0.25%, and 0.5%) were mixed with 2% Cs hydrogel and
clotting time of Cs-Ca hydrogel was evaluated. The duration of clotting was less in the 2% Cs-0.25 Ca (157+/-
9s) and 2%-0.5% Ca (146+/-7) hydrogel. W h e n 0. 5 % w e r e u s e d i n t h e c cytocompatibility study,
the percentage of viable cells decreased significantly. T h e 2% Cs-0.25% PA-0.25% Ca hydrogel showed
the least clotting time of 112+/-12s.Th combination of 2% Cs-0.25% PA-0.25% Ca resulted in rapid blood
clot formation due to the combined effects of interaction with cell membrane of red blood cells (RBC) and
platelets, resulting in aggregation and formation of a clot. The in vivo studies on rat liver and femoral artery
hemorrhage models revealed that 2% Cs-0.25% PA-0.25% Ca composite hydrogel had the potential to form
stable blood clots in the shortest time with minimal blood loss, surpassing the performance of commercial
hemostatic agents. This suggests that the developed injectable Cs-PA-Ca composite hydrogel could act on
multiple mechanisms of hemostasis and facilitate rapid bleeding control even without the need for external
compression. Composite hydrogel is a potential promising option for controlling bleeding during surgeries,
trauma, or other bleeding emergencies.
Rafetley R et al. [13] investigated the utilization of natural collagen in tissue engineering for orthopedic
applications. Collagen has inherent bioactivity; however, its mechanical strength and degradation rate are
limited. The paper explored the use of combining collagen with other materials such as chitosan, to improve
the mechanical and biological properties of the scaffold. The researchers evaluated the efficacy of bovine-
derived collagen to salmon skin-derived collagen and assessed their ability to support mesenchymal stem
cell proliferation and differentiation. The scaffold’s architecture and mechanical properties were evaluated,
and the compressive strength, swelling ratio, degradation rate, pore size, porosity, and water uptake capacity

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 were quantified. Bovine tendon collagen and marine salmon skin collagen were employed to create a range
of collagen-based scaffolds either alone or as composites with chitosan derived from crustacean shells. The
scaffolds were cultured in serum containing media for 28 days at 37 degrees Celsius. A significant reduction in
pore size to 74.7+/-2.3 micrometer was observed, when the chitosan to collagen ratio was 50:50, but there
was no significant difference in pore size in bovine-derived collagen scaffolds containing chitosan t ratios
of 100:0, 90:10, and 75:25 respectively. Pore size was the smallest in the salmon-derived collagen groups in
the scaffold with a collagen to chitosan ratio of 100:0, but it increased dramatically as additional chitosan
was added.

Fig. 6. Effect of chitosan incorporation on scaffold morphology. Bovine and salmon-derived collagen scaffolds
containing chitosan at ratios of 100:0, 90:10, 75:25 and 50:50 was assessed for pore size, porosity, and porous
architecture. The incorporation of chitosan into a bovine-derived collagen scaffold causes a decrease in pore size while
the opposite occurs with salmon-derived collagen scaffold groups (A). Data plotted shows mean ± standard deviation
(n = 3) and p < 0.05, ⁄⁄ p < 0.01 and ⁄⁄⁄ p < 0.001. Percentage porosity was calculated as a relative ratio of scaffold density
to the density of solid collagen and chitosan. All scaffolds had mean porosity of >97% (B).

Overall, the bovine-derived collagen scaffold outperformed the salmon derived collagen scaffolds in all tests,
with larger pore sizes and higher percentages of porosity (Fig 6 A and B). Furthermore, the number of cells
on the collagen chitosan scaffold made from salmon were 50 % lower than that of bovine (90:10 and 75:25). As
a result, the bovine derived collagen scaffold with a 75:25 was deemed the best scaffold for bone and cartilage
tissue engineering.
Sun Pi et al. [14] studied wound healing in the presence of bacterial infections, which can lead to chronic
wounds due to antibiotic resistance. To enhance wound healing, the researchers recommended using a
multifunctional nanocomposite hydrogel (CS-HCA-Icps) containing chitosan, catechol-grafted chitosan (CS-
HCA), and curcumin-Fe3+ coordination nanoparticles (Icps, Cur- Fe3+). The hydrogel had anti-bacterial,
antioxidant and anti-inflammatory properties and responded to the wound’s acidic microenvironment. They
proposed that this hydrogel could be a potential technique for safe and efficient wound healing, particularly
bacterial infections were present. The hydrogel was synthesized by mixing chitosan, catechol-grafted, and
curcumin-Fe3+ coordination nanoparticles. The researchers used Fourier transformation infrared
spectroscopy, scanning electron microscopy, and rheological measurements. They also evaluated the
hydrogel’s properties such as antioxidant activity, antibacterial activity and biocompatibility using in vitro
and in vivo experiments. Finally, they tested the hydrogel’s efficacy in promoting wound healing using a

