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Wound-healing effects of human dermal components with gelatin dressing

Article  in  Journal of Biomaterials Applications · November 2017

DOI: 10.1177/0885328217741758

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Soft Tissues and Materials
Journal of Biomaterials Applications
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Wound-healing effects of human dermal Reprints and permissions:
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components with gelatin dressing DOI: 10.1177/0885328217741758
journals.sagepub.com/home/jba

Hyun-Jun Jang1, Yu-mi Kim1, Bo-Young Yoo2 and

Young-Kwon Seo1

Abstract
There have been numerous investigations regarding various types of dressings and artificial dermis of solid form, yet
limited research and development on paste types, such as hydrogels with dermal powder, have ensued. In this study, we
compared the in vivo wound healing effects of gelatin paste containing dermal powder to a collagen type I/chondroitin
6-sulfate (coll/chondroitin) sponge and gelatin alone, after 48 days post grafting, in a skin wound rat model. In the dermis
powder/gelatin paste-treated group, wound area contraction was minimized 50%, while in the gelatin and coll/chondro-
itin sponge groups, the initial area contracted 83–85% and 79–85%, respectively. Histological analysis revealed the
wounds treated with dermal powder/gelatin were associated with many fibroblasts, which infiltrated the wound bed,
as well as thick collagen bundles that were arranged in dendritic arrays, resembling normal skin. Furthermore, in contrast
to the gelatin- and coll/chondroitin sponge-treated groups, the powder/gelatin paste-treated wounds exhibited an abun-
dance of elastic fibers (Victoria blue staining) and extensive formation of blood vessels around the dermis (CD31
staining). Therefore, the dermis powder/gelatin paste not only renders convenience to users but also has prominent
wound-healing effects on full-thickness wounds.

Keywords
Dermis powder, gelatin, paste dressing, skin regeneration, wound healing

