Macromolecular Nanotechnology

The role of rheology of polymer solutions in predicting nanober

formation by electrospinning
R. Roic
a
, J. Pelipenko
a
, P. Kocbek
a
, S. Baumgartner
a
, M. Beter-Rogac
b
, J. Kristl
a,
a
Faculty of Pharmacy, University of Ljubljana, Ljubljana, Slovenia
b
Faculty of Chemistry and Chemical Technology, University of Ljubljana, Ljubljana, Slovenia
a r t i c l e i n f o
Article history:
Received 21 December 2011
Received in revised form 16 April 2012
Accepted 3 May 2012
Available online 16 May 2012
Keywords:
Nanobers
Electrospinning
Interfacial rheology
Complex uids
Chitosan
Alginate
a b s t r a c t
Electrospun polymer nanobers are gaining increasing importance in tissue engineering,
wound dressing and drug delivery. Here, we present a thorough rheological study of poly-
mer solutions in the bulk and at the interface to nd correlations between those properties
and the electrospinnability of the solutions and the morphology of the resultant nanobers.
Our results indicate that blended solutions of chitosan or alginate with poly(ethylene
oxide) (PEO) are appropriate for electrospinning when they form conductive, unstructured
uids displaying plasticity, rather than elasticity, in the bulk and at the interface. The inter-
facial rheological parameters are three orders of magnitude lower than those in the bulk.
We demonstrate for the rst time that interfacial, rather than bulk, rheological parameters
show improved correlation and can be used to predict the success of the electrospinning
process. Using the interfacial parameters of samples with homologous compositions, dif-
ferent groups of solutions can be identied that form smooth nanobers. However, rheo-
logical parameters of the bulk and at the interface provide complimentary information.
The bulk parameters are determined by polymer concentration and directly affect jet ini-
tiation, while the interfacial behaviour determines the continuation of the jet and bre for-
mation. We propose that interfacial parameters are indispensible tools for the design of
electrospinning experiments.
2012 Elsevier Ltd. All rights reserved.
1. Introduction
Polymer nanobers represent an emerging class of
biomimetic nanostructures that have shown tremendous
promise as tissue scaffolds, modern wound dressings and
advantageous drug delivery systems [13]. The main reason
for the increased interest is the unique ability of nanobers
to mimic and organise biological microenvironments, sup-
plement damaged or diseased tissue and stimulate tissue
regeneration [46]. To date, three processing techniques,
self-assembly, phase separation and electrospinning, have
been developed, with the last method showing the greatest
promise [1,7]. To prepare nanobers by electrospinning,
high voltage is applied to a polymer solution, whereupon
a charged jet is ejected from the needle and then undergoes
extensive stretching and thinning during a rapid solvent
evaporation stage. While the jet travels towards the
grounded collector, polymer bres are formed [8]. The elec-
trospinning process is governed by a variety of forces
including the Coulomb force between the charges on the
jet surface, the electrostatic force due to the external electric
eld, the viscoelastic force of the solution, the surface ten-
sion, the gravitational force, and the frictional force due to
air drag [9].
More than one hundred polymers have been investi-
gated for the design of electrospun nanobers, with poly-
mers of natural origin being generally favoured [4,5].
Among those nanobers containing chitosan and alginate
(Fig. 1) have demonstrated promising properties as tissue
0014-3057/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.eurpolymj.2012.05.001

Corresponding author. Address: Faculty of Pharmacy, University of

Ljubljana, Askerceva cesta 7, 1000 Ljubljana, Slovenia. Tel.: +386 1 47 69
521.
E-mail address: julijana.kristl@ffa.uni-lj.si (J. Kristl).
European Polymer Journal 48 (2012) 13741384
Contents lists available at SciVerse ScienceDirect
European Polymer Journal
j our nal homepage: www. el sevi er . com/ l ocat e/ eur opol j
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engineering constructs due to their biocompatibility,
biodegradability, safety and hydrophilicity [5,1012].
Additionally, chitosan suppresses an inammation re-
sponse during healing and expresses antimicrobial activity
[13].