8
rat model.
The paper’s findings revealed that the multifunctional nanocomposite hydrogel (Cs-HCA-Icps) has
excellent wound healing characteristics. The hydrogel exhibited antioxidant, antibacterial, and anti-
inflammatory activities and was sensitive to the wound’s acidic microenvironment. The invitro and in vivo
tests revealed that the hydrogel was biocompatible. The hydrogel enhanced wound healing in the rat model,
especially in the presence of bacterial infections. The paper concludes that the multifunctional
nanocomposite hydrogel (CS-HCA-Icsp) is a promising bioactive material for promoting infected wound
tissue repair and regeneration. The hydrogel has anti-bacterial antioxidant, and anti- inflammatory
properties, and is responsive to the acidic microenvironment of the wound. Hydrogel was found to be
biocompatible and effective in promoting wound healing in vitro and in vivo experiments. The researchers
suggested that hydrogel could be a potential clinical treatment for infected wounds.

3. Applications of Polyurethane in wound healing and tissue regeneration

Sergei et al. [15] researched on the use of polyurethane (PU) as a versatile construction material in various
applications, including medical applications due to its unique properties. They focused on modifying PU
with a polymeric biocide called poly hexamethylene guanidine hydrochloride (PHMG-Cl) to create
antibacterial polymer composites with controlled biocide release. The elasticity and hydrophobicity of the
modified PU films were improved, and the biocide release was gradual over time. According to the paper,
the modified PU can be used in medical applications such as wound dressing and surgical drapes where local
antimicrobial activity is required. Antibacterial polymer biocide PHMG-Cl was prepared with solvent
casting method. The modified polymer films were tested for their tensile strength, elasticity, hydrophobicity,
thermal and thermophysical properties, and biocide release rate. The paper also conducted a microbiological
study to evaluate the antibacterial activity of modified PU films. The paper used standard methods for each
test, and the measurements were performed in triplicate for each polymer film.

