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Application of electrospinning techniques for the production of
tissue engineering scaffolds: A Review
Article    in  Europe an Scientific Journal · Januar y 2014
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European Scientific Journal May 2014 edition vol.10, No.15 ISSN: 1857 – 7881 (Print) e – ISSN 1857- 7431
265 APPLICATION OF ELEC TROSPINNING
TECHNIQUES FOR THE PRODUCTION OF
TISSUE ENGINEERING SCA FFOLDS: A REVIEW

Md. Mahabub Hasan
Asst. Prof., Primeasia University, Dhaka, Bangladesh
A.K.M. Mashud Alam
Chairman, Bangladesh University of Business and Technolgy,
Dhaka, Bangladesh
Khandakar Abu Nayem
Sr. Lecturer, Primeasia University, Dhaka, Bangladesh

Abstract
Electrospinning of nanofibers with diameters that fall into nanometer
range have attracted growing attention in recent years due to the ability to
produc e scaffolds with nanoscale properties. Scaffolds from nanofibers for
tissue engineering are large field. The goal of this review is to demonstrate
an obj ective and ove rall picture of current research work on e lectrospinning
technique, where different types of polymers are used for produc ing tissue
engineering scaffolds. The target is also to describe the theory of different
electrospinning process and di scussing the applications and i mpacts of
electrospinning on the field of tissue engineering.

Keywords: Electrospinning, Tissue Engineering, Nanofiber, Scaffold

Introduction
The nonw oven indus try generally considers nanofibers as ‘having a
diameter of less than one micron, a lthoug h the National Science Founda tion
(NSF) defines nanofibers as having at least one dimension of 100 n anometer
(nm) or less.
Nanofibers are the new class of materials, which are used for several
value added applications such as medical textile, personal care, filtration,
barrier, composite, insulation a nd e nergy storage. Due to the special
properties of nanofibers they are suitable for a wide range of applications
from medical to consumer products and indus trial to high-tech applications
for aerospace, energy storage, fuel cells, information t echnology, drug

European Scientific Journal May 2014 edition vol.10, No.15 ISSN: 1857 – 7881 (Print) e – ISSN 1857- 7431
266 delivery system and most imp ortantly in the field of tissue engineering
(Raghavendra R H et al. 2005) .
Tissue engineering combines the design pr inciples of living
organisms and modern engineering with the development of viable
substitutes of hum an tissues such as muscle, skin, cartilage, bon e, even
cardiovascular and neuronal structures. The field is developed via scaffolds
implementing with a variety of bioactive mo lecules to balance cel l
proliferation and di fferentiation. S caffolds are usually produc ed for the
following purposes:
• To provide cell attachment, proliferation and m igration
• To assist in the growth of three-dimensional tissue and or gans
• Deliver and retain cells and bi ochemical factors
• Enable diffusion of vital cell nutrients
• Exert certain mechanical and biological influences to modify the
behavior of the cell phase.

Tissue engineering scaffolds
Tissue en gineering scaffolds are defined as three-dimensional
structures that assist in the tissue engineering process by providing a site for
cells to attach, proliferate, differentiate and secrete an extra-cellular matrix,
eventually leading to tissue formation. It is also possible to guide cells into
forming a neo-tissue of predetermined three-dimensional shape and size by
optimiz ing the scaffold structure to get the desired cellular activities. Tissue
engineering scaffolds can be either permanent or temporary in nature,
depending on t he application a nd the function of the neo-tissue. Usually
temporary scaffolds are made from biodegradable pol ymers, such as
polyglycolic acid, which degrade within the body to leave a purely biological
neo-tissue (Freed, LE. et al, 1994) . On the other hand, P ermanent scaffolds
remain within the body, working with ingrown tissue to form a
polymeric/biological composite (Matsumoto H et al., 2001) .

Electrospinning for tissue engineering application
Electrospinning is a simple and cost-effective method to
produc e scaffolds with an inter-connected pore structure and fiber diameters
in the sub-micron range compared to self-assembly and pha se separation
techniques. The field of tissue engineering is thought to capitalize upon t hese
features for the produc tion of 2D or 3D scaffolds.

