Advanced Review

Tissue scaffolds for skin wound

healing and dermal reconstruction
S. P. Zhong,1 Y. Z. Zhang2 and C. T. Lim1,3,4∗

One of the major applications of tissue-engineered skin substitutes for wound

healing is to promote the healing of cutaneous wounds. In this respect, many
important clinical milestones have been reached in the past decades. However,
currently available skin substitutes for wound healing often suffer from a range
of problems including wound contraction, scar formation, and poor integration
with host tissue. Engineering skin substitutes by tissue engineering approach has
relied upon the creation of three-dimensional scaffolds as extracellular matrix
(ECM) analog to guide cell adhesion, growth, and differentiation to form skin-
functional and structural tissue. The three-dimensional scaffolds can not only
cover wound and give a physical barrier against external infection as wound
dressing, but also can provide support both for dermal fibroblasts and the overlying
keratinocytes for skin tissue engineering. A successful tissue scaffold should exhibit
appropriate physical and mechanical characteristics and provide an appropriate
surface chemistry and nano and microstructures to facilitate cellular attachment,
proliferation, and differentiation. A variety of scaffolds have been fabricated
based on materials ranging from naturally occurring ones to those manufactured
synthetically. This review discusses a variety of commercial or laboratory-
engineered skin substitutes for wound healing. Central to the discussion are
the scaffolds/materials, fabrication techniques, and their characteristics associated
with wound healing. One specifically highlighted emerging fabrication technique
is electrospinning that allows the design and fabrication of biomimetic scaffolds
that offer tremendous potential applications in wound healing of skin .  2010 John
Wiley & Sons, Inc. WIREs Nanomed Nanobiotechnol 2010 2 510–525

S kin is the largest organ of the body in vertebrates

and plays a crucial role in many functions, such as
protecting against external insults, fluid homeostasis,
sensory detection, and self-healing. Skin is composed
of thin and high cellular epidermis and relatively
acellular dermis of collagen-rich extracellular matrix
(ECM).1 The epidermis mainly consists of layers
of keratinocytes separated from the dermis by
the basement membrane. The epidermis having a
∗ Correspondence to: ctlim@nus.edu.sg
1
Division of Bioengineering, National University of Singapore,
Singapore
2 Department of Bioengineering, College of Chemistry, Chemical
Engineering and Biotechnology, Donghua University, Shanghai
201620, China
3 Research Centre of Excellence in Mechanobiology, National
University of Singapore, Singapore
4 Department of Mechanical Engineering, National University of
Singapore, Singapore
DOI: 10.1002/wnan.100
high impermeability is the outermost barrier of
our body controlling water loss and protecting
the body from external insults. The dermis is
composed of the predominating ECM [collagen,
elastin, and glycosaminoglycans (GAGs)] and the
fewer cellular constituents of mainly fibroblasts. This
layer mainly provides the physical strength as well
as flexibility to skin and supports the extensive
vasculature, lymphatic system, and nerve bundles.2
Damage or loss of the integrity of skin caused
by skin or cutaneous wound may impair the skin
functions at various extents ranging from significant
disability to even death.3,4 Skin wound can arise
from mechanical trauma, surgical procedures, reduced
blood circulations, burns, or aging.5 Most skin
wounds can heal naturally, but additional surgery
necessitates immediate coverage using skin substitutes
to aid repair and regeneration when extensive or
irreversible damages to skin are caused.6 Many
natural skin substitutes such as xenografts, allografts,

510  2010 Jo h n Wiley & So n s, In c. Vo lu me 2, September/Octo ber 2010

 WIREs Nanomedicine and Nanobiotechnology
and autografts have been used for wound healing.
However, these naturally derived skin substitutes
cannot accomplish skin regeneration due to limited
donor sites, risk of infection, slow healing, and
association with the formation of scar.7,8
It is most urgent for skin wound therapy to
provide the outmost barrier, i.e., epidermal coverage
in order to prevent infection, reduce water/blood loss,
and control pain. However, an epidermal coverage
alone often fails to restore the structure and functions
of the skin and many of the problems such as
fragility of the graft, wound contraction, and scar
formation often occur.9–11 Increasingly, apparent
evidences demonstrate that effective wound healing
always necessitates the presence of the dermis layer
in the skin substitutes.12,13 The re-epithelialization
of keratinocytes and take rate are promoted through
dynamic dermal–epidermal interactions, and hence the
graft exhibits higher resistance to wound contraction
and scarring. The restoration of dermis requires
three-dimensional scaffolds to provide elasticity and
strength to the epidermal graft and feed the
keratinocytes in epidermal layer. Hence, one crucial
factor in skin tissue engineering is the construction of
a tissue scaffold as template to guide restructuring of
cells and subsequent host infiltration of the skin graft.