However, pure chitosan and alginate, as well as most
natural polymers, are difcult or impossible to electrospin
into brous structures due to their limited water solubility,
ionic character, three-dimensional network due to hydro-
gen bonds, coupled with high solution viscosity at low con-
centrations [5,8]. These limitations can be overcome by
blending them with non-ionogenic linear exible poly-
mers, such as poly(ethylene oxide) (PEO) and poly(vinyl
alcohol) (PVA) which signicantly improve the natural
polymers spinnability [14,15]. Moreover, such blends have
added advantages over the electrospun nanobers of pure
natural polymers because their mechanical properties, bio-
compatibility and antibacterial effects are drastically en-
hanced [16]. In our study, chitosan and alginate were
blended with PEO, which was chosen due to its solubility
in water, thereby avoiding the use of organic solvents dur-
ing production.
Extensive research has been conducted to determine
the spinnability of particular polymer solutions, but the
mechanisms of bre formation are still not completely
understood. In general, a solution is spinnable if the poly-
mer jet remains unbroken until dry nanobers are formed.
Numerous studies have shown that the optimal electros-
pinning conguration and operational conditions differ
drastically from one polymer to another, with process
parameters (applied electric eld, needle-to-collector dis-
tance, ow rate, type of collector) and solution properties
(viscosity, surface tension, conductivity) being the main
factors inuencing the transition of a polymer solution into
ultrane bres [10,17,18]. Barnes et al. [1] demonstrated a
linear relationship between electrospun polymer solution
concentrations (in a range from 2% to 30%) and bre
diameter for poly(glycolic acid), poly(lactic acid), their
blends or copolymers, poly(caprolactone) and gelatine. Be-
cause polymer type and concentration are the main
parameters dening the rheological behaviour of the poly-
mer solutions, it would be reasonable to expect that those
are the determining factors in bre formation. Thus, rheol-
ogy is the spotlight of the present study.
The majority of published papers on electrospun nanof-
ibers routinely examine the viscosity of the polymer solu-
tions; with most of the collected data suggesting that
higher viscosity favours smoother nanober formation
[16,17,19,20]. Other rheological parameters, such as the
elastic (G
0
) and plastic (G
0 0
) modulus, are rarely considered.
A few studies have examined the role of elasticity of the
polymer solutions [21,22]. The authors hypothesised that
elastic forces resist the bending of the jet and hinder a
jet from breaking up. Su et al. [22] also concluded that -
brous structures actually originated from the elasticity of
the liquid jet out of the needle during the electrospinning
and that greater elasticity resulted in a beadless nanober
structure. Even more rarely reported in the literature is the
role of interfacial rheology in the electrospinning process,
although the importance of this feature becomes obvious
when considering the increase in surface area-to-volume
ratio of the electrospun jet during the thinning process,
where jet diameter decreases and the area increases by
several orders of magnitude. One previously published
study [21] has shown that spinnable solutions have an
interfacial G
0 0
that is 10 times greater than that of solutions
from which bres could not be prepared.
In this context, a deeper understanding of the rheolog-
ical parameters of polymer solutions used for nanober
formation is required. It cannot be neglected that viscosity
in bulk is important in toto, but because the majority of
processes in the electrospinning mechanism actually takes
place at the interfaces, it would not be surprising that
interfacial rheology may determine the docking site of
the complex jet formation. Therefore, understanding how
and when interfacial rheology properties dominate the
process should be considered as an integral part of poly-
mer solution management for electrospinning.
The aim of the present research is to present a thorough
rheological study of polymer solutions in the bulk and at
the interface in an effort to nd correlations between those
properties and the electrospinnability of the solutions and
the morphology of the resultant nanobers. Our hypothe-
sis is that the viscoelasticity of the interfacial layer of the
solution is a better predictive parameter for jet formation
relative to bulk parameters. This analysis will permit fur-
ther interpretation of the results, which are essential for
jets that do not break up and that form uniform nanobers.
The conclusions of this study will be generally applicable
for investigations of nanober design and production.
2. Materials and methods
2.1. Materials
The chitosan used was low-viscous chitosan (viscosity
of a 1% solution in acetic acid at 20 C 6 200 mPas) from
Fig. 1. Chemical structure of chitosan (A) and alginate (B). Chitosan is a
linear polymer composed of randomly distributed b-(1-4)-D-glucosamine
and N-acetyl-D-glucosamine, whereas alginate is formed by blocks of a-
(1-4)-D-mannuronic acid and a-(1-3)-L-guluronic acid.
