Materials Science and Engineering C 45 (2014) 659–670

Contents lists available at ScienceDirect

Materials Science and Engineering C

journal homepage: www.elsevier.com/locate/msec

Biodegradable polymers for electrospinning:

Towards biomedical applications
Dan Kai a,1, Sing Shy Liow a,1, Xian Jun Loh a,b,c,⁎
a
Institute of Materials Research and Engineering (IMRE) Agency for Science, Technology and Research (A*STAR), 3 Research Link, Singapore 117602, Singapore
b
Department of Materials Science and Engineering, National University of Singapore, 9 Engineering Drive 1, Singapore 117576, Singapore
c
Singapore Eye Research Institute, 11 Third Hospital Avenue, Singapore 168751, Singapore

a r t i c l e i n f o a b s t r a c t

Article history: Electrospinning has received much attention recently due to the growing interest in nano-technologies and the
Received 22 January 2014 unique material properties. This review focuses on recent progress in applying electrospinning technique in pro-
Received in revised form 2 April 2014 duction of biodegradable nanofibers to the emerging field of biomedical. It first introduces the basic theory and
Accepted 21 April 2014
parameters of nanofibers fabrication, with focus on factors affecting the morphology and fiber diameter of biode-
Available online 28 April 2014
gradable nanofibers. Next, commonly electrospun biodegradable nanofibers are discussed, and the comparison of
Keywords:
the degradation rate of nanoscale materials with macroscale materials are highlighted. The article also assesses
Electrospun nanofibers the recent advancement of biodegradable nanofibers in different biomedical applications, including tissue engi-
Biodegradable polyesters neering, drug delivery, biosensor and immunoassay. Future perspectives of biodegradable nanofibers are
Tissue engineering discussed in the last section, which emphasizes on the innovation and development in electrospinning of
Drug delivery hydrogels nanofibers, pore size control and scale-up productions.
Biosensors © 2014 Elsevier B.V. All rights reserved.

1. Introduction To date, over 100 types of natural and synthetic polymers were
electrospun into nanofibers [7]. Popular materials includes: collagen,
Electrospinning was first being introduced in early 1930s for fabrica- elastin, fibrinogen, alginates, polyesters, polyurethanes and their blends
tion of nanofibers as filter materials and textile yarns. Since 1990s, after etc [8]. Nanofibers that are biodegradable and biocompatible have ad-
Reneker et al. demonstrated the feasibility to produce electrospun vantages in a few aspects: they metabolize into biocompatible degrada-
nanofibers from many polymers, the number of publications about tion products in human body; therefore, second surgery for implant
electrospinning has grown exponentially [1,2]. removal is unnecessary. Degradation profile of the nanofibers is tunable
Electrospinning received much attention for biomedical applications to match with the tissue regeneration time frame. Ideally nanofibers
mainly due to the growing interest in nano-technologies and the unique should degrade at the same pace as new tissue grows. Although degrada-
material properties. Electrospinning is an inexpensive and simple tion of polymers in vitro and in vivo is comprehensively studied, the
method to create nanoscale polymer fibers with diameter range from degradation mechanisms of polymer nanofibers are still under-explored.
3–5000 nm [3]. Nanofibers are suitable to mimic biological environment Based on relevant US patents filed in recent years, most of the appli-
because they are in the same scale as biological molecules. In fact, cations nanofibers are in the field of biomedical prosthesis predomi-
nanomaterials like particles, fibrous morphologies or other complex nantly blood vessels and grafts. Specifically, biodegradable polymer
forms, have shown improved interactions with cells, for example, selec- nanofibers showed promising perspective in cosmetic, life science and
tive endocytosis, adhesion and orientation [4–6]. In addition, large sur- tissue engineering scaffolds, in laboratory scale. More efforts are expect-
face area to volume ratio (SVR) of these structures provides the ed in future to scale-up these nanofibers into industrials scale.
nanofibrous mat high pore volumes with different pore sizes. These In this review, we report brief theory and parameters of electrospinning
pores facilitate the loading of bioactive molecules and transportation process, types of biodegradable nanofibers and assess recent advance-
of nutrients and waste. These outstanding properties enable the poly- ment of biodegradable nanofibers in different biomedical applications.
mer nanofibers become an important class of biomaterial.
2. Fabrication of electrospun biodegradable nanofibers

⁎ Corresponding author at: Institute of Materials Research and Engineering (IMRE), 3 2.1. Theory and parameters of electrospinning
Research Link, Singapore 117602, Singapore.
E-mail addresses: lohxj@imre.a-star.edu.sg, XianJun_Loh@scholars.a-star.edu.sg
(X.J. Loh). Electrospinning is attractive thanks to the simplicity and inexpen-
1
These authors contributed equally to this work. sive nature of setup. The basic setup for electrospinning is shown in

http://dx.doi.org/10.1016/j.msec.2014.04.051
0928-4931/© 2014 Elsevier B.V. All rights reserved.
660 D. Kai et al. / Materials Science and Engineering C 45 (2014) 659–670