Over the past three decades, extraordi-
nary advances and improved understanding in
cell/molecular biology have led to achievements in
skin tissue regeneration for wound healing.1 Tissue
engineering skin, based on the concept of a cell–matrix
construct, has been a significant advancement in the
field of wound healing. A number of products are
commercially available and many others are in devel-
opment. Over 200,000 patients have been treated
with tissue-engineered skin products.14–17 Examples
of skin substitutes may be acellular or cellular. Acel-
lular products only contain the matrix based on
natural or synthetic materials with the required phys-
ical and chemical structure allowing easy access to
host cells during wound healing. Cellular products
with/without matrix include epidermal cell sheets,
epidermal constructs, dermal replacements, and bilay-
ered skin equivalents.16,17 However, currently none of
these is fully satisfactory and can replace native skin
autograft and allograft permanently.
Based on the recent developments and an exten-
sive understanding of the wound healing process, this
article reviews tissue scaffolds used in commercial skin
grafts and some of the new developments occurring
at the laboratory stage. The discussion will center on
the processing technique, characteristics, and mate-
rials of tissue scaffolds involving wound repair or
healing. Novel biomimetic scaffolds fabricated using
Vo lu me 2, September/Octo ber 2010  2010 Jo h n Wiley & So n s, In c.
Tissue scaffolds for skin wound healing
electrospinning and advantages of nanofibers when
used as wound cover or matrices for tissue repair and
regeneration are emphasized.
CURRENT MATERIALS AND
SCAFFOLDS IN WOUND HEALING
Collagen-Based Matrices
Collagen is a major ECM protein of the dermal
layer of skin, and it is very logical that the
early studies on using biodegradable polymers for
wound dressing or skin tissue engineering focused
on using collagen. The structural and functional
characteristics of collagen similar to native ECM
stimulated the use of collagen matrices for wound
healing.18 A number of collagen-based dressings
containing a variety of carriers/combining agents
in the form of gel, sheet, lattice, or sponge are
commercially available now (Promogram , Johnson
& Johnson, and Puraply , Royce Medical). Many
of them have been used under various clinical
conditions as a temporary covering for ulcers and
burns with favorable results being achieved.19 Other
engineered acellular dermis, cellular epidermis/dermis,
or bilayered skin equivalents involving collagen-based
materials have been developed as biological skin
substitutes for wound healing.
A naturally derived dermis traded as AlloDerm
(LifeCell Corporation) has a porous and fibrillar struc-
ture similar to native dermis prepared using lyophiliza-
tion technique after decellularization treatment as
shown in Figure 1A and B.20,21 AlloDerm has been
used as autograft in the resurfacing of burn wound
reconstruction due to its human origin.22 Another
acellular natural product (Integra , Integra Life Sci-
ences) was engineered using a porous composite of
bovine collagen-chondroitin-6-sulfate (8 wt%) with
an outer silicone covering.23,24 The outer silicone layer
(Figure 1C25 ) serves as a temporary barrier preventing
water loss and microbial invasion, and this layer can
be removed 2–3 weeks after wound replacement upon
vasularization of the dermis. The porous collagen lay-
ers (Figure 1D),26 manufactured by lyophilization of
a dispersion of collagen, crosslinking, and sterilization
by chemical means, were designed to act as template
for host fibroblast infiltration and capillary growth.
Integra has been widely used for split-thickness skin
grafts or wound closure on severe burn wounds when
donor sites are limited.27 But Integra faces the prob-
lems of high cost, the need for two surgeries, and the
risk of infection complications.28 Recently, inclusion
of skin cells (keratinocytes, fibroblasts, or both) with
Alloderm or Integra acellular dermis has been
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 Advanced Review
(a)
1B
(c)
performed to architect one cellular bilayered skin
equivalent in some studies.29–31 Fine capillary for-
mation and accelerated healing rate can be achieved
on the treatment of full-thickness wounds. However,
poor ingrowth of dermal fibroblasts was found in
some fashions of Alloderm due to the denser struc-
ture of collagen fibrils.32,33
One of most advanced product of Apligraf
(Figure 2A,7 organogenesis) has been developed based
(a)
FIGURE 2 | Bilayered skin substitutes: (A) Apligraf (Reprinted with permission from Ref 7. Copyright 2008 Elsevier). (B) Comparison of
Apligraf and natural human skin; Apligraf exhibits similar structure as the natural skin. (Reprinted with permission from Ref 36. Apligraf ).
512
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(b)
FIGURE 1 | Acellular skin substitutes:
50 µm (A) appearance of Alloderm native tissue
(Reprinted with permission from Ref 20.