R. Roic et al. / European Polymer Journal 48 (2012) 13741384 1375
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Fluka (Buchs, Switzerland). Sodium alginate (viscosity of a
2% solution in water at 25 C 6 250 mPas) and poly(ethyl-
ene oxide) (PEO) (Mw = 400,000 g/mol) were supplied by
SigmaAldrich Chemie GmbH, Germany. Acetic acid was
provided by Merck (Darmstadt, Germany).
2.2. Preparation of the polymer solutions
Solutions of pure polymers were prepared separately
and then mixed in order to obtain a single blended solu-
tion. A 3% (w/w) chitosan solution in 2% (w/w) acetic acid
and a 3% (w/w) PEO solution in distilled water were used
to prepare blends with chitosan:PEO mass ratios of
100:0, 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70,
20:80, 10:90, 93:7, 97:3 and 0:100, with a nal polymer
concentration of 3% (w/w). A 4% (w/w) alginate solution
and a 4% (w/w) PEO solution in distilled water were mixed
to gain the same mass ratios described above, with a nal
polymer concentration of 4% (w/w). The blends were stir-
red at 20 rpm with a magnetic stirrer for 24 h to obtain
homogenous solutions at equilibrium. Previous studies
guided our choice of total mass concentration for the poly-
mers in the nal blended solution. Those studies showed
that the optimal total mass concentration for chitosan-
blended solutions is 3% (w/w) and for alginate-blended
solutions is 4% (w/w) [14,23,24].
2.3. Electrospinning of the polymer solutions
The electrospinning setup is schematically presented in
Fig. 2. The polymer solution was placed in a 20 ml plastic
syringe tted with a metallic needle with an inner diameter
of 0.8 mm. A syringe pump (Model R-99E, RazelTM) was
used to feed at a constant rate. High voltage at the needle
was achieved by connection to a voltage generator (model
HVG-P60-R-EU, Linari Engineering s.r.l., Italy) capable of
generating voltages in the range of 060 kV. For each solu-
tion, a variety of parameters were tested for the electrospin-
ning process, including applied voltage (1030 kV), solution
ow rate (0.13 ml/h) and needle-to-collector distance (10
20 cm). The optimal applied voltage for both polymers was
found to be 25 kV with a needle-to-collector distance of
17 cm. The optimal ow rate was determined to be
1.8 ml/h for chitosan:PEO and 0.4 ml/h for alginate:PEO
blends.
2.4. Characterisation of the polymer solutions
The rheological properties in the bulk and at the inter-
face as well as the conductivity and surface tension were
determined for all polymer solutions.
Rotational and oscillatory rheological tests were per-
formed using a Physica MCR 301 rheometer (Anton Paar,
Graz, Austria) with a cone-plate measuring system CP50-2
(cone radius 24.981 mm, cone angle 2.001, sample volume
1.15 ml) at a constant temperature 25.0 0.1 C. Rotational
tests were usedto determine the viscosity, whichfor a cone-
plate measuring systemis calculated as g s
c
=
_
c, where
_
c is
the shear rate and s
C
the shear stress. Oscillatory tests were
performed to dene the elastic and loss modulus, which are
calculated as G
0
= (s
a
/c
a
)cosd and G
0 0
= (s
a
/c
a
)sind, where
s
a
is the shear stress, c
a
is the deformation and d is the phase
shift angle [25,26].
The shear rate during the rotational tests ranged from 2
to 100 s
1
. The oscillatory shear measurements were per-
formed at an amplitude within the linear region (ampli-
tude 10%) with a frequency from 0.2100 s
1
.
Interfacial rheological tests were carried out using the
same rheometer with interfacial cell, with a biconical mea-
suring system (cone radius 34.171 mm, cup radius
40.00 mm, cone angle 4.973, approximate sample volume
104 ml), where the shear rate was set between 2 and
100 s
1
and the frequency in the oscillatory measurement
was in the range of 0.2100 s
1
(amplitude 1%). The system
was positioned at the interface, and the sample was delib-
erately kept in direct contact with the ambient
atmosphere.
