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

and autografts have been used for wound healing. electrospinning and advantages of nanofibers when
However, these naturally derived skin substitutes used as wound cover or matrices for tissue repair and
cannot accomplish skin regeneration due to limited regeneration are emphasized.
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 CURRENT MATERIALS AND
provide the outmost barrier, i.e., epidermal coverage SCAFFOLDS IN WOUND HEALING
in order to prevent infection, reduce water/blood loss,
and control pain. However, an epidermal coverage
Collagen-Based Matrices
alone often fails to restore the structure and functions Collagen is a major ECM protein of the dermal
of the skin and many of the problems such as layer of skin, and it is very logical that the
fragility of the graft, wound contraction, and scar early studies on using biodegradable polymers for
formation often occur.9–11 Increasingly, apparent wound dressing or skin tissue engineering focused
evidences demonstrate that effective wound healing on using collagen. The structural and functional
always necessitates the presence of the dermis layer characteristics of collagen similar to native ECM
in the skin substitutes.12,13 The re-epithelialization stimulated the use of collagen matrices for wound
healing.18 A number of collagen-based dressings
of keratinocytes and take rate are promoted through
containing a variety of carriers/combining agents
dynamic dermal–epidermal interactions, and hence the
in the form of gel, sheet, lattice, or sponge are
graft exhibits higher resistance to wound contraction
and scarring. The restoration of dermis requires commercially available now (Promogram , Johnson
three-dimensional scaffolds to provide elasticity and & Johnson, and Puraply , Royce Medical). Many
strength to the epidermal graft and feed the of them have been used under various clinical
keratinocytes in epidermal layer. Hence, one crucial conditions as a temporary covering for ulcers and
factor in skin tissue engineering is the construction of burns with favorable results being achieved.19 Other
a tissue scaffold as template to guide restructuring of engineered acellular dermis, cellular epidermis/dermis,
cells and subsequent host infiltration of the skin graft. or bilayered skin equivalents involving collagen-based
Over the past three decades, extraordi- materials have been developed as biological skin
nary advances and improved understanding in substitutes for wound healing.
cell/molecular biology have led to achievements in A naturally derived dermis traded as AlloDerm
skin tissue regeneration for wound healing.1 Tissue (LifeCell Corporation) has a porous and fibrillar struc-
engineering skin, based on the concept of a cell–matrix ture similar to native dermis prepared using lyophiliza-
construct, has been a significant advancement in the tion technique after decellularization treatment as
field of wound healing. A number of products are shown in Figure 1A and B.20,21 AlloDerm has been
commercially available and many others are in devel- used as autograft in the resurfacing of burn wound
opment. Over 200,000 patients have been treated reconstruction due to its human origin.22 Another
with tissue-engineered skin products.14–17 Examples acellular natural product (Integra , Integra Life Sci-
of skin substitutes may be acellular or cellular. Acel- ences) was engineered using a porous composite of
lular products only contain the matrix based on bovine collagen-chondroitin-6-sulfate (8 wt%) with
natural or synthetic materials with the required phys- an outer silicone covering.23,24 The outer silicone layer
ical and chemical structure allowing easy access to (Figure 1C25 ) serves as a temporary barrier preventing
host cells during wound healing. Cellular products water loss and microbial invasion, and this layer can
with/without matrix include epidermal cell sheets, be removed 2–3 weeks after wound replacement upon
epidermal constructs, dermal replacements, and bilay- vasularization of the dermis. The porous collagen lay-
ered skin equivalents.16,17 However, currently none of ers (Figure 1D),26 manufactured by lyophilization of
these is fully satisfactory and can replace native skin a dispersion of collagen, crosslinking, and sterilization
autograft and allograft permanently. by chemical means, were designed to act as template
Based on the recent developments and an exten- for host fibroblast infiltration and capillary growth.
sive understanding of the wound healing process, this Integra has been widely used for split-thickness skin
article reviews tissue scaffolds used in commercial skin grafts or wound closure on severe burn wounds when
grafts and some of the new developments occurring donor sites are limited.27 But Integra faces the prob-
at the laboratory stage. The discussion will center on lems of high cost, the need for two surgeries, and the
the processing technique, characteristics, and mate- risk of infection complications.28 Recently, inclusion
rials of tissue scaffolds involving wound repair or of skin cells (keratinocytes, fibroblasts, or both) with
healing. Novel biomimetic scaffolds fabricated using Alloderm or Integra acellular dermis has been

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Advanced Review

(a) (b)

FIGURE 1 | Acellular skin substitutes:

1B 50 µm (A) appearance of Alloderm native tissue
(Reprinted with permission from Ref 20.
Copyright 2010, (B) SEM
(c) (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
brs it 03.html).

performed to architect one cellular bilayered skin on the pioneer work on the cellular skin equivalent
equivalent in some studies.29–31 Fine capillary for- involving collagen.7,34 Briefly, dermal fibroblasts are
mation and accelerated healing rate can be achieved seeded in suspension mixed with bovine type I collagen
on the treatment of full-thickness wounds. However, solution and fibroblast–collagen interaction results in
poor ingrowth of dermal fibroblasts was found in the rearrangement and increased density of collagen
some fashions of Alloderm due to the denser struc- fibrils with a dermal–lattice equivalent formed in
ture of collagen fibrils.32,33 4–6 days. Keratinocytes cultured onto the surface
One of most advanced product of Apligraf of the dermal layer are exposed to an air–liquid
(Figure 2A,7 organogenesis) has been developed based interface to promote maturation and cornification

APLIGRAF Human skin

(a) (b)