the substitutes. Likewise, the efficacy of acellular

Introduction
Many factors assist wound closure, from macromol-
ecular growth factors to constructed tissues, such as
the ApligrafÕ bioartificial skin. Consequently, numer-
ous approaches have been developed to accomplish
skin regeneration for burns, donor wounds, ulcers,
and other similar injuries. A porous collagen sponge
graft, containing chondroitin 6-sulfate (CS), was used
on severely burned patients as an artificial dermis sub-
stitute.1 In a clinical study, dermal substitutes com-
posed of collagen sponge inoculated with cultured
allogenic fibroblasts achieved good results in 138
out of 145 cases, including patients with deep
dermal burns, donor wounds, traumatic skin defects,
pressure ulcers, and skin ulcers.2 Truong et al.3
evaluated human skin-derived (AlloDermÕ ,
DermalogenTM, and an acellular dermal matrix) and
synthetic (Dermagraft-TC, IntegraÕ ) matrices in an
animal model of wound healing and observed that
AlloDermÕ and the acellular dermal matrix produced
less contraction and the thickest dermal layer among
dermal particles, regarding wound contraction,
epithelialization, and histological evaluation, was
demonstrated in a rat model of skin defect.4 Besides
natural compounds, polymer scaffolds, such as poly-
glycolic acid and polyglactin-910 combined with cul-
tured fibroblasts, have also revealed benefits as
dermal substitutes.5
Recently, the performance of paste-type dressing
materials was investigated in various in vitro experi-
ments and clinical trials. For instance, IodosorbÕ , a
hydrophilic starch powder containing iodine, proved
suitable as a dressing in chronic venous ulcer patients,
1
Department of Medical Biotechnology (BK21 Plus team), Dongguk
University, Seoul, Republic of Korea
2
Medical & Scientific Affairs Team, CGBIO Research Center,
Seoul,Republic of Korea
Corresponding author:
Department of Medical Biotechnology (BK21 Plus team), Dongguk
University, Seoul, Republic of Korea
Email: bioseo@dongguk.edu
2 Journal of Biomaterials Applications 0(0)
absorbing exudate and particulate matter from the
surface of granulating wounds to suppress bacterial
growth.6 Another paste-type dressing material, com-
posed of polyethylene glycol-400, poly 2-OH ethyl
methacrylate, and silver sulfadiazine (l–3%), enabled
effective antimicrobial drug delivery in second-degree
burn patients.7 Molan and Cooper8 prepared a sugar
paste as a dressing material to enhance antibacterial
effects in wounds and ulcers, and Lyall9 formulated
a bismuth iodoform paste for primary closure,
with resorbable sutures to reduce bacterial infections
at surgical wounds. Additionally, Murugan and
Ramakrishna10 developed a composite bone paste
based on hydroxyapatite nanocrystals and chitosan
(a natural polysaccharide), by a wet chemical method,
to promote bone regeneration with high resorbability.
The aforementioned dressing materials of paste type
have not only functional effects but also convenient
handling.
Although numerous investigations, regarding the
various types of dressings and artificial dermis substi-
tutes of the solid form, have been conducted, research
and development of paste-type hydrogels and dermal
powders have received little attention. Therefore, we
sought to evaluate the wound-healing effects of gelatin
paste dressings containing dermal powder. In this work,
we investigated the regeneration of full-thickness skin
wounds treated with gelatin solutions, collagen type
I/CS (coll/chondroitin) sponges and an acellular
dermis powder/gelatin paste. After 48 days, wound
contraction, blood evaluation and hematoxylin and
eosin (H&E), CD31, and Victoria blue staining were
performed for analyses of collagen, blood vessels,
and elastin.
Materials and methods
Preparation of coll/chondroitin sponges, gelatin
sponges, and dermis powder/gelatin sponges
for in vitro experiments
For the in vitro cell culture, all materials were