The presented values of viscosity in bulk and at the
interface are taken at the lowest measured shear rate (for
bulk viscosity at 2 s
1
, while the rst measured point for
interfacial viscosity was at 9 s
1
), where the load of the
system only minimally inuences viscosity. The oscillatory
tests were compared at low (0.2 s
1
), medium (3.98 s
1
)
and high frequency (100 s
1
). However, the results of G
0
and G
0 0
presented here are taken only at the lowest fre-
quency because these values show the most dramatic dif-
ferences between the blended solutions.
Conductivity of solutions was measured at room tem-
perature using an Iskra Conductivity meter MA 5964 (Iskra,
Ljubljana, Slovenia) with an electrode conductivity con-
stant of 0.7265 cm
1
.
Surface tension was measured at 25 0.5 C by the plate
method with a Processor Tensiometer K-12, Version 5.05
(Kruss Gmbh, Hamburg). For each solution, average values
of parameters from at least three measurements are
reported.
2.5. Characterisation of nanobers
The diameter and morphology of the electrospun nanof-
ibers were examined using a 235 Supra 35VP-24-13 high-
resolution scanning electron microscope, SEM (Carl Zeiss,
Fig. 2. A sheme illustrating the basic principles of the electrospinning
process.
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Germany) operated at an accelerating voltage of 1 kV with
a secondary detector; no conductive coating layer was ap-
plied before imaging. The obtained images were used to
determine the bre diameter using ImageJ 1.44p software
(NIH, USA) by measuring 50 bres chosen randomly.
Fourier transform infrared (FTIR) spectroscopic analysis
was used to qualitatively characterise the interactions be-
tween chitosan, or alginate, and PEO. FTIR spectra were
collected with a Nexus FT-IR Nicolet spectrophotometer
at a resolution of 4 cm
1
, with signal averaging over 250
scans in each interferogram over the range of 500 to
4000 cm
1
.
3. Results and discussion
Nanobers of chitosan or alginate combine the benets
of the polymer properties with improved physical struc-
ture, offering an advantageous approach for wound healing
and tissue engineering. However, both natural polymers
have proven to be challenging to electrospin. Therefore,
the present paper is aimed at developing a rational expla-
nation for the effect of rheological properties of polymer
solutions on the electrospinning process. The results are
divided into three parts: rst, evaluation of chitosan:PEO
and alginate:PEO solution properties, with an emphasis
on solution rheology; second, the morphology of nano-
bers formed by the electrospinning process; and third, pre-
diction of nanober formation from the rheological
properties of polymer solutions.
3.1. Conductivity and surface tension of polymer solutions
The conductivity and surface tension of solutions, as a
function of the amount of chitosan and alginate in the
blends, are shown in Fig. 3. The results clearly show that
the surface tension of blends remained nearly unchanged
regardless of the solution composition, whereas the con-
ductivity correlated with the proportion of chitosan or algi-
nate in the blends. Conductivity correlated with the
polymer concentration because both natural polymers dis-
play polyelectrolyte properties in aqueous media, i.e.,
chitosan is positively charged in acetic acid, and alginate
is negatively charged in distilled water. The results of sur-
face tension experiments may be explained by the fact that
there was always the same total mass of polymer in the
blends, which spreads uniformly across the surface.
3.2. Rheology of polymer solutions in bulk
The main emphasis of the present research was to study
rheological properties as a function of solution composi-
tion. Rotational and oscillatory rheological measurements
were performed on all chitosan:PEO and alginate:PEO
blended solutions as well as on solutions of the pure com-
ponents. Rotational tests were used to determine the vis-
cosity of the solutions as a function of shear rate, while
oscillatory tests were performed to determine dynamic
moduli, i.e., storage and loss moduli. The storage, or elastic,
modulus (G
0
) is a measure of the elasticity of a material and
represents the materials ability to store energy. By
contrast, the loss, or viscous, modulus (G
0 0
) relates to the
ability of the material to dissipate energy, which is used
to change the materials structure and is lost as heat.
Fig. 4 shows the results obtained from rotational tests,
where changes in viscosity as a function of shear rate for
blended solutions with different compositions are pre-
sented. The shape of the viscosity curves clearly shows
decreasing viscosity values with increasing shear rate,
indicating that all solutions behave as non-Newtonian
(pseudoplastic), shear-thinning uids. The data also show
that the viscosity curves of the chitosan:PEO blends all
lay between the curves for the pure components over the
entire compositional range, indicating that miscibility of
the blends occurs and that compatible, one-phase blends
are formed.
