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Research Article
A Combination of Coconut Fiber Suture and Tamarind Seed
Gel with Dehydrated Human Amnion Membrane for Wound
Surgery in Rats
Raghu Babu Pothireddy ,
1
Angeline Julius,
2
Manu Thomas Mathai,
1
Ganesh Lakshmanan,
3
and Beimnet Asfaw Hailemariam
4
1
Department of Zoology, Madras Christian College, Affiliated to University of Madras, Chennai 600059, Tamil Nadu, India
2
Centre for Materials Engineering and Regenerative Medicine, Bharath Institute of Higher Education and Research,
Chennai 600126, Tamil Nadu, India
3
Department of Anatomy, Asan Memorial Dental College and Hospital, Chennai 603105, Tamil Nadu, India
4
Institute of Biotechnology, Addis Ababa University, Addis Ababa, Ethiopia
Correspondence should be addressed to Raghu Babu Pothireddy; raghubabu_84@yahoo.com and Beimnet Asfaw Hailemariam;
beimnet.asfaw@aau.edu.et
Received 8 July 2021; Accepted 10 August 2021; Published 19 August 2021
Academic Editor: Ravichandran M
Copyright ©2021 Raghu Babu Pothireddy et al. is is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited.
Today, there are over 2,000 different biomaterials used for various medical applications, but none of these biomaterials are 100%
compatible with all human beings. Coconut fiber is widely available but has not been tested as a safe natural alternative for sutures.
Immature coconut fiber is nonabsorbable and is effective for cuts and open wounds when used in combination with dehydrated human
amnion membrane (dHAM). Immature coconut fiber, tamarind seed polysaccharide (TSP), and dHAM were prepared to test their
combinational effect on wound healing in rats. TSP enhanced cell viability, proliferation, and migration in human skin cells and cured
wounds both individually and in combination with dHAM. An antibiotic-free combination of the human amniotic membrane with
intact epithelium, tamarind seed polysaccharide, and immature coconut fiber provided faster wound healing. Significantly higher wound
healing was seen on the 11
th
day based on an initial 10 mm biopsy punch surgery in Wistar rats compared to control groups. Histological
studies revealed thickened dermis edges with more neutrophil infiltration. Collagen deposition in the dermis was homogeneous across
the excised skin tissue in the test group, again attesting to the utility of this procedure. is research signifies the use of TSP gel together
with the amnion membrane representing a “smart patch” with wound healing potential, which would encourage further research on the
smart patch made using a combination of plant and animal biological materials.
1. Introduction
Wound healing is a natural and complex process of tissue
recovery of injured tissues involving growth factors and
cytokines, released at the injured site. Delayed or impaired
wound healing may occur due to several reasons like chronic
medical conditions and medications that inhibit the healing
process [1]. Medicinal plants with wound healing properties
have been used to treat acute and chronic wounds for the
past three decades [2, 3]. Among the world population, 70 to
80% depend on medicinal plants for the management of
various ailments since ancient time [3]. Wound dressings
made of pectin and collagen enhance wound healing but are
highly expensive [4]. Identification of potent and effective
natural compounds for wound healing would benefit in the
management of wounds in a cost-effective manner.
e present research relates to the use of plant materials
and biological membrane together as biocompatible bio-
materials for wound healing. Coconut fibers are available
plenty in India and are used for different purposes. e scope
of this research is to come up with this novel use of immature
coconut fiber along with other novel combinations of
Hindawi
Advances in Materials Science and Engineering
Volume 2021, Article ID 8122989, 12 pages
https://doi.org/10.1155/2021/8122989
biomaterials for cut and open wound healing studies. Co-
conut fiber of green coconuts is immature and tough because
of the presence of lignin [5] and the presence of biode-
gradable hemicellulose and cellulose that contribute to
wound healing [6, 7]. e coconut fiber material has never
been thought of as a suture, nor has been used as a cheaper,
safer, economically viable, and easily available suture ma-
terial to date. is green, alternative, nonabsorbable suture
(Indian Patent no. 298076) is effective when compared with
commercial nonabsorbable sutures such as prolene, silk, and
nylon.