prepared in solid form. Coll/chondroitin sponges were
made by dissolving type I atelocollagen powder
(Bioland, Korea) in 10 mM HCl at 10 mg/mL, which
was coprecipitated by the dropwise addition of CS
(Sigma Chemical Company, St. Louis, MO, USA)
while stirring using a homogenizer. The CS was dis-
solved in 10 mM HCl at 10 mg/mL; the amount
added was approximately 5 mg/50 mg of the total dis-
persed solids. The resulting collagen–CS solution
was placed in a freezer at 80 C for 6 h, and then
lyophilized with a freeze-dryer (Samwon Freezing
Engineering Co., Pusan, Korea) at 80 C for 48 h,
yielding a coll/chondroitin sponge. The sponges
were subsequently incubated in 20 mL of 40%
(vol/vol) ethanol, containing 50 mM 2-morpholi-
noethane sulfonic acid (MES; Fluka Chemie, Buchs,
Switzerland; pH 5.5), at room temperature for 30 min.
Next, the coll/chondroitin sponges were immersed
in 20 mL of 40% (vol/vol) ethanol, containing
50 mM MES (pH 5.5), 24 mM 1-ethyl-3-(3-dimethyl
aminopropyl) carbodiimide (Fluka Chemie), and
5 mM N-hydroxysuccinimide (Fluka Chemie) for 12 h
at room temperature. After a complete reaction, the
sponges were washed twice in 0.1 M Na2HPO4
(pH 9.0) for 12 h. Lastly, the sponges were again
washed twice with 1 M NaCl for 6 h, then in 2 M
NaCl for 2 days, and finally rinsed with distilled
water and lyophilized.11
To prepare a gelatin sponge, 2% aqueous gelatin
solutions were stirred at about 2000 r/min for 30 min
at room temperature, then lyophilized. Gelatin solu-
tions and gelatin/dermis powder pastes were placed in
a freezer at 80 C for 6 h and were then lyophilized at
80 C for 48 h.12
The dermis powder/gelatin scaffold was made by
lyophilization of CG pasteÕ , which is composed of gel-
atin and dermal powder (CG Bio, Gyeonggi-do,
Korea), and placed in a freezer at 80 C for 6 h.
They were then lyophilized at 80 C for 48 h, yielding
a gelatin.
Cell culture
The fibroblasts (ATCCÕ , PCS-201-012TM) were
cultured in Dulbecco’s modified Eagle’s medium
(Welgene, Korea) supplemented with 10% heat-
inactivated fetal bovine serum (Welgene), 50 U/mL
penicillin, and 50 mg/mL streptomycin (Hyclone,
USA). The cells were incubated in 5% CO2 at 37 C.
Cells were cultured in 100-mm-diameter tissue culture
plates and seeded in three different dermal substitutes.
Scanning electron microscopy
The morphologies of the three materials were investi-
gated by scanning electron microscopy. The specimens
were fixed in a mixture of 4% glutaraldehyde and 2%
formaldehyde in 0.2 M phosphate buffer (monosodium
phosphate/dipotassium phosphate mixture, pH 7.4) at
room temperature for 2 h. After thorough rinsing with
0.175 M phosphate buffer, the specimens were
immersed in 0.2 M phosphate buffer containing 2%
osmium tetroxide for 2 h. The samples were dehydrated
in hexamethyldisilazane (E-3100, Bio-Rad, Hercules,
CA, USA), sputter-coated with gold-palladium (E-
5400, Bio-Rad), and observed under a scanning elec-
tron microscope (XFlash Detector 410-M, Bruker,
Germany) at 20 kV.11
Journal of Biomaterials Applications
In vivo experiments
In vivo experiments were performed on Sprague-
Dawley rats. The animals were anesthetized with
an intraperitoneal injection of ZoletilÕ and RompunÕ ,
followed by hair removal using a clipper. The dorsal
region was chosen to restrict the rats’ access to their
wounds; animals were also individually housed to pre-
vent cross-access to the lesions. Each rat received two
back wounds with operating scissors: one on the left
and one on the right side, each measuring 20 mm in
diameter and bilaterally distant from the spine at
approximately 1 cm. The rats were divided into three
groups (n ¼ 10 per group) and received the following
treatments: a 2% gelatin solution (group A), a coll/
chondroitin sponge (group B), and a gelatin paste con-
taining dermal powder (group C) (Figure 1).
To ensure that the biomaterial stayed housed in the