When compared to neat PU, the modified PU films had higher elasticity and hydrophobicity. The biocide
was released gradually over time, and the modified PU films exhibited high antibacterial activity with 2%
and 3% polymeric biocide. The material can be considered both biocide release and contact active system,
and the latter mechanism of antibacterial activity probably dominates, especially at low PHMG-Cl content
of 2% and 3%. This suggests that the modified PU can be used in medical applications such as wound
dressings and surgical drapes where local antimicrobial activity is required. This concludes that modifying
PU resin with polymeric biocide PHMG-Cl is an efficient approach to prepare new promising antimicrobial
activity when contained 2% and 3% polymeric biocide. The biocide release and contact active system, and
the latter mechanism of antibacterial activity probably dominates especially at low PHMG-Cl content of
2% and 3%. Therefore, the modified PU can be used in medical applications such as wound dressings and
surgical drapes where local antimicrobial activity is required.
The increasing demand for bone replacement and the limitations of available grafts were explored by
Nilawar et al. [16]. They proposed that bone tissue engineering offers promising alternative for grafts, and
the tissue scaffold is a key ingredient that provides a suitable microenvironment for cells to attach,
proliferate, and differentiate to generate the desired tissue. The biomaterial utilized to build the scaffold
should offer a combination of several properties such as degradation, biocompatibility, and differential
ability. The methods used in this study involve the synthesis of two different polyurethanes from olive oil,
with optional polyethylene glycol (PEG) incorporation, as well as physical, mechanical, and thermal
characterization of the polymers created. The synthesized polymers were compressed molded into flat two-
dimensional substrates by compression molding and salt leached into three-dimensional porous structure.
The prepared polyurethanes were also evaluated in vitro for their cytocompatibility and their ability to
support the osteogenic differentiation of pre-osteoblasts. It was concluded that the synthesized olive oil-
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based polyurethanes are potential biomaterials for building scaffolds with customizable degradation and
mechanical properties for bone tissue engineering applications. The incorporation of PEG into the polymer
network improved the hydrophilicity and degradation rate of the polyurethanes, but adversely affected
osteogenic differentiation. The synthesized polyurethanes can be fabricated into 2D substrates and 3D
scaffolds via solvent casting, compression molding, and salt leaching methods.
Wong D et al. [17] studied on the use of a synthetic biomaterial called quadral hexamethylene diisocyanato
(QHM) polymer for tendon repair and regeneration. The researchers aimed to enhance the bioactivity and
strength of the QHM polymer by immobilizing a protein called growth and differentiation factor-7 (GDF 7),
which promotes the differentiation of tenocytes (cells that form tendons) and tendon-like tissue formation.
The study used rat and mouse models for in vitro and in vivo experiments. The researchers discovered that
QHM polymers were cell-compatible and could be coated with fibrin to immobilize GDF-7, resulting in the
formation of tendon-like tissue. In vivo and in vitro experiments were conducted to investigate the use of
QHM polymer for tendon repair and regeneration. They investigated the effects of GDF-7 immobilization
on tenocyte development and the creation of tendon-like tissue formation in mouse and rat models.
Subcutaneous implantation of the QHM polymer in mice and rotator cut injury models in rats were used in the
investigation. Histology, immunohistochemistry, and polarized light microscopy were utilized to analyze the
tissue samples.

The results of the research revealed that HM polymer was cell-compatible and could be coated with fibrin
to immobilize GDF-7 protein. In mouse subcutaneous and rat rotator cut injury models, GDF immobilization
increased tenocyte-like development in vitro and tendon-like tissue formation in vivo. They concluded that
the strategy of employing GDF-7 immobilized QHM polymer is effective for biological enhancement of
synthetic biomaterials in rotator cut repair and regeneration to generate bioactive and robust tendon like
grafts. The study demonstrated that the use of QHM polymer coated with fibrin and immobilized with GDF-
7 protein is effective for promoting tenocyte-like differentiation and tendon-like tissue formation in vitro
and in vivo. The authors concluded that this strategy can be used for biological augmentation of synthetic
biomaterials to achieve bioactive and strong tendon-like grafts in rotator cuff repair and regeneration.
Li M et al. [18] conducted study on the significance of wound dressings in the medical field and every life. It
emphasizes the need for wound dressings to have good biocompatibility and non-toxicity, as well as various
functions to aid in wound healing. The research work mentions the benefits of wet wound dressings over
dry dressings in promoting biosynthetic activities of the skin. in vivo Preparation of electroactive
polyurethane films, evaluation of mechanical and shape memory properties, testing their biocompatibility
and antioxidant capacity, investigating their effect on the cell proliferation, and assessing their in vivo wound
healing properties. Researchers used nuclear magnetic resonance, Fourier transform infrared
spectroscopy, and gel permeation chromatography to investigate the chemical characterization of PCL PEG-
AT copolymers, Additionally, an agar diffusion test was performed to evaluate the antibacterial activity of
VCM-loaded films and tested the hydrophilicity of all copolymer films by water contact angel
measurement and swelling experiment.
The paper’s findings revealed that the electroactive polyurethane films have excellent mechanical and shape
memory properties, as well as good biocompatibility, and antioxidant capability. The films considerably
enhanced the proliferation of L929 cells and significantly promoted in vivo wound healing. The antibacterial
activity was also demonstrated by the VCM-loaded films. Nuclear magnetic resonance, Fourier transform
infrared spectroscopy, and gel permeation chromatography were used to evaluate the chemical properties of
PCL_PEG_AT copolymers. All copolymer films were also tested for hydrophilicity.