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267

Fig.1: Schematic diagram of an electrospinning process (Nandana et al. 2010)

Three basic compone nts as shown in figure 1 are required to setup the
electrospinning process: a syringe with a metal spinneret of small diameters,
a high voltage supplier and a rotating collector. A high voltage in the range
of 10-50 kV is applied to create an electrically charged jet of polymer
solution or melt out of the spinneret in the electrospinning process. A cone-
shaped of the polymer solution dr oplet directed to the counter electrode is
formed unde r the high voltage (Deitzel JM et al. 2001, R eneker DH et al.
2007) . The droplet on the spinneret is slowly stretched as the voltage
increasing and if the increase of voltage is continued, a jet is formed from the
deformed droplet, which moves towards the counter electrode and be comes
narrower in the process as in figure 2 (Theron S A et al. 2005) . It is thought
that the electrospinning gives us the impression of being a very simple and
easily controlled technique for the produc tion of nanofibers. But, actually the
process is very intricate. That’s why, it is also described as the interaction of
several physical instability processes (Reneker DH et al. 2002, Theron SA et
al. 2004) . Advancement of this technique in bot h micro and nano fabrication
have pow ered the field of tissue engineering in many aspect. More than 100
raw materials can be used in this technique.

Fig.2: Schematic diagram of a taylor cone formation with increase in applied voltages
(Wang HS et al. 2009)
Spinneret
Taylor Cone
Ejected

Critical

Increasing Applied

Syringe Polymer solution
Spinneret Collector
Fibers
High Voltage

European Scientific Journal May 2014 edition vol.10, No.15 ISSN: 1857 – 7881 (Print) e – ISSN 1857- 7431
268 Electrospinning of functional polymeric nanofibers and their application
Natural polymer scaffold
Most of the research on e lectrospinning of natural pol ymers is
focused on bi opolymers, be cause these naturally occurring polymers
normally exhibit better biocompatibility and low toxicity than other
polymers. C omplicated solvent systems are us ually required f or
electrospinning of natural biopolymers, such as hexafluoroisopropyl alcohol
for collagen (Matthews JA et al. 2002, Rho K S et al. 2006) or gelatin (Li M
et al. 2005) and formic acid for silk fibroin (Min BM et al. 2004, W ang H et
al. 2005) . Nmethylmorpholine (NMO)/water or N, N-dimethyl acetamide
mixed solvent is used for electrospinning of cellulose (Kulpinski P et al.
2005, K im CW et al. 200 5). The silk fibroin is biocompatible and its pow der
is useful as a substance for growing or activating epidermal cells. Silk fibroin
nanofiber mat from electrospinning is effective in regeneration of damaged
periodical tissues and the correspond ent membrane could guide bone tissue
regeneration (Chung C.P et al. 2006) .
Chitosan is a copolymer of N-acetyl-glucosamine and N -glucosamine
units. It is produc ed by deacetylation of naturally occurring chitin, which is
extracted from shellfish sources. The term chitosan is used when the
percentage of N-acetyl-glucosamine units is lower than 50% (Jayakumar, R
et al. 2008) . Chitosan is accessible for various established pr ocessing
technologies (e.g. freeze-drying, freeze-gelation) and has been us ed to
produc e films, gels as well as porous sponge -like scaffolds, because of its
solubility in dilute acids. Chitosan has become a frequently applied material
as regenerative medicine and biomaterials research including orthopedics,
periodont ology, drug delivery systems, wound healing applications and
tissue engineering since last two decades (Manjubala, I et al. 2008, H su S. H
et al. 2004) .
Collagen is the most abundant protein family in the body, which has
been extensively used for in vitro and in vivo tissue engineering. In many
popul ar tissues, pol ymers of type I and type III collagen are the principal
structural elements of the extracellular matrix (Parry and Craig, 1988). There
are various types of collagens, which can be isolated from a variety of
sources. More than 80% of collagens in the body consist of mainly type I, II
& III and share similar features in all species. Collagen is highly conserved,
relatively non-immunoge nic and ha s been us ed in a variety of tissue
engineering applications (Matthews JA et al. 2002 ). The function of collagen
is to provide structural suppor t to the tissue, in which it is present and at the
same time it is also known to sequester many factors required for tissue
regeneratopm amd maintenance. Therefore, it is also considered as ‘ideal’
scaffold material in the tissue engineering field (Boland E R et al. 2004) . The
classification of a solvent, which di ssolved collagen at sufficient

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269 concentrations to accomplish electrospinning and the volatile nature of the
solvent for rapid dr ying of electrospun m ats are the key issues for
electrospinning of collagen. For the first time electrospinning of collagen is
attempted by using type I collagen (calf skin) dissolved in hexafluro-2-
propanol (HFP) and characterization with scanning electron microscopy
(SEM) and transmission e lectron m icroscopy (TEM) is carried out . The
electrospun collagen scaffolds have been applied for wound d ressings and
prelimin ary vascular tissue engineering as it provides an in vitro method to
create a p reformed, nanofibrous collagen scaffold that closely mimic s the
native collagen network (Matthews et al. 2003, B oland et al., 2004; Shields
et al., 2004; Rho et al., 2006) .