Copyright 2010 emedicine.com), (B) SEM
(d) micrograph of the surface (Reprinted with
permission from Ref 24. Copyright 1981
Elsevier). (C) Integra template with
silicon layer in situ (Reprinted with
permission from Ref 25. Copyright 2006
Elsevier). and (D) Porous sponge-like
collagen–glycosaminoglycans structure of
Integra . (Reprinted with permission
from Ref 26. Copyright 2007
http://www.ilstraining.com/idrt/idrt/
brs it 03.html).
on the pioneer work on the cellular skin equivalent
involving collagen.7,34 Briefly, dermal fibroblasts are
seeded in suspension mixed with bovine type I collagen
solution and fibroblast–collagen interaction results in
the rearrangement and increased density of collagen
fibrils with a dermal–lattice equivalent formed in
4–6 days. Keratinocytes cultured onto the surface
of the dermal layer are exposed to an air–liquid
interface to promote maturation and cornification
APLIGRAF Human skin
(b)
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 WIREs Nanomedicine and Nanobiotechnology
of the stratified epidermal layer, which can support
long-term maintenance of the fibroblasts.35 Much like
human skin, Apligraf contains key structure and
components similar to human skin (Figure 2B36 ). The
vascularization and well integration of Apligraf into
the wound bed was observed within 14 days and an
improved healing was found for chronic venous leg
ulcers and diabetic foot ulcers.37–39 Orcel (Ortec
International) is another bilayered cellular product
using porous collagen sponge with one side coated
with collagen gel. Dermal fibroblasts are cultured on
and within the porous sponge side of the collagen
matrix and keratinocytes are seeded on the gel-
coated side and thus ingrowth of keratinocytes can
be prevented.40 Accelerated healing rate and reduced
scar formation were achieved with this graft compared
with conventional therapy with Biobrane-L.41 But
there are limited clinical trial data available on this
graft compared with Apligraf .
Major forms of collagen dressings or matrices
for skin tissue engineering are hydrogel, sponge,
and lattice. Initial collagen gel such as floating
or anchored gel was introduced to form dermal
equivalent to investigate the effect of cellular force on
collagen contraction, which was linked to subsequent
wound contraction.42 The culture work and clinical
procedures necessitated the use of denser collagen such
as lattice or sponge with higher viscosity and strength.
Collagen lattice condensed from fibroblast-populated
collagen solution was used to mimic the in vivo dermal
layer. However, there was up to 20% contraction
of lattice even after transplantation, which inhibited
effective wound covering and closure. Besides, the
decreased synthesis of collagen was due to the cell
confinement.43,44 Dehydrated collagen sponges are
normally produced by freeze drying a dispersion of
collagen gel or acidic solution.45 Collagen sponge
is solid with stronger mechanical properties than
hydrated collagen gel, and sponge implant has been
efficient in the use for recovery of skin, and various
types of artificial skin in the form of sponge were
developed.41,46,47
However, collagen-based scaffold or dressing
often has poor biostability and low mechanical prop-
erties and wound contraction easily occurs.42,48,49
Extensive modifications by crosslinking treatment or
combining other natural or synthetic polymers, such
as ECM, GAGs, chitosan, polycaprolactone (PCL),
and PLGA are used to form intra- and interchain
bridges between/within collagen molecules with or
without external crosslinking agents.50 As a result, the
increased mechanical properties can prevent the colla-
gen matrix from contracting during the culture period
and the wound healing process and the increased
Vo lu me 2, September/Octo ber 2010  2010 Jo h n Wiley & So n s, In c.
Tissue scaffolds for skin wound healing
biostability can also extend the clinical durability of
the graft. Besides, incorporation of ECM components
into collagen such as fibronectin, elastin, and GAGs
plays an important role in guiding the cell physiol-
ogy and behavior in wound healing.13,51,52 Especially,
fibronectin plays an integral role in the attachment of
keratinocytes and re-epithelialization.52 GAGs such as
hyaluronic acid and chondroitin 6-sulfate have multi-
functional facilitators for wound healing and inhibit
wound contraction as well.53 Combining chitosan,
as one analog to GAGs, with collagen can inhibit
its degradation, improve its mechanical strength, and
accelerate its wound healing.53
Recently, Seo et al.54 developed a novel rein-
forced collagen scaffold used for constructing one
artificial dermis layer. The collagen sponge was rein-
forced by incorporation of collagen-knitted mesh
fabricated by stacking multilayered collagen thread
as shown in Figure 3.54 Continuous collagen thread
was fabricated by extrusion of collagen acid solution
into a coagulation bath and multilayered knitted col-
lagen mesh was fabricated by cross joining methods.
The study showed that significantly increased ten-
sile strength was achieved on the reinforced collagen
sponge compared with a typical collagen sponge, and
rapid and firm cell adhesion to the surface and consid-
erable cell infiltration into the pores were observed.