The paper concluded that the electroactive shape memory film dressings with multiple functions favorable to
wound healing process significantly promoted the in vivo wound healing of full-thickness skin wound
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compared to non-electroactive film and commercial dressing. The PCL-PEG-AT12 dressing was found to
be the most effective in promoting wound healing. The ability to promote wound healing was further
strengthened with the antibacterial activity of vancomycin, indicating that they are excellent candidates as
bioactive wound dressings for cutaneous skin wound healing.

4. Applications of Silver in wound healing and tissue regeneration

The paper by He C et al. [19] described the development of a wound dressing that has antibacterial
properties and is compatible with cells. The dressing was designed to prevent external microbe adherence
and invasion, destroying invading microorganisms, and encourage cell growth. The dressing was created
by electrospinning, and silver nanoparticles were added to enhance antibacterial activity. The dressing was
examined in vitro and in vivo, and the results suggest that the sandwich structure design could lead to the
development of improved wound dressings for clinical use. The sandwich-structured wound dressing was
created using electrospinning by the researchers. The dressing was made primarily of polycaprolactone
(PCL) and gelatin (Gel). To offer antibacterial action, silver nanoparticles (Ag NPs) were incorporated into
the middle layer of the dressing. The dressing was characterized using various techniques, including
scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and X-ray
diffraction (XRD). In vitro experiments were conducted to evaluate the antibacterial activity and
biocompatibility of the dressing, and in vivo experiments were conducted using a rat wound model.
Statistical analyses were performed using SPSS version 21.

According to the findings, the sandwich-structured wound dressing with Ag NPs exhibited good
antibacterial activity and biocompatibility. In vitro experiments showed that the dressing containing 10%
Ag NPs had better antibacterial activity than the dressings containing 1% and 5 % Ag NPs. The
biocompatibility of the dressing was much better than that of the commercial silver sulfadiazine (SSD)
dressing. In vivo experiments using a rat wound model showed that the dressing loaded with 10 % Ag NPs
resulted in a significantly faster wound healing rate than the SSD dressing and gauze. The results indicate
that the sandwich structure design could lead to the development of improved wound dressings for clinical
use. The research concluded that the sandwich-structure wound dressing with Ag NPs has good
antibacterial activityand biocompatibility. The dressing was designed to prevent the adhesion and invasion
of external microorganisms., kill invading microorganisms, and promote cell growth. The results of in vitro
and in vivo experiments suggest that the sandwich structure design could lead to the development of
improved wound dressings for clinical use. This suggests that the dressing has potential for use in actual
clinical practice.

Ahsan et al. [20] used a green approach to synthesize silver nanoparticles from cabbage extract. The
nanoparticles created are placed onto PVA hydrogels patches. The effectiveness of these patches in
promoting wound healing and reducing inflammation was evaluated through in vitro and in vivo
experiments. The size and morphology of the nanoparticles were analyzed using SEM and UV-Vis’s
spectroscopy. The researchers utilized a green approach to synthesize silver nanoparticles from cabbage
extract. These nanoparticles were then loaded onto PVA hydrogel patches. The effectiveness of these
patches in promoting wound healing and reducing inflammation is evaluated through in vitro and in vivo
experiments. The size and morphology of the nanoparticles were analyzed using SEM and UV-Vi’s
spectroscopy. The results showed that the silver nanoparticles loaded PVA hydrogel patches synthesized
through a green approach had significant potential in promoting wound healing and reducing inflammation.
The in vitro experiments showed that the patches have antibacterial activity against E. coli and S Aureus.
The in vivo experiments on rats also showed that the patches were effective in promoting wound healing
and reducing inflammation. The synthesized nanoparticles were found to be spherical in shape with an
average size of 20-30 nm. The research concluded that the green synthesis of silver nanoparticles using
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cabbage extract is an effective and eco-friendly method. The synthesized nanoparticles were found to be
effective in promoting wound healing and reducing inflammation when loaded onto PVA hydrogel patches.
The in vitro and in vivo experiments showed that the patches have antibacterial activity against E. coli and
S aureus. The study suggests that these patches have potential for use in wound healing applications.