Synthetic homopolymer scaffold
Polycaprolactone (PCL) and pol ylactide (PLA) are biocompatible
and bi odegradable polymers, which can be synthesized by living ring-
opening polymerization. T hey have been firstly electrospun i nto nanofibers
as scaffolds in tissue engineering due to their biomedical properties
(Bognitzki M et al. 2007 , Chen F.J et al. 2006) . The shape-ability and fiber
surface morphology is difficult to design to fulfill the mechanical shapes and
sizes, thoug h electrospinning can be carried out easily.

Synthetic polymer scaffold
For modifying functional polymer materials, electrospun of block
copolymers are of great interest. It is also possible to generate new materials
of desired properties. The performance of electrospun f iber mat based on
copolymers can be significantly improved in comparison with that of
homopolymers, if the materials are properly implemented. For example, a
biopolymer namely PLGA (copolymer of PGA and P LA) is popul ar and
well-studied system that has been broadly used as electrospun s caffolds for
biomedical applications. The mechanical properties and degradation of
produc ed fiber is quite different from the original i.e. Polyglycolic acid
(PGA) and P LA hom opolymers. The nanofibrous PGLA scaffolds generally
degrade faster than the regular casting film with the same dimensions and
composition, m ainly because of the nanofiber surface properties and the high
water adsorption a bility of the material (Zong X et al. 2002) . Another
example for biomedicine application is copolymer of P(LA-CL), which is
synthesized from the copolymerization of Lactide and c aprolactone. The
degradation a nd p roperties of the copolymer is between those of the two
homopolymers (PLA and P CL). The functionality and pot ential use of
produc ed P(LA-CL) has been investigated by several groups (Kwon IK et al
2005, M o XM et al. 200 4, Xu CY et al. 2003) . Other functional nanofibers
are manufactured b y electrospinning of environmental respons ive
copolymers, such as pH – respons ive copolymer, thermal-responsive

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270 copolymer etc. For example, the gel of a monodisperse triblock copolymer
consisting of poly (methyl methacrylate-blockpoly [2-(diethylamino) ethyl
methacrylate]-block-polymethyl methacrylate) has been shown pH-
respons ive behavior (Topham PD et al. 2006) .

Synthetic extracellular matrices
One of the main obj ects of tissue engineering is to develop the design
of polymeric scaffold with specific mechanical and bi ological properties
simila r to native extracellular matrix (ECM) in order to modulate cellular
behavior (Langer R et al. 1993) . A vast majority of the cells are in contact
with the ECM in vivo, which is composed of a network of nano-meter-sized
proteins and glycosaminoglycans. The intricate complexities of this spatial
and t emporal environment dynamically influence phe notypic and ot her
cellular behavior by providing indirect and di rect informational signaling
cues (Xu, C et al. 2004) . For example, the presence of an organized collagen
type I ECM for integral binding is required for the development of
osteoprogenitor cells towards mature osteoblasts in case of bone (Behonick
D.J et al. 2003 ). The interactions between cells and E CM can modulate
cellular activities such as mig ration, proliferation, differentiation and
secretion of various hormones and g rowth factors (Franceschi R.T et al.
1992) . Thus, the more closely the in vi vo e nvironment (i.e. chemical
composition, m orphology, surface functional groups) can be recreated, the
success is more for the tissue engineering scaffold (Lan C.W et al. 2003) .
Tissue engineering scaffolds work as temporary ECMs unt il repair or
regeneration oc curs (Li W.J et al. 2002, M o X.M et al. 2004 a nd Smith L.A
et al. 2004) .

Composite scaffolds
Electrospinning can also be used to produc e composite scaffolds. For
example, a scaffold with layers can be created by sequentially spinning
different polymer solutions. Each layer can be tailored for specific cell
adhesion and could be potentially beneficial for zonal articular cartilage or
arterial vessel repair (Kidoaki S et al. 2005) . Boland e t al. have showed
smooth muscle cell infiltration into a multi-layered scaffold of collagen types
I and III and elastin, when cultured in a rotary cell culture system.
Alternatively, two or more polymer solutions can be spun concurrently,
resulting in a scaffold with mixed types of fibers (Boland, E .D et al. 2004).
Collagen types I and III could be spun in this manner to create a scaffold
better mimic s their in vivo ratios.