Although the abovementioned modification and
improvement have been achieved, using collagen-
based polymers for wound healing still creates
problems such as wound contraction and scarring.
Other concerns discouraging collagen applications
include the high cost of pure collagen, variability
in the physicochemical and degradation properties
depending on source and processing.55 Some other
natural or synthetic polymers have been attempted to
overcome these concerns.56,57
Other Natural Polymers
Some other natural polymers including chitosan,
fibrin, elastin, gelatin, and hyaluronic acid have been
investigated for wound healing. Although some of
their dressing products have been commercialized,
their cellular skin grafts are only in the laboratory
stage and further clinical data are needed for FDA
approval or commercialization.
Chitosan is another natural polymer most
widely used next to collagen in wound healing due
to its many advantages including biocompatibility,
biodegradability, hemostatic activity, and antibacte-
rial properties.58–60 Chitosan can stimulate collagen
synthesis and well incorporate into fibroblast growth
factor due to the electrostatic function of chitosan,
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 Advanced Review
(a) (b)
(c) (d)
which can enhance the wound healing rate.61,62
Chitosan materials can be easily fabricated into differ-
ent structures under various process conditions. The
films, gels, or sponges of chitosan have recently been
investigated for use in full-thickness burn wounds.63
However, chitosan has rapid biodegradability espe-
cially in acid environment that is often formed in
wound healing.64 Various crosslinking treatments
have been used to increase the structural stability of
chitosan.56 Also, it is highly desirable to incorporate
chitosan into another one or two different polymers to
balance its advantages and disadvantages.65,66 Large
number of amino groups in chitosan plays a significant
role in bridging and crosslinking between/within chi-
toscan and/or other polymers such as collagen, gelatin,
and GAGs.67 The increased crosslinking efficiency
may improve the biostability and mechanical proper-
ties of scaffolds. Ma casted chitosan film onto collagen
sponge to form one bilayer structure for skin tissue
engineering.68 This chitosan layer with denser struc-
ture can enhance the functions of the artificial dermis
with increased controlled fluid loss and antibacterial
activity. However, some disadvantages of chitosan
such as unsatisfactory mechanical properties, severe
shrinkage, and deformation after drying discouraged
its application in tissue engineering field.69
Fibrin glues in suspension, gel, membrane, or
sheet form have been used as a dressing or co-culture
system for delivering keratinocytes to treat burn
wound. Rapid re-epithelialization of keratinocytes
514  2010 Jo h n Wiley & So n s, In c.
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FIGURE 3 | Optical
microscopic photographs of
collagen thread mesh (A),
reinforced collagen sponge
with collagen mesh (B), SEM
micrographs of collagen
sponge reinforced by collagen
mesh (C), and the fibroblasts
growing and filling the
collagen mesh (D). (Reprinted
with permission from Ref 54.
Copyright 2008 Springer).
and well organization of fibroblasts within fibrin
matrices were found in culture models and good take
rate was achieved for subsequent wound healing.70–73
Coating of synthetic polymer surface using fibrin
glue was used to produce more suitable scaffolds
for civilizing keratinocytes with higher strength and
acceptable biocompatibility.74
Gelatin as one protein derived from collagen has
been investigated as matrices in the dehydrated form
such as sponge or film for skin tissue engineering,
and promoted epithelialization and granulation tissue
formation were shown in wound healing.75 But this
material has the limitation of low mechanical strength
and is effectively used only as incorporated component
with others polymers to promote or modify the
biological or mechanical properties.76–78
Hyaluronic acid or hyaluronan-based scaf-
folds have been studied due to its excellent
biocompatibility.79 One non-woven fleece of 100%
benzyl-esterified derivative of hyaluronic acid was
already marked as Hyaff (Fidia Advanced Biopoly-
mers) and its efficacy used for cultured epidermal
graft (Laserskin ) or dermal graft (Hyalograft 3D)
was demonstrated in extensive burns.80,81 Fibrob-
last–keratinocyte composite based on this Laserskin
membrane with the promoted three dimension can
produce skin equivalent, and promising in vitro results
suggested the potential application for burns and
chronic wounds.82
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 WIREs Nanomedicine and Nanobiotechnology
In summary, the success from the use of natural
biodegradable cell matrices has been encouraging
and continues to facilitate broader use in the
future. However, many problems occurring with
natural polymers include low mechanical strength,
shrinkage/contraction, difficulty in handling, and
risks of immunological rejection.57 Use of synthetic
polymer or incorporating synthetic polymer into
natural polymer may be favorable when higher
mechanical properties and more flexible fabrication
ability of synthetic polymers are highly desirable.12
Synthetic Polymer Scaffolds
Generally, synthetic polymers possess lot-to-lot uni-
formity and have less expensive and more reliable
sources of raw material. They can be tailored using
various fabrication techniques to provide a wide range
of physical properties. Although some work indicated
that acceptable epithelialization of keratinocytes can
be achieved on synthetic polymer surface, currently
no successful epidermal graft using synthetic poly-
mers has been achieved due to their limited cellular
recognition and tissue compatibility.83 The studies
using synthetic polymers for skin substitutes are
already focusing on the combination with natural
polymers used as matrices for temporary dressing,
epidermal/dermal cell carriers, or full-thickness skin
equivalent.