lesion after the procedure, the wounds were bandaged
with TegadermTM film from 3 M (St Paul, MN, USA);
the bandage is permeable to oxygen but acts as a bar-
rier to liquids and bacteria. Specimens were harvested
at 8 and 48 days post grafting; animals were
euthanized in a CO2 chamber, and the lesions were
removed with a scalpel, including approximately
20 mm of adjacent skin. Tissues were stored in 4%
phosphate-buffered paraformaldehyde, for subsequent
histological analyses.13
Evaluation of wound contraction
The wound sizes were calculated at 8 and 48 days post
grafting. The wound area was photographed and quan-
tified using Leopard imaging software (Leopard
Imaging Inc., Fremont, California). The wound con-
traction rate was calculated as the percentage of the
original wound area, according to the following equa-
tion: original wound area (%) ¼ actual wound area/ori-
ginal wound area  100%.
Histological evaluation
Histological analyses were conducted at 8 and 48 days
post grafting. The sampled wounds were fixed in 4%
paraformaldehyde, embedded in paraffin, sectioned and
stained with H&E and Masson’s trichrome.14
Immunohistochemistry
The skin samples were fixed in 4% buffered formalde-
hyde solution and embedded in paraffin for the prepar-
ation of 4-mm sections. Sections were then dewaxed and
dehydrated, and antigen retrieval was performed by
boiling in 10 mM sodium citrate buffer, pH 6.0, for
10 min in a microwave oven. Sections were preincu-
bated with normal serum for 10 min; rabbit serum
3
was used for monoclonal antibodies, while swine serum
was used for polyclonal antibodies. Preincubation was
followed by incubation with a mouse anti-rat CD31 pri-
mary antibody (Abcam catalog no. ab28364) for 1 h.
Sections were subsequently rinsed in phosphate-buffered
saline (PBS) and incubated with a biotin-labeled second-
ary antibody for 30 min. After washing in PBS, sections
were incubated with streptavidin-biotin-horseradish per-
oxidase complex for 1 h. Sections were rinsed with PBS,
followed by visualization with 3,30 -diaminobenzidine
(0.1 mg/mL, 0.02% H2O2).15
Analysis of leukocytes in blood
The animals were anesthetized, and whole blood sam-
ples, collected via cardiac puncture, were treated with
EDTA (1 mg/mL). A total blood cell count was then
conducted. The hematological parameters were auto-
matically assessed using HoribaÕ ABX equipment,
and reticulocyte analyses were performed manually.16
Results
In vitro analysis
Fourteen days after human fibroblasts were cultured in
a gelatin sponge (Figure 2(a) and (d)), dermal powder/
gelatin scaffold (Figure 2(c) and (g)), and coll/chondro-
itin sponge (Figure 2(b) and (f)), H&E staining results
established that most cells had attached to the surface
of the gelatin-containing matrices, while most of the
cells had infiltrated into the middle of the coll/chondro-
itin scaffold (Figure 2(b) and (f)).
Masson’s trichrome staining was performed to visu-
alize the formation of collagen. Collagen fibers
(stained blue) were deposited along the entire scaffold
(Figure 2(f) and (g)), indicating that the cells had fully
infiltrated into the scaffold. The scaffold itself consisted
of collagen as its major component, whereas the other
scaffolds only included gelatin (Figure 2(d)). Scanning
electron micrographs of a cultured fibroblast scaffold
cross section revealed the morphology of the fibro-
blasts (white arrow) and cell growth in the scaffold
(Figure 2(h–j)). Moreover, the scaffold structure and
extracellular matrix (ECM) components secreted (red
arrow) by the cells could be observed.
Wound contraction
The wound areas were imaged and quantified at 18 and
48 days post grafting (Figure 3). At 18 days post graft-
ing, that wound areas had contracted 85% in the
untreated group, 85% in the gelatin group, 76% in
the coll/chondroitin sponge group, and 51% in the
dermis powder/gelatin paste group compared to the
4 Journal of Biomaterials Applications 0(0)