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271 Techniques used for electrospinning
Coaxial electrospinning
It has been investigated that nanofibers required the functionalizing
agents (for example, biomolecules, such as enzymes, proteins, drugs, viruses
and bacteria) for keeping them in the fluid environment to maintain their
functionality. In order to meet their requirement, core-shell nanofibers are
manufactured by a modified electrospinning pr ocess, such as coaxial
electrospinning. Figure 3 shows the basic setup for coaxial electrospinning
and the fabrication pr ocess of common core-shell nanofibers. Depending on
the setup of electrospinning, two syringes feed inter-separated and coaxial
‘inner fluid’ and ‘outer fluid’ to spinneret. The electrospinning liquid is
drawn out from spinneret and forms a ‘compoun d taylor cone’ with a core-
shell structure unde r the application of high voltage (Loscertales, et al.
2002) .
If the ap propriate technical parameters are selected, core-shell
nanofibers can be produced with high precision from a hug e variety of
materials by coaxial electrospinning process. Han et al. 2008 patented
recently the composite nanofiber with a polycarbona te (PC) shell and a
polyurethane (PU) core. The nonw oven fabric or membrane with the
composite fiber combines the filtration efficiency of the PC with the
mechanical characteristics, such as elasticity of PU. The membrane or fabric
is useful in filters and aviation dr esses or clothing. Extraordinary fiber
structures could be formed through coaxial electrospinning of special
polymers, such as the ‘nanocables’ can be formed with electrically
conduc tive polyhexylthiophe ne as the core and Polyethylene oxide (PEO) as
the insulating shell (Sun Z et al. 2003) .
Coaxial electrospinning is not limited to the production of core-shell
nanofibers with a continuous core. Core-shell droplet can also be generated
by coaxial electrospinning. A coaxial jet of hydrophilic polymer (outside)
and a hydrophobi c liquid (inner) is electrospun, w hich produc es beaded
fibers, encapsulating the hydrophobi c liquid into these beads (Díaz JE et al.
2006)

Fig.3: The basic setup for coaxial electrospinning (Zhang Y et al. 2004)
High voltage DC

inlet of the outer

inner and out er

core shell

collecto
cork
inlet of
the inner

European Scientific Journal May 2014 edition vol.10, No.15 ISSN: 1857 – 7881 (Print) e – ISSN 1857- 7431
272 In this case, the beads are regularly distributed along the fibers and
their sizes exhibit a uniformly distribution. T he outer liquid flow rate control
the bead to bead distance and fiber diameter, while the bead diameter can be
maintained by controlling the inner liquid flow rate. Under the appropriate
condition, it is possible to produc e hollow fibers via coaxial electrospinning
(Xia Y et al. 2006) . Depending on e vaporation of the solvent, the core
polymer precipitates on the wall of the previously formed shell. Hollow
nanofibers with functionalized inner and outer surfaces are directly
fabricated by coaxial electrospinning (Li D et al. 2005) . Now, it is thought
that this technique provides a unique protocol for manufacturing complex
catalyst system, drug delivery and filtration.

Modified electrospinning processes
Electrospinning procedure is further modified to accommodate the
needs of ma terials for biomedical applications. Dual syringe reactive
electrospinning as shown in figure 4 is one of such modifications (Ji J et al.
2006) . The cross-linking reaction oc curs simultaneously during the
electrospinning process using a dual syringe mixing technique.

Fig.4: Schematic illustration of a modified electrospinning setup (Ji J et al. 2006)

Zhong et al. 2006 describes the fabrication of alligned collagen
nanofibrous scaffolds. The electrospinning apparatus used by them is shown
in figure 5. T he structure and in vitro properties of these scaffolds are
compared with a random collagen scaffold.
Highly por ous 3D nanofibrous scaffold us ing PCL is made by
electrospinning with the help of auxilliary electrode and chemical blowing
agent (BA) by Kim et al. 2007.
cross-inker Teflon tubing
collector HA/PEO
High voltage
power supply
Water
Extraction
HA nanofibrous scaffold Lyophilization
Syringe pump#1 Syringe pump#2

European Scientific Journal May 2014 edition vol.10, No.15 ISSN: 1857 – 7881 (Print) e – ISSN 1857- 7431
273

Fig.5: Fiber alignment during electrospinning (Agarwal et al. 2008)