(a)
100 µm
FIGURE 4 | Polyglactin mesh (A),
Dermagraf as received from a pack
(B), dermal fibroblasts cultured on (c)
polyglactin mesh (C), and
collagen-hybridized polyglactin mesh
(D), collagen promotes adhesion and
growth of fibroblasts after 5 days of
culture (A, C, D reprinted with
permission from Ref 86. Copyright
2004 Expert Review of Medical
Devices, and B reprinted with
permission from Ref 86. Copyright
2004 Expert Review of Medical
Devices).
Vo lu me 2, September/Octo ber 2010  2010 Jo h n Wiley & So n s, In c.
Tissue scaffolds for skin wound healing
Polyurethane (PU) has been extensively studied
as materials for wound dressing due to its semiperme-
ability. PU wound dressings are impermeable to bacte-
ria and water but permeable to gas with moist environ-
ment desirable for wound healing to occur. PU-based
membranes have been commercially available for
wound dressing (Tegaderm, 3M Health Care and
OpSite, Smith & Nephew). The products are cost-
effective for covering small-sized, split-thickness skin
graft, but limited adherence to wound bed was found.
Nylon material, one widely used material in suture, is
also widely used for wound cover in the form of nylon
mesh (Biobrane, Dow B. Hickam, Inc.). Collagen
or collagen-derived peptide is coated to improve cell
adherence or tissue biocompatibility. These wound
dressings have been used as a standard control of cell
carrier to compare their in vitro biocompatibility with
those of some natural or synthetic dressings.84
A knitted polyglycolic acid/polylactic acid
(PGA/PLA or namely Polyglactin) mesh as typically
shown in Figure 4A85 was cultured with human
neonatal fibroblasts leading to the development
of one commercial product of cryopreserved
Dermagraft (Advanced Tissue Sciences) (Figure
4B).86 Dermagraft has been used successfully to treat
full-thickness diabetic foot ulcers. The knitted PLGA
meshes support homogenous cell distribution and
(b)
(d)
68.7 µm 68.7 µm
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 Advanced Review
withstand the cell contractile force (Figure 4C). How-
ever, the PLGA scaffold has the problems of surface
hydrophobicity and limited cell adhesion. One modi-
fied knitted PLGA mesh hybridized with collagen has
been investigated as a three-dimensional culture sys-
tem for skin tissue engineering85 or some other tissues
construction.87,88 These web-like collagen sponges
can facilitate seeding, uniform distribution, and
ECM synthesis of dermal fibroblasts (Figure 4D).85
TransCyte , formerly known as dermagraft-TC, is a
cell-seeded Biobrane product, composed of a semiper-
meable silicone membrane and newborn human
fibroblast cells cultured on a collagen-coated nylon
mesh. But it is only a temporary covering due to the
poor biodegradability of nylon.89,90
Many other synthetic polymers used in
restorable sutures or wound covering have been
investigated as matrix materials for dermal substi-
tutes or full-thickness skin equivalent. They include
PCL, poly(l-lactide) (PLLA), PGA, and copoly-
mer poly(ethyleneglycolterephthalate)-poly (butylenes
terephthalate).12,91 However, limited clinical success
was achieved because synthetic polymers often have
lower rates of cell attachment and proliferation due
to the limited biological signals. The combination of
two or more different polymers including natural or
synthetic is necessary to produce suitable scaffolds for
skin regeneration when the advantages or disadvan-
tages of each material are well balanced. Although
the overall research is encouraging, our knowledge
and understanding of the skin substitute and wound
healing involving biology and material science still
need to be improved. In the past 10 years, the devel-
opment of nanotechnology involving fabrication and
characterization methods has encouraged the research
community to investigate the potential of applying
nanofibrous scaffolds in the tissue engineering field.
ELECTROSPUN BIOMIMETIC
NANOFIBROUS SCAFFOLDS
FOR WOUND HEALING
Characteristics of Electrospinning and
Electrospun Scaffolds
As we know, native ECM in dermis layer is
composed of collagen nanoscale fibers and it provides
structural integrity and mechanical strength to
skin tissues. Thus, a biologically inspired scaffold
fabrication approach for wound healing is to create
ECM analogs composed of nanoscale fibers with
mimicking structure and functions to native ECM.92,93
There are many fiber fabrication methods such
516  2010 Jo h n Wiley & So n s, In c.