Xyloglucans of Tamarindus indica L. have been currently
explored for its property of wound healing, individually or in
combination to heal wounds by enhancing cell viability,
proliferation, and migration in human skin keratinocytes
[8]. Xyloglucans are polysaccharides, which are the main
constituents of the tamarind seed kernel and are rich in
xylose and galactoxylose substituents. Due to their me-
chanical properties, they have a wide application in hydrogel
production, films, and as drug delivery agents for slow drug
delivery [9]. Xyloglucan is abundantly found in plant cell
walls, contains (β1⟶4)-linked d-glucan substituted with
xylose, possesses mucoadhesive properties mainly due to the
mucin-like structure, and belongs to the group of poly-
saccharides, referred to as hemicelluloses [10]. e
mucoadhesive property of xyloglucan has permitted its use
as an adhesive with antimicrobial property to prevent
bacterial adherence and invasion [11]. Xyloglucans when
introduced into nanofibrillated cellulose (NFC) through
adsorption and presorption to strengthen the NFC revealed
highest adsorption, reinforcement, enhancement of cell
growth, and proliferation for wound healing [12].
Hemicellulose films have proved to be haemostatic,
absorptive, and bactericidal and have shown effective epi-
thelial wound healing in leukaemia patients with herpes
zoster infections [13]. A natural hydrogel from honey in
combination with polyvinyl pyrrolidone, polyethylene gly-
col, and agar solution showed a significant wound healing
effect compared to the control groups. e hydrogel dem-
onstrated histopathologically confirmed reduction in wound
size and has been recommended for burn injuries due to a
high fluid absorption rate [14]. Furthermore, a porous
hydrogel (size: 32.8–101.6 μm) from a mixture of chitosan
and xyloglucan with good mechanical properties has en-
hanced the properties of chitosan with the addition of
xyloglucan, without affecting its antimicrobial activity for
wound dressing [15]. Since xyloglucans have shown positive
effects on wound healing [16], hydrogels of xyloglucans
could exhibit wound healing action and also act as a vector
for slow drug delivery to aid healing. is work focuses on
extraction, identification of polysaccharide consisting of
xyloglucan from the kernel of tamarind, preparation of the
wound gel by crosslinking with epichlorohydrin, and pro-
viding a platform for the intervention of efficacious and cost-
effective wound healing agent.
e human amniotic membrane (HAM) has been
proved to be an excellent source of material for wound
therapy [17], since it induces reepithelialization meanwhile
processing antiangiogenic and antimicrobial properties. e
human amniotic membrane (HAM) lacks immunogenicity
and acts as a substrate for growth, adhesion, and migration
[18]. e wound healing ability of HAM accounts for the
presence of growth factors such as EGF, KGF, and HGF to
aid wound healing [19]. e three biomaterials used for the
study are biowastes, which were used positively for wound
healing, and this research could cause a great impact on the
identification of novel biomaterials that could work in
combination to provide high healing efficiency.
is study employs plant and animal biomaterials to
treat cut and open wounds. Figure 1 illustrates the prepa-
ration of immature coconut fiber, tamarind seed polysac-
charide, and dehydrated human amnion membrane
(dHAM) to test wound healing in rats. e use of plant and
human tissue combinations for wound healing and man-
agement had made this research novel in its attribute that
has not been performed or reported before.
2. Materials and Methods
2.1. Preparation and Evaluation of Physical Parameters of
Immature Coconut Fiber
2.1.1. Preparation of Immature Coconut Fiber Suture.
e fiber of green coconuts was removed from the shell of
the nut and was soaked in water for 24 to 48 hours to allow
the fiber to be separated into strands. e fiber strands are
then soaked into 70% isopropyl alcohol for decolourization
for 5 hours and dried in a hot air oven between 40 and 50°C
for 1 hour.
2.1.2. Determination of Tensile Strength of Coconut Fiber
Using Universal Testing Machine (UTM). e thickness of
each fiber was measured (in diameter) using a dial thickness
gauge. e average diameter of the fibers (n�3) was noted to
determine the tensile strength. Each fiber was inserted into
the universal testing machine and ensured that the ends were
gripped symmetrically so that the tension force was dis-
tributed uniformly over the cross section. e load cell value
was set to zero, and the speed of the moving grip was 10 mm/
min. Changes in the test length were noted throughout the
test and were continued until the break of the test sample.
ree samples of thin and thick immature coconut fibers
were taken in comparison with prolene and silk sutures [20].
