Indian Journal of Fibre & Textile Research

Vol. 37, June 2012, pp. 127-132

Calcium alginate/PVA blended nano fibre matrix for wound dressing
K Tarun & N Gobi
a
Department of Textile Technology, Anna University, Chennai 600 025, India
Received 1 April 2011; revised received and accepted 7 June 2011
The nanocomposite web of calcium alginate and polyvinyl alcohol with varying proportion has been prepared by
electrospinning and its application for wound healing is examined. Scanning electron microscope and Fourier transform
infrared spectroscopy have been used to characterize the morphology and molecular fingerprint. The water vapour
transmission rate of the blended nanofibre composite web having maximum calcium alginate content is found to be
2725.8 g/m
2
/2h, which helps in maintaining a moist environment over the wound surface. The wound healing capacity of the
nanofibre has been evaluated using a rat model. When the wound is covered with nanofibre, it forms gel due to the presence
of calcium alginate and filled with new epithelium without any harmful reactions.
Keywords: Calcium alginate, Nanofibre, Polyvinyl alcohol, Wound healing

1 Introduction
Electrospinning is a renowned technique for
generation of polymeric nanofibres that has very large
surface area to volume ratio
1
. Nanofibres obtained
from electrospinning has diverse applications in
nanocomposites such as scaffolding, biomedical and
drug delivery system
2
.
Polyvinyl alcohol (PVA) is a water soluble
polyhydroxy polymer, which has good mechanical
properties, chemical resistance and biological
properties. PVA can be cross-linked with biopolymers
to improve mechanical properties and anti-water
solubility
3,4
. PVA has been electrospun into
nanofibres with diameters ranging from 100nm to
1000 nm. Studies showed that lower concentration of
the solution will form fibres with beads and with
increment in solution concentration, the form changed
from beaded fibre to smooth and uniform fibres
5
.
Calcium alginate is an anionic linear
polysaccharide extracted from brown seaweed. It is
composed of 1-4 -L-Guluronic acid (G) and -D-
Mannuronic acid. Calcium alginate is biodegradable
and has a broad application in the field of textiles,
food and biomedical application. The alginates form
firmer structure with calcium ions, producing calcium
alginates which form stronger gels on wound surface
compared to that of sodium alginate
6
.
Wound healing will start from predictable sequence
of events that has a cascade effect until the wound is
bridged by a scar tissue. It is accepted that rapid
wound healing occurs in the moist environment and
the present wound care products provide these
conditions. A chronic wound fails to heal because the
excessive production of exudates causes maceration
of healthy skin tissue around the wound and prevents
wound healing
7,8
.
Many wound dressing materials are available and
they are being investigated. The use of hydrogel will
provide a rapid wound healing by creating a moist
wound surface that reduces the buildup of necrotic
tissue. Sodium alginate and polyvinyl alcohol based
hydrogel showed an improved healing rate of artificial
wounds in rats. Alginates are highly absorbent and gel
forming agents which has haemostatic properties. In
contact with the wound surface, the wound exudates
convert calcium into sodium salt, forms gel, and gets
absorbed by the body. The latter facilitates the
removal of alginate dressing by dissolution
9
.
Electrospinning of pure alginate solution is not
possible
10
. For polyvinyl alcohol, continuous fibres
were successfully electrospun from PVA solutions of
7, 8 and 9%. Blended solutions of polyvinyl alcohol
and sodium alginate have been electro spun into
nanofibres having 2 wt% sodium alginate and 7, 8 and
9 wt% polyvinyl alcohol. PVA and sodium alginate
solutions were blended in the ratios of 100:0, 70:30,
50:50 and 30:70. The results showed that, uniform
nanofibres were generated, when polyvinyl alcohol
___________________________
a
To whom all the correspondence should be addressed.
E-mail: gobsnn@gmail.com
INDIAN J. FIBRE TEXT. RES., JUNE 2012


128
proportion in the blend was increased
11
. The
maximum concentration of sodium alginate
electrospun was ~4 wt%
12
. Electrospinning of
polymer blends have combined properties of
individual polymer. For example, poor mechanical
performance of biopolymer is overcome by including
a synthetic polymer that has good mechanical
properties, biodegradability and biocompatibility
13
.
There have been studies about calcium alginate
hydrogels which shows poor mechanical properties
14
,
but no report has been found in the literature
regarding the blend of calcium alginate and PVA.
The present study is therefore undertaken to
prepare calcium alginate/PVA blended nanofibre
matrix for its use in wound dressing. The study has
been carried out in two parts. The first part involves
the examination of PVA and calcium alginate blend
while the second part involves study on the rate of
wound healing on the wound surface.