Fig. 1(a) [9]. There are 3 elementary components to complete the pro- “electrospraying” happens instead of “electrospinning” when low
cess: a capillary tube as a reservoir for polymer solution, a high- molar mass polymers are used. Electrospraying results in small droplets
voltage power supply, and a metallic collector. During the spinning pro- due to instable jet formation. The spinnability of the polymer depends
cess, high voltage (5–15 kV) is applied between a needle capillary end on the onset of chain entanglement between polymer chains, and it is
and a collector. The polymer solution is electrically charged. At the nee- varied for different polymers. For example, chitosan in acetic acid solu-
dle tip, the polymer solution deforms from a spherical pendant droplet tion, with low molar mass of 30 kDa, it formed fragile fibers with many
to a conical shape, known as “Taylor cone”. As the electric field is stron- beads; with medium molar mass of 106 kDa, beads-free, uniform and
ger than the surface tension of the polymer solution, the jet is ejected continuous fibers (130 nm) were obtained [18].
from the cone surface. As the jet travels, the solvent evaporates in the The effect of surface tension of polymer solution on size or morphol-
air, together with the stretching and acceleration of the polymer jet, ogy of nanofibers is controversial. It depends on different polymer and
leading to the extreme thin polymer fibers deposition on the collector solvent systems. Doshi and Reneker reported that by reducing surface
[10]. Electrical bending instability occurs when the distance from the tension of the polymer solution, beads-free fibers can be obtained. Zuo
tip to collector is sufficiently long; in case of a short distance, the jet is et al. reported the fiber morphologies of poly(hydroxybutyrate-co-
typically straight. Fig. 1(b) shows the instability of polymer jet captured valerate) (PHBV) in different solvent systems [19]. Smooth fibers were
by high speed video [11]. Under the action of electric field, polymer jets obtained when the surface tension of the solvent is lowered by adding
experience the bending instability primarily due to mutual repulsion of alcohol. However, a lower surface tension is not always suitable for
the excess electric charges carried by electrospun jets. electrospinning. For example, acetone and dimethyacetamide (DMAc)
The electrospinning process and the formation of polymer fibers have surface tension of 23.7 and 32.4 dyne/cm, respectively. Liu and
are affected by many parameters. Spinnability, fiber diameters, fiber Hsieh studied electrospinning of cellulose acetate and reported that
uniformity, fiber alignment, defects control (e.g. beads, junctions, and using neither acetone nor DMAc alone can produce fiber free of beads.
pores), and other properties are tunable by changing these parameters, Only using a mixture of acetone and DMAc, beads-free fibers are
(1) substrate-related parameters (polymer concentration, viscosity, obtained [20].
molecular weight, surface tension); and (2) apparatus-related parame-
ters (flow rate and electric field). 2.1.2. Apparatus-related parameters
Lower feeding rate (also known as flow rate of the polymer solution)
2.1.1. Substrate-related parameters leads to smaller diameter of the fibers [13]. On the other hand, high
Most studies agreed that polymer viscosity the main determinants of feeding rate results in more beads formation. Based on the results
fiber diameter and morphology. Increased viscosity due to high polymer reported by Zuo et al., as the feeding rate increases, more solution is
molar mass or concentration can result in larger fiber diameters [2,12]. ejected from the needle tip [19]. The drying and evaporation of the
And also, beading is less likely to form, and more uniform fiber struc- solvent is less effective before the fiber reached the collector.
tures are observed [13,14].The relationship between polymer concen- An increased in applied electric field typically resulted in reduced
tration and fiber diameter and morphology of biodegradable polymers fiber diameter due to more stretching of the polymer solution. For
including poly (DL-lactide-co-glycolide) (PLGA) (50:50), poly(DL-lactic PLGA, increase in voltage (from 0.375–1.0 kV/cm) resulted in significant
acid) (PDLA), poly(L-lactide) (PLLA), gelatin, and dextran were reported reduction in fiber diameter, then the change of diameter was not signif-
in recent studies [9,13–17]. However, if the viscosity is too high, the flow icant when the voltage is further increased [9]. In addition, increasing
of the polymer solution may be hindered and the droplet dries at the tip. applied voltage typically leads to more beads formation, for example
On the other hand, if the viscosity is too low, fiber jet may break into in PDLA [14], chitosan [18] and gelatin [16]. However, for PHBV, higher
droplets due to the lack of chain entanglement. For example, when voltage leads to formation of beads-free PHBV fibers [19].
the concentration of PLGA in (THF + DMF) was 0.10 g/mL or less,
beads and droplets were obtained instead of nanofibers [9]. Ki et al. 2.2. Biodegradable polymers for electrospinning (synthetic/natural/blends)
studied the gelatin nanofibers. In the range of polymer concentration
of 8–12 wt. %, fiber diameter is exponentially increased with increasing The success of electrospun nanofibers that based on a wide range of
polymer concentration. In other words, the change of fiber diameter vs. biodegradable and biocompatible materials has been reported in recent
polymer concentration is nonliner. Uniform and beads-free gelatin fi- reviews [21,22], including natural proteins such as collagen, gelatin, silk,
bers (76–169 nm) were obtained [16]. chitosan and alginate; synthetic polymers such as polyglycolide (PGA),
Polymer molar mass affects viscosity of polymer solution. Typically, poly(ε-caprolactone) (PCL), PLA and their copolymers P(LLA-CL) and
low molar mass polymers lead to bead formation, while high molar PLGA that have been approved by FDA for clinical use; and their blends
mass polymers form fibers with larger diameters [10]. In addition, [8]. Fig. 2 displays the biological, mechanical and physiochemical

Fig. 1. (a) Schematic drawing of elementary setup for electrospinning. (Reprinted with permission from John Wiley and Sons [9]) (b) Typical bending instability of the jet during
electrospinning captured by high speed video. (Reprinted with permission from Elsevier [11]).
 D. Kai et al. / Materials Science and Engineering C 45 (2014) 659–670 661

Fig. 2. Biological, mechanical and physiochemical properties of commonly studied biodegradable natural and synthetic polymer (reprinted with permission from Elsevier [8]).