Figure 1. Overall process of the in vivo experiment. Wounds were treated with gelatin (a), collagen type I/chondroitin 6-sulfate
sponge (b), or dermal powder/gelatin paste (c). Biopsies were performed at 18 and 48 days post grafting.

Figure 2. Representative images of H&E staining (a–c), Masson’s trichrome staining (d–g), and scanning electron micrographs (h–j)
14 days after in vitro culturing. Fibroblasts were cultured in gelatin (a, d, h), collagen type I/chondroitin 6-sulfate sponge (b, f, i), or
dermal powder/gelatin scaffolds (c, g, j). Original magnification: 200 (a–c), 40 (d–g), and 800 (h–j). Dots in Figure 2(a–c) indicate
fibroblasts. Red arrows (h–j) indicate the extracellular matrix components. White arrows (h–j) indicate the fibroblasts. H&E: hema-
toxylin and eosin.

initial areas. Similarly, at 48 days post grafting, the gelatin paste group compared to the initial areas.
wound areas were contracted 80% in the untreated Consequently, there were no significant differences
group, 83% in the gelatin group, 79% in the coll/chon- between the untreated and gelatin-treated groups,
droitin sponge group, and 49% in the dermis powder/ whereas the wounds in the remaining cohorts had
Journal of Biomaterials Applications
Figure 3. Images of the wound site and wound area at 18 and 48 days post grafting. (a) Untreated, (b) gelatin-treated, (c) collagen
type I/chondroitin 6-sulfate sponge-treated, (d) dermal powder/gelatin paste-treated, and (e) initial areas.
contracted four-fold compared to the initial area.
However, there was evidence of wound contraction
inhibition in those treated with the dermis powder/gel-
atin paste, as the remaining wound area was only half
contracted relative to the original area. These results
suggest that treatment with a dermis powder/gelatin
paste was approximately two times more effective
than the other treatments.
Immunohistological analyses of in vivo experiments
At 48 days post grafting, the untreated group had
a thick spinous layer with hyperkeratosis in their epi-
dermal layer, as severe wound contraction promoted
keratinocyte migration (Figure 4(a)). Treatment with
gelatin and coll/chondroitin sponges also showed a
thick spinous layer and hyperkeratosis (Figure 4(b)
and (c)). Hyperkeratosis, with a thick spinous layer
on the margin, was also observed in the dermis
powder/gelatin paste group; however, keratinocytes
had not yet migrated to the center of the wound,
due to a relatively larger wound area compared to the
other experimental groups (Figure 4(d)). The untreated
group had the least number of fibroblasts and most
number of monocytes and neutrophils observed in
the dermis layer (Figure 4(a)). The gelatin- and coll/
chondroitin sponge-treated groups seem to have
5
slightly increased fibroblast infiltration, with observed
monocyte, neutrophil, and neovascularization
(Figure 4(b) and (c)), while the dermal powder/gelatin
paste group had observably outnumbered fibroblast
infiltration with monocyte, macrophage, and neovascu-
larization (Figure 4(d)).
Masson’s trichrome staining results indicated differ-
ences in collagen fiber structure, whereby the untreated
and gelatin-treated groups presented thin collagen fibers
(Figure 4(f) and (g)), while the coll/chondroitin sponge-
treated group had a slightly parallel compact fiber array
and thicker collagen bundle (Figure 4(f–h)). The dermis
powder/gelatin paste group had the thickest collagen
bundles, which were arranged in dendritic arrays, resem-
bling normal skin (Figure 4(i) and (j)).
Victoria blue staining, which stains elastic fibers
blue, demonstrated few elastic fibers in the untreated
and gelatin- and coll/chondroitin sponge-treated
groups, in contrast to the dermis powder/gelatin paste
group, which displayed the greatest abundance of elas-
tic fibers (Figure 5(a–d)). Also, CD31 staining indicated
that few blood vessels had formed in the untreated and
gelatin-treated groups (Figure 5(f) and (g)), while more
were observed in the coll/chondroitin sponge-treated
group (Figure 5(h)). Moreover, copiousness blood ves-
sels were noticed in the dermis powder/gelatin paste-
treated group (Figure 5(i)).
6 Journal of Biomaterials Applications 0(0)

Figure 4. Representative images of H&E (a–e) and Masson’s trichrome staining (f–j) at 48 days post-grafting. (a, f) Untreated, (b, g)
gelatin-treated, (c, h) collagen type I/chondroitin 6-sulfate-treated, (d, i) dermal powder/gelatin paste-treated, (e, j) normal skin.
Original magnification: 100 (a–e) and 200 (f–j). H&E: hematoxylin and eosin.

Figure 5. Representative images of Victoria blue (a–e) and CD31 staining (f–j) at 48 days post grafting. (a, f) Untreated, (b, g) gelatin-
treated, (c, h) collagen type I/chondroitin 6-sulfate sponge-treated, (d, i) dermal powder/gelatin paste-treated, (e, j) normal skin.
Original magnification at 100 (a–e) and 40 (f–j). Arrows on (a–e) indicate the elastic fibers, while those on (f–j) indicate the blood
vessels.

Table 1. Leukocyte values of (rats) treated with gelatin, collagen sponge, or gelatin paste at 18 and 48 days post grafting.