The analysis dened the differences in rheological
behaviour between the blended solutions, since the shape
of the curves are dependent on the solution composition.
Solutions with a higher percentage of chitosan or alginate
show a greater decrease in effective viscosity with increas-
ing shear rate relative to solutions rich in PEO. For exam-
ple, the viscosity of a 90:10 solution containing 10% PEO
varied from 2.1 to 1.0 Pas with shear rate, while a 10:90
solution containing 10% chitosan varied from 0.6 to
0.4 Pas. These results lead to the conclusion that the
pseudoplasticity of the solutions increases with an increas-
ing proportion of polyelectrolyte in the blends. The pro-
nounced shear-thinning effect can be related to the
Fig. 3. Surface tension and conductivity of chitosan:PEO (A) and algi-
nate:PEO (B) blends as a function of solution composition. The total
amount of polymer in chitosan:PEO is 3%, and in alginate:PEO, the
amount is 4%.
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structure of the polymer chains in the solutions. Without
an external load, each single macromolecule can be found
in the shape of a three-dimensional coil because this is
lowest energy state. During the shear process, however,
the molecules are more or less oriented parallel to the
direction of shear, resulting in elongation, which lowers
their ow resistance and results in a decrease in the bulk
viscosity.
The results presented in Fig. 5 show the viscoelastic
properties of the chitosan:PEO (Fig. 5A) and alginate:PEO
(Fig. 5B) blended solutions as a function of their composi-
tion. It is observed that viscosity and G
0 0
increase monoton-
ically with an increase in the proportion of chitosan in the
blends. Generally this trend is true for alginate:PEO blends
as well, although the slope is not as extreme. Additionally,
G
0 0
, as well as its slope, is always much greater than G
0
. The
differences between the G
0
and G
0 0
values for the blends in-
crease as the chitosan or alginate content increases and are
generally higher for the chitosan relative to the alginate
samples. Notably, the viscosity, G
0
and G
0 0
of pure chitosan
is higher than those of pure PEO (bulk viscosity of chitosan
2.1 Pas, PEO 0.4 Pas), explaining the observation of lower
values in the blends with higher PEO content. However,
the experimental values are not simple proportional sums
of the individual viscosities. This nding may be attributed
to the fact that the polysaccharides are able to form inter-
actions between individual polymer chains.
In order to assign intermolecular interactions between
ionogenic and nonionogenic polymers, FTIR spectroscopy
was used. The FTIR spectra were suitable for studying the
spectral changes caused by polymerpolymer interactions
by measuring changes in the relative intensities of vibra-
tional bands of the dried blended material versus the pure
Fig. 4. Viscosity curves showing the shear-thinning behaviour for chito-
san:PEO (A) and alginate:PEO (B) blended solutions obtained using a
rotational cone-plate measuring system.
Fig. 5. Viscosity, storage (G
0
) and loss (G
0 0
) moduli as a function of
chitosan:PEO (A) and alginate:PEO (B) blended solution compositions as
determined using a cone-plate measuring system with a viscosity
measured at a shear rate of 2 s
1
and dynamic moduli measured at a
frequency 0.2 s
1
.
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components. Blended samples exhibited most of the char-
acteristics of the pure components (chitosan and PEO),
with an additional broad band at approximately
3350 cm
1
(Fig. 6). On the other hand, we are aware that
our samples contain certain amount of water, which can
contribute to formation of the same peak at 3350 cm
1
.
Similar changes were observed on the FTIR spectra of algi-
nate, PEO and their blends (results not shown).
The presence of PEO in blends changes the inter- and in-
tra-molecular interactions of the chitosan and alginate
chains by interacting with the backbone of the natural
polymer and disrupting the self-association of the chains.
Murray [27] reported that such inter-polymer interactions
result in the formation of a compact structure such that the
hydrodynamic volume is smaller than the volume sum of
individual macromolecules, and this modulation is physi-
cally manifested as a decrease in the solution viscosity.
The effect of reduced viscosity in solutions with a high pro-
portion of alginate can be attributed to the fact that in the
present study, alginate forms a less ordered structure due
to its stronger repulsive forces between charged groups,
as proven by the observation of higher conductivity shown
in Fig. 3.