Multi-channel coaxial electrospinning
The experimental setup of multi-fluidic compound -jet electrospinning
is shown in figure 6. Various metallic capillaries with varying outer diameter
and inner diameter are arranged at the several vertexes of an equilateral
triangle. Then the bundle of capillaries is inserted into a plastic with gaps
between individual inner capillaries and outer syringe. Two immis cible
viscous liquids are fed separately to the three inner capillaries and out er
syringe in an appropriate flow rate. A 20% Polyvinylpyrrolidone (PVP)
ethanol solution is used as outer liquid, while a nondissolution pa raffin oil is
selected as inner liquid. Then a high vol tage between three inner metallic
capillaries and a metallic plate coated with a piece of aluminum foil acted as
counter electrode and provide the driving and controlling for the
electrospinning. The immiscible inner and out er fluids (red for paraffin oil
and bl ue for Ti(OiPr)4 solution) are carried out separately from individual
capillaries. With the appropriate high vol tage application, a whipping
compound f luid jet is formed unde r the spinneret and then a fibrous
membrane is collected on the aluminium foil (Fengyu Li et al. 2010 ).

Fig.6: Schematic illustration of a) multi-channel coaxial electrospinning and b) fiber shaping
Syringe pum p
Syringe
Coverslip
High voltage source
Rotating wheel
Inner

Outer

a)
Solvent
volatilizatio
Tube
solidificatio

b)

European Scientific Journal May 2014 edition vol.10, No.15 ISSN: 1857 – 7881 (Print) e – ISSN 1857- 7431
274 Wound dr essing
Electrospinning can be applied to pr oduc e a scaffold with more
homogeneity besides meeting ot her requirements like oxygen permeation
and pr otection of wound from infection and de hydration for use as wound-
dressing materials. Various types of synthetic and na tural polymers like
carboxyethyl chitosan/PVA (Khanam N et al. 2007) , collagen/chitosan (Chen
JP et al. 2008) , silk fibroin, ABA type poly(dioxanone co-L-lactide)-block-
poly(ethylene glycol) block copolymer (Kim HY et al. 2004) have been
electrospun t o suggest them for wound -dressing applications. Further,
wound -dressing material is produc ed by electrospinning as shown in figure 7
of PVA/AgNO 3 aqueous solution into non -woven webs and then treating the
webs by heat or UV radiation (Hong KH et al. 2007)

Fig.7: Two stream electrospinning (Hong Y et al. 2008)

Effects of various parameters on electrospinning.
The el ectrospinning process is affected by many parameters,
classified broadly into polymer solution parameters, process parameters and
ambient parameters. The parameters of polymer solution m ay be viscosity,
conduc tivity, molecular weight, surface tension and pr ocess parameters
include applied electric field, tip to collector distance an d flow rate. Each of
the parameters significantly affect the fibers morphology obtained as a result
of electrospinning and by proper manipulation of these parameters, it is
possible to get the nanofibers of desired morphology and di ameters (Chong
et al., 2007) . Finally, ambient parameters encompass the hum idity and
temperature of the surroundings, which play a significant role in determining
the morphology and di ameter of electrospun na nofibers (Li and Xia, 2004 ).

PLGA/tet solution
Capillary A Syringe pump A
X axis raster
Rotation Metal collection rod
Capillary B PEUU solution
Syringe pump B

European Scientific Journal May 2014 edition vol.10, No.15 ISSN: 1857 – 7881 (Print) e – ISSN 1857- 7431
275 Conclusion
Electrospinning is a very simple and versatile method f or creating
polymer based high functional and hi gh performance nanofibers. Tthoug h,
electrospinning is first described ove r 70 years ago, acceptability of the
technique has increased dramatically in the past 10 years due to the rising
interest in nanoscale properties of the materials. This technique allows for
the produc tion of polymer fibers with diameters on the nanometer scale.
Recently, electrospinning has gained popul arity with the tissue
engineering community as a potential means of produc ing scaffolds. The
objective of this review is to describe briefly the theory behind the technique,
observe the effect of changing the parameters on f iber morphology and
discuss the application and impact of electrospinning on the field of tissue
engineering. In the future it is important to apply this technique for the
produc tion of nanofibers from different types of polymers.
Electrospinning matrices are able to suppor t the attachment and
proliferation of a wide variety of cell types. Using innova tive collectors and
spinning techniques, scaffolds with aligned fibers, different compositions,
improved mechanical properties, varying degradation rates or functional
properties can be devedope d. N evertheless, despite the comprehensive
experimental and theoretical studies illustrating the ability to control fiber
formation, c oncerns with fiber diameter uniformity still need to be addressed.
In summary, electrospinning is an attractive and promising approach for the
preparation of functional nanofibers due to its wide applicability to materials,
low cost and hi gh produc tion rate.

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