2.1.3. Skin Holding Effect of Coconut Fiber, Prolene, and Silk
Sutures in Rats. Sprague Dawley (SD) rats (14 numbers)
were used for the study, and they were anesthetized with
ketamine and xylazine and acclimatized for 7 days. Animals
were randomly divided into two groups, with 7 animals in
each group. Group 1 was tested with thin coconut fiber in
comparison with the prolene suture. Group 2 was tested with
thick coconut fiber in comparison with silk suture.
Two 3.5 cm long parallel full-thickness skin incisions
were made under aseptic conditions on the back of the
experimental rat. e incisions were closed immediately by 4
simple sutures (Figure 2). Rats were sacrificed by carbon
dioxide inhalation. e skin wounds were removed from the
2Advances in Materials Science and Engineering
body after 24, 48, 72, 96, 120, 144, and 168 h (n�1/group/
time point). Histopathological analyses were performed
using hematoxylin-eosin, Azur, PAS, and van Gieson stained
slides.
2.2. TSP Gel Wound Healing Ability in Wistar Rats. Male
Wistar albino rats (90 days old) weighing around 200 g to
250 g were used for the study. e animals (n�3) were fed
with standard laboratory diet in the pellet form, and the rats
had access to drinking water and libitum. Under intra-
muscular injections of a combination of ketamine (40 mg/kg
body weight (b.w) and xylazine (15 mg/kg b.w), the dorsal
aspect of the rats was shaved. An excision punch biopsy was
done passing through both sides of the lifted midline,
achieving two 8 mm diameter excision wound side by side to
its spine below the neck region in the dorsal aspect [21, 22].
One side of the excision was treated with the prepared TSP
gel (approximately 0.25 ml of thawed gel twice daily for 5
days) and the other side was not treated (control).
Postcreation of the wound, the animals were given a
broad-spectrum antibiotic, amoxicillin (0.001 mL/kg b.w,
intramuscularly, single dose), and anti-inflammatory/anal-
gesic agent, piroxicam (3 mg/kg b.w, intramuscularly daily
for 3 days), and monitored for any signs of active infection
for the first two days. At the end of the study period (after 7
days), the animals were euthanized using a gas chamber
filled with isofluorane fumes. e wound area with the
surrounding tissue was excised to its full depth and fixed in
10% neutral buffered formalin and processed for routine
histopathology.
2.3. Combined Wound Surgery with the Prepared Biomaterials
in the Rat Model. e wound was created using a 10 mm
biopsy punch on animals used for the study. One of the
excised wounds was treated with the prepared biomaterials
(application of approximately 0.25 ml TSP gel on wound
area with dHAM placed on top and sutured with immature
coconut fiber). Another excision wound was untreated
(control) in Wistar rats of group 1 (n�3). Similarly, one
excised wound of group 2 animals (n�3) was treated with
TSP gel, applied on the surface of the wound area (ap-
proximately 0.25 ml) with dHAM placed on top and sutured
Preparation of
Immature thick and
thin coconut fiber
Tamarind seed kernel powder
Tensile strength testing
Histological studies
in rat model
(Immature Coconut
fiber, prolene and
silk sutures)
Extraction of TSP[30]
Identification of Xyloglucan
FTIR NMR TGA
TSP cross-linking with Epichlorohydrin
MTT Assay
Collection of the placental
sample
Preparation of Amnion
from chorian [39]
Gamma sterilization
Animal studies
(Biopsy punch)-
Wound measurement
and Histological
studies
Animal studies
(Biopsy punch)-
Wound
measurement and
Histological
studies
Figure 1: Combination therapy for cut and open wounds. TSP: tamarind seed polysaccharide; FTIR: Fourier-transform infrared spec-
troscopy; NMR: nuclear magnetic resonance; TGA: thermogravimetric analysis; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylte-
trazolium bromide.
Figure 2: SD rat sutured with immature coconut fiber and silk
suture.
Advances in Materials Science and Engineering 3
with immature coconut fiber. e other wound incision was
treated with the commercial silicone gel membrane and
sutured with the commercial silk suture.