properties of commonly studied biodegradable natural and synthetic [4–6]. The aligned fiber is formed when the fiber is collected on a rotat-
polymers. ing mandrel [30], by external electric field [10] or magnetic field [31].
Courtney et al. prepared aligned polyurethane-urea fibers using rotating
2.2.1. Macroscale vs. nanoscale, which one degrades faster? mandrel [30]. Fig. 3 presents various degree of fiber alignment in
It is reasonable that degradation behavior of nanofibers is different correspond to the speed of the mandrel rotation. The aligned fibrous
from their respective polymers in macroscales (e.g. polymer films, pel- membrane exhibited stress-strain curves showing anisotropicity and
lets). The structure of the nanofibers (e.g. 3D scaffolds, non-woven modulus in the range of soft tissue, as illustrated in Fig. 4. Xie et al. dem-
mat, membranes etc.) plays important role in determining the degrada- onstrated that radically aligned PCL fibers can induce faster cellular mi-
tion profile of nanofibers. Compare nanofibrous scaffold with polymer gration and population than random fibers [32]. The radically aligned
films, electrospun nanofibrous scaffolds have higher surface area to vol- fibers were fabricated by application of external electric field at the O-
ume ratio (SVR), and thus higher porosity. This facilitates the diffusion ring collector.
of degradation products, resulting in faster rate of degradation. Semi-
crystalline polymers nanofibers including PGA and PCL were reported 2.3.2. Core/shell fibers
showing this observation. Core/shell nanofibers show advantages in delivery of delicate drug
The degradation profile of PGA nanofibers was rapid without an in- in a sustained way and preventing decomposition or fast degradation
duction period (first stage of degradation: water penetration or swelling of labile compounds. The core/shell nanofibers can be fabricated via
without mass loss and morphology change) [23]. In vitro degradation coaxial electrospinning from two immiscible solutions. The basic setup
showed 60 % weight loss in 20 days. PGA nanofibers degrade faster of the coaxial electrospinning is essentially similar with the typical
than microfibers and pellets because in PGA nanofibers, higher SVR al- electrospinning except the introduction of the spinneret that consists
lows higher water penetration into the highly crystalline matrix. Similar of an inner capillary tube. Zhang et al. reported core/shell nanofibers
observation is reported for PCL nanofibers [23–25]. Thinner nanofibers made of gelatin-core and PCL-shell [33]. Their results showed that the
showed faster diminishing trend in mechanical strength than nanofi- overall diameters of the core/shell nanofibers are increased as the
bers with larger diameters [26]. polymer concentration of the core material increases in the range of
In contrary, the porosity of the electrospun nanofibrous scaffold re- 7.5–15 w/v %.
duces the autocatalytic effect resulted from accumulation of acidic deg-
radation products, leading to slower degradation [27,28]. Amorphous 3. Electrospun nanofibers in biomedical application
poly (DL-lactide-co-glycolide) (PLGA) (50:50) polymer films samples
(0.5 mm) degrades faster than their nanofibers (550 nm) because of To date, the various applications of electrospun nanofibers have
autocatalysis effect within the polymer bulk [29]. been widely expanded due to their advantages. For example,
electrospun fibrous membranes possess high surface to volume ratio,
2.3. Advanced electrospinning high porosity and tunable physic-mechanical properties, as polymer
solutions and process parameters can be easily adjusted to obtain de-
2.3.1. Aligned fibers sired fiber morphology and mechanical properties. In addition, a wide
Several studies have shown that the anisotropic structure and to- range of synthetic and natural polymers are able to be electrospun
pography of aligned fibers not only resulted in anisotropic mechanical into nanofibers. From a biological perspective, the native extracellular
properties mimicking the ECM, but also shown improved interactions matrix (ECM) of human tissues and organs is composed of a network
with cells, for example, selective endocytosis, adhesion and orientation of micro/nano-scaled protein and glycosaminoglycan fibers, which
662 D. Kai et al. / Materials Science and Engineering C 45 (2014) 659–670

Fig. 3. Morphology changes of electrospun polyurethane-urea fibers as mandrel rotational velocity changes (a) random (b) 0.3 (c) 1.5 (e) 4.5 (f) 9.0 (g) 13.8 m/s (Reprinted with permis-
sion from Elsevier [30]).

provide support to resident cells and regulate cellular activities. With 3.1.1. Nanofibers for skin tissue engineering
the similar fibrous architecture of native ECM, electrospun nanofibers Electrospinning technique has greatly accelerated the development
have been broadly utilized in biomedical applications, such as tissue of innovative grafting scaffolds for skin TE. For wound healing applica-
engineering, drug delivery, cosmetics and biosensors. tion, the high porosity of electrospun nanofibers could provide more
structural space for accommodation of the grafted cells, facilitate cell
proliferation and migration and improve oxygen exchange, nutrient de-
3.1. Tissue engineering livery and exudates extrusion. One the other hand, the small pore size of
nanofibrous scaffolds is able to limit would wound infection and dehy-
As an interdisciplinary technique, Tissue engineering (TE) uses three dration during wound healing. Additionally, the tunable mechanical
basic components (cells, scaffolds and biomolecules) to develop properties of electrospun nanofibers could retain mechanical integrity
biofunctional substitutes for restore and maintenance of tissue function. between TE grafts and host tissue, and also prevent wound contraction
However, it is still a big challenge to design an ideal scaffold that mimics during implanting.
the structure and biofunctions of the native ECM, and the capability of Various natural and synthetic polymers have been electrospun into
possessing the ECM-like nanofibrous structure is an essential consider- nanofibrous scaffolds for skin TE. Natural polymers, such as collagen,
ation in rational design of TE scaffolds. Electrospun nanofibers, due to gelatin, silk, chitosan and fibrinogen, have been fabricated into nanofi-
their nanofibrous structure and high surface area to volume ratio, bers for wound healing, and cell culture results showed that those nano-
have shown to favor the adhesion, proliferation and differentiation of fibers could favor the attachment and proliferation of keratinocytes or
various cells, and serve as promising scaffolds for tissue regeneration. fibroblasts [34–37]. Among all these natural polymers, collagen type I,