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

WIREs Nanomedicine and Nanobiotechnology Tissue scaffolds for skin wound healing

of the stratified epidermal layer, which can support biostability can also extend the clinical durability of
long-term maintenance of the fibroblasts.35 Much like the graft. Besides, incorporation of ECM components
human skin, Apligraf contains key structure and into collagen such as fibronectin, elastin, and GAGs
components similar to human skin (Figure 2B36 ). The plays an important role in guiding the cell physiol-
vascularization and well integration of Apligraf into ogy and behavior in wound healing.13,51,52 Especially,
the wound bed was observed within 14 days and an fibronectin plays an integral role in the attachment of
improved healing was found for chronic venous leg keratinocytes and re-epithelialization.52 GAGs such as
ulcers and diabetic foot ulcers.37–39 Orcel (Ortec hyaluronic acid and chondroitin 6-sulfate have multi-
International) is another bilayered cellular product functional facilitators for wound healing and inhibit
using porous collagen sponge with one side coated wound contraction as well.53 Combining chitosan,
with collagen gel. Dermal fibroblasts are cultured on as one analog to GAGs, with collagen can inhibit
and within the porous sponge side of the collagen its degradation, improve its mechanical strength, and
matrix and keratinocytes are seeded on the gel- accelerate its wound healing.53
coated side and thus ingrowth of keratinocytes can Recently, Seo et al.54 developed a novel rein-
be prevented.40 Accelerated healing rate and reduced forced collagen scaffold used for constructing one
scar formation were achieved with this graft compared artificial dermis layer. The collagen sponge was rein-
with conventional therapy with Biobrane-L.41 But forced by incorporation of collagen-knitted mesh
there are limited clinical trial data available on this fabricated by stacking multilayered collagen thread
graft compared with Apligraf . as shown in Figure 3.54 Continuous collagen thread
Major forms of collagen dressings or matrices was fabricated by extrusion of collagen acid solution
for skin tissue engineering are hydrogel, sponge, into a coagulation bath and multilayered knitted col-
and lattice. Initial collagen gel such as floating lagen mesh was fabricated by cross joining methods.
or anchored gel was introduced to form dermal The study showed that significantly increased ten-
equivalent to investigate the effect of cellular force on sile strength was achieved on the reinforced collagen
collagen contraction, which was linked to subsequent sponge compared with a typical collagen sponge, and
wound contraction.42 The culture work and clinical rapid and firm cell adhesion to the surface and consid-
procedures necessitated the use of denser collagen such erable cell infiltration into the pores were observed.
as lattice or sponge with higher viscosity and strength. Although the abovementioned modification and
Collagen lattice condensed from fibroblast-populated improvement have been achieved, using collagen-
collagen solution was used to mimic the in vivo dermal based polymers for wound healing still creates
layer. However, there was up to 20% contraction problems such as wound contraction and scarring.
of lattice even after transplantation, which inhibited Other concerns discouraging collagen applications
effective wound covering and closure. Besides, the include the high cost of pure collagen, variability
decreased synthesis of collagen was due to the cell in the physicochemical and degradation properties
confinement.43,44 Dehydrated collagen sponges are depending on source and processing.55 Some other
normally produced by freeze drying a dispersion of natural or synthetic polymers have been attempted to
collagen gel or acidic solution.45 Collagen sponge overcome these concerns.56,57
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 Other Natural Polymers
types of artificial skin in the form of sponge were Some other natural polymers including chitosan,
developed.41,46,47 fibrin, elastin, gelatin, and hyaluronic acid have been
However, collagen-based scaffold or dressing investigated for wound healing. Although some of
often has poor biostability and low mechanical prop- their dressing products have been commercialized,
erties and wound contraction easily occurs.42,48,49 their cellular skin grafts are only in the laboratory
Extensive modifications by crosslinking treatment or stage and further clinical data are needed for FDA
combining other natural or synthetic polymers, such approval or commercialization.
as ECM, GAGs, chitosan, polycaprolactone (PCL), Chitosan is another natural polymer most
and PLGA are used to form intra- and interchain widely used next to collagen in wound healing due
bridges between/within collagen molecules with or to its many advantages including biocompatibility,
without external crosslinking agents.50 As a result, the biodegradability, hemostatic activity, and antibacte-
increased mechanical properties can prevent the colla- rial properties.58–60 Chitosan can stimulate collagen
gen matrix from contracting during the culture period synthesis and well incorporate into fibroblast growth
and the wound healing process and the increased factor due to the electrostatic function of chitosan,

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Advanced Review

(a) (b)

FIGURE 3 | Optical
(c) (d) 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).

which can enhance the wound healing rate.61,62 and well organization of fibroblasts within fibrin
Chitosan materials can be easily fabricated into differ- matrices were found in culture models and good take
ent structures under various process conditions. The rate was achieved for subsequent wound healing.70–73
films, gels, or sponges of chitosan have recently been Coating of synthetic polymer surface using fibrin
investigated for use in full-thickness burn wounds.63 glue was used to produce more suitable scaffolds
However, chitosan has rapid biodegradability espe- for civilizing keratinocytes with higher strength and
cially in acid environment that is often formed in acceptable biocompatibility.74
wound healing.64 Various crosslinking treatments Gelatin as one protein derived from collagen has
have been used to increase the structural stability of been investigated as matrices in the dehydrated form
chitosan.56 Also, it is highly desirable to incorporate such as sponge or film for skin tissue engineering,
chitosan into another one or two different polymers to and promoted epithelialization and granulation tissue
balance its advantages and disadvantages.65,66 Large formation were shown in wound healing.75 But this
number of amino groups in chitosan plays a significant material has the limitation of low mechanical strength
role in bridging and crosslinking between/within chi- and is effectively used only as incorporated component
toscan and/or other polymers such as collagen, gelatin,
with others polymers to promote or modify the
and GAGs.67 The increased crosslinking efficiency
biological or mechanical properties.76–78
may improve the biostability and mechanical proper-
Hyaluronic acid or hyaluronan-based scaf-
ties of scaffolds. Ma casted chitosan film onto collagen
folds have been studied due to its excellent
sponge to form one bilayer structure for skin tissue
biocompatibility.79 One non-woven fleece of 100%
engineering.68 This chitosan layer with denser struc-
ture can enhance the functions of the artificial dermis benzyl-esterified derivative of hyaluronic acid was
with increased controlled fluid loss and antibacterial already marked as Hyaff (Fidia Advanced Biopoly-
activity. However, some disadvantages of chitosan mers) and its efficacy used for cultured epidermal
such as unsatisfactory mechanical properties, severe graft (Laserskin ) or dermal graft (Hyalograft 3D)
shrinkage, and deformation after drying discouraged was demonstrated in extensive burns.80,81 Fibrob-
its application in tissue engineering field.69 last–keratinocyte composite based on this Laserskin
Fibrin glues in suspension, gel, membrane, or membrane with the promoted three dimension can
sheet form have been used as a dressing or co-culture produce skin equivalent, and promising in vitro results
system for delivering keratinocytes to treat burn suggested the potential application for burns and
wound. Rapid re-epithelialization of keratinocytes chronic wounds.82

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WIREs Nanomedicine and Nanobiotechnology Tissue scaffolds for skin wound healing

In summary, the success from the use of natural Polyurethane (PU) has been extensively studied
biodegradable cell matrices has been encouraging as materials for wound dressing due to its semiperme-
and continues to facilitate broader use in the ability. PU wound dressings are impermeable to bacte-
future. However, many problems occurring with ria and water but permeable to gas with moist environ-
natural polymers include low mechanical strength, ment desirable for wound healing to occur. PU-based
shrinkage/contraction, difficulty in handling, and membranes have been commercially available for
risks of immunological rejection.57 Use of synthetic wound dressing (Tegaderm, 3M Health Care and
polymer or incorporating synthetic polymer into
OpSite, Smith & Nephew). The products are cost-
natural polymer may be favorable when higher
effective for covering small-sized, split-thickness skin
mechanical properties and more flexible fabrication
ability of synthetic polymers are highly desirable.12 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
Synthetic Polymer Scaffolds mesh (Biobrane, Dow B. Hickam, Inc.). Collagen
Generally, synthetic polymers possess lot-to-lot uni- or collagen-derived peptide is coated to improve cell
formity and have less expensive and more reliable adherence or tissue biocompatibility. These wound
sources of raw material. They can be tailored using dressings have been used as a standard control of cell
various fabrication techniques to provide a wide range carrier to compare their in vitro biocompatibility with
of physical properties. Although some work indicated those of some natural or synthetic dressings.84
that acceptable epithelialization of keratinocytes can A knitted polyglycolic acid/polylactic acid
be achieved on synthetic polymer surface, currently
(PGA/PLA or namely Polyglactin) mesh as typically
no successful epidermal graft using synthetic poly-
shown in Figure 4A85 was cultured with human
mers has been achieved due to their limited cellular
recognition and tissue compatibility.83 The studies neonatal fibroblasts leading to the development
using synthetic polymers for skin substitutes are of one commercial product of cryopreserved
already focusing on the combination with natural Dermagraft (Advanced Tissue Sciences) (Figure
polymers used as matrices for temporary dressing, 4B).86 Dermagraft has been used successfully to treat
epidermal/dermal cell carriers, or full-thickness skin full-thickness diabetic foot ulcers. The knitted PLGA
equivalent. meshes support homogenous cell distribution and

(a) (b)

100 µm

FIGURE 4 | Polyglactin mesh (A),

Dermagraf as received from a pack
(B), dermal fibroblasts cultured on (c) (d)
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
68.7 µm 68.7 µm