Postcreation of the wound, the animals were given a
broad-spectrum antibiotic, amoxicillin (0.001 mL/kg b.w,
intramuscularly, single dose), and anti-inflammatory/anal-
gesic agent, piroxicam (3 mg/kgb.w, intramuscularly daily
for 3 days), and monitored for any signs of active infection
for the first two days. At the end of the study period (after 11
days), the animals were euthanized using a gas chamber
filled with isofluorane fumes. e wound area with the
surrounding tissue was excised to its full depth and fixed in
10% neutral buffered formalin and processed for routine
histopathological examinations.
3. Results and Discussion
3.1. Analysis of Immature Coconut Fiber
3.1.1. Mechanical Testing of Immature Coconut Fiber.
e tensile strength of the immature coconut fiber was tested
using the universal testing machine (UTM).Table 1 lists the
parameters for tensile strength estimation.
3.1.2. Histopathological Examination of Sutures in Wistar
Rats. e skin sections from all the groups revealed spurs of
epithelial cell migration towards the wound edges in the
epidermal layer and acute neutrophilic infiltration in the
dermis and presence of necrotic myofibers of the injured
skeletal muscles in the deepest part of the wounds from day 1
to day 3 with similar severity grades.
On days 4 to 6, the epithelial cell proliferation resulted in
a thickened epidermal layer. While in the dermis, neutro-
philic infiltrations were largely replaced by macrophages
along with the formation and invasion of granulation tissue.
Maximal neovascularization and collagen production were
observed in all three sutured skin samples.
On day 7, the epidermal layer recovered its normal
thickness (re-epithelialization) and differentiation with self-
keratinisation for the immature coconut fiber treated groups
equal to the silk and prolene treated groups. e dermal layer
revealed the remodelling phase with the presence of diffused
and organized collagen fibers with granulation tissue for-
mation (Figures 3–6).
3.2. TSP Gel Wound Healing Ability in Wistar Rats. TSP gel
application on an 8 mm wound on the right side of the
animals (n�3) resulted in gradual wound reduction com-
pared to the nontreated control. e wound site measure-
ment of the control and the treated on the 7
th
day reveal
exceptional wound healing property of TSP. e treated site
had its size reduced to 3.5 mm (±0.13) on an average, while
the nontreated control had a wound size of 6.5 mm (±0.22),
which is almost double the size of the TSP treated site,
revealing the wound healing ability of TSP. Difference be-
tween the two groups was tested using the students t-test and
was found to be statistically significant (p<0.001), revealing
the wound healing ability of TSP (Figure 7).
3.3. Histological Investigation. e epidermis of the control
animals was thickened at its cut edges. e dermis close to
the excision area showed rich polymorph nuclear infiltra-
tion. A demarcation line was formed, which separated the
necrotic slough tissue from viable tissue. Mild fibroblast
proliferation was noted in the dermis region beneath the
wound. Neovascularisation in the form of capillary blood
vessel formation was noted. However, new collagen for-
mation was minimally seen (Figure 8(a)).
In TSP gel treated animals, the wound edges are ap-
proximated and the dermis edges are thickened with more
polymorph nuclear infiltration. Fibroblast proliferation is
well noted with collagen deposition noted along the wound
area. New blood vessel formation was well marked
(Figure 8(b)).
3.4. Combination erapy Involving Natural Biomaterials for
Wound Management. Group I animals (n�3), 10 mm
wounds, sutured with immature coconut fiber with prepared
dHAM and TSP gel in combination, had better healing after
11 days. Profound wound reduction was observed in the
treated wound area with a measurement of 2 mm (±0.10),
differing much with the nontreated control measuring 6 mm
(±0.10) (Figure 9(c)) Difference between the two groups was
tested using the Student t-test and was found to be statis-
tically significant (p<0.001). Following the study, the an-
imals were euthanized, and the wound area was processed
for histopathological examinations.