2 Materials and Methods
2.1 Materials
Calcium alginate [(C
6
H
7
Ca
1/2
O
6
)
n
, MW-1170.93
g/mol] was used for the study. Polyvinyl alcohol
[(C
4
H
6
O
2
)
n,
35-50 cps, 500g] was used to blend with
calcium alginate. Sodium carbonate [(Na
2
CO
3
),
MW- 105.99 g/mol, 500 g] was used as a solvent to
dissolve calcium alginate.

2.2 Methods
2.2.1 Electrospinning
Calcium alginate solutions were prepared through
mild stirring using sodium carbonate as a solvent to
dissolve calcium alginate. Since maximum
concentration of sodium alginate electrospun was
4 wt%, in this experiment also, calcium alginate
concentration was kept 4 wt%. On the other hand,
polyvinyl alcohol solution was prepared through
stirring using water as a solvent. The concentration of
polyvinyl alcohol was ~7 wt%. The separately
prepared PVA (7 wt %) and calcium alginate (4 wt %)
solutions were blended in different proportions
such as 90:10, 80:20 and 70:30 (PVA/calcium
alginate). The blended solutions were taken
individually and fed into 5 mL syringe fitted with a
needle tip. A DC voltage of 16-18 kV (Gamma
High Voltage Apparatus, USA) was applied
between the syringe tip and the collector covered
with aluminum foil. Droplet emerged from the tip
of the needle was split due to the repulsive force set
at the needle. During this process, the solution
evaporated and polymeric fibres deposited on the
aluminum foil in the form nonwoven fibrous mat.

2.2.2 Characterization
The morphology of the nanofibres was analyzed
using scanning electron microscope (HITACHI
S-3400 SEM) at a voltage of 10-15 kV. The diameter
of the nanofibres was determined at 5 different points.
Fourier transform infrared spectroscopy
(FTIR- TENSOR 27) was used to identify the change
in molecular structure of PVA and alginate. It was
also used to identify the structure of calcium alginate
and PVA blended nanofibre web. The spectra were
scanned between 500 cm
-1
and 4000 cm
-1
.

2.2.3 Moisture Vapour Transmission Rate and Porosity
Balakrishnan et al.
14
, determined the moisture
vapour permeability of a nanofibrous material by
measuring moisture vapour transmission rate (MVTR)
across the material. With the diameter of 35 mm, the
materials were mounted on the mouth of cylindrical
beakers containing 10 mL water with negligible water
vapour transmittance. The material was fastened using
a teflon tape across the edges to prevent any water
vapour loss across the boundary and finally the
assembly was kept at 37C and 35% relative humidity
in an incubator. The assembly was weighed at regular
intervals of time and weight loss versus time plot was
constructed. From the slope of the plot, MVTR was
calculated using the following formula:

WVTR =
Slope24
A
g/m
2
/day

where A is the test area of the sample in m
2
.
The water stability of the nanofibre matrix was
analyzed by dipping the nanofibre web in the water
and examining it by scanning electron microscopy.
The fibre length (L) in the piece of mat was calculated
using the following equation
14
:

L =
2
( /2)
m
d D


where m is the mass of the fibre in the piece of mat
(3 cm 3cm); d , the density of the fibre; and D , the
mean fibre diameter. Using the above linear
relationship, mean pore diameter was calculated
following the formula
15
as shown below:

MPD = -0.39 (FL/1000) + 2.02,
TARUN & GOBI:

CALCIUM ALGINATE/PVA BLENDED NANO FIBRE MATRIX FOR WOUND DRESSING


129
where MPD is the logarithm of mean pore diameter
(m); and FL , the logarithm of fibre length (m).

2.2.4 Antibacterial Activity
The antibacterial activity of nanofibre matrix was
determined by disc diffusion method. Nanofibrous
matrix having diameter of 35 mm was taken.
According to AATCC 147, 1.3 g of nutrient broth was
dissolved in 100 mL of distilled water that was taken
in the conical flask. The conical flask was plugged
with the cotton and then sterilized. After sterilization,
slant bacteria was put inside the conical flask and
plugged with cotton. The conical flask was kept inside
the incubator for 24 h at 37C. Another conical flask
with 100 mL distilled water was taken with 2.4 g of
agar and 1.3 g of nutrient broth. This flask was also
cotton plugged and sterilized. A negligible quantity of
nutrient was poured in the petