TISSUE ENGINEERING: Part B

Volume 18, Number 2, 2012

ª Mary Ann Liebert, Inc.
DOI: 10.1089/ten.teb.2011.0390

Fabrication of Large Pores in Electrospun Nanofibrous

Scaffolds for Cellular Infiltration: A Review

Shaoping Zhong, Ph.D.,1 Yanzhong Zhang, Ph.D.,2 and Chwee Teck Lim, Ph.D.1,3,4

In the past decade, considerable effort has been made to construct biomimetic scaffolds from electrospun nano-
fibers for engineering different tissues. However, one of the major concerns with electrospun nanofibrous scaffolds
is that the densely arranged architecture of fibers and small pores or voids between fibers hinder efficient cellular
infiltration or prevent three dimensional (3D) cellular integration with host tissue in vivo after implantation. To
overcome this problem, many concepts or strategies applicable during the electrospinning or post-electrospinning
procedures have been proposed to enlarge pore size of electrospun scaffolds. This article addresses the issues of
pore geometry and cellular infiltration of electrospun scaffolds, and first reviews the fabrication solutions/ap-
proaches applied to achieve larger micropores in electrospun mats. The evidence and potential for fostering
cellular infiltration using these improved porous scaffolds are then discussed. Finally, it is hoped that this will
enable us to better exploit viable technologies or develop new ones for constructing ideal nanofibrous architecture
for fulfilling specific tissue engineering needs.

Introduction In the design and fabrication of tissue engineering scaf-

folds, pore size or porosity have been one of the most

E lectrostatic spinning or electrospinning has gained

tremendous attention in the past decade as it can pro-
duce ultrafine nanoscale fibers that are of interest to both the
important criteria to consider. The necessity and significance
of the pores of the scaffolds have been well recognized in
many experimental works. For example, direct osteogenesis
research community as well as industry.1,2 The electrospin- could be formed on porous scaffolds, while no new bone
ning technique exhibits the unique features of simplicity, occurred on the solid particles in bone regeneration.19
affordability, wide range of materials selection, and high Increased porosity and pore size are reported to facilitate cell
flexibility in controlling diameters and spatial orientation. seeding and diffusion throughout the scaffolds, which is
Electrospun fibers are notable for their ultrafine diameter associated with increased vascularization, bone ingrowth,
and large surface area-to-volume ratios, which enable them and osteointegration.20–23 Although controversy exists in the
to be tailored for a multitude of applications. The extremely literature regarding the optimum pore size efficient for dif-
high surface area-to-volume ratios and high porosity are ferent cell ingrowth or tissue regeneration, a broad consensus
advantages in promoting interfacial transport or interaction has been reached that pore size of the order of few tens to a
in areas such as catalysis and energy storage.3–5 Recent few hundred micrometers is necessary to provide adequate
studies have also demonstrated the potential applications of space for cells diffusion and ingrowth.23 With regard to po-
electrospun constructs in tissue engineering and regenerative rosity, higher porosity of scaffolds is desirable to provide
medicine.1,6–8 The nanofibrous dimensions of electrospun better diffusion of nutrient and waste as long as reasonable
nanofibers make it possible to emulate the structural char- mechanical properties can be preserved. The mechanical
acteristics of native extracellular matrix (ECM), which has strength of scaffolds is impaired with increasing pore size
been linked to the enhanced cell–matrix interactions and and porosity and, hence, extremely high porosity and large
ECM synthesis.9,10. The cell-populated electrospun constructs pore size can be undesirable in many cases.24
have been employed in a number of tissue engineering ap- Although the effect of pore size and porosity of scaffolds
plications including vascular, bone, skin, cornea, neuron, on cell growth has been well identified in many traditional
tendon, and ligament.11–18 scaffold designs, they form key challenges in the use of

1
Department of Bioengineering, National University of Singapore, Singapore.
2
Department of Bioengineering, College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai, P.R.
China.
3
Department of Mechanical Engineering, National University of Singapore, Singapore.
4
Mechanobiology Institute, National University of Singapore, Singapore.

77
78 ZHONG ET AL.

electrospun mats as tissue engineering templates. Small wide variety of fiber geometries including typical circular
pores formed by the electrospun nanofibers can be an ad- cross-sections, branched fibers, and flat or other geometric
vantage in some applications such as skin wound dressings ribbons have been fabricated, the pores or voids formed by
and endothelium regeneration where barrier features are these fibers exhibited quite similar shapes with irregularly
desirable or cellular infiltration is not needed,7 but a small polygonal frame in a two-dimensional plane as shown in
pore is always a disadvantage for tissue engineering appli- Figure 1A (our own unpublished image). The pores at three
cations where sufficiently large pore size is required to allow different layers exhibit different polygonal shapes as high-
for efficient 3D cell ingrowth.8 This article addresses the lighted by three different colors of blue, green, and white
concerns of pore geometry and cellular infiltration in elec- from top to bottom. The pore size of the lower layer is
trospun tissue scaffolds, and provides an overview on the smaller than that of the upper layer due to the separation of
various fabrication approaches used to achieve larger pores the underlying fibers. Therefore, the pores of an electrospun
or higher porosity in the electrospinning field, some of which nonwoven fibrous scaffold appear to be ‘‘open’’ and the pore
have been applied to encourage cellular infiltration in elec- size decreased with increasing fiber layers.
trospun mats for biomedical applications. The pore size and its distribution are traditionally deter-
mined by experimental intrusion or extrusion types (e.g.,
mercury porosimetry or bubble point method) based on the
Pore Size and Porosity of Electrospun
principle of capillary flow, in which the porous material will
Fibrous Structures
allow a fluid to pass when the pressure applied on the sys-
Pore characteristic is one of the main structural features for tem exceeds the capillary force of attraction of the fluid in the
evaluating the performance of any nonwoven web. The size largest pore.27 The diameter of polygonal shapes (Fig. 1B) is
and shape of the pores play an important role in the perme- well defined in the capillary flow as a circular opening di-
ability, filtration, and mechanical properties,25 and a certain ameter of the right cylinder that will pass through the web as
pore size is associated with promoting cell growth and ECM shown in Figure 1C.26,28 Therefore, the measured ‘‘charac-
synthesis.26 The unique pore features of electrospun nanofi- teristic’’ pore diameter by the porosimetry method, which
bers such as high bulk porosity (up to 80%), fully inter- assumes a circular cross-section of the polygonal pore, could
connected pore structures, and a wide open surface encourage not be thought of as exact pore diameters, but as an equiv-
nutrient and vapor transport desirable for cell growth. How- alent size of average fiber-to-fiber distance for a given po-
ever, there is currently little understanding of the pore char- lygonal pore. This measured value of the pore diameter is
acteristics of nonwoven networks and effective and accurate only comparative in nature and cannot reflect pore shape
characterization of pore size, shape, and porosity. features. Determining pore size using porosimetry for elec-
In a conventional electrospinning process, random pack- trospun structures remains controversial as a conventionally
ing of fibers during fiber deposition typically produces a theoretical approach defined the pores with closed volume,
nonwoven web of randomly oriented fibers. Although a while electrospun fibers form open pores.29 The pore

FIG. 1. Electrospun fibrous

structure and pore geometry.
(A) Typical three-dimensional
morphology of electrospun
nanofibers (AFM image), (B)
typical pore morphology for
the polygonal geometry of
nonwoven web, (C) equiva-
lent diameter of polygonal
pore, (D) cells growing un-
derneath the layers of the
electrospun fibers.42 Color
images available online at
www.liebertonline.com/teb
FABRICATION OF LARGE PORES IN ELECTROSPUN NANOFIBROUS SCAFFOLDS 79

measurements can be roughly estimated from SEM images pores even smaller than the size of the cell.8,33 The enlarged
or more accurately measured using image analysis tech- void space created due to the fast dissolution of the natural
niques to directly determine the pore sizes.29,30 Using image polymers may encourage cell migration into the scaffold
analysis techniques for preprocessing and digitizing images interior during in vitro cell culture.34,36 Investigations of the
can provide more comprehensive and accurate measure- effect of fiber diameter and pore size on cellular infiltration
ments of pore characteristics such as pore size, shape, ori- showed that electrospun scaffolds with fiber diameters larger
entation, and boundary locations.25,30 But this method can than 1 mm forming large pore sizes of tens of micrometers
analyze only superficial pores and thus is not applicable for a were able to permit cell infiltration, while smaller diameters
thick nonwoven fibrous mat with high fiber density. There- of less than 1 mm inhibited infiltration significantly for most
fore, current methods still contain inherent disadvantages types of cells.26,37 Currently the dimensions of ultrafine fibers
with regard to pore measurement. with diameters of approximately 100–500 nm form pore sizes
One issue that may easily be ignored is that the loose of only 1–10 mm, which is smaller than the normal cell size of
packing of nanofibers makes them susceptible to environ- about 10–15 mm.38 Further experiments demonstrated that
mental or external influences such as mechanical deformation only small nanoparticles ( < 300 nm) can penetrate through
and liquid soaking. For example, most of the electrospun the electrospun meshes while larger particles ( > 1 mm) will be
nanofibers cannot bear the high pressure used in some por- trapped in the meshes.39
osimetry measurements such as the mercury intrusion meth- Using dynamic seeding or a culture system such as perfu-
od,31 and thus, dimensional compression of samples may be sion seeding flow, vacuum pressure, or a bioreactor system
induced during measurement and the pore or porosity could was reported to foster cell penetration/diffusion into the
be impaired. Also when using the scaffolds in liquids such as scaffold interior.26,40–42 However, the small pores, to a great
culture medium or aqueous treatment of scaffolds, shrinking extent, still limit cellular infiltration into the scaffolds. To ad-
of layers and swelling of the fibers will easily occur, which can dress this limitation, various approaches have been introduced
lead to decreased pore size or porosity of scaffolds. These in recent years to produce larger pores and higher porosity,
structural or geometric changes should be considered in the which could facilitate cellular infiltration, and these studies
use of this kind of nonwoven fabric. have suggested the potential application of these spacious
microporous fibrous structures for biomedical applications.
Cellular Infiltration Phenomena
Recent Progress in Enlarging Pore Size
of Electrospun Scaffolds
of Electrospun Scaffolds
Cellular infiltration into the scaffold interior is always of
Here, we cover new and emerging approaches to produce
significance in terms of producing 3D cell–scaffold constructs
advanced electrospun tissue engineering scaffolds for ap-
and enhancing the integration between host tissue and
plications where larger pores or higher porosity are needed.
scaffold after implantation.32 Although an electrospun na-
These approaches can be classified into several categories as
nofibrous structure has been considered promising scaffolds
follows: salt/polymer leaching and wet electrospinning, us-
to support cell attachment and growth, cellular infiltration
ing bath, ice crystal, laser irradiation, electric matrix, and
into electrospun mats has been a noted problem. There are
nanofibrous and microfibrous composite, among others. In
only a few studies that address the cellular infiltration phe-
this review, the fabrication processes and concepts involved
nomena, but they nevertheless provide further understand-
will also be presented to allow researchers to choose any
ing of cell behavior and guidance in designing improved
suitable method or develop new potential methods for con-
biomimetic nanofibrous scaffolds for target applications. Li
structing nanofibrous architectures for their specific needs.
et al. reported the first observation of cell infiltration into the
electrospun scaffold interior as shown in Figure 1D.33 The
Salt/polymer leaching
fibroblasts migrated through the pores and grew underneath
the layers of the electrospun poly(lactic-co-glycolic acid) Salt leaching with or without other methods such as sol-
(PLGA). Zhang et al. reported that bone marrow stromal cells vent casting and gas foaming has been widely used to fab-
can grow and migrate into the electrospun polycaprolactone ricate scaffolds for tissue engineering applications.43 This
(PCL) and blended PCL/gelatin scaffolds, and the blended technique is based on the principle that salt particles can first
PCL/gelatin electrospun scaffold displayed over twice the be dispersed into a polymer solution followed by the disso-
infiltration depth compared to the pure PCL one.34 In an- lution of the salt to then form a highly porous scaffold. The
other study by Shabani et al., stem cells showed very high scaffolds with controlled pore size and porosity can be pre-
cell infiltration into the poly(ether sulfones) (PES) electro- pared by varying the salt size and salt/polymer ratio.44 Lee
spun nanofibers grafted with collagen.35 These preliminary et al. reported the first combination of salt leaching/gas
investigations indicated that appropriate hydrophilicity and foaming with an electrospinning technique by delaminating
biochemical signals could promote cell–matrix interaction the electrospun poly(L-lactic acid) (PLLA) mesh sparsely
and as a result, cell migration into the scaffold interior could with NH4HCO3/NaCl salt particles.45 Another electrospin-
be facilitated. ning method reported by Nam et al.46 used one sheath sur-
However, more and more investigations indicated that the rounding a spinning needle (Fig. 2A) to incorporate the NaCl
physical structure plays a significant role in facilitating cell particles into the electrospun PCL nanofibers at intervals
migration and cell–matrix integration. Many studies sug- between certain electrospinning periods. Using these two
gested that the loose packing of electrospun nanofibers may methods, the layer-by-layer nanofiber constructs containing
allow the cells to push the surrounding fibers aside and tiny salt particles (Fig. 2B) were fabricated and mechanically
hence be able to penetrate into the electrospun mat through compressed and molded into desirable shapes. The
80 ZHONG ET AL.