Group II animals (n�3) with 10 mm wound excisions
treated with dHAM, TSP gel, and immature coconut fiber
(Figure 10(a)) were compared with 10 mm wound excisions,
treated with silicone gel membrane and silk suture as the
positive control. Wound measuring 1.5 mm (±0.17) at the
test site (dHAM + TSP gel + coconut fiber) in comparison
with the positive control (silicone gel membrane + Silk su-
ture) after 11 days of treatment reveal the wound healing
potency of the biomaterials tested (Figure 10(c)). ere is no
statistical difference between the groups (p>0.05)..
3.5. Histopathology Investigation of Group I Samples.
Control animals exhibited thickened epidermis at its cut
edges (Figure 11(a)). e dermis close to the excision area
showed rich polymorph nuclear infiltration. A demarcation
line was formed, which separated the necrotic slough tissue
from the viable tissue. Mild fibroblast proliferation was
noted in the dermis region beneath the wound. Neo-
vascularisation in the form of capillary blood vessel for-
mation was noted. However, new collagen formation was
minimally seen.
In test (dHAM + TSP + coconut fiber treated) animals, the
wound edges were thickened with high epithelialization features
(Figure 11(b)). e PMNL infiltration was seen in clusters and
evenly dispersed across the wound area. Fibroblast proliferation
was high in the dermis region with added neovascularisation
across the dermis and also in the underlying subcutaneous
matrix. e dermis edges are thickened with more polymorph
nuclear infiltration. Circular clusters of collagen deposition in
the dermis are noted all over the excised skin tissue.
4Advances in Materials Science and Engineering
3.6. Histopathology Investigation of Group II Samples. In the
test (dHAM + TSP gel + coconut fiber) treated animals, the
wound edges are thickened with high epithelialization fea-
tures. e PMNL infiltration was seen in clusters and evenly
dispersed across the wound area. Fibroblast proliferation
was high in the dermis region with added neovascularisation
across the dermis and also in the underlying subcutaneous
matrix. e dermis edges are thickened with more poly-
morph nuclear infiltration. Collagen deposition in the
dermis is homogeneous and was noted all over the excised
skin tissue (Figure 12(a)).
In the positive control (silicone gel membrane + silk
suture) treated animals, the edges were in close approxi-
mation with a great reduction in wound space. Healing is
hastened with a good amount of collagen deposition all over
the excision space. PMNL infiltration has started to clear off
with signs of thickened epithelium development
(Figure 12(b)).
e complex and coordinated process of wound
healing involves different factors and steps and requires
additional care to prevent the worsening of the wound
and abnormal scar development. ough traditional
therapies for wound care have shown beneficial effects,
there remain certain challenges that require novel ther-
apeutic approaches. Wound closure techniques have
evolved initially from suture materials comprising of
absorbable and nonabsorbable properties [23].
Noncontaminated and small skin wounds are ideally
sealed by topical skin adhesives or glues that are cost-
effective to prevent further infection. Topical skin ad-
hesives are also proved to be effectively used along with
sutures. Since the degree of healing depends on the af-
fected area, therapeutic process, and compatible material
used for treatment, interventions on combination ther-
apy for wound management would be a better option to
contain and treat wounds in a multidirectional per-
spective [24].
Large size wounds pose a serious problem and, prefer-
ably, an autograft is installed at the wound site. Minimal or
lack of graft tissue for treatment had resulted in the use of
allograft human amnion/chorion tissue as an alternative to
autografts, which could modulate inflammation and en-
hance healing of tissues, thus promoting wound healing.
Bioavailability of factors of wound healing and the increased
shelf life of the naive and immunomodulatory human
amnion membrane have been major reasons for its clinical
use [25].
Our multidirectional research has employed a combi-
national treatment approach for cut and open wound
management, combining the use of TSP gel, the human
amnion membrane, and the novel immature coconut fiber
suture.
ough topical therapy is common in wound man-
agement, our objective was to provide the best natural
Table 1: Parameters for tensile strength estimation.
Material Average diameter (mm) Average length Rate (Speed) (mm/min) Tensile strength (N)
Coconut thin fiber (Non absorbable) 0.126 18 cm to 25 cm 10 1.63
Prolene (Non absorbable) monofilament 0.099 45cm 10 1.96
Coconut thick fiber (Non absorbable) 0.248 18 cm to 25 cm 10 7.40
Silk braided (Non absorbable) 0.225 45 cm 10 20.12
(a) (b) (c) (d)
(e) (f ) (g)
Figure 3: Histopathological indications of immature coconut fiber (multifilament) sutured skin area sections. (a) 24 hrs—hematoxylin and
eosin (H and E) 10x, necrosis of epidermal and dermal cells, mild neutrophilic infiltration. (b) 48 hrs—Giemsa 10x, epithelial cell migration
and moderate neutrophilic infiltration. (c) 72 hrs—H and E 10x, moderate epidermal proliferation, mild granulation tissue invasion. (d)
96 hrs—H and E 10x, epidermal layer thickening, moderate granulation tissue formation. (e) 120 hrs—H and E 10x, moderate epidermal
keratinization, mild collagen proliferation and granulation tissue. (f) 144 hrs—van Gieson 10x, moderate amount of diffuse collagen
deposition with granulation tissue formation. (g) 168 hrs—PAS 10x, moderate amount of new blood vessels with granulation tissue.
Advances in Materials Science and Engineering 5
alternative to the available treatment options involving
synthetic materials in wound treatment. Porous silicone
membranes play a dual role, serving as epidermal barriers
and as a scaffold for delivering therapy to the affected
area. Collagen-based silicone gel sheet, comprising of a
porous silicone sheet coated with collagen, had been
proved to heal different grades of the wound in several
studies and decreased hypertrophic scarring when ap-
plied to surgical wounds [26]. Treatment using topical
silicon sheets date back to the early 1980s, where silicone
sheets were used to treat hypertrophic and keloids scars
[27]. Studies indicate the improvement of hypertonic and
keloid scars in 85% of the cases treated with silicone gel
sheet [28] though reported with skin irritation, a well-
known side effect. dHAM allografts have been employed
to heal wounds without any complications or rejection
even in elderly individuals. e usage of dHAM further
can rule out inconveniences and pain, mainly due to the
anti-inflammatory properties of the membrane and its
action as a barrier covering the nociceptors [17].
Our research uses three different biomaterials for
wound therapy, each having its own medicinal value
contributing to the wound healing effect. Our novel study
investigates and evaluates the use of combination therapy
(a) (b) (c) (d)
(e) (f) (g)
Figure 4: Histopathological indications on immature coconut fiber (monofilament) sutured skin area sections. (a) 24 hrs—H and E 10x,
necrosis of epidermal and dermal cells with scab formation and severe neutrophilic infiltration. (b) Epithelial cell migration and pro-
liferation with severe neutrophilic infiltration. (c) 72 hrs—H and E 10x, mild epidermal proliferation with granulation tissue invasion. (d)
96 hrs—H and E 10x, epidermal layer thickening, mild granulation tissue formation. (e) 120 hrs—H and E 10x, severe epidermal kera-
tinization, moderate invasion of granulation tissue. (f) 144 hrs—van Gieson 10x, organized collagen proliferation. (g) 168 hrs—PAS 10x,
moderate amount of neovascularization, granulation tissue formation.
(a) (b) (c) (d)
(e) (f) (g)
Figure 5: Histopathological indications on braided silk sutured skin area sections. (a) 24 hrs—H and E 10x, necrosis of epidermal and
dermal cells with scab formation and mild neutrophilic infiltration. (b) 48 hrs—Giemsa 10x, epithelial cell migration with moderate
neutrophilic infiltration. (c) 72 hrs—H and E 10x, epidermal proliferation, mild granulation tissue invasion. (d) 96 hrs—H and E 10x,
epidermal layer thickening, mild granulation tissue formation. (e) 120 Hrs—H and E 10x, severe epidermal keratinization, moderate
collagen production. (f) 144hrs—van Gieson 10x, diffuse moderate collagen production. (g) 168 hrs—PAS 10x, severe collagen production,
moderate neovascularization in the dermis.
6Advances in Materials Science and Engineering
in wound healing and management of cut and open
wounds.
Immature coconut fiber, both thin and thick, had satisfying
skin holding capacity equal to prolene, a monofilament suture,
and silk, a multifilament suture. Neovascularisation, collagen
production, and re-epithelialization were observed with the
recovery of the epidermal layer to its normal thickness on the 7
th
day of the study in all three suture treated groups.