3.3. Interfacial rheology of polymer solutions
A detailed investigation of the rheological parameters of
the bulk may not be sufcient when dealing with complex
processes, such as electrospinning, Interfacial parameters
differ from the bulk due to reorganisation of the polymer
molecules at the interface itself. Until now, interfacial rhe-
ological properties were mostly studied in surfactant solu-
tions, foams and emulsions [2729]; their use for
characterising polymer blends for electrospinning is stud-
ied here for the rst time.
The results depicted in Fig. 7 showchanges of interfacial
viscosity as a function of the applied shear rate for chito-
san:PEO and alginate:PEO blended solutions, indicating
their interfacial pseudoplasticity. A wide gap can be clearly
observed between the interfacial viscosity curves of
blends, while viscosity curves in the bulk followed sequen-
tially subsequently (Fig. 4). Notably, the effect is much
more obvious with chitosan than alginate (in Fig. 7).
The results of the interfacial measurements presented
in Fig. 8 show the same parameter trends as was observed
in the bulk. However, the values of the interfacial rheolog-
ical parameters are three orders of magnitude lower than
those observed in the bulk. The interfacial viscosity and
loss modulus have signicantly higher values as compared
to the storage modulus. Nonetheless, an impressive differ-
ence is established in the slopes of the plots for interfacial
viscosity and G
0 0
as a function of the solution composition,
a trend not observed for G
0
(Fig. 8A). The values are much
more signicant in the case of chitosan as compared to
alginate (Fig. 8A and B).
In the case of chitosan, the slope in the rst region in-
cludes solutions having a chitosan proportion in the blends
of 10% or less, the second region includes chitosan content
of up to 60%, and solutions with higher chitosan content
are placed into the third region.
According to the results from the alginate:PEO blends,
the solutions having concentrations of alginate up to 30%
are in the rst region and those up to 60% are in the second
region. With 70% alginate, the parameters start to decrease,
although division into three regions remains consistent be-
cause the parameter values remain above those of the
other two regions.
3.4. Electrospinning of polymer solutions
Because the success of the electrospinning process is
strongly correlated to the properties of the solutions from
which the nanobers are prepared, variation between the
morphology of the obtained nanobers was expected.
Nanobers from blended solutions and those obtained
from pure chitosan or alginate and PEO were prepared
using the same process parameters in order to directly Fig. 6. FTIR spectra of pure chitosan, PEO and the chitosan:PEO blend.
R. Roic et al. / European Polymer Journal 48 (2012) 13741384 1379
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correlate the solution characteristics to the spinnability
and morphology of the products.
Electrospinning pure chitosan and alginate solutions
(without any addition of PEO) resulted in the formation
of polymer droplets on the collector without any nanober
formation as is seen in Fig. 9. A proposed explanation is the
inability of jet formation during electrospinning due to the
high bulk viscosity and pseudoplastic structure of the pure
solutions. Generally, there is a critical viscosity value (close
to 1.5 Pas) required to allow for electrospinning because
below these values, improved results are observed as
shown in Fig. 8 and 9. In addition to the viscosity, chitosan
and alginate are polyelectrolytes that are charged in solu-
tion. Strong repulsive forces between ionogenic groups
within the polyelectrolyte backbone impede the formation
of continuous bres.
The spinnability of chitosan and alginate was achieved
by the addition of PEO in different ratios as suggested by
others [14,15]. The morphologies of a series of the electro-
spun samples were analysed by SEM, with representative
micrographs shown in Fig. 8. It can be clearly seen that
the composition of the spinning solutions had a signicant
effect on the morphology of the nanobers. Improved
Fig. 7. Interfacial viscosity as a function of shear rate for chitosan:PEO (A)
and alginate:PEO (B) blended solutions. Measurements were performed
using a biconical measuring system presented schematically above.
Fig. 8. Interfacial viscosity, storage (G
0
) and loss (G
0 0
) moduli as a function
of solution composition of chitosan:PEO (A) and alginate:PEO (B) blends.
Solutions are categorised in three groups according to spinnability and
nanober morphology. Conditions: biconical measuring system, viscosity
at a shear rate of 9 s
1
and dynamic moduli at a frequency of 0.2 s
1
.