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Advanced Review

withstand the cell contractile force (Figure 4C). How- as fiber drawing, template synthesis, temperature-
ever, the PLGA scaffold has the problems of surface induced phase separation, molecular self-assembly,
hydrophobicity and limited cell adhesion. One modi- and electrospinning.94 However, only electrospinning
fied knitted PLGA mesh hybridized with collagen has has emerged as an efficient technique to produce
been investigated as a three-dimensional culture sys- a variety of polymeric nanofibers.95 Electrospinning
tem for skin tissue engineering85 or some other tissues generally involves the application of an electrostatic
construction.87,88 These web-like collagen sponges force between polymer solution (or melt) kept in
can facilitate seeding, uniform distribution, and a syringe or pipette and a counter metal electrode
ECM synthesis of dermal fibroblasts (Figure 4D).85 (collector plate) kept at a certain distance from
TransCyte , formerly known as dermagraft-TC, is a the polymer solution as shown in Figure 5A. The
cell-seeded Biobrane product, composed of a semiper- electrospinning technique has gained attention and
meable silicone membrane and newborn human popularity in the last 10 years due to an increased
fibroblast cells cultured on a collagen-coated nylon interest in nanoscale properties and technologies.
mesh. But it is only a temporary covering due to the One major attractive feature of electrospinning is
poor biodegradability of nylon.89,90 the simplicity and inexpensive nature of its setup.
Many other synthetic polymers used in Electrospun scaffolds provide high surface area to
restorable sutures or wound covering have been volume ratio, which has been proved to promote
investigated as matrix materials for dermal substi- cell–matrix interaction at the nanoscale.96,97
tutes or full-thickness skin equivalent. They include This high surface area to volume ratio of
PCL, poly(l-lactide) (PLLA), PGA, and copoly- electrospun scaffolds facilitates oxygen permeability
mer poly(ethyleneglycolterephthalate)-poly (butylenes and allows fluid accumulation, which are highly
terephthalate).12,91 However, limited clinical success desirable in the wound healing course. Besides, the
was achieved because synthetic polymers often have pores in non-woven form of electrospun scaffolds
lower rates of cell attachment and proliferation due (normally 1–10 µm) are small enough to prevent
to the limited biological signals. The combination of bacterial penetration.95,102 Much attention has been
two or more different polymers including natural or drawn to the use of electrospun scaffolds as wound
synthetic is necessary to produce suitable scaffolds for dressing or cellular carrier for skin substitutes
skin regeneration when the advantages or disadvan- made of a variety of materials including collagen,
tages of each material are well balanced. Although gelatin, fibrinogen, chitosan, PU, PCL, PLA, and
the overall research is encouraging, our knowledge PLGA,103–106 or some blends of them.107,108 Current
and understanding of the skin substitute and wound work using electrospun nanofibrous membranes as
healing involving biology and material science still medical dressings or skin regeneration for wound
need to be improved. In the past 10 years, the devel- healing is still at an early stage.
opment of nanotechnology involving fabrication and
characterization methods has encouraged the research
community to investigate the potential of applying Electrospun Natural and Synthetic
nanofibrous scaffolds in the tissue engineering field. 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
ELECTROSPUN BIOMIMETIC (HFP) as shown in Figure 5B. Structural and bio-
NANOFIBROUS SCAFFOLDS logical characterizations were conducted to confirm
FOR WOUND HEALING that the structural and biological properties of native
collagen were maintained after solvent dissolution
Characteristics of Electrospinning and and electrospinning. Subsequently many research
Electrospun Scaffolds works extended the application of electrospun col-
As we know, native ECM in dermis layer is lagen to several tissue engineering fields such as skin,
composed of collagen nanoscale fibers and it provides cornea, and blood vascular system.109–112 Electro-
structural integrity and mechanical strength to spun collagen was found to be the most biomimetic
skin tissues. Thus, a biologically inspired scaffold nanofibrous scaffolds due to its structure and origin
fabrication approach for wound healing is to create closely similar to the native ECM of dermis. Several
ECM analogs composed of nanoscale fibers with studies already demonstrated that collagen nanofi-
mimicking structure and functions to native ECM.92,93 brous matrices exhibited excellent biocompatibility to
There are many fiber fabrication methods such dermal fibroblast and keratinocytes, and significantly

516  2010 Jo h n Wiley & So n s, In c. Vo lu me 2, September/Octo ber 2010

WIREs Nanomedicine and Nanobiotechnology Tissue scaffolds for skin wound healing

(a) (b)

High voltage Polymer


Polymer jet


FIGURE 5 | Electrospinning and

electrospun nanofibers. (A) Schematic
diagram of the electrospinning setup,
(B) collagen nanofibers, (reprinted with 100 nm

permission from Ref 99. Copyright 2007 John

Wiley & Sons, Inc.), scale bar = 100 nm,
(e) (d)
(C) PVA/AgNO3 nanofiber, (reprinted with
permission from Ref 98. Copyright 2002 Nanofiber scaffold

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 0.2 µm
Ref 101. Copyright 2007 Elsevier).

accelerated wound healing and inhibited wound con- organization, and excellent barrier desirable for
traction (compared with collagen sponge) were found wound healing were discussed.104,116 However these
in early-stage wound healing.103,109 However, one studies suggested that combination with synthetic
recent report provided very direct evidences that over- polymers (PU or PCL) is highly desirable to provide
whelming denaturation of collagen into gelatin occurs higher mechanical properties of scaffold.
with those commonly used fluorinated alcohols such Some other natural polymers including silk
as HFP or 2,2,2-trifluoroethanol (TFE) as the dis- fibroin, chitin/chitosan, fibrinogen, or their blends
solving solvent for collagen.113 Considering the high have been electrospun as wound dressing or cellular
cost of collagen material, further investigations are matrix, and efficient cell attachment, growth, or
needed to clarify the effect of collagen denaturation infiltration has been demonstrated in in vitro
on the biological activity and structural mechanical culture,117–119 but limited clinical trial has been
integrity, which are highly desirable in mimicking reported. Recently, hemoglobin and myoglobin have
native ECM. been electrospun as wound dressing aimed to
Gelatin, as an alternative protein to collagen, is promote oxygen delivery to the healing tissue using
commercially available at significantly lower cost and hemoglobin’s functional analog of red blood cells.
preserves many merits of collagen such as biological Increased oxygen delivery was deemed to increase
origin, biodegradability, and biocompatibility. Gelatin the metabolic rate and infectious resistance of the
was successfully electrospun into nanofibers using wound, and thus wound healing was supposed to be
several solvents such as TFE, HFP, and formic accelerated.120
acid.104,114,115 Potential application of electrospun Natural electrospun nanofibers normally require
gelatin as wound dressing and optimal fiber density crosslinking treatment to stabilize the structures of
to provide high cell viability, and optimal cell nanofibers before in vitro or in vivo use. However,