The research by Ovais M et al. [21] investigated the problem of antibiotic resistance in the treatment of
bacterial infection and the potential of silver metal as an antimicrobial agent. However, the use of silver ions
has shown side effects. Nanotechnology can potentially overcome these challenges by using silver
nanoparticles. They presented a sustainable supramolecular chemistry approach to make stabilized silver-
peptide composite nanoparticles that have promising activity against Escherichia coli and Staphylococcus
aureus at an in vitro level and could cure wound disinfections in-vivo. The composite nanoparticles show
excellent biocompatibility, and no side effects were observed in liver enzymes and organs in a
histopathology study. The research used a sustainable supramolecular chemistry approach to make
stabilized silver-peptide composite nanoparticles under mild UV light exposure. The photochemical method
is a green approach that reduces the toxic effects of chemicals used in traditional synthetic strategies. The
composite nanoparticles were tested for their antimicrobial activity against Escherichia coli and
Staphylococcus aureus at an in-vitro level and their ability to cure wound disinfections in-vivo.

The composite nanoparticles made using the sustainable supramolecular chemistry approach demonstrated
promising activity against Escherichia coli and Staphylococcus aureus at an in-vitro level and could cure
wound disinfections in-vivo. The composite nanoparticles also showed excellent biocompatibility, and no
side effects were observed in liver enzymes and organs in a histopathology study. This leads to the conclusion
that the sustainable supramolecular chemistry approach used to make stabilized silver-peptide composite
nanoparticles is a promising strategy for developing novel nanomaterials as potential antibiotic agents of the
future. The composite nanoparticles demonstrated excellent antimicrobial activity against Escherichia coli
and Staphylococcus aureus at an in-vitro level and could wound disinfections in-vivo. The composite
nanoparticles also showed excellent biocompatibility and no side effects were observed in liver enzymes
and organs in the histopathology study.

Pal S et al. [22] researched the potential of bacterial cellulose (BC) as a wound healing material. BC is a
nanofibrous network structure produced by the fermentation of Acetobacter xylinum that has excellent
mechanical properties, moldability, and biocompatibility. It can be functionalized with a wide range of
nanomaterials, making it suitable for various applications. The research focused on the functionalization of
BC with silver nanoparticles (AgNPs) to create an antimicrobial membrane for wound healing. The AgNPs
were synthesized inside the BC network using a green synthesis method and were found to have a narrow
size distribution. The resulting composite showed high antibacterial activity against Escherichia coli and did
not release significant amounts of silver even after long soaking times. They utilized the green synthesis
method to prepare silver nanoparticles (AgNPs) inside the bacterial cellulose (BC) network. An ethanol-
water mixture was used as the solvent, and the BC pellicles were irradiated with UV light for 15 minutes.
The treated samples were then washed with ethanol and distilled water, followed by autoclaving and storage
in deionized water. The resulting Ag/BC composite was characterized using various techniques such
as X-ray diffraction and morphological analysis. The antibacterial activity of the composite was evaluated
using disk diffusion and growth dynamics methods against Escherichia coli.

A green synthesis approach was used to successfully synthesis silver nanoparticles (AgNPs) inside the
bacterial cellulose (BC) network... Even after extensive soaking times, the resultant Ag/BC composite
demonstrated strong antibacterial activity against Escherichia coli and did not release significant amounts of
silver. The composite was found to have a narrow size distribution of AgNPs, and the BC network showed
a highly crystalline nature. The AgNPs were photochemically deposited onto the BC gel network as well as
chemically bonded to the cellulose fiber surfaces. The results suggest that the Ag/BC composite could be
effective in wound-healing treatments. The paper concluded that the bacterial cellulose (BC) functionalized

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with silver nanoparticles (Ag NPs) could be a promising material for wound-healing treatment. Using a
green manufacturing process, the Ag NPs were successfully deposited on the BC matrix, and the resulting
composite demonstrated high antibacterial activity against Escherichia coli. The slow release of silver from
the composite suggests that it could be used for long-term wound healing. The highly porous cellulose
pellicle, as a substrate, favored the diffusion of the silver ion and the hydroxyl group-rich cellulose fiber
surface accelerated the deposition of ionic silver, which turned into metallic silver when exposed to UV
radiation.