Fig. 4. (a) Schematic of the native pulmonary valve (PV) leaflet. (b) The biaxial tension vs. strain curves of the aligned fibrous membrane and the native PV. PD: preferred direction, CD:
cross-preferred direction. (Reprinted with permission from Elsevier [30]).
 D. Kai et al. / Materials Science and Engineering C 45 (2014) 659–670 663

composed of two α1 chains and one α2 chain, is a particular excellent including PGA, PLA, PCL and their copolymers, are commonly used for
candidate for skin TE scaffolds, because as the main component of skin and other TE because of their favorable mechanical and biodegrad-
human skin ECM, it forms a 3D network fibrillar structure (with fiber di- able properties. Kumber et al. fabricated PLGA electrospun nanofibers
ameter of 50–500 nm) to modulate cell attachment, proliferation and with different fiber diameters (150–6000 nm), and they found that
differentiation in skin tissue [38]. Rho et al. electrospun collagen type I PLGA nanofibers with fiber diameters in the range of 350–1100 nm
nanofibers with an average diameter of 460 nm and demonstrated showed improved cell proliferation and spreading of human skin fibro-
that normal human keratinocytes exhibited improved cell adhesion blasts and significantly upgraded the expression of collagen type III gene
and spreading rate on collagen type I nanofibers compared to other [41]. To investigate the relationship between the degradation properties
protein-coated nanofibers (fibronectin, bovine serum albumin and lam- of nanofibers and their efficacy for dermal regeneration, PLA and PLGA
inin) [39]. In addition, the animal study showed that early-state healing with different lactide/glycolide mole fractions (85:15, 75:25 and
in the collagen nanofiber groups was promoted with the absence of sur- 50:50) were mixed and electrospun into nanofibers [42]. The in vivo
face tissue debris, prominent capillary and fibroblast proliferation. study showed that PLLA nanofibers remained stable after 12 months
Vatankhah et al. developed cellulose acetate/gelatin electrospun nanofi- of implantation whereas nanofibers of PLGA 85:15, 75:25 lost 50% of
bers to mimic the composition of dermis ECM (a complex combination their original masses after 4 and 3 months, respectively. Among all the
of proteins and polysaccharides), and they confirmed that electrospun tested nanofibers, electrospun PLGA (85:15, 75:25) were demonstrated
cellulose acetate/gelatin 25:75 nanofibers showed distinct adherency to be favorable biodegradable scaffolds for dermal replacement as be-
features and high proliferation of human dermal fibroblasts [40]. How- side supporting the growth of keratinocyte, fibroblast and endothelial
ever, the poor resistance of enzymatic degradation and weak mechani- cell, their degradation rate could match the healing rate in defected tis-
cal properties are two major drawbacks of using natural polymers in TE. sue (Fig. 5). However, the hydrophobic surface and the lack of cell-
On the other hand, multiple biodegradable synthetic polymers, recognition signals limit the application of synthetic polymers. Recently,

Fig. 5. H&E images of PLLA (a–c), PLGA 85:15 (D–F), PLGA 75:25 (g–l) following implantation into the flank of adult male Wistar rats at the time points indicated (2 weeks to 1 year).
Implanted scaffold has been labelled as (ES), with underlying muscle (M) and skin (S). Scale bar = 1 mm (Reprinted with permission from Elsevier [42]).
664 D. Kai et al. / Materials Science and Engineering C 45 (2014) 659–670