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Advanced Review

choosing efficient crosslinking for electrospun natural the scaffolds suitable for tissue engineering skin with
polymer needs to proceed with extra care as aqueous a good balance of mechanical strength and biological
degradation or swelling of natural polymer in the properties. In our laboratory, Zhang et al. showed
early stage of crosslinking easily damages the pores of that PCL/collagen nanofibers in core–shell form
matrix or impairs the porosity of the electrospun mats. fabricated using coaxial core/shell electrospinning
As suggested in several studies, glutaraldehyde vapor (Figure 5D) exhibited higher biological affinity than
crosslinking is a highly efficient method, and nanoscale the PCL nanofibers simply coated with collagen.100
dimension and porous structure of collagen, gelatin, or More recently, PCL/gelatin nanofibrous scaffold
fibrinogen nanofibers can be preserved.105,114,121,122 and layered dermal reconstitution were evaluated
Electrospun synthetic polymers such as PU, for wound healing. PCL/gelatin was electrospun
PCL, PLLA, and poly (vinyl phenol) have been onto one commercial PU membrane of Tegaderm
proposed for wound dressing application in wound (Tegaderm, 3M Medical) as shown in Figure 5E.
healing.101,102,116 Early studies using electrospun Tegaderm membrane can facilitate clinical handling,
synthetic polymers for wound dressing, together provide a protective barrier to external infections as
with some electrospun natural polymers, have synthetic epidermis, and control water loss. Significant
created wound healing scaffolds with such inherent cell adhesion, growth, and infiltration achieved
properties of the electrospun material. Controlled on the PCL/gelatin produced a fibroblast-populated
evaporative water loss, excellent oxygen permeability, three-dimensional dermal analog. This cost-effective
and promoted fluid drainage have been revealed on composite could be promising wound dressing or
most of the electrospun mats used. In some recent tissue scaffolds for skin construct.
studies, the addition of drugs into nanofibers as The potential application for PLGA/dextran
controlled release system has been investigated for was investigated as tissue scaffolds for skin tis-
wound dressing to provide the required protection and sue engineering.129 A complete cell biological
pain management. Some antibiotic or antibacterial response of dermal fibroblasts was tested on
drugs/components such as cefazolin, liodocaine, electrospun PLGA/dextran nanofibers. The results
mupirocin, or Ag particles have been electrospun showed that favorable fibroblasts interacted with
into synthetic nanofibers in the blended form the scaffolds and resembled a dermal-like archi-
(Figure 5C), and increased or controlled antimicrobial tecture. Aimed to overcome poor infiltration of
or antibiotic ability desirable for wound healing has fibroblasts into electrospun mats, a novel three-
been achieved with a sustained controlled release rate dimensional multilayered cell–nanofiber constructs of
of drugs.99,115,123–125 These drug delivery systems dermis was fabricated by architecting layer-by-layer
using electrospun nanofibers exhibit pain relief and of nanofibers–cells alternated using electrospinning
extended antibacterial activity with great potential of nanofibers of PCL/collagen and seeding human dermal
the applications in wound healing. fibroblasts (Figure 6).130 Dermal tissue or bilayered
For the use of electrospun pure synthetic poly- skin equivalent was produced by continuous cul-
mer in skin tissue engineering, only limited synthetic turing of fibroblast/fiber-layered constructs or inclu-
polymers such as PLLA, PLGA, blended poly(dl- sion of keratinocytes seeded onto the dermal layer.
lactide), and poly(ethylene glycol) having acceptable Several other composites including PLGA/collagen,
hydrophilicity have been reported for the potential PLLA/gelatin, and PVA/chitosan have been electro-
application in skin tissue scaffolding.106,126,127 It is spun and their characteristics oriented for the appli-
widely accepted that the incorporation of natural poly- cation in skin tissue engineering were discussed.98,118
mer into electrospun synthetic nanofibers is desirable These scaffolds showed their efficacy for wound cov-
in skin tissue engineering to promote the biological erage or cell growth biocompatibility, but further
activity of scaffolds. Composite polymeric nanofibers clinical studies are needed.
can be electrospun using mixed solution,107 using Previous work using this nanofabrication tech-
dual or co-electrospinning method in separate solvent nique already suggested great potential of applying
systems111 or coaxial core/shell electrospinning.100 nanofibrous scaffolds for wound dressing or skin
Nanofibrous composite of PCL/collagen was tissue engineering. However, very limited in vivo
electrospun, as the one in early model of composite applications for skin repair and regeneration have
used to support dermal fibroblasts. Heather et al. been reported. More effort is still needed to seek
investigated the association of mechanical strength FDA approval for use and to prove their effectiveness
and biological affinity of blended PCL/collagen in repairing and regenerating skin. Electrospun
scaffolds with various ratios of PCL/collagen.128 scaffolds have been extensively used in vitro to study
Minimal addition of PCL (10%) to collagen produces cell–scaffold interactions, and further understanding

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WIREs Nanomedicine and Nanobiotechnology Tissue scaffolds for skin wound healing

Step 1: Fiber spinning (1 layer fiber) Step 2: Cell seeding (1 layer cell)

FIGURE 6 | Schematic Fiber

illustration of layer-by-layer dermal Cell
reconstruction using nanofibrous Cell
scaffold. (Reprinted with Fiber
permission from Ref 130. Copyright Cell
2009 Mary Ann Liebert, Inc.
Publishers). Repeat step 1 & step 2 until desirable layers of fibers and cells On site layer-by-layer tissue generation

of the cellular response of nanofibrous scaffolds might Although the potential risks and benefits of com-
ultimately lead to the success in tissue repair or regen- posite scaffolds need to be further evaluated in clinical
eration of skin. 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
CONCLUSIONS AND FUTURE natural and synthetic polymers and can result in a new
OUTLOOK biomaterial with appropriate biocompatibility and
mechanical, physical, and chemical properties. Elec-
Tissue scaffolds have been demonstrated to be useful trospinning can produce new type nanofibrous scaf-
in wound dressing or skin tissue engineering in a folds for wound dressing or skin tissue engineering.
variety of wound conditions. Tissue scaffolds can not This electrospun type scaffold exhibits an appropriate
only can cover the wound and provide a physical porous structure that can mimic the collagenous struc-
barrier, but can also can offer a cellular skin ture of native dermis. The electrospun PCL/gelatin
with excretive biological components to stimulate scaffolds developed in our laboratory exhibit excellent
re-epithelialization and formation of granulation biocompatibility and biodegradability. The low cost
tissue. A variety of scaffolds fabricated based on and existing FDA approval for use of PCL and gelatin
materials ranging from naturally occurring materials and their proven efficiency in tissue engineering con-
and those manufactured synthetically have been tribute to the ease of FDA approval for clinical use and
reviewed in this article. Collagen-based materials commercialization of the final artificial skin using this
have been investigated extensively and significant scaffold. In future, the directions in scaffold design-
success has been achieved in a variety of commercial ing would be to incorporate molecules such as ECM,
or laboratory-engineered skin substitutes for wound growth factors, or cytokines associated with enhanc-
healing. However, many problems associated with ing wound healing into the right scaffolds. A relatively
the current scaffolds and artificial skin substitutes, clearer understanding of the approaches involving
which include contraction, delayed vascularization, material design and cellular/molecular process has
high cost, and scarring, have been emerging and they been achieved.
vary in their severity. It is hoped that a combination However, there are still many challenges
of appropriate biomaterials and techniques can collectively faced by bioengineers, cell biologists, and
overcome many of these problems in order to eliminate clinicians, and further development in this area will
or reduce the risk of failure in skin graft. require ongoing interactions and collaborations.93

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Advanced Review

New technologies are being developed to provide hope proved that stem cells can contribute to skin reconsti-
for future regenerative skin technologies. For example, tution in skin wounds.134,135 The limited regenerative
stem cell technology is making new milestones with capacity of epidermal keratinocytes could be over-
advances in the efficiency of cell culturing. Stem cells come by using self-renewing keratinocyte stem cells.
Locating hair follicle stem cells can bring possible
are defined by their capacity of self-renewal and mul-
hope of forming hair follicle in future bioengineered
tilineage differentiation, which make them attractive
skin products.132,134,135 Stem cell techniques will bring
in treating a broad spectrum of human diseases using new breakthroughs in skin tissue engineering. The for-
regenerative medicine approaches.131,132 Significant mation of a new generation of skin substitutes with
advances have been made in identifying and locating complete functions and structures of native skin fully
the stem cells in the skin,133 and several studies have restored can potentially be achieved in the next decade.