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morphological changes are observed with decreasing
amounts of chitosan or alginate in the polymer
blends, resulting in a decrease in the values of their rheo-
logical parameters and conductivity as well. The highest
Fig. 9. Representative SEM images of nanobers obtained from blended solutions of chitosan:PEO (above) and alginate:PEO (below) at mass ratio (A/a)
10:90, (B/b) 20:80, (C/c) 30:70, (D/d) 40:60, (E/e) 50:50, (F/f) 60:40, (G/g) 70:30, (H/h) 80:20, (I/i) 90:10 at 25 kV, needle-to-collector distance 17 cm and
ow rate for chitosan 1.8 ml/h and alginate 0.4 ml/h. Total polymer concentration is 3% (w/w) for chitosan blends and 4% (w/w) for alginate blends.
R. Roic et al. / European Polymer Journal 48 (2012) 13741384 1381
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investigated chitosan:PEO or alginate:PEO ratio resulted in
no nanober formation (Fig. 9I and i). At an 80:20 ratio,
beads with rare nanobers were formed (Fig. 9H and h),
and at the ratio of 70:30, a structure of short bres, which
extend between the beads, was observed for both poly-
mers (Fig. 9G and g). Electrospinning of other solutions re-
sulted in the formation of three-dimensional nanober
networks. In the case of chitosan, smooth nanobers with
no beads were formed when the content of chitosan in
the blends was 10% or less (Fig. 9A), whereas solutions
with 30% alginate content still produced nanobers with-
out beads (Fig. 9ac). Nanobers with a bead-on-string
morphology were produced when the content of chitosan
in the blend was 40% or less (Fig. 9BF) and at 4060% in
the case of alginate (Fig. 9df). In both cases, a higher
amount of polyelectrolyte triggers the formation of a high-
er number of beads. Therefore, increasing the amount of
PEO in blended solutions and relatively decreasing the
amount of polyelectrolyte causes a progressive transfor-
mation of the electrospun samples morphology from
nanoparticles to bead-on-string structures to nanobers.
The mean diameter of the obtained nanobers was in
the range of 15130 nm for chitosan and from 30
120 nm for alginate. In general, the diameter of the electro-
spun nanobers depended on the polymer concentration,
3% or 4%, with variability caused by the proportion of both
polymers in the blends. The nanobers grew thinner as the
content of chitosan or alginate increased. The reason for
obtaining a reduced diameter in spite of having higher
solution viscosity is due to the charge density in the
ejected jet, which imposed higher elongation forces lead-
ing to thinner bre formation, in agreement with previ-
ously published data [14,17].
3.5. Prediction of nanober formation
The morphology of prepared electrospun products is
difcult to predict before completing all of the experimen-
tal procedures and analyses, including SEM, which requires
a lot of time. Therefore, determining the key parameters
necessary to predict electrospinning success from solution
properties is essential. Regarding the electrospinning
mechanism, solution viscoelasticity profoundly inuences
jet formation and jet stability, which is crucial for success.
The solution has to maintain appropriate viscoelasticity in
order to survive stretching, acceleration and whipping.
The analysis of nanober morphology in relation to rhe-
ological properties in the bulk suggests that polymer solu-
tions from which nanobers are produced should be
conductive, unstructured shear-thinning uids and express
both plastic and elastic behaviour. The plasticity of the
blends has to be greater than elasticity, and only a proper
relation between these two features enable jet formation
and stabilisation. Elasticity, however, has to be as low as
possible, but still present, to enable jet initiation. The for-
mation of the electrospun jet from spinnable (Supplemen-
tary 1) and unspinnable (Supplementary 2) solution was
recorded and can be seen in supplement. The underlying
explanation is that the increased elastic force increases
the tendency of the jet to contract, which prevents jet
initiation and elongation. Our ndings downplay the
importance of the elastic response of the polymer chains
for jet stabilisation and support an absence of any signi-
cant bulk elasticity in shear. The Regev research group
[21] has obtained similar results. Our results are not in
accordance with the traditional explanation for jet stabili-
sation, which places an emphasis on the role of the elastic
response over the plastic response [21,22]. Nevertheless,
during the drying of the jet when the overall polymer con-
centration increases, the proportion of elastic and plastic
moduli changes (G
0 0
/G
0
decreases with higher overall poly-
mer concentration). An important additional cause for
unsuccessful nanober formation from solutions contain-
ing a higher proportion of charged natural polymer is the
growing repulsive forces, which viscous forces are not able
to overcome. It was determined that the minimum neces-
sary proportion of spinnable carrier in the blends is 40%,
which is equal to a concentration of 1.2% (w/w) in the case
of chitosan and 1.6% (w/w) in the case of alginate.