more researches are focusing on composite nanofibrous scaffolds as showed that human MSCs on PLCL/collagen nanofibers showed neuro-
they possess both the physical properties of synthetic polymers and bio- nal morphology, with multipolar elongations and expressed higher
activity of natural polymers. For example, PLCL/collagen nanofibers level of neuronal specific proteins such as neurofilament 200 and nestin,
were fabricated by electrospinning process, and compared to pure compared to pure PLCL nanofibers[68]. Beside collagen, laminin and gel-
PLCL nanofibers, PLCL/collagen nanofibers not only increased the atin also showed positive effects on neural growth and differentiation
proliferation rate of human MSCs, but also promoted the epidermal dif- [69–71].As the direction of axon growth is a crucial consideration in
ferentiation of the stem cells [38]. Similarly, some other degradable neural regeneration, aligned nanofibers have been widely used in this
composite nanofibers, such as PLGA/dextran, PCL/gelatin, PLCL/gelatin, field to promote neurite outgrowth and linkage to neighboring cells.
PLCL/fibrinogen have been developed and fabricated for skin tissue Yang et al. found that aligned PLLA electrospun nanofibers would pro-
engineering, and promising results were obtained [37,43–47]. mote the elongation and neurite outgrowth of neonatal mouse cerebel-
As prevention of an infection during wound healing is crucial for skin lum C17.2 stem cells parallel to the direction of aligned nanofibers
regeneration, a wide range of anti-bacterial nanofibers incorporated [72].Similarly, Xie et al. reported that the aligned PCL nanofibers not
with antibacterial agents (metallic, inorganic and organic)increasingly only enhance the differentiation of embryonic stem cells (ESCs) into
emerge and they show effective treatments for both the Gram- neural lineages but also direct the neurite outgrowth [73]. Furthermore,
positive and the Gram-negative bacteria present during wound healing. fiber diameter has been demonstrated as another factor to influence
Silver is the most commonly used metallic antibacterial agent, which neural stem cells (NSCs) behavior. He et al. evaluated the effects of
displays a wide range of biocidal activity and a low bacterial resistance. PLLA nanofibers with different fiber dimension on morphology of neo-
In wound healing, silver shows the capability of reducing surface in- natal mouse cerebellum C17.2 stem cells, and results showed that
flammation, increasing surface calcium and inducing epithelialization wider cell spreading with decreasing fiber diameter, and the small di-
[48,49]. Jin et al. fabricated PLCL based antibacterial electrospun nanofi- ameter nanofibers (b 500 nm) stimulated filopodia-like extensions of
bers by blending with different concentrations (0.25–0.75 wt.%) of sil- the cells, while the fibers with a relatively large diameter (N 700 nm) re-
ver nitrate (AgNO3) for skin TE [50]. Results showed that the sulted in a round morphology after 1 day of culture [74]. Christopherson
antibacterial activity of the nanofibers against Staphylococcus aureus et al. fabricated electrospun polyethersulfone (PES) fiber meshes with
and salmonella enterica was detected and the activity increased with different fiber diameters (283 ± 45 nm, 749 ± 153 nm and 1452 ±
the increasing concentration of silver nanoparticles in the nanofibers. 312 nm) to investigate the impact of fiber diameter on the differentia-
Similarly, some other polymers, including PLA, PLGA, PVA, polysulfone, tion of adult rat hippocampal-derived NSCs, and results showed that
beta-cyclodextrin and polyurethane (PU), were also electrospun into NSCs cultured on smaller diameter fibers differentiated preferentially
nanofibers and blended or coated with silver nanoparticles to induce into oligodendrocyte precursors, while the NSCs preferentially differen-
antibacterial properties [51–57]. However, in spite of its excellent anti- tiated into neuronal precursors on the larger diametered (i.e. 749 nm)
bacterial properties, sliver, like many other metals, could result in irrita- fibers (Fig. 6) [75].
tion and bind to DNA preventing to replication, causing cell death and Recently, the application of electrical stimulation (ES) in nerve TE has
hindering the healing processing. On the other hand, inorganic mate- become an emerging approach to promote neurite growth and neural dif-
rials, such as titania, have been incorporated into polymeric electrospun ferentiation, and electrically conductive nanofibers have been developed
nanofibers, which showed antimicrobial activity against multiple bacte- as a crucial substrate for ES. Therefore, conductive polymers, such as poly-
rial growth. Yan et al. prepared electrospun PU nanofibers with in-situ pyrrole (PPy), polyaniline (PANi), poly(3,4-ethylenedioxythiophene)
generated TiO2 as a wound dressing, and the PU/TiO2 nanofibers exhib- (PEDOT) and even carbon nanotubes, have been incorporated into
ited antibacterial efficiency against Pseudomonas aeruginosa and Staph- nanofibers during electrospinning. It is reported that conductive
ylococcus aureus [58]. Later on, electrospun silk fibroin/TiO2 nanofibers polymer-contained nanofibers could enhance the proliferation of
were fabricated and results showed that the nanofibers not only have nerve cells. Rat NSCson PLLA/PANi scaffolds showed higher prolifera-
good hemocompatibility and cytocompatibility, but also exhibited anti- tion than those on PLLA nanofibers after 8 days of cell culture [76].
bacterial activity against Escherichia coli under UV irradiation [59]. Similarly, NSCs on PANi/PCL/gelatin nanofibers exhibited enhanced
Moreover, carbon nanotubes were demonstrated to be highly cytotoxic cell proliferation and neurite outgrowth compared to PCL/gelatin nano-
to bacteria and kill microbes on contact. Schiffman has found that even fibers under ES [77]. In another study, random and aligned PPy-coated
at a low concentration (0.1 to 1 wt.%) of single-walled carbon nano- PLGA nanofibers were fabricated and seed with rat PC12 cells [78].
tubes in electrospun polysulfone nanofibers, loss of bacteria (Escherichia Under the stimulation of 10 mV/cm potential, PC12 cells exhibited
coli) was observed [60]. In addition, inspired by nature, some native an- 40–50% longer neurites and 40–90% more neurite formation compared
tibacterial agents, including shikonin, alkannin, fusidic acid, chitosan, to those cells without ES. Moreover, cells on aligned conductive nanofi-
lysostaphin and cinnamaldehyde, have been developed and incorporat- bers showed longer neurites and more neurite-bearing cells than those
ed into electrospun nanofiber to provide biocidal activity for wound on random nanofibers, indicating that both ES and topographical guid-
healing [61–66]. ance affect neurite growth for nerve regeneration.

3.1.2. Nanofibers for nerve tissue engineering 3.1.3. Nanofibers for cardiac tissue engineering
Other interesting application of electrospun nanofibers is for nerve To date, heart disease has become the first leading cause of death all
regeneration. The aim of nerve TE is to develop effective neural guid- over the world, especially in developed countries, and cardiac TE gained
ance conduits for bridging gaps in damaged peripheral or central neu- great attention recently as it promises to revolutionize the treatment of
rons, and the function of neural TE scaffolds should be directing patients with end-stage heart failure and provide new solutions to the
axonal sprouting and promoting the diffusion of neurotrophic factors. serious problems of heart donor shortage [79,80]. Electrospun nanofi-
Electrospun nanofibers are suitable materials for nerve TE as their struc- bers have been considered high-attentive scaffolds for cardiac TE be-
ture not only mimics the neural fibrous ECM, but also provides substrate cause their tunable mechanical properties and orientation of fibers are
topographical guidance to direct neural cells growth[67]. significant for myocardial regeneration [81]. Electrospun PCL nanofibers
It is reported that the properties of electrospun nanofibers (chemical were fabricated and seeded with rat cardiomyocytes (CMs), and the
components, diameter, orientation) could affect the proliferation, mor- cells attached on the scaffolds and started beating after 3 days, and
phology and differentiation of neural cells. The presence of natural poly- the expression of cardiac-specific proteins such as α-myosin heavy
mers in nanofibers is important for the growth of neural cells. chain, connexin43 and troponin I were detected after 14 days
Prabhakaran et al. demonstrated the advantage of collagen in neuronal [82].Thick cardiac grafts were created by overlapping up to 5 layers of
differentiation of MSCs on electrospun nanofibers, and the results the cell-nanofiber membranes[83]. After 1 week of culture in vitro,
 D. Kai et al. / Materials Science and Engineering C 45 (2014) 659–670 665

Fig. 6. Immunofluorescence analysis of rat NSCs cultured on TCP and nanofibers with different fiber diameters. Quantification of staining results is shown (a) with corresponding repre-
sentative images of cells on each substrate (b–d). All images captured 200×, with scale bar = 100 μm. Circled cells on 283-nm fiber mesh are cells stained double positive for RIP and Tuj-1
(e). Example of statistically unquantifiable 1452-nm mesh is shown in (f) (Reprinted with permission from Elsevier [75]).