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as fiber drawing, template synthesis, temperature-
induced phase separation, molecular self-assembly,
and electrospinning.94 However, only electrospinning
has emerged as an efficient technique to produce
a variety of polymeric nanofibers.95 Electrospinning
generally involves the application of an electrostatic
force between polymer solution (or melt) kept in
a syringe or pipette and a counter metal electrode
(collector plate) kept at a certain distance from
the polymer solution as shown in Figure 5A. The
electrospinning technique has gained attention and
popularity in the last 10 years due to an increased
interest in nanoscale properties and technologies.
One major attractive feature of electrospinning is
the simplicity and inexpensive nature of its setup.
Electrospun scaffolds provide high surface area to
volume ratio, which has been proved to promote
cell–matrix interaction at the nanoscale.96,97
This high surface area to volume ratio of
electrospun scaffolds facilitates oxygen permeability
and allows fluid accumulation, which are highly
desirable in the wound healing course. Besides, the
pores in non-woven form of electrospun scaffolds
(normally 1–10 µm) are small enough to prevent
bacterial penetration.95,102 Much attention has been
drawn to the use of electrospun scaffolds as wound
dressing or cellular carrier for skin substitutes
made of a variety of materials including collagen,
gelatin, fibrinogen, chitosan, PU, PCL, PLA, and
PLGA,103–106 or some blends of them.107,108 Current
work using electrospun nanofibrous membranes as
medical dressings or skin regeneration for wound
healing is still at an early stage.
Electrospun Natural and Synthetic
Nanofibers for Wound Healing
Feasibility of electrospinning collagen was first
reported by Bowlin et al.98 using calf skin type I col-
lagen dissolved in 1,1,1,3,3,3-hexafluoro-2-propanol
(HFP) as shown in Figure 5B. Structural and bio-
logical characterizations were conducted to confirm
that the structural and biological properties of native
collagen were maintained after solvent dissolution
and electrospinning. Subsequently many research
works extended the application of electrospun col-
lagen to several tissue engineering fields such as skin,
cornea, and blood vascular system.109–112 Electro-
spun collagen was found to be the most biomimetic
nanofibrous scaffolds due to its structure and origin
closely similar to the native ECM of dermis. Several
studies already demonstrated that collagen nanofi-
brous matrices exhibited excellent biocompatibility to
dermal fibroblast and keratinocytes, and significantly
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 WIREs Nanomedicine and Nanobiotechnology
(a)
High voltage
Collector
FIGURE 5 | Electrospinning and
electrospun nanofibers. (A) Schematic
diagram of the electrospinning setup,
(B) collagen nanofibers, (reprinted with
permission from Ref 99. Copyright 2007 John
Wiley & Sons, Inc.), scale bar = 100 nm,
(e)
(C) PVA/AgNO3 nanofiber, (reprinted with
permission from Ref 98. Copyright 2002
ACS), (D) core–shell polycaprolactone/
collagen nanofiber with the insert of the
coaxial spinner, (reprinted with permission
from Ref 100. Copyright 2005 ACS), and
(E) polycaprolactone/gelatin nanofiber
scaffold collected with Tegaderm Tegaderm
wound dressing
membrane. (Reprinted with permission from
Ref 101. Copyright 2007 Elsevier).
accelerated wound healing and inhibited wound con-
traction (compared with collagen sponge) were found
in early-stage wound healing.103,109 However, one
recent report provided very direct evidences that over-
whelming denaturation of collagen into gelatin occurs
with those commonly used fluorinated alcohols such
as HFP or 2,2,2-trifluoroethanol (TFE) as the dis-
solving solvent for collagen.113 Considering the high
cost of collagen material, further investigations are
needed to clarify the effect of collagen denaturation
on the biological activity and structural mechanical
integrity, which are highly desirable in mimicking
native ECM.
Gelatin, as an alternative protein to collagen, is
commercially available at significantly lower cost and
preserves many merits of collagen such as biological
origin, biodegradability, and biocompatibility. Gelatin
was successfully electrospun into nanofibers using
several solvents such as TFE, HFP, and formic
acid.104,114,115 Potential application of electrospun
gelatin as wound dressing and optimal fiber density
to provide high cell viability, and optimal cell
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Tissue scaffolds for skin wound healing
(b)
Polymer
solution
Polymer jet
(c)
100 nm
(d)
Nanofiber scaffold
0.2 µm
organization, and excellent barrier desirable for
wound healing were discussed.104,116 However these
studies suggested that combination with synthetic
polymers (PU or PCL) is highly desirable to provide
higher mechanical properties of scaffold.