1. Falanga V, Faria K, Robert L, Joseph V. Bioengineered 12. Dai NT, Williamson MR, Khammo N, Adams EF,
skin constructs. Principles of Tissue Engineering. 3rd Coombes AGA. Composite cell support mem-
ed. Burlington: Academic Press; 2007, 1167–1185. branes based on collagen and polycaprolactone
2. Pomahac B, Svensjo T, Yao F, Brown H, Eriksson E. for tissue engineering of skin. Biomaterials 2004,
Tissue engineering of skin. Crit Rev Oral Biol Med
1998, 9:333–344. 13. Ruszczak Z, Effect of collagen matrices on der-
mal wound healing. Adv Drug Deliv Rev 2003,
3. Tsuruta D, Green KJ, Getsios S, Jones JCR, The bar-
rier function of skin: how to keep a tight lid on water
loss. Trends Cell Biol 2002, 12:355–357. 14. Jones I, Currie L, Martin R. A guide to biological skin
substitutes. Br J Plastic Surg 2002, 55:185–193.
4. Kaustabh G, Clark RAF, Lanza R, Langer R,
Vacanti J. Wound repair. Principles of Tissue Engi- 15. Whitaker MJ, Quirk RA, Howdle SM, Shakesheff
neering. 3rd ed. Burlington: Academic Press; 2007, KM. Growth factor release from tissue engi-
1149–1166. neering scaffolds. J Pharmacy Pharmacol 2001,
5. Allgower M, Schoenberger GA, Sparkes BG. Burning
the largest immune organ. Burns 1995, 21:S7–S47. 16. Mansbridge J. Tissue-engineered skin substitutes. Exp
Opin Biol Therapy 2002, 2:25–34.
6. Balasubramani M, Kumar TR, Babu M. Skin substi-
tutes: a review. Burns 2001, 27:534–544. 17. Ehrenreich M, Ruszczak Z. Update on tissue-
engineered biological dressings. Tissue Eng 2006, 12:
7. Price R, Anthony E, Myers S, Navsaria H. Tissue engi- 2407–2424.
neering for skin transplantation. Tissue Engineering.
Burlington: Academic Press; 2008, 507–532. 18. Sai KP, Babu M. Collagen based dressings—a review.
Burns 2000, 26:54–62.
8. Clark RAF, Ghosh K, Tonnesen MG. Tissue engineer-
ing for cutaneous wounds. J Invest Dermatol 2007, 19. Rastogi S, Modi M, Sathian B. The efficacy of collagen
127:1018–1029. membrane as a biodegradable wound dressing mate-
rial for surgical defects of oral mucosa: a prospective
9. Williamson JS, Snelling CFT, Clugston P, Macdonald study. J Oral Maxillofac Surg 2009, 67:1600–1606.
IB, Germann E. Cultured epithelial autograft: five
years of clinical experience with twenty-eight patients. 20. Schwade ND. Implants, Soft Tissue, AlloDerm.
Annual Meeting of the Trauma-Association-of- 2008. Available at:
Canada, Toronto, Canada, 1994. article/879007-print. Accessed October 20, 2009.

10. Carsin H, Ainaud P, Le Bever H, Rives J-M, Lakhel A, 21. Sun WQ, Gouk SS. Aging of a regenerative biologic
Stephanazzi J, Lambert F, Perrot J. Cultured epithelial scaffold (AlloDerm Native Tissue Matrix) during stor-
autografts in extensive burn coverage of severely trau- age at elevated humidity and temperature. Tissue Eng
matized patients: a five year single-center experience Part C Methods 2009, 15:23–31.
with 30 patients. Burns 2000, 26:379–387. 22. Kearney JN. Clinical evaluation of skin substitutes.
11. Hafemann B, Ensslen S, Erdmann C, Niedballa R, Burns 2001, 27:545–551.
Zühlke A, Ghofrani K, Kirkpatrick CJ. Use of a 23. Geiger M, Li RH, Friess W. Collagen sponges for
collagen/elastin-membrane for the tissue engineering bone regeneration with rhBMP-2. Adv Drug Deliv
of dermis. Burns 1999, 25:373–384. Rev 2003, 55:1613–1629.

520  2010 Jo h n Wiley & So n s, In c. Vo lu me 2, September/Octo ber 2010

WIREs Nanomedicine and Nanobiotechnology Tissue scaffolds for skin wound healing

24. Burke JF, Yannas IV, Quinby WC, Bondoc CC, Hardin-Young J, Human Skin G, et al. Rapid heal-
Jung WK. Successful use of a physiologically accept- ing of venous ulcers and lack of clinical rejection with
able artificial skin in the treatment of extensive burn an allogeneic cultured human skin equivalent. Arch
injury. Ann Surgery 1981, 194:413–428. Dermatol 1998, 134:293–300.
25. Anthony ET, Syed M, Myers S, Moir G, Navsaria H. 38. Curran MP, Plosker GL. Bilayered bioengineered skin
The development of novel dermal matrices for cuta- substitute (Apligraf (R))—a review of its use in the
neous wound repair. Drug Discov Today Ther Strateg treatment of venous leg ulcers and diabetic foot ulcers.
2006, 3:81–86. Biodrugs 2002, 16:439–455.
26. Product makeup/histology. 2007, Available at: 39. Sabolinski ML, Alvarez O, Auletta M, Mulder G, it 03.html. Parenteau NL. Cultured skin as a ‘smart material’
Accessed October 20, 2009. for healing wounds: experience in venous ulcers.
27. Fitton AR, Drew P, Dickson WA. The use of a bilam- Biomaterials 1996, 17:311–320.
inate artificial skin substitute (Integra (TM)) in acute 40. Eisenberg M, Llewelyn D. Surgical management of
resurfacing of burns: an early experience. Br J Plastic hands in children with recessive dystrophic epider-
Surg 2001, 54:208–212. molysis bullosa: use of allogeneic composite cultured
28. Chou TD, Chen SL, Lee TW, Chen SG, Cheng TY, skin grafts. Br J Plastic Surg 1998, 51:608–613.
Lee CH, Chen TM, Wang HJ. Reconstruction of 41. Still J, Glat P, Silverstein P, Griswold J, Mozingo D.
burn scar of the upper extremities with artificial skin. The use of a collagen sponge/living cell composite
Plastic Reconstruct Surg 2001, 108:378–384. material to treat donor sites in burn patients. Burns
29. Kumar S, Tran J-L, Hadeed J, Bellavance E, Strande 2003, 29:837–841.
L, Eydelman R, Matthews M, Marra S, Hewitt C. 42. Auger FA, Rouabhia M, Goulet F, Berthod F,
Capillary formation in bioengineered human skin con- Moulin V, Germain L. Tissue-engineered human
structs (BHSC) designed to study burn injury. J Am skin substitutes developed from collagen-populated
Coll Surgeons 2004, 199(Suppl 1):65–65. hydrated gels: clinical and fundamental applications.
30. Segal N, Andriani F, Pfeiffer L, Kamath P, Lin N, Med Biol Eng Comput 1998, 36:801–812.
Satyamurthy K, Egles C, Garlick JA. The basement 43. Ho G, Barbenel J, Grant MH. Effect of low-level
membrane microenvironment directs the normaliza- laser treatment of tissue-engineered skin substitutes:
tion and survival of bioengineered human skin equiv- contraction of collagen lattices. J Biomed Optics
alents. Matrix Biol 2008, 27:163–170. 2009, 14.
31. Cuono C, Langdon R, McGuire J. Use of cul- 44. Berthod F, Hayek D, Damour O, Collombel C. Colla-
tured epidermal autografts and dermal allografts as gen synthesis by fibroblasts cultured within a collagen
skin replacement after burn injury. Lancet 1986, sponge. Biomaterials 1993, 14:749–754.
1:1123–1124. 45. O’Brien FJ, Harley BA, Yannas IV, Gibson L. Influ-
32. Ojeh NO, Frame JD, Navsaria HA. In vitro charac- ence of freezing rate on pore structure in freeze-
terization of an artificial dermal scaffold. Tissue Eng dried collagen-GAG scaffolds. Biomaterials 2004,
2001, 7:457–472. 25:1077–1086.
33. Ng KW, Khor HL, Hutmacher DW. Skin tissue engi- 46. Powell HM, Boyce ST. Wound closure with EDC
neering part II - the in vitro evaluation of natural and cross-linked cultured skin substitutes grafted to
synthetic 3-D matrices as dermal substrates. In: Reis athymic mice. Biomaterials 2007, 28:1084–1092.
RL, Román US, eds. Biodegradable Systems in Tis- 47. Kumar S, Tran JL, Hadeed J, Bellavance E, Strande
sue Engineering and Regenerative Medicine. London: L, Eydelman R, Matthews M, Marra S, Hewitt C.
CPC Press; 2005, 529–547. Triphala incorporated collagen sponge–a smart bio-
34. Bell E, Ehrlich HP, Buttle DJ, Nakatsuji T. Living tis- material for infected dermal wound healing. J Surgical
sue formed in vitro and accepted as skin-equivalent Res 2008. In Press, Corrected Proof.
tissue of full thickness. Science 1981, 211:1052–1054. 48. Ono I, Tateshita T, Inoue M. Effects of a colla-
35. Wilkins LM, Watson SR, Prosky SJ, Meunier SF, Par- gen matrix containing basic fibroblast growth factor
enteau NL. Development of a bilayered living skin on wound contraction. J Biomed Mater Res 1999,
construct for clinical applications. Biotechnol Bioeng 48:621–630.
1994, 43:747–756. 49. Berry DP, Harding KG, Stanton MR, Jasani B,
36. The science behind ApligrafR . Available from: http:// Ehrlich HP. Human wound contraction: collagen is apligraf/how organization, fibroblasts, and myofibroblasts. Plastic
works/the science behind apligraf.html. Reconstruct Surg 1998, 102:124–131.
37. Falanga V, Margolis D, Alvarez O, Auletta M, Mag- 50. Park S-N, Lee HJ, Lee KH, Suh H. Biological char-
giacomo F, Altman M, Jensen J, Sabolinski M, acterization of EDC-crosslinked collagen-hyaluronic