morphologic and electrical communication between the intimately was detected in the aligned cardiac nanofibrous graft by the higher ex-
adhered layers was established, and synchronized contraction was pression of cardiac markers, such as GATA4, Nkx2.5 and MEF2C.
also observed. Stable and homogeneous PLGA/gelatin/elastin nanofibers Some conductive nanofibers have also been used in cardiac TE. The
were electrospun for cardiac TE, and the nanofibers were found to co-electrospinning of PANi with gelatin was carried out to obtain con-
promote H9c2 rat cardiac myoblasts proliferation and assist MSCs to ductive nanofibers and H9c2 rat cardiac myoblast cells were seeded to
penetrate into the center of scaffolds [84]. The mechanical properties investigate their potential for cardiac TE [93]. Results showed that the
of cardiac scaffolds are crucial for the function of cardiac cells. PGS/ concentration of PANi influenced the morphologies of cells seeded on
fibrinogen core/shell nanofibers were fabricated with comparable stiff- them. PPy/PCL/gelatin conductive nanofibers were electrospun by
ness of native myocardium, and Neonatal CMs cultured on nanofibers blending different concentration of PPy into PCL/gelatin, and cell assay
expressed high level of cardiac specific marker proteins, such as α- results showed that the proliferation of CMs on conductive nanofibers
actinin, Troponin, β-myosin heavy chain and connexin 43 [85]. increased when the PPy concentration was 15%, whereas the prolifera-
Cell alignment is also found in myocardial tissues and aligned ECM tion rate reduced when PPy reached 30% [94].PLA/carbon nanotube
plays an important role in the behavior and function of cardiac cells nanofibers were electrospun as a conductive platform to direct MSCs
in vivo, and therefore aligned nanofibers have been widely reported to differentiation towards a CM lineage under electrical stimulation [95].
direct the cell morphology and guide cell orientation in cardiac TE Under ES, cells reoriented perpendicular to the direction of the electric
[86–88]. Aligned biodegradable non-woven PLGA nanofibrous mem- field and adopted an elongated morphology. Moreover, an upgrade in
branes were electrospun to provide topographical cues for isotropic or a range of cardiac markers, such as cardiac myosin heavy chain,
anisotropic growth of neonatal rat CMs, and cell orientation and Nkx2.5, GATA-4, cardiac troponin T and connexin 43 were detected
elongation were enhanced on aligned nanofibers [89]. Electrospun after 10 days.
polymethylglutarimide (PMGI) nanofibrous meshes were fabricated to
guide the orientation of CMs and aligned cell growth was guaranteed 3.1.4. Nanofibers for bone tissue engineering
when the distance between fibers is below 30 μm [90].In another Unlike soft tissues in our body, bone is a physically hard, rigid and
study, both electrospun PCL and PCL/gelatin nanofibers were fabricated strong connective tissue, which microscopically contains relatively
and found to be able to guide the orientation of rabbit CMs [91]. Yet, small number of cells within abundant ECM in the form of collagen
greater cell alignment was observed on the aligned PCL/gelatin nanofi- nanofibers and stiffening inorganic substrate, such as hydroxyapatite
bers compared to that on PCL nanofibers, indicating that both scaffold (HAp). Therefore, the unique bone ECM is an organic-inorganic
topography and constituents influence cell orientation. To mimic the nanofibrous composite, in which osteocytes are able to perform good
cell alignment of myocardium and enhance cardiac differentiation, a functions and biological roles. To develop bone ECM mimicking scaf-
tissue engineered cardiac graft was generated by simultaneously folds, electrospun composite nanofibers of degradable polymers and
electrospinning elastic PU nanofibers and electrospraying MSCs [92]. calcium phosphate are subject of substantial investigations for bone
By controlling the processing parameters, the obtained tissue constructs TE. Ngiam et al. fabricated mineralized polymeric nanofibrous compos-
possessed the fibrous and anisotropic structure, and mechanical re- ites by soaking electrospun PLLA or PLLA/collagen nanofibersin calcium
sponse similar to native myocardium. Enhanced cardiac differentiation chloride solution and disodium phosphate solution alternately, and they
666 D. Kai et al. / Materials Science and Engineering C 45 (2014) 659–670