FIG. 2. Salt/polymer leaching for en-

larging pore size. (A) Schematic showing
an electrospinning setup with the intro-
duction of salt crystals in the surrounding
sheath, (B) multilayered PCL nanofibers
and NaCl salts construct, (C) micropores
formed after salt leaching of the sample in
(B), (D) homogeneous HA/collagen na-
nofibers and NaCl salts construct, (E)
microporous scaffolds after leaching of
the sample in (D), (F) the electrospun
PCL/PEO nanofibers, (G) the PCL scaf-
folds after PEO leaching. (A, B, C) from
Nam et al.,46 (D, E) from Kim et al.,47 (F,
G) from Baker et al.51 HA, hyaluronic
acid; PCL, polycaprolactone; PEO, poly-
ethylene oxide. Color images available
online at www.liebertonline
.com/teb

microsized pores with the same size of salt particles (ranging after salt leaching.48 However, electrospinning of a homo-
from 20 to 300 mm) were created between the layers of elec- geneous polymer solution containing microsized salt parti-
trospun meshes after salt leaching (Fig. 2C). cles has not been reported, likely due to the easy salt blocking
Homogeneous hyaluronic acid (HA)/collagen nanofibers of the small needle, which could lead to difficulty in forming
and NaCl salt construct (Fig. 2D) were produced by simul- continuous fibers during the electrospinning process.
taneously depositing salt particulates as a porogen during Using a similar concept of salt leaching, selectively
electrospinning as reported by Kim et al.47 An automatic vi- leaching one polymer phase out from bi/multicomponent
bration-controlled sieving method was used to simulta- electrospun constructs has been used to increase the pore
neously sieve the salts during the electrospinning of the size or porosity.49 Water-soluble polymers of poly(ethylene
nanofibers. This resultant nanofibers/salt composite can then oxide) (PEO) or gelatin have been incorporated with other
be cut into the desired shape and size. Also, acceptable water-insoluble or slow-degrading polymers such as poly-
dimensional shrinking did occur after salt leaching, but the urethane (PU), PCL, or PLLA, which will be left as the bulk
structural integrity of scaffolds with microsized pores was material after leaching the soluble polymers.50–52 Bi/multi-
still maintained (Fig. 2E). Mixed electrospinning of PCL component electrospun materials were fabricated using
solution containing nanosized CaCO3 salt particles was re- multilayered or mixed electrospinning and, subsequently,
ported to create the nanosized pores in the electrospun fibers polymer leaching was used to create less dense electrospun
FABRICATION OF LARGE PORES IN ELECTROSPUN NANOFIBROUS SCAFFOLDS 81

meshes (Fig. 2F and G).51 The secondary micropore size bers (Fig. 3B). In contrast, remarkably larger pores of 1–
created was dependent on the structure or form of the sac- 500 mm and 3D open structures were achieved in the work of
rificed fiber phase entangled in electrospun constructs and it Leong et al. (Fig. 3C and D). The limited enlargement of pore
is obvious that microfibers of sacrificial polymer can produce size in the former work may be attributed to the inappro-
a very limited increase in porosity and pore size. priate use of the drying method and the voids of ice crystals
Significant cellular infiltration has been reported for the were difficult to preserve using a vacuous desiccator at room
salt-induced microporous electrospun fabrics as compared temperature as reported. Further in vitro and in vivo experi-
with the very minimal cellular infiltration observed on the ments in the work of Leong et al. indicated that cellular in-
conventional electrospun fabrics in the work of Nam et al.46 filtration into the scaffolds of up to 50 mm was observed as
However, only a marginal improvement in cellular infiltra- compared to no cellular infiltration in the conventional
tion was observed on the polymer leaching electrospun electrospun scaffolds. However, for this method, the homo-
mats,53 which may be attributed to the rather limited in- geneity of pore size was difficult to achieve along the depth
crease in pore size created by this method as observed in of very thick electrospun mats.
Figure 2G. In addition, there are some concerns regarding
the leached scaffolds that include some slight swelling of Wet electrospinning using bath collector
fibers and scaffolds, irregularly shaped pores, and decreased
A liquid reservoir collector such as coagulation bath, when
pore interconnectivity.14 Also this technique does not work
combined with the electrospinning technique, termed ‘‘wet
well as there is difficulty in leaching the salt/polymer em-
electrospinning,’’ has been used to collect electrospun na-
bedded too deeply in the scaffolds.
nofibers before being drawn into a rotating roller to produce
continuous and bundle yarns.56 Applying a coagulation bath
Ice crystals for electrospinning collection device
to induce the dispersion effect on the bulk density of nano-
Ice crystals formed at low temperature had been used to fibrous structure was reported by Ki et al.57 and Yokoyama
provide a systematic topographical pattern by controlling et al.58 Electrospun nanofibers spraying toward the bath loop
the cooling environmental parameters (temperature and on the liquid surface subsequently were dispersed in de-
humidity). Using ice crystals as an electrospinning collection creased bulk density and deposited onto the metal electrode
device was first reported by Simonet et al.54 This technique bottom with the resultant wet foam (Fig. 4A). The highly
was termed cryogenic electrospinning in another similar porous sponge was formed after freeze-drying, while one
study by Leong et al.55 In these two studies, electrospun fi- dense membrane layer was produced using the conventional
bers were collected onto ice crystals formed on the surface of electrospinning method (Fig. 4B and C). The dispersion effect
a chilled ground drum (Fig. 3A) with a hollow cavity to of the liquid bath, which decreased the bulk density of the
allow the loading/flowing of the cold source such as dry ice nanofibers, was demonstrated in these two studies. The
and liquid nitrogen. After drying the samples, the ice parti- varying density and porosity of the nanofibers can be con-
cles can be removed and macrosized pores or voids are trolled by varying the bath solvents, which resulted in
formed between fibers. Although the technique was de- varying surface tensions as demonstrated by Yokoyama
scribed in detail for the optimization of the porosity and pore et al.58 Different shapes of nanofibrous foam can be easily
size by Simonet et al., the increase in pore size was limited molded by using different shaped vessels. Increased cellular
and the morphology of the electrospun fibers exhibited very ingrowth in these 3D electrospun foam scaffolds has been
similar morphology to that of conventional electrospun fi- confirmed in vitro.57 However, some major problems using

FIG. 3. Cryogenic electrospinning for

forming large pores. (A) Cooling drum
with ice crystals formed on the surface,
(B) electrospun poly(ether urethane)
(PEU) scaffold fabricated using cryogenic
electrospinning in (A), (C) electrospun
PLLA scaffolds using cryogenic electro-
spinning, (D) higher magnified image of
the PLLA scaffolds in (C). (A, B) From
Simonet et al.,54 (C, D) from Leong et al.55
Color images available online at
www.liebertonline.com/teb
82 ZHONG ET AL.

this method are the difficulties in choosing suitable solvents

and drying the samples for desirable pore size.

Laser/UV irradiation
The laser has become an invaluable tool in processing and
treating biological tissues due to its precision in the removal
of materials.59 Laser structuring of electrospun fiber meshes
was first used to locally ablate electrospun PCL nanofibers
by Choi et al.60 A laser with high intensity and ultrafast ir-
radiance was scattered onto the target surface of electrospun
fibers. Ultrafast heating, melting, and evaporation of the fi-
bers induced by high laser energy can produce voids in the
fibers. Thus, desirable geometric patterns such as grooves or
matrix pores as shown in Figure 5A61 and B60 were fabri-
cated by controlling process parameters such as the power,
pulse, scanning rates, and orientation. Remarkable melting
FIG. 4. Disperse effect of liquid bath in wet electrospinning of the remaining fibers could be prevented and its minimal
for creating a macroporous scaffold. (A) Schematic showing detrimental effect on cell growth was demonstrated. In-
the wet electrospinning with a liquid bath, (B) conventional creased cellular penetration was demonstrated in another
electrospun nanofiber membrane, (C) electrospun sponge study producing patterned macrosized pores using laser
foam using wet electrospinning. (B, C) From Ki et al.57
photolithography.62 Dong et al.63 demonstrated the potential
application of using photolithography with one porous mask

FIG. 5. Laser structuring of electrospun

nanofibrous scaffolds. (A) Groove pat-
terned structure by laser, (B) matrix with
microhole created by laser (inset is the
magnified view of a hole), (C) schematic
diagram of UV photolithography, (D)
electrospun PLGA nanofibers after 1 h UV
photolithography, (E, F) SMCs cultured
on UV photolithography-treated P(LLA-
CL) nanofiber after 1 day (E), and after 20
days (F). (A, B) From Choi et al.,60 (C, D,
E, F) from Dong et al.63 UV, ultraviolet;
PLGA, poly(lactic-co-glycolic acid); SMC,
smooth muscle cell. Color images avail-
able online at www.liebertonline
.com/teb
FABRICATION OF LARGE PORES IN ELECTROSPUN NANOFIBROUS SCAFFOLDS 83

to fabricate a microporous patterned nanofiber structure (Fig. but lack the high surface area-to-volume ratio of the nano-
5C). The shallow holes (Fig. 5D) were etched on the nanofi- fibers.64 Combining both nanofibers and microfibers into
brous mesh under the porous mask due to the UV-induced composites has been suggested to overcome these short-
degradation of fibers. An in vitro culture study indicated that comings. Double layered nanofibers and microfibers were
cells preferred to migrate into the holes after 20 days of in- prepared by electrospinning nanofibers on top of an exist-
cubation (Fig. 5F) as compared with few cells observed in the ing microfibers layer, which could be made by electro-
holes for 1-day culture (Fig. 5E). Hence this microporous spinning as well65 or other techniques such as the extruding
structure appears to promote cellular migration or infiltration method.66 One commercial double-layered ultraweb is
and allow for cell–matrix integration. Using laser scattering to shown as Figure 6A, with the nanofibers electrospun onto
ablate or degrade fibers could be a promising tool to produce the surface of spunbond nonwoven microfibers; this has
specific microenvironments to guide the cellular behavior for been widely used to remove pollutants from air or water in
both fundamental biological studies and biomedical applica- a wide range of filtration applications.67 A multilayered
tions. However, this laser microstructuring method has sev- structure can be fabricated by sequentially alternating
eral disadvantages including a lack of interconnectivity of the the electrospinning of nanofibers and microfibers (Fig. 6B)65
micropores/grooves and impaired structural or mechanical or folding or rolling the double-layered mesh.66 Homo-
integrity and potential cytotoxicity of a degrading or sintering geneously mixed nanofibers and microfibers can be fabri-
scaffold induced by the laser structuring. cated using mixed electrospinning with two spinnerets to
simultaneously spin a wide range of different sized fibers.49
However, significantly higher cellular infiltration was
Combination of nanofibers and microfibers
reported in these kinds of nanofibers and microfibers only
It is well known that electrospun nanofibers have the in- under flow perfusion conditions, and the small pore
herent drawback of being an extremely dense structure with dimensions of nanofibers formed in the composite still
small pore size, while microfibers exhibit a larger pore size, hindered cells from infiltrating.65

FIG. 6. Nanofibers combined

with microfibers for enlarging
pore size. (A) One commercial
product of electrospun nanofibers
on the microfibers developed by
Donaldson, (B) multilayered na-
nofibers and microfibers by alter-
native electrospinning, (C)
schematic showing an electro-
spinning setup for producing PCL
nanofibers coated onto single PLA
microfibers, (D) nanofiber-coated
microfibers (inset shows a micro-
fiber coated with nanofibers), (E)
chondrocytes seeded on the mi-
crofibers coated with nanofibers
(D), after 2 weeks of culture. (A)
From Fang et al.,67 (B) from Pham
et al.,65 (C, D, E) from Thorvalds-
son et al.68 PLA, poly(lactic acid).
Color images available online
at www.liebertonline.
com/teb
84 ZHONG ET AL.