Comparison table 1

Chemical Mechanical Thermal Rheological Biological Citation

properties properties properties properties
PEG2k (1): g- Inhibition:70 % [11]
Cs Antimicrobial
activity: 2% and
above
2 % Cs:0.25 Clotting [12]
%, PA:0.25 % time:112+/-12s
Least blood loss
CS: HCA- Toughness: Kill ratio:63.97 [13]
ICPS 3.18 KJ/m3 % against E. coli
at 0.4 % and S. aureus
ICPS
Bovine Porosity- 2.5: fold greater [14]
collagen: above 99 % no. of cells
chitosan Bulk comp
modulus:0.
92 to 1.09
KPa
PU/PHMG-Cl Elasticity: Highest zone of [15]
:2-4 % at least 20 inhibition at 4 %
% increase

PEG TGA: Swelling [16]

polyurethane 283 ratio-93 %.
degrees Neat PU-
Celsius 11 %
UVQHM Tensile Cell [17]
strength: attachment:218
12-74 MPa X
Tensile Cell
Modulus: proliferation:14
0.6-2.7 X
MPa
PCl-PEG- Swelling [18]
AT12 film ratio-60 %
Contact
angle-66o

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10 % Ag NP Inner layer Inhibition zone [19]
contact diameter-2.26+/-
angel-23+/- 0.57mm
4.7o
Outer
contact
angel-
129.1+/-
5.2o
AgNP-BC-HP Wound Minimum [20]
healing inhibition conc:
area on the E. coli:6.3
last day- µg/mL
0.00+/- S. aureus-12.5
0.00cm2 µg/mL

Ag/FFssFF Wound reduction [21]

nanocomposite area-3 fold on
day 9
Survival rate-2.9
% for E. coli and
2.1 % for S.
aureus
Ag/BC at 10-2 E. coli cell no. [22]
M AgNO3 (*108 CFU/mL)-
0.2 for 25-100
hrs.
Inhibition radius-
6.5 mm

5. Conclusions
Tissue engineering is a promising field that regenerates damaged tissues using cells. Scaffolds, and bioactive
chemicals. These scaffolds should be morphologically, biochemically, and mechanically like ECM to
promote cell adhesion, proliferation, and differentiation. Various natural and synthetic polymers, along with
some metals have been used for preparation of the scaffolds. Each material has its own advantages and
disadvantages, and combining the various materials will bring together the best of each material to create
an ideal scaffold or hydrogel for wound healing and tissue regeneration. Furthermore, the development of
smart polymers has opened new pathways for medication administration and healing by mimicking changes in
pH, temperature, and biochemical reactions. One of the important structures for smart hydrogels is that it
may be utilized as an injectable in-situ forming hydrogel for any internal injuries, which is a less invasive
technique. Hence, according to the target tissue for treatment, the appropriate biomaterials and method of
scaffold fabrication can be designed.

14
6. Mini Proposal

Fig. 3. Proposed combination of chitosan, polyurethane, and silver nanoparticles to create an ideal
scaffold/graft for wound healing and tissue regeneration.
Combining various biocompatible materials for wound healing has been an interest in researchers for the
past decade. To acquire the specified qualities of the e scaffold/hydrogel, two or more polymers/ materials
have been combined. Chitosan is a great natural polymer due to its strong mechanical properties as well as
biocompatibility; but, because of its non-tunable polymer properties, the erosion of the polymer that
ultimately controls drug delivery is not as manageable. Polyurethane, on the other hand, has tunable erosion
properties that allow the polymer to degrade over a range of days, weeks, and months, depending on the severity
of wound. Although both chitosan and polyurethane have excellent wound healing properties, neither can
protect the wound from bacterial infections. Hence, silver nanoparticles can serve as an excellent adjuvant in
providing antiseptic properties along with the wound healing properties. Silver nanoparticles alone are not
able to create a 3D mesh structure to support tissue regeneration and wound healing. Thus, combining all
three materials in one scaffold or graft would essentially combine the best of all the materials and lead
to a novel material that not only has superior wound healing and tissue regeneration properties, but also does
not require the addition of a separate antiseptic to the system due to its own antiseptic properties. This novel
material has the potential to be applied to the skin or injected for internal damage. Thus, a combination of
these materials should be evaluated in future to create the ideal scaffold/graft for wound healing and tissue
regeneration.

15
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