A (No C (Collagen D (Dermal powder/

treatment) B (Gelatin) sponge) gelatin paste) E (Normal skin) Normal
(N ¼ 2) (N ¼ 5) (N ¼ 5) (N ¼ 5) (N ¼ 2) range

Day 18 5.77  2.78 7.28  3.24 7.06  1.62 9.17  2.28 9.99  1.65 4.9–16.5
Day 48 5.46  1.81 8.78  2.71 9.42  2.14 7.90  2.98 10.55  3.74

Analysis of leukocytes Discussion

The blood samples taken from the animals were eval-
uated for signs of inflammatory or immune responses
(Table 1). A comparison of the leukocyte values in the
experimental groups to the control verified that
the number of leukocytes in each group was within
the normal range, suggesting the absence of significant
inflammatory or immune responses at both 18 and 48
days post grafting.
Various dermal substitutes have been developed for the
regeneration of skin, such as AlloDermÕ , XenoDermÕ ,
IntegraÕ , and CymetraÕ , and these applications have
all been reported with autologous split-thickness skin
grafts (STSGs). However, skin regeneration surgery in
the absence of proper dermal substitutes was found to
have poor aesthetic results, and autologous STSGs
have been known to result in donor-site morbidity.
Journal of Biomaterials Applications
Therefore, numerous tissue engineering techniques
have cultured porous scaffolds with fibroblasts as an
artificial dermis for skin grafting, as fibroblasts release
various chemokines and produce structural proteins
like collagen and elastic fibers, which are responsible
for the tensile strength of skin and elastic recoil of the
dermal skin.17,18 ApligrafÕ , a skin-equivalent product
composed of fibroblasts and keratinocytes, is con-
sidered to contain key structures and components simi-
lar to human skin.19,20 However, its cost and storage
limit its applications compared to acellular products.
Gelatin is one of the proteins derived from collagen
and is known to promote epithelialization, granulation,
and tissue formation; it is often used with other poly-
mers, such as chitosan and hyaluronic acid, to improve
or modify biological or mechanical properties.21 Choi
and colleagues22 investigated the wound-healing effects
of gelatin-alginate sponges as a wound dressing mater-
ial with drug-delivering abilities. Similarly, Kawai and
colleagues23 impregnated gelatin microspheres with
basic fibroblast growth factor into an artificial dermis
for skin regeneration. Gelatin is currently used as the
main delivery material for many dressings and healing
components, yet gelatin dressings have relatively little
wound-healing effects compared to other ECM compo-
nents, such as collagen.
ECM components secreted by fibroblasts into the
acellular dermis, including collagen type I, IV and
VII, and elastin, all have notable wound-healing
effects.3,24 Seo and colleagues11 cultured fibroblasts on
collagen sponges for 30 days, removed the cells from
the sponge, and retained the newly secreted ECM and
cytokines. When the acellular artificial dermis was com-
pared to a bioartificial dermis, the ECM and cytokines
secreted by the fibroblasts displayed similar wound-
healing effects to fibroblasts contained within the col-
lagen sponge. Also, there were no significant differences
in the wound contraction, angiogenesis, collagen for-
mation, and basement membrane repair, between the
fibroblast-containing bioartificial dermis and acellular
artificial dermis.11 Zuo and colleagues4 reported that
when used as a dermal regeneration template, a particu-
late acellular dermal matrix facilitated maturation of
the thick collagen bundle structure, as well as improved
the gap rate between collagen bundles, in the regener-
ated dermis compared to the controls. In another study,
a micronized allogeneic dermal matrix (CymetraÕ ) was
successfully used with saline in the treatment of ulcers,
by promoting cellular migration, tissue integration, and
fibroblast infiltration, while decreasing infection, ero-
sion, and rejection in recalcitrant sinus tracts.25 The
findings suggested that the dermal powder itself had
wound-healing effects comparable to the bioartificial
dermis, and the form of the dermal powder affected
the wound-healing outcome.25
7
As a wound heals, the underlying contractile
connective tissue shrinks in size to bring the wound
margins together, but excessively contracted wounds
can lead to physical deformity, functional limitations,
and poor aesthetic outcomes because of irregularly