To overcome this problem, another combination of nano- livery. But these two studies report neither specific applica-
fibers and microfibers was developed by electrospinning tion nor cellular infiltration. In one more recent study, some
nanofibers onto single microfibers.68 As shown in Figure 6C, patterned metal collectors such as wire (Fig. 7C) or sieves
one single microfiber was wound through the electrospin- were used to achieve macropores in the electrospun scaffolds
ning jet direction onto a rotating wheel and electrospun na- (Fig. 7D).72 The more sparse nanofibers can be formed in the
nofibers could homogeneously deposit onto the microfiber. wire pores (as shown in the inset of Fig. 7D). The increased
The collected nanofiber-coated microfibers could be manu- cellular infiltration was evidenced by culturing 3T3 fibro-
ally molded into any shape and any scaffold porosity (Fig. blasts in the study.
6D). Good cellular infiltration and spreading along the na-
nofiber-coated microfibers were observed (Fig. 6E), while Other approaches
only one confluent layer of cells on the surface was formed
In addition to the above-mentioned approaches, several
on the nanofibrous mesh. This new form of nanofibrous and
other techniques have also emerged to improve the pore size
microfibrous scaffolds could provide both appropriate sur-
and porosity. The addition of a chemical blowing agent
face properties and a structural environment for cell growth.
into the electrospinning solution was reported by Kim et al.73
and the microsized pores were formed after the scaffolds
Electric field for controlling deposition of nanofibers
were blown after exposure to 100C. However, sintering
Varying electrode configurations to form specific electric or melting of the nanofibers may occur under this tempera-
field distribution can control the deposition of the electro- ture. In our laboratory work, water drops were sprayed in-
spinning jet. Using this concept, auxiliary parallel electrodes termittently onto the layers of electrospun PCL with a
have been widely applied to fabricate aligned electrospun high hydrophobicity during the electrospinning process
nanofibers.69 One early study reported the use of various (unpublished data). Large pores and high porosity can
patterned conductive protrusion collectors for controlling the be achieved after freeze-drying of the electrospun fibers. In
fiber deposition to form different fiber patterns (Fig. 7A).70 one strategy of cellular microintegration without changing
This kind of grid-like patterned collector can form a hetero- the porous structure of electrospun nanofibers, some
geneously distributed electric force field. Competition of at- attempts to integrate cells into electrospun fibers have
traction imposed by contiguous protrusions may lead to the been reported recently, which included a coelectrospray/
higher density of fibers on shorter sidelines between pro- electrospinning cellular solution and polymer solution,40
trusions, while the relatively sparse crossing orientation of encapsulated cellular microthreads using coaxial electro-
fibers was formed between longer adjacent protrusions. The spinning,74 multilayered cell/fibers composite by alternating
different deposited densities on the different regions formed the electrospinning of nanofibers, and seeding cells.75 Good
a regularly patterned structure with protruded or depressed distribution of cells can be achieved using these methods, but
topography as shown in Figure 7B, which shows three- cell survivability, residual solvent toxicity, and difficulty in
dimensional features of electrospun nanofibers. One matrix maintaining a sterile environment are some potential con-
with metallic protruded needles similar to the grid-like pat- cerns. In another study, layered hydroelectrospinning using
tern was used to collect electrospun nanofibers and a similar a coagulation bath was reported by Tzezana et al. to fabri-
patterned appearance of electrospun nanofibers was fabri- cate multilayered nanofibers in which cells may assemble.76
cated.71 This regular pattern can produce micropores or The feasibility of assembling cells into the nanofibers can be
regular 3D structures, which can be useful in many fields improved and the residual solvent can be minimized with
such as tissue engineering, catalysis, sensing, and drug de- the assistance of a coagulation bath.

FIG. 7. Using an electric field to form a

patterned electrospun nanofibrous structure.
(A) Grid-like patterned protrusion collector,
(B) grid-like patterned morphology of elec-
trospun nanofibers collected using the collec-
tor in (A), (C) metal wire net collector, (D)
patterned nanofibers collected using the col-
lector in (C). (A, B) From Zhang et al.,70 (C, D)
from Vaquette et al.72
FABRICATION OF LARGE PORES IN ELECTROSPUN NANOFIBROUS SCAFFOLDS 85

Conclusions cular graft for blood vessel tissue engineering. Tissue Eng
11(9/10), 1575–1589, 2005.
This article reviews some recently emerging fabrication 12. Yoshimoto, H., et al. A biodegradable nanofiber scaffold by
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Disclosure Statement 21. Wake, M.C., Patrick, C.W., and Mikos, A.G. Pore morphol-
ogy effects on the fibrovascular tissue-growth in porous
No competing financial interests exist.
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Chwee Teck Lim
Mater 4(3), 035004, 2009.
Department of Bioengineering
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