noticed that bone-like nano-HAp was successfully deposited on both to blended nanofibers. Moreover, the addition of PEG into PCL nanofi-
nanofibers, whereas the formation of nano-HAp on PLLA/collagen nano- bers accelerated protein release and helped to preserve up to 5 % of
fibers was faster and more uniform than on PLLA nanofibers. Moreover, the initial biological activity of the protein in the coaxial nanofibers
the nano-HAp deposited nanofibers showed enhanced capture efficacy [108].
of human fetal osteoblast cells within 20 min[96]. Later on, a novel Although electrospun nanofibers, especially coaxial nanofibers,
bone scaffold was developed by simultaneous electrospraying of show their advantages in drug delivery, several limitations still need
nano-HA on electrospun gelatin nanofibers, and the spin/spray gela- to be addressed for their wider application. Initial burst release of drug
tin/HA nanofibrous composite showed higher mechanical properties from nanofibers is still one of major issues especially when the drug
and promoted the cell proliferation, alkaline phosphatase (ALP) activity loading is high, as drug molecules tend to aggregate near the surface
and mineralization of osteoblast cells compared to pure gelatin nanofi- of fibers [109]. Recently, it was reported that the incorporation of
bers [97]. superhydrophobic agent into nanofibers would be an effective way to
Stem cell-based therapy for bone regeneration has aroused interest reduce burst release at early stage and prolong the sustained release
of many scientists, and among all stem cells, MSCs have been widely of drug. Highly hydrophobic electrospun PCL nanofibers loading with
used in bone TE due to its easy availability, self-renewing ability and po- a model bioactive agent (SN-38) were fabricated by adding varying
tential of osteogenic differentiation. Incorporation of nano-HAp into amounts (0–50%) of poly(glycerol monostearate-co-ε-caprolactone)
electrospun nanofibers has been also shown to promote cell adhesion (PGC-C18) as a hydrophobic polymer dopant [110]. Results showed
and proliferation and even enhance osteogenic differentiation of that the release rate of SN-38 highly depending on the content of
MSCs. Lee et al. reported that human MSCs cultured on PLGA/HAp com- PGC-C18 and the apparent contact angle of the fiber, and the fibers
posite nanofibers exhibited enhanced ALP activity, upgraded expression with higher hydrophobicity resulted in slower release rate, as the air
of osteogenic genes and increased calcium mineralization compared to layers trapped within the hydropholic fibrous meshes would restrict
those on pure PLGA nanofibers [98]. Similarly, Peng et al. found that water penetration. As a result, PCL nanofibers with high content of
chitosan/HAp nanofibrous scaffolds not only supported better cell at- PGC-C18 (30 and 50%) exhibited less than 10% release over 60 days.
tachment and proliferation of mouse MSCs, but also promoted the oste- Besides control of initial burst release, the release of multiple drugs
ogenic differentiation by up-regulating osteogenic gene expression, without interfering the release kinetics of each other is another issue
even in the absence of osteogenic supplementation [99]. Different con- to be solved. Therefore, study on nanofiber/particle electrospun com-
centrations (0–50%) of nano-HAp were blended into PCL to fabricate posite for drug delivery is emerging fast, and different particle, including
electrospun PCL/HAp nanofibers for bone TE, and results showed that PLGA nanoparticles, alginate microspheres and chitosan microspheres
MSCs on nanofibers showed increased cell proliferation rate and en- were successfully incorporated into electrospun nanofibers [111–114].
hanced osteogenic differentiation capability (ALP activity and minerali- With this technique, different drugs could be easily loaded into nano-
zation extent) with the increasing concentration of HAp in PCL particles or nanofibers, especially hydrophobic drugs together with hy-
nanofibers. Beside HAp, some other calcium salts, including beta- drophilic drugs. Xu et al. encapsulated BSA (as a hydrophilic model) into
tricalcium phosphate, calcium carbonate, and even calcium phosphate chitosan microspheres, dissolved benzoin (as a hydrophobic model
cements, have been incorporated into polymeric nanofibers for bone drug) into PLLA solution, and fabricated PLLA fiber/ chitosan micro-
TE, indicating that polymer/bioceramics nanofibrous composites are sphere composites by electrospinning to investigate the dual release
promising substrates for accelerating bone regeneration [100–102]. of these two drugs[114].Results showed that chitosan microspheres dis-
persed uniformly in the nanofibers, and the hydrophilic BSA had a
3.2. Drug delivery short-term release while hydrophobic benzoin had a relative longer
and sustained release. On the other hand, mesoporous silica nanoparti-
The aim of designing a drug delivery system is to enable to control cles (MSNs), due to their large specific surface area, tunable mesoporous
drug release towards alleviating medical conditions at a defined rate structure and facile surface fictionalization, have emerged as promising
over a definite period [103]. Electrospun nanofibers have shown their drug delivery carriers and they have been incorporated into electrospun
advantages in the field of drug delivery due to their high surface area- nanofibers as dual drug release system (Fig. 7) [115,116]. Song et al. de-
volume ratio with interconnected pores in the fibers, which ensure bet- veloped dual drug-loaded PLGA/MSNs electrospun composites, where
ter dissolution rate and high therapeutics take-up. Furthermore, the rate PLGA nanofibers were loaded with hydrophilic drug, fluorescein and
of drug release can be tailored for various applications by easily tuning MSNs were loaded with hydrophobic rhodamine B [117]. Results
relevant nanofiber properties, such as fiber diameter, porosity, and showed that separate and distinct profiles of two individual drugs
drug binding mechanisms [104]. To date, numbers of drugs and biomol- were observed, indicating that there was no interaction between two
ecules, including genes, proteins and enzymes, have been successfully drugs. Moreover, most of the fluorescein was fast released during the
incorporated into electrospun nanofibers, mainly by two approaches: first 324 h, while rhodamine B exhibited a sustained release behavior.
blending electrospinning and coaxial electrospinning [105]. Compared These studies have confirmed that the nanofiber/particle composite sys-
to blended nanofibers in which the polymer and biomolecules are tem is able to promote sustained and independent release of multiple
mixed, the coaxial nanofibers encapsulate biomolecules inside of the drugs.
polymers with a core-shell structure, leading to a reduced initial burst
release and longer release period. PLCL nanofibers containing tetracy- 3.3. Biosensor and immunoassay
cline hydrochloride (TCH) were fabricated by blending and coaxial
electrospinning, and in vitro drug release study showed that 60–80% Biosensors, which are analytical devices for the detection of biologi-
of loaded TCH was released within the first 5 h for the blended nanofi- cal components (analytes), have been widely utilized for environmen-
bers, whereas the burst release from coaxial electrospun nanofibers tal, food and clinical application. As detection of gases and biological
was reduced to only 5–10% followed b stable and sustained release analytes normally is at low amount and concentration, sensitivity and
[106]. Later on, the same group reported that loading bone morphoge- limit-of-detection (LOD) play very important roles in the function of
netic protein 2 (BMP-2) and dexamethasone into the core of PLCL/ biosensors. Recently, researchers have recognized the advantage of in-
collagen nanofibers by coaxial electrospinning would lead to a controlled creasing surface area of the detector substrate to increase the sensitivity
release rate of the two proteins compared to those from blended nanofi- and LOD of biosensors without increasing the amount of overall sample
bers [107]. Bovine serum albumin (BSA) as a model protein was incorpo- required, and therefore nanomaterials with extremely high surface area
rated into PCL nanofibers by blending and coaxial electrospinning. to volume ratio, which could increase the number of binding sites avail-
Coaxial nanofibers exhibited better sustained release profiles compared able for biological recognition element immobilization, are being
 D. Kai et al. / Materials Science and Engineering C 45 (2014) 659–670 667