dense collagenous tissue.26 De Vries and colleagues20
investigated allogenic collagenous matrices in a
human punch biopsy wound model, to reduce scar for-
mation and wound contraction. Native collagen matri-
ces with elastin were highlighted for their ability to
decrease wound contraction and contribute to dermal
regeneration, in contrast to collagen matrices coated
with fibronectin or hyaluronic acid or without coat-
ing.20 In this regard, wound contraction can be con-
sidered one of the more important factors of a
successful dressing material.
In the current study, wound contraction was mini-
mized in the dermis powder/gelatin paste-treated
group, suggesting the presence of various ECM com-
ponents that facilitated skin regeneration, considering
the severely contracted wound areas of the gelatin- and
coll/chondroitin sponge-treated groups. Histological
analysis revealed the in vitro cultured fibroblasts
seeded on gelatin sponges (Figure 2(a) and (d)) and
dermis powder/gelatin scaffolds (Figure 2(c) and (g))
had merely attached to the surface of the networks.
Contrastingly, the fibroblasts seeded on coll/chondro-
itin sponges had migrated into the middle of the sponge
matrix and secreted ECM components (Figure 2(b) and
(f)). In contrast to the in vitro experiments, the in vivo
results showed good healing of the wounds treated with
the dermis powder/gelatin paste (Table 2), as various
types of cells participate during the wound-healing pro-
cess, including inflammatory cells, which secrete a
broad variety of chemotactic signals.27
Collagen fibers also function as a framework for the
dermis; secretion of new collagen fibers around
the wound area is considered one of the main factors
in the wound-healing process.26 In the dermis powder/
gelatin paste-treated group, fibroblasts had infiltrated
the wound bed (Figure 4(d)), where they secrete colla-
gen fibers.1 Moreover, the thick collagen bundles were
arranged in dendritic arrays, similar to the controls, in
agreement with the results of Zuo and colleagues.4
For rapid wound closure, a functional scar and new
blood vessel formation are critical components of
wound healing. During the wound-healing process,
endothelial cells digest and penetrate the underlying
vascular basement membrane, invade the ECM
stroma, and form tube-like structures, which continue
to extend, branch, and create networks, pushed by
endothelial cell proliferation from the rear and pulled
by chemotaxis from the front.28–31 The studies above
supported the notion that the dermis powder/gelatin
paste-treated wounds were the most abundantly stained
8 Journal of Biomaterials Applications 0(0)
Table 2. Histochemistry and immunohistochemistry staining results.
A (No
treatment) B (Gelatin)
Day 48 (N ¼ 2) (N ¼ 5)
Epidermis regeneration þþ þþ
Collagen fiber þ þ
Elastic fiber – –
Blood vessels – –
with CD31, among the treatments (Figure 5(i)). Elastic
fibers play a key role in supporting the mechanical
properties of skin and reducing wound contraction
and scar formation.32–34 It is not clear whether the
Victoria blue stained–elastin fibers were newly gener-
ated during the wound-healing process or pre-existed
in the dermal powder. Regardless, the more elastic
fibers existing in the tissue, the more the wound healing
results would resemble normal skin tissue.
Conclusion
Various techniques and biomaterials have been developed
for wound healing, such as membranes, hydrogels, and
powder types. However, the convenience and nature of
the dressing material used is as important as the material
itself, in which the application of appropriate types of
materials to wounds can improve wound-healing results.
Moreover, the dermal powder was shown to have notable
effects on wound healing, owing to various ECM com-
ponents, which facilitated fibroblast infiltration, collagen
bundle production, and elastic fiber and blood vessel for-
mation. Therefore, the dermis powder/gelatin paste not
only renders convenience to users but also displays prom-
inent wound-healing effects on full-thickness wounds.
Declaration of conflicting interests
The author(s) declared no potential conflicts of interest with
respect to the research, authorship, and/or publication of this
article.
Funding
The author(s) received no financial support for the research,
authorship, and/or publication of this article.
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