However, a much more signicant discovery was made
based on interfacial rheology. The results clearly show the
dependence of the morphology on the solution composi-
tions, subdivided into regions as dened previously
(Fig. 8). From solutions processed in the rst region,
smooth, uniform and bead-free nanobers are formed,
whereas the solutions in the second region form beaded
nanobers, and only droplets with few nanobers form
in the solutions in the third region. In the case of alginate,
the breaks between different regions are less distinct, but
the analysis revealed that the quality of the obtained elec-
trospun nanobers is better and variations between bres
prepared from different blends are smaller. To conclude,
the production of nanobers from solutions with smaller
variation in interfacial parameters is easier because the
proper solution parameters can now be quickly achieved.
We discovered that in the case of chitosan, the formation
of three-dimensional nanober networks was enabled
from the solutions past the gap in interfacial viscosity
(from approximately 9 to 18 mPas).
Inimitable correlation between the interfacial viscosity
and the plastic modulus is not surprising when considering
the basic physical mechanism of electrospinning. During
the process, the electrospun jet thins dramatically from
800 lm (diameter of the needle) to a few nm (Fig. 10),
causing much more dominant role for surface properties.
In the region of the Taylor cone of ejected jet bulk rheolog-
ical properties play a more important role due to the larger
diameter and volume, whereas thinning of the jet causes
reduced importance of the bulk rheological parameters
and increased the signicance of interfacial rheological
parameters. Notably, the thinner the jet is, the greater
the surface area, and therefore, the greater the importance
of the interfacial characteristics. Moreover, due to solvent
evaporation, which occurs at the interface, a concentration
gradient of polymer molecules is formed, further increas-
ing the interfacial effect.
Additionally, much greater sensitivity of interfacial rhe-
ology enables the determination of even small differences
between solutions, which is also the reason why bulk rhe-
ological parameters in chitosan:PEO and alginate:PEO
show continuous increase, while in the interfacial results
the denition of distinctive slopes is possible. Moreover,
1382 R. Roic et al. / European Polymer Journal 48 (2012) 13741384
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interfacial shear rheology is so far the best rheological
method which allows the determination of real dilatational
material properties of the interface at extension rates that
occur during actual electrospinning process [21,28,30]. All
aforementioned facts support the conclusion that the cor-
relation between the rheological characteristics of the
interface and nanober morphology is far more distinctive,
obvious and evident than are the correlations with rheo-
logical properties in the bulk or any other solution
characteristic.
4. Conclusion
The present study explored correlations between the
rheological parameters of solutions and electrospinnabili-
ty. The rheological results indicate that blended solutions
of polyelectrolyte and nonionogenic polymer for electros-
pinning are effective when they are conductive, unstruc-
tured uids. The blends should show greater viscous
(plastic) properties over elastic behaviour in the bulk as
well as at the interface. For the rst time, we proved that
interfacial rheological parameters correlate much better
with the outcomes of, and predict the success of, the elec-
trospinning process. Based on interfacial parameters, dif-
ferent groups of solutions can be dened, predicting
where smooth nanobers form. Because the prediction
was proven for two distinct natural polymers, the conclu-
sions may be extended to other polymer blends containing
polyelectrolytes. Still, rheological parameters in bulk and
at the interface have to be taken as complementary. Bulk
properties are largely determined by polymer concentra-
tion and thus allow for the prediction of jet and bre for-
mation, while the interfacial properties enable the
prediction of jet continuation. We are sure that interfacial
parameters will be indispensable tools in the design of
polyelectrolyte solutions for electrospinning in the future.
Acknowledgment
The authors are grateful to the Slovenian Research
Agency for nancial support of this research work: P1 -
0189, J1 - 4236, 1000 - 09 - 310085 and 1000 - 11
310213.
Appendix A. Supplementary data
Supplementary data associated with this article can be
found, in the online version, at http://dx.doi.org/10.1016/
j.eurpolymj.2012.05.001.
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