Tamarind seed xyloglucan of tamarind seed kernel
powder act as a drug vehicle and influence cell viability, cell
migration, and gene expression of human skin keratinocytes
and fibroblasts [8]. e use of noncarcinogenic TSP in the
(a) (b) (c) (d)
(e) (f) (g)
Figure 6: Histopathological indications on Prolene suture skin area sections. (a) 24 Hrs—H and E 10x, necrosis of epidermal and dermal
cells, severe neutrophilic infiltration. (b) 48 hrs—Giemsa 10x, epithelial cell migration, severe neutrophilic infiltration. (c) 72hrs—H and E
10x, moderate amount of epidermal proliferation and thickening. (d) 96 hrs—H and E 10x, epidermal layer thickening, mild granulation
tissue formation. (e) 120 hrs—H and E 10x, severe epidermal keratinization, moderate granulation tissue formation. (f ) 144 hrs—van Gieson
10x, diffuse fibroblast proliferation in the dermis. (g) 160 hrs—PAS 10x, severe collagen production, maximal neovascularization in the
dermis.
(a) (b)
(c) (d)
Figure 7: TSP gel wound healing ability in Wistar rats. (a) TSP gel application site (right side), (b) wound site reduction at TSP gel applied
site (after 7 days), (c) wound site measurement of control (6.5 mm) at 7
th
day, and (d) wound site measurement of TSP gel applied site
(3.5 mm) at 7
th
day. Values are expressed as mean ±SE (n�3), where p≤0.001.
Advances in Materials Science and Engineering 7
drug delivery system accounts for its mucoadhesive property
and drug holding ability [29].
e gel-like consistency of TSP, when mixed with water,
add additional advantage to its wound healing property in
retaining its characteristics during treatment [30]. It also acts
as a carrier in drug delivery as reported by several studies
[31, 32]. Its slow drug-delivering action ensures proper and
timely delivery with its elasticity, mimicking a scaffold that
would benefit in gripping of the treatment site. Additionally,
its bioadhesive nature has been exploited in the development
of polymeric films in the treatment of candida vaginitis using
nystatin as the drug [31]. Its high drug holding nature has
facilitated its use as carriers to substantiate the sustained
release of drugs.
A combination of immature coconut fiber, dHAM, and
TSP gel healed wounds much faster than the nontreated
control, with wound measurements 2 mm (±0.10) and 6 mm
(±0.10), respectively. Similarly, a 10 mm wound treated with
(a)
(b)
Figure 8: (a) Control (untreated) and (b) TSP gel treated: photomicrographs of H &E-stained images showing the wound edges and wound
crater. Epithelialization is marked by a red arrow and collagen formation by green arrows.
(a) (b)
(c) (d)
Figure 9: Group I animals tested with a combination of dHAM, TSP gel, and immature coconut fiber compared with untreated (control).
(a) dHAM surgery on the right side with TSP gel and coconut fiber suture for 10 mm biopsy punch diameter wound. (b) After 11 days,
wound reduction site on the right side is better than the nontreated control. (c) Wound site measurement (6 mm) at the nontreated control
site after 11 days. (d) Wound site measurement (2 mm) at the test site (dHAM +TSP gel+coconut Fiber) after 11 days. Values are expressed
as mean ±SE (n�3), where p≤0.001.
8Advances in Materials Science and Engineering
(a) (b)
(c)
Figure 10: Group II animals tested with a combination of dHAM, TSP gel, and immature coconut fiber compared with silk suture and
silicone gel membrane (positive control). (a) dHAM + TSP gel + coconut fiber (Left side) and silicone gel membrane + silk suture (right
side). (b) After 11 days, the test sample on the left side showed healing in comparison with completely healed positive control on the right
side. (c) Wound site measurement (1.5mm) at the test site (dHAM +TSP gel+coconut fiber) after 11 days. e silicone gel membrane +silk
suture site (positive control) was completely healed after 11 days, and hence no measurement was taken. Values are expressed as mean ±SE
(n�3), where p≤0.05).
(a)
(b)
Figure 11: (a). Control (untreated) and (b) test (dHAM +TSP gel+coconut fiber): photomicrographs of H and E-stained images showing
the wound edges and wound crater. Epithelialization is marked by a red arrow and collagen formation by green arrows.