Some other natural polymers including silk
fibroin, chitin/chitosan, fibrinogen, or their blends
have been electrospun as wound dressing or cellular
matrix, and efficient cell attachment, growth, or
infiltration has been demonstrated in in vitro
culture,117–119 but limited clinical trial has been
reported. Recently, hemoglobin and myoglobin have
been electrospun as wound dressing aimed to
promote oxygen delivery to the healing tissue using
hemoglobin’s functional analog of red blood cells.
Increased oxygen delivery was deemed to increase
the metabolic rate and infectious resistance of the
wound, and thus wound healing was supposed to be
accelerated.120
Natural electrospun nanofibers normally require
crosslinking treatment to stabilize the structures of
nanofibers before in vitro or in vivo use. However,
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 Advanced Review
choosing efficient crosslinking for electrospun natural
polymer needs to proceed with extra care as aqueous
degradation or swelling of natural polymer in the
early stage of crosslinking easily damages the pores of
matrix or impairs the porosity of the electrospun mats.
As suggested in several studies, glutaraldehyde vapor
crosslinking is a highly efficient method, and nanoscale
dimension and porous structure of collagen, gelatin, or
fibrinogen nanofibers can be preserved.105,114,121,122
Electrospun synthetic polymers such as PU,
PCL, PLLA, and poly (vinyl phenol) have been
proposed for wound dressing application in wound
healing.101,102,116 Early studies using electrospun
synthetic polymers for wound dressing, together
with some electrospun natural polymers, have
created wound healing scaffolds with such inherent
properties of the electrospun material. Controlled
evaporative water loss, excellent oxygen permeability,
and promoted fluid drainage have been revealed on
most of the electrospun mats used. In some recent
studies, the addition of drugs into nanofibers as
controlled release system has been investigated for
wound dressing to provide the required protection and
pain management. Some antibiotic or antibacterial
drugs/components such as cefazolin, liodocaine,
mupirocin, or Ag particles have been electrospun
into synthetic nanofibers in the blended form
(Figure 5C), and increased or controlled antimicrobial
or antibiotic ability desirable for wound healing has
been achieved with a sustained controlled release rate
of drugs.99,115,123–125 These drug delivery systems
using electrospun nanofibers exhibit pain relief and
extended antibacterial activity with great potential of
the applications in wound healing.
For the use of electrospun pure synthetic poly-
mer in skin tissue engineering, only limited synthetic
polymers such as PLLA, PLGA, blended poly(dl-
lactide), and poly(ethylene glycol) having acceptable
hydrophilicity have been reported for the potential
application in skin tissue scaffolding.106,126,127 It is
widely accepted that the incorporation of natural poly-
mer into electrospun synthetic nanofibers is desirable
in skin tissue engineering to promote the biological
activity of scaffolds. Composite polymeric nanofibers
can be electrospun using mixed solution,107 using
dual or co-electrospinning method in separate solvent
systems111 or coaxial core/shell electrospinning.100
Nanofibrous composite of PCL/collagen was
electrospun, as the one in early model of composite
used to support dermal fibroblasts. Heather et al.
investigated the association of mechanical strength
and biological affinity of blended PCL/collagen
scaffolds with various ratios of PCL/collagen.128
Minimal addition of PCL (10%) to collagen produces
518  2010 Jo h n Wiley & So n s, In c.
www.wiley.com/wires/nanomed
the scaffolds suitable for tissue engineering skin with
a good balance of mechanical strength and biological
properties. In our laboratory, Zhang et al. showed
that PCL/collagen nanofibers in core–shell form
fabricated using coaxial core/shell electrospinning
(Figure 5D) exhibited higher biological affinity than
the PCL nanofibers simply coated with collagen.100
More recently, PCL/gelatin nanofibrous scaffold
and layered dermal reconstitution were evaluated
for wound healing. PCL/gelatin was electrospun
onto one commercial PU membrane of Tegaderm
(Tegaderm, 3M Medical) as shown in Figure 5E.
Tegaderm membrane can facilitate clinical handling,
provide a protective barrier to external infections as
synthetic epidermis, and control water loss. Significant
cell adhesion, growth, and infiltration achieved
on the PCL/gelatin produced a fibroblast-populated
three-dimensional dermal analog. This cost-effective
composite could be promising wound dressing or
tissue scaffolds for skin construct.
The potential application for PLGA/dextran
was investigated as tissue scaffolds for skin tis-
sue engineering.129 A complete cell biological
response of dermal fibroblasts was tested on
electrospun PLGA/dextran nanofibers. The results
showed that favorable fibroblasts interacted with
the scaffolds and resembled a dermal-like archi-
tecture. Aimed to overcome poor infiltration of
fibroblasts into electrospun mats, a novel three-
dimensional multilayered cell–nanofiber constructs of
dermis was fabricated by architecting layer-by-layer
of nanofibers–cells alternated using electrospinning
nanofibers of PCL/collagen and seeding human dermal
fibroblasts (Figure 6).130 Dermal tissue or bilayered
skin equivalent was produced by continuous cul-
turing of fibroblast/fiber-layered constructs or inclu-
sion of keratinocytes seeded onto the dermal layer.