desirable large surface area to volume ratio, electrospun nanofibers

can easily be functionalized through the incorporation of doping agents
during spinning or through surface modification after spinning.
Fast glucose sensors with good sensitivity and selectivity are always
highly demanded as glucose detection is remarkably vital to the patients
suffering from diabetes. As glucose oxidase (GOD) is highly sensitive and
selective to glucose and shows good stability in a large range of pH, this
enzyme has been widely used to fabricate glucose biosensors. Recently,
GOD has been successfully immobilized in/on various electrospun nano-
fibers for glucose detection. Ren et al. fabricated electrospun PVA/GOD
nanofibers as a glucose biosensor, and chronoamperometric measure-
ments showed that nanofibrous enzymatic electrodes exhibited a
rapid response (1 s) and a higher response current (l A level) to glucose
in the normal and diabetic level, and the linear response range (from 1
to 10 mM) and the LOD (0.05 mM) of the biosensor also meet the
requirements of glucose detection in medical diagnosis[120]. Later on,
a single ZnO nanofiber-based glucose biosensor was developed by
functionalizing an individual electrospun ZnO nanofiber on a gold
electrode with GOD by physical adsorption (Fig. 8) [121]. Electrochem-
ical measurements revealed that the biosensor showed a high and re-
producible sensitivity of 70.2 μAcm−2 mM− 1within a response time
of less than 4 s, as well as a linear range from 0.25 to 19 mM with a
low LOD of 1 μM. Moreover, the biosensor exhibited a good anti-
interference ability and favorable stability over relatively long-term
storage (more than 4 months).Although numbers of new strategies
have been explored to design advanced glucose enzyme sensors, insuf-
ficient stability due to the intrinsic nature of the enzyme limits the ap-
plication of GOD based sensors. Therefore, recently more attentions
Fig. 7. SEM (a), TEM (b) and UC luminescence (c) images of PCL/gelatin/silica nanoparti- have been paid on developments of non-enzymatic glucose sensing,
cles composite (Reprinted with permission from [115]. Copyright (2013) American Chem-
and various carbon, metals (Au, Pt, Ni and Cu) and their oxides have
ical Society).
been exploited as electrode materials to construct enzyme-free glucose
sensors. For example, CuO nanofibers were prepared by electrospinning
integrated within analytical systems to allow for the detection of low and subsequent thermal treatment processes for glucose non-
concentrations of analytes without any complicated amplification pro- enzymatic detection, and the biosensor showed a high sensitivity
cesses[118,119]. Due to their unique characteristics, electrospun nanofi- (431.3 μA cm− 2 mM− 1), fast response (about1 s) and long-term
bers have gained great attention in biosensor application. Besides their stability [122]. Similarly, Co3O4 nanofiber-based glucose sensor was

Fig. 8. a) Schematic diagram of the modified gold electrode and the mechanism of the glucose sensing on the modified electrode. (b) Cyclic voltammograms of the bare and modified gold
electrode without and with 100 μM glucose in pH 7.0 PB solution. (c) Cyclic voltammograms of the biosensor in PB solution (pH 7.0) containing 100 μM glucose at a scan rate of (a) 100 mV,
(b) 80, (c) 50, and (d) 20 mV s−1 (Reprinted with permission from [121]. Copyright (2010) American Chemical Society).
668 D. Kai et al. / Materials Science and Engineering C 45 (2014) 659–670

fabricated by electrospinning and subsequent calcination, and the bio- their large surface area, have been demonstrated as advanced
sensor exhibited a fast response time (less than 7 s), a high sensitivity substrates for biosensors based on biotin-streptavidin immobilization.
of36.25 μA cm−2 mM−1, good reproducibility and selectivity, and a de- Li et al. successfully incorporated biotin into PLA nanofibers through
tection limit of 0.97 μM (S/N = 3) [123]. In addition, its application in electrospinning and the streptavidin immobilized on PLA nanofibrous
the detection of glucose inhuman blood serum sample showed agreed substrates could capture a biotinylated DNA probe [124]. On the
results with those obtained from commercial glucose meter, indicating other hand, Yang et al. electrospun poly(dimethylsiloxane) (PDMS)/
that electrospun nanofibers show great potential applications in the de- poly(methyl methacrylate) (PMMA) nanofibers as substrates for pro-
velopment of biosensors for glucose detection. tein microarrays, and the results of immunoassays demonstrated the
Electrospun nanofibers incorporating a binder have been also used superior performance of PDMS/PMMA nanofibers, where the LOD of
as a substrate for biosensor assay. In most immunoassay, streptavidin is nanofibrous substrates was 32 times lower than that on nitrocellulose
utilized as substrate surface, which could be easily conjugated with a membrane[125].
biotinylated biorecognition agent for the detection of specific target In research and clinical settings, Enzyme-linked immunosorbent
analytes, and recently, electrospun nanofibrous membranes, due to assay (ELISA) is the current gold standard immunoassay due to its

Fig. 9. Simultaneous analysis of immunoassay and enzyme-based assay using micropatterned PS/PSMA nanofibers. (a) Illustration of IgG-immobilized nanofibers that were
micropatterned with enzyme-entrapped PEG hydrogel and detection logic, (b) fluorescence image of micropatterned PS/PSMA nanofibers that reacted with solution containing glucose,
Amplex Red and FITC-labeled anti-IgG (scale bar = 500 mm), and (c) fluorescence intensity from the fiber and hydrogel region in the micropatterned nanofibers that reacted with three
different samples (Reprinted with permission from Royal Society of Chemistry [128].
 D. Kai et al. / Materials Science and Engineering C 45 (2014) 659–670 669

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