Vo lu me 2, September/Octo ber 2010  2010 Jo h n Wiley & So n s, In c. 521

Advanced Review

acid matrix in dermal tissue restoration. Biomaterials 64. Madihally SV, Matthew HWT. Porous chitosan
2003, 24:1631–1641. scaffolds for tissue engineering. Biomaterials 1999,
51. Doillon CJ, Silver FH. Collagen-based wound dress-
ing: effects of hyaluronic acid and firponectin on 65. Kellouche S, Martin C, Korb G, Rezzonico R,
wound healing. Biomaterials 1986, 7:3–8. Bouard D, Benbunan M, Dubertret L, Soler C,
Legrand C, Dosquet C. Tissue engineering for full-
52. Doillon CJ, Silver FH, Berg RA. Fibroblast growth on
thickness burns: a dermal substitute from bench
a porous collagen sponge containing hyaluronic acid
to bedside. Biochem Biophys Res Commun 2007,
and fibronectin. Biomaterials 1987, 8:195–200.
53. Ding CM, Zhou Y, He YN, Tan WS. Perfusion seed-
66. Liu HF, Mao JS, Yao KD, Yang GH, Cui L, Cao YL.
ing of collagen-chitosan sponges for dermal tissue
A study on a chitosan-gelatin-hyaluronic acid scaffold
engineering. Process Biochem 2008, 43:287–296.
as artificial skin in vitro and its tissue engineer-
54. Seo YK, Youn HH, Park CS, Song,KY, Park JK. Rein- ing applications. J Biomater Sci Polymer Ed 2004,
forced bioartificial dermis constructed with col- 15:25–40.
lagen threads. Biotechnol Bioprocess Eng 2008, 67. Mao JS, liu HF, Yin YJ, Yao KD. The properties of
13:745–751. chitosan-gelatin membranes and scaffolds modified
55. Lee CH, Singla A, Lee Y. Biomedical applications of with hyaluronic acid by different methods. Biomate-
collagen. Int J Pharm 2001, 221:1–22. rials 2003, 24:1621–1629.
56. Adekogbe I, Ghanem A. Fabrication and charac- 68. Ma L, Gao C, Mao Z, Zhou J, Shen J, Hu X, Han C.
terization of DTBP-crosslinked chitosan scaffolds Collagen/chitosan porous scaffolds with improved
for skin tissue engineering. Biomaterials 2005, biostability for skin tissue engineering. Biomaterials
26:7241–7250. 2003, 24:4833–4841.
57. Woei K, Hutmacher DW, Schantz JT, Seng C, 69. Cestari AR, Vieira EFS,Pinto AA, Lopes ECN. Mul-
Too HP, Chye T, Phan TT, Teoh SH. Evaluation of tistep adsorption of anionic dyes on silica/chitosan
ultra-thin poly(epsilon-caprolactone) films for tissue- hybrid: 1. Comparative kinetic data from liquid-
engineered skin. Tissue Eng 2001, 7:441–455. and solid-phase models. J Colloid Interface Sci 2005,
58. Ma J, Wang H, He B, Chen J. A preliminary in vitro
study on the fabrication and tissue engineering appli- 70. Meana A, Iglesias J, Del Rio M, Larcher F,
cations of a novel chitosan bilayer material as a Madrigal B, Fresno MF, Martin C, San Roman F,
scaffold of human neofetal dermal fibroblasts. Bio- Tevar F. Large surface of cultured human epithelium
materials 2001, 22:331–336. obtained on a dermal matrix based on live fibroblast-
containing fibrin gels. Burns 1998, 24:621–630.
59. Horn MM, Martins VCA, Plepis AMD. Interaction
of anionic collagen with chitosan: effect on thermal 71. Sun T, Haycock J, MacNeil S. In situ image analysis
and morphological characteristics. Carbohydr Polym of interactions between normal human keratinocytes
2009, 77:239–243. and fibroblasts cultured in three-dimensional fibrin
gels. Biomaterials 2006, 27:3459–3465.
60. Taravel MN, Domard A. Collagen and its interac-
tion with chitosan II influence of the physiochemi- 72. Ronfard V, Broly H, Mitchell V, Galizia JP,
cla characteristics of collagen. Biomaterials 1995, Hochart D, Chambon E, Pellerin P, Huart JJ. Use
16:865–871. of human keratinocytes cultured on fibrin glue in the
treatment of burn wounds. Burns 1991, 17:181–184.
61. Chung LY, Schmidt RJ, Hamlyn PF, Sagar BF,
Andrews AM, Turner TD. Biocompatibility of poten- 73. Hunyadi J, Farkas B, Bertenyi C, Olah J, Dobozy A.
tial wound management products: fungal mycelia as Keratinocyte grafting: a new means of transplantation
a source of chitin/chitosan and their effect on the for full-thickness wounds. J Dermatol Surg Oncol
proliferation of human F1000 fibroblasts in culture. 1988, 14:75–78.
J Biomed Mater Res 1994, 28:463–469. 74. Khor HL, Ng KW, Htay AS, Schantz JT, Teoh SH,
62. Mizuno K, Yamamura K, Yano K, Osada T, Saeki S, Hutmacher DW. Preliminary study of a polycapro-
Takimoto N, Sakurai T, Nimura Y. Effect of chitosan lactone membrane utilized as epidermal substrate.
film containing basic fibroblast growth factor on J Mater Sci Mater Med 2003, 14:113–120.
wound healing in genetically diabetic mice. J Biomed 75. Tanaka A, Nagate T, Matsuda H. Acceleration of
Mater Res A 2003, 64A:177–181. wound healing by gelatin film dressings with epider-
mal growth factor. J Vet Med Sci 2005, 67:909–913.
63. Shi C, Zhu Y, Ran X, Wang M, Su Y, Cheng T.
Therapeutic potential of chitosan and its deriva- 76. Deng C-M, He L-Z, Zhao M, Yang D, Liu Y. Biolog-
tives in regenerative medicine. J Surgical Res 2006, ical properties of the chitosan-gelatin sponge wound
133:185–192. dressing. Carbohydrate Polym 2007, 69:583–589.

522  2010 Jo h n Wiley & So n s, In c. Vo lu me 2, September/Octo ber 2010

WIREs Nanomedicine and Nanobiotechnology Tissue scaffolds for skin wound healing

77. Wang T-W, Sun J-S, Wu H-C, Tsuang Y-H, Wang 90. Amani H, Dougherty WR, Blome-Eberwein S. Use of
W-H, Lin F-H. The effect of gelatin-chondroitin transcyte? and dermabrasion to treat burns reduces
sulfate-hyaluronic acid skin substitute on wound heal- length of stay in burns of all size and etiology. Burns
ing in SCID mice. Biomaterials 2006, 27:5689–5697. 2006, 32:828–832.
78. Lee SB, Kim YH, Chong MS, Hong SH, Lee YM. 91. Beumer GJ, van Blitterswijk CA, Bakker D, Ponec M.
Study of gelatin-containing artificial skin V: fabrica- Cell-seeding and in vitro biocompatibility evaluation
tion of gelatin scaffolds using a salt-leaching method. of polymeric matrices of PEO/PBT copolymers and
Biomaterials 2005, 26:1961–1968. PLLA. Biomaterials 1993, 14:598–604.
79. Price RD, Berry MG, Navsaria HA. Hyaluronic acid: 92. Lanza RP, Langer R, Vacanti J. Principles of Tis-
the scientific and clinical evidence. J Plastic Recon- sue Engineering. 2nd ed. New York: Academic Press;
struct Aesthetic Surg 2007, 60:1110–1119. 2000.
80. Harris PA, di Francesco F, Barisoni D, Leigh IM, 93. Griffith L, Naughton G. Tissue engineering: cur-
Navsaria HA. Use of hyaluronic acid and cultured rent challenges and expanding opportunities. Science
autologous keratinocytes and fibroblasts in extensive 2002, 295:1014.
burns. Lancet 1999, 353:35–36. 94. Kumbar SG, James R, Nukavarapu SP, Laurencin CT.
81. Hollander DA, Soranzo C, Falk S, Windolf J. Exten- Electrospun nanofiber scaffolds: engineering soft tis-
sive traumatic soft tissue loss: reconstruction in sues. Biomed Mater 2008, 3.
severely injured patients using cultured hyaluronan- 95. Huang ZM, Zhang YZ, Kotaki M, Ramakrishna S.
based three-dimensional dermal and epidermal auto- A review on polymer nanofibers by electrospinning
grafts. J Trauma Injury Infection Crit Care 2001, and their applications in nanocomposites. Compos
50:1125–1136. Sci Technol 2003, 63: 2223–2253.
82. Zacchi V, Soranzo C, Cortivo R, Radice M, Brun P, 96. Tan W, Krishnaraj R, Desai TA. Evaluation of nanos-
Abatangelo G. In vitro engineering of human skin-like tructured composite collagen-chitosan matrices for
tissue. J Biomed Mater Res 1998, 40:187–194. tissue engineering. Tissue Eng 2001, 7:203–210.
83. Vacik J, Dvorankova B, Michalek J, Pradny M, 97. Zhang RY, Ma PX. Synthetic nano-fibrillar extracel-
Krumbholcova E, Fenclova T, Smetana K. Cultiva- lular matrices with predesigned macroporous archi-
tion of human keratinocytes without feeder cells on tectures. J Biomed Mater Res 2000, 52:430–438.
polymer carriers containing ethoxyethyl methacry-
late: in vitro study. J Mater Sci Mater Med 2008, 98. Matthews JA, Wnek GE, Simpson DG, Bowlin GL.
19:883–888. Electrospinning of collagen nanofibers. Biomacro-
molecules 2002, 3:232–238.
84. Chua AWC, Ma DR, Song IC, Phan TT, Lee ST,
Song C. In vitro evaluation of fibrin mat and Tega- 99. Hong KH. Preparation and properties of electrospun
derm (TM) wound dressing for the delivery of ker- poly (vinyl alcohol)/silver fiber web as wound dress-
atinocytes—implications of their use to treat burns. ings. Polymer Eng Sci 2007, 47:43–49.
Burns 2008, 34:175–180. 100. Zhang YZ, Venugopal J, Huang ZM, Lim CT,
85. Chen G, Sato T, Ohgushi H, Ushida T, Tateishi T, Ramakrishna S. Characterization of the surface
Tanaka J. Culturing of skin fibroblasts in a thin biocompatibility of the electrospun PCL-collagen
PLGA-collagen hybrid mesh. Biomaterials 2005, nanofibers using fibroblasts. Biomacromolecules
26:2559–2566. 2005, 6:2583–2589.
86. Marston WA. Dermagraft (R), a bioengineered human 101. Chong EJ, Phan TT, Lim IJ, Zhang YZ, Bay BH.
dermal equivalent for the treatment of chronic non- Evaluation of electrospun PCL/gelatin nanofibrous
healing diabetic foot ulcer. Exp Rev Med Devices scaffold for wound healing and layered dermal recon-
2004, 1:21–31. stitution. Acta Biomater 2007, 3:321–330.
87. Chen G, Sato T, Sakane M, Ohgushi H, Ushida T, 102. Khil MS, Cha DI, Kim HY, Kim IS, Bhattarai N.
Tanaka J, Tateishi T. Application of PLGA-collagen Electrospun nanofibrous polyurethane membrane as
hybrid mesh for three-dimensional culture of canine wound dressing. J Biomed Mater Res B Appl Biomater
anterior cruciate ligament cells. Mater Sci Eng C 2004, 2003, 67B:675–679.
24:861–866. 103. Powell HM, Supp DM, Boyce ST. Influence of elec-
88. Sato T, Chen G, Ushida T, Ishii T, Ochiai N, trospun collagen on wound contraction of engineered
Tateishi T. Tissue-engineered cartilage by in vivo skin substitutes. Biomaterials 2008, 29:834–843.
culturing of chondrocytes in PLGA-collagen hybrid 104. Powell HM, Boyce ST. Fiber density of electrospun
sponge. Mater Sci Eng C 2001, 17:83–89. gelatin scaffolds regulates morphogenesis of dermal-
89. Mansbridge J, Robert L, Robert L, Joseph V. Tissue- epidermal skin substitutes. J Biomed Mater Res A
engineered skin products. Principles of Tissue Engi- 2008, 84A:1078–1086.
neering. 3rd ed. Burlington: Academic Press; 2007, 105. McManus MC, Boland ED, Koo HP, Barnes CP,
1201–1213. Pawlowski KJ, Wnek GE, Simpson DG, Bowlin GL.

Vo lu me 2, September/Octo ber 2010  2010 Jo h n Wiley & So n s, In c. 523

Advanced Review

Mechanical properties of electrospun fibrinogen and its effect on the adhesion and spreading of nor-
structures. Acta Biomater 2006, 2:19–28. mal human keratinocytes and fibroblasts in vitro.
106. Kumbar SG, Nukavarapu SP, James R, Nair LS, Biomaterials 2004, 25:1289–1297.
Laurencin CT. Electrospun poly(lactic acid-co- 118. Noh HK, Lee SW, Kim J-M, Oh J-E, Kim K-H,
glycolic acid) scaffolds for skin tissue engineering. Chung C-P, Choi S-C, Park WH, Min B-M. Electro-
Biomaterials 2008, 29:4100–4107. spinning of chitin nanofibers: degradation behavior
107. Venugopal JR, Zhang YZ, Ramakrishna S. In vitro and cellular response to normal human keratinocytes
culture of human dermal fibroblasts on electrospun and fibroblasts. Biomaterials 2006, 27:3934–3944.
polycaprolactone collagen nanofibrous membrane. 119. Wnek GE, Carr ME, Simpson DG, Bowlin GL. Elec-
Workshop on the Regulatory Process for Pediatric trospinning of nanofiber fibrinogen structures. Nano
Mechanical Circulatory Support Devices, Washing- Lett 2003, 3:213–216.
ton, DC, 2006. 120. Barnes CP, Sell SA, Boland ED, Simpson DG,
108. Chong EJ, Phan TT, Lim IJ, Zhang YZ, Bay BH, Bowlin GL. Nanofiber technology: designing the next
Ramakrishna S, Lim CT. Evaluation of electrospun generation of tissue engineering scaffolds. Adv Drug
PCL/gelatin nanofibrous scaffold for wound healing Delivery Rev 2007, 59:1413–1433.
and layered dermal reconstitution. 2nd Symposium 121. Zhong SP, Teo WE, Zhu X, Beuerman RW, Ramakr-
on Biological Materials Science held at the 2006 TMS ishna S, Yung LYL. An aligned nanofibrous collagen
Annual Meeting, San Antonio, TX, 2006. scaffold by electrospinning and its effects on in vitro
109. Rho KS, Jeong L, Lee G, Seo B-M, Park YJ, Hong S- fibroblast culture. J Biomed Mater Res A 2006,
D, Roh S, Cho JJ, Park WH, Min B-M. Electrospin- 79A:456–463.
ning of collagen nanofibers: effects on the behavior of 122. Zhang S, Huang YQ, Yang XP, Mei F, Ma Q,
normal human keratinocytes and early-stage wound Chen GQ, Ryu S, Deng XL. Gelatin nanofibrous
healing. Biomaterials 2006, 27:1452–1461. membrane fabricated by electrospinning of aque-
110. Sell SA, McClure MJ, Garg K Wolfe PS, Bowlin GL. ous gelatin solution for guided tissue regeneration.
Electrospinning of collagen/biopolymers for regener- J Biomed Mater Res A 2009, 90A:671–679.
ative medicine and cardiovascular tissue engineering. 123. Thakur RA, Florek CA, Kohn J, Michniak BB. Elec-
Adv Drug Delivery Rev 2009, 61:1007–1019. trospun nanofibrous polymeric scaffold with targeted
111. Min BM, You Y, Kim JM, Lee SJ, Park WH. For- drug release profiles for potential application as
mation of nanostructured poly(lactic-co-glycolic wound dressing. Int J Pharm 2008, 364:87–93.
acid)/chitin matrix and its cellular response to nor- 124. Katti DS, Robinson KW, Ko FK, Laurencin CT.
mal human keratinocytes and fibroblasts. Carbohydr Bioresorbable nanofiber-based systems for wound
Polym 2004, 57:285–292. healing and drug delivery: optimization of fabrication
112. Yoo JJ, Liu J, Soker S, Komura M, Lim G, Atala A, parameters. J Biomed Mater Res B Appl Biomater
Stitzel J. Electrospinning fabrication of collagen- 2004, 70B:286–296.
based scaffolds for vascular tissue engineering. 125. Duan YY, Jia J, Wang SH, Yan W, Jin L,
Experimental Biology Meeting, San Francisco, CA, Wang ZY. Preparation of antimicrobial poly(epsilon-
2006. caprolactone) electrospun nanofibers containing
113. Zeugolis DI, Khew ST, Yew ESY, Ekaputra AK, silver-loaded zirconium phosphate nanoparticles.
Tong YW, Yung LYL, Hutmacher DW, Sheppard C, J Appl Polymer Sci 2007, 106: 1208–1214.
Raghunath M. Electro-spinning of pure collagen 126. Blackwood KA, McKean R, Canton I, Freeman CO ,
nano-fibres—just an expensive way to make gelatin? Franklin KL, Cole D, Brook I, Farthing P, Rimmer S,
Biomaterials 2008, 29:2293–2305. Haycock JW, et al. Development of biodegradable
114. Zhang YZ, Venugopal J, Huang ZM, Lim CT, electrospun scaffolds for dermal replacement. Bio-
Ramakrishna S. Crosslinking of the electrospun materials 2008, 29:3091–3104.
gelatin nanofibers. Polymer 2006, 47:2911–2917. 127. Cui WG, Zhu XL, Yang Y, Li XH, Jin Y. Evaluation
115. Rujitanaroj P-O, Pimpha N, Supaphol P. Wound- of electrospun fibrous scaffolds of poly(DL-lactide)
dressing materials with antibacterial activity from and poly(ethylene glycol) for skin tissue engineer-
electrospun gelatin fiber mats containing silver ing. Mater Sci Eng C Mater Biol Appl 2009,
nanoparticles. Polymer 2008, 49:4723–4732. 29:1869–1876.
116. Gu SY, Wang ZM, Ren J, Zhang CY. Electrospinning 128. Powell HM, Boyce ST. Engineered human skin fab-
of gelatin and gelatin/poly(L-lactide) blend and its ricated using electrospun collagen-PCL blends: mor-
characteristics for wound dressing. Mater Sci Eng C phogenesis and mechanical properties. Tissue Eng A
Mater Biol Appl 2009, 29:1822–1828. 2009, 15:2177–2187.
117. Min B-M, Lee G, Kim SH, Nam YS, Lee TS, 129. Pan H, Jiang H, Chen W. Interaction of dermal fibrob-
Park WH. Electrospinning of silk fibroin nanofibers lasts with electrospun composite polymer scaffolds

524  2010 Jo h n Wiley & So n s, In c. Vo lu me 2, September/Octo ber 2010

WIREs Nanomedicine and Nanobiotechnology Tissue scaffolds for skin wound healing

prepared from dextran and poly lactide-co-glycolide. 133. Webb A, Li A, Kaur P. Location and phenotype
Biomaterials 2006, 27:3209–3220. of human adult keratinocyte stem cells of the skin.
130. Yang XC, Shah JD, Wang HJ. Nanofiber enabled Differentiation 2004, 72:387–395.
layer-by-layer approach toward three-dimensional
134. Coraux C, Hilmi C, Rouleau M, Spadafora A, Hin-
tissue formation. Tissue Eng A 2009, 15:945–956.
nrasky J, Ortonne JP, Dani C, Aberdam D. Reconsti-
131. Behfar A, Terzic A. Mesenchymal stem cells: engineer- tuted skin from murine embryonic stem cells. Curr
ing regeneration. Clin Trans Sci 2008, 1:34–35. Biol 2003, 13:849–853.
132. Metcalfe AD, Ferguson MWJ. Skin stem and progen-
itor cells: using regeneration as a tissue-engineering 135. Schluter H, Kaur P. Bioengineered human skin from
strategy. Cell Mol Life Sci 2008, 65:24–32. embryonic stem cells. Lancet 2009, 374:1725–1726.

Vo lu me 2, September/Octo ber 2010  2010 Jo h n Wiley & So n s, In c. 525