Several other composites including PLGA/collagen,
PLLA/gelatin, and PVA/chitosan have been electro-
spun and their characteristics oriented for the appli-
cation in skin tissue engineering were discussed.98,118
These scaffolds showed their efficacy for wound cov-
erage or cell growth biocompatibility, but further
clinical studies are needed.
Previous work using this nanofabrication tech-
nique already suggested great potential of applying
nanofibrous scaffolds for wound dressing or skin
tissue engineering. However, very limited in vivo
applications for skin repair and regeneration have
been reported. More effort is still needed to seek
FDA approval for use and to prove their effectiveness
in repairing and regenerating skin. Electrospun
scaffolds have been extensively used in vitro to study
cell–scaffold interactions, and further understanding
Vo lu me 2, September/Octo ber 2010
 WIREs Nanomedicine and Nanobiotechnology
Step 1: Fiber spinning (1 layer fiber)
FIGURE 6 | Schematic
illustration of layer-by-layer dermal
reconstruction using nanofibrous
scaffold. (Reprinted with
permission from Ref 130. Copyright
2009 Mary Ann Liebert, Inc.
Publishers). Repeat step 1 & step 2 until desirable layers of fibers and cells
of the cellular response of nanofibrous scaffolds might
ultimately lead to the success in tissue repair or regen-
eration of skin.
CONCLUSIONS AND FUTURE
OUTLOOK
Tissue scaffolds have been demonstrated to be useful
in wound dressing or skin tissue engineering in a
variety of wound conditions. Tissue scaffolds can not
only can cover the wound and provide a physical
barrier, but can also can offer a cellular skin
with excretive biological components to stimulate
re-epithelialization and formation of granulation
tissue. A variety of scaffolds fabricated based on
materials ranging from naturally occurring materials
and those manufactured synthetically have been
reviewed in this article. Collagen-based materials
have been investigated extensively and significant
success has been achieved in a variety of commercial
or laboratory-engineered skin substitutes for wound
healing. However, many problems associated with
the current scaffolds and artificial skin substitutes,
which include contraction, delayed vascularization,
high cost, and scarring, have been emerging and they
vary in their severity. It is hoped that a combination
of appropriate biomaterials and techniques can
overcome many of these problems in order to eliminate
or reduce the risk of failure in skin graft.
Vo lu me 2, September/Octo ber 2010
Tissue scaffolds for skin wound healing
Step 2: Cell seeding (1 layer cell)
Fiber
Cell
Cell
Fiber
Cell
On site layer-by-layer tissue generation
Although the potential risks and benefits of com-
posite scaffolds need to be further evaluated in clinical
trials, it is acceptable that they offer a new significant
direction for the treatment of wounds. Composite of
natural and synthetic polymers provides a compro-
mised solution for overcoming the shortcomings of
natural and synthetic polymers and can result in a new
biomaterial with appropriate biocompatibility and
mechanical, physical, and chemical properties. Elec-
trospinning can produce new type nanofibrous scaf-
folds for wound dressing or skin tissue engineering.
This electrospun type scaffold exhibits an appropriate
porous structure that can mimic the collagenous struc-
ture of native dermis. The electrospun PCL/gelatin
scaffolds developed in our laboratory exhibit excellent
biocompatibility and biodegradability. The low cost
and existing FDA approval for use of PCL and gelatin
and their proven efficiency in tissue engineering con-
tribute to the ease of FDA approval for clinical use and
commercialization of the final artificial skin using this
scaffold. In future, the directions in scaffold design-
ing would be to incorporate molecules such as ECM,
growth factors, or cytokines associated with enhanc-
ing wound healing into the right scaffolds. A relatively
clearer understanding of the approaches involving
material design and cellular/molecular process has
been achieved.
However, there are still many challenges
collectively faced by bioengineers, cell biologists, and
clinicians, and further development in this area will
require ongoing interactions and collaborations.93
 2010 Jo h n Wiley & So n s, In c. 519
 Advanced Review
New technologies are being developed to provide hope
for future regenerative skin technologies. For example,
stem cell technology is making new milestones with
advances in the efficiency of cell culturing. Stem cells
are defined by their capacity of self-renewal and mul-
tilineage differentiation, which make them attractive
in treating a broad spectrum of human diseases using
regenerative medicine approaches.131,132 Significant
advances have been made in identifying and locating
the stem cells in the skin,133 and several studies have
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