Advances in Materials Science and Engineering 9
the biological preparation had rapid healing, having a
wound measurement of 1.5 mm (±0.17), compared to the
completely healed wound treated with the positive control,
i.e., silk suture with silicone gel membrane. e rigidity,
thickness, and the direct pasting of the silicone gel sheet
(positive control) during surgery might account for better
healing compared to the individual application of test
materials (TSP gel and thin dHAM). TSP acts as a carrier for
the transport of growth factors and cytokines from the
dHAM, and the hydrating potential acts as a shield for
preventing skin irritation. Histopathological examinations
revealed high epithelialization and thickened wound edges
and were similar to the positive control with collagen de-
position all over the excised skin tissue. Our study proves the
treatment efficiency of the biological preparation comprising
of dehydrated human amnion membrane, TSP gel, and fiber
suture in wound healing. Animal study results clearly in-
dicate the potency of natural biomaterials in wound healing,
resembling treatment with commercial and synthetic
biomaterials.
Conventional tissue adhesive patches serve wound
management and fixation of medical devices. Tissue adhe-
sives, butylcyanoacrylate and octylcyanoacrylate, were not
efficient in decreasing the wound closure time when com-
pared with the tissue bandages [33]. In contrast to the
conventional patches, multifunctional smart skin adhesive
patches serve multiple functions of being thin, flexible, and
incorporate monitoring technology [34]. Smart patches with
capabilities of preventing wound infections and the pro-
motion of tissue remodelling are of high value. Recently,
smart patches consisting of biomass chitosan microneedle
array with responsive drug delivery with the application of
hydrogel has been proved beneficial in wound healing [35].
Wound patches with artificial intelligence have wide ap-
plication in various disciplines, especially it can be used to
monitor and promote wound healing [36]. On the other
hand, advanced, multifunctional, next generation smart
bandages that could deliver and monitor oxygen in the
wound site are under research to be made available as a low-
cost alternative for quick healing [37].
Smart hydrogel wound patches can act as a carrier for
drug delivery with a combination of drugs and also as a
wound healing indicator when incorporated with modified
pH indicator dyes to monitor the tissue healing process by
the colour transition of the hydrogel patch [38]. is re-
search signifies the use of TSP gel together with the amnion
membrane representing a “smart patch” with wound healing
potential, which would encourage further research on the
smart patch made using a combination of plant and animal
biological materials.
4. Conclusion
e natural novel combination of biomaterials (dHAM, TSP
gel, and immature coconut fiber suture) showed better
wound healing ability than nontreated controls and closely
similar wound healing activity to commercial biomaterials in
animal studies. Natural biomaterials were tested individually
and also in combination and compared with commercial
biomaterials used in wound management. e materials are
safer and are easily available and can be greatly used by the
medical/veterinary community in the near future.
(a)
(b)
Figure 12: (a) Test (dHAM + TSP gel + coconut fiber) and (b) positive control (silicone gel membrane with silk suture): photomicrographs
of H and E-stained images showing the wound edges and wound crater. Epithelialization is marked by a red arrow and collagen formation by
green arrows.
10 Advances in Materials Science and Engineering
e unresolved wound treatment challenges can be
solved by using different treatment approaches involving
natural substances as substitutes or adjuvant therapy in
current wound care procedures. A “smart patch” consisting
of natural wound healers is the need of the hour, and the
combination of plant and animal biological materials would
improve the search for novel natural materials for wound
healing.
5. Limitations
e length of the immature coconut fiber suture could not be
more than 25 cm, while commercial sutures are available in
different lengths. Treatment procedures involved the ap-
plication of 0.25 ml TSP gel to the wound and can be tried
with different volumes for its best use and effectiveness.
Data Availability
Data used to support the findings of this study are available
from the corresponding author upon request.
Conflicts of Interest
e authors declare that they have no conflicts of interest.
Acknowledgments
e authors thank Madras Christian College, affiliated to
University of Madras for providing research facilities. De-
partment of Scientific and Industrial Research (DSIR),
Government of India, provided funds for coconut fiber
research: DSIR/tepp/861/2010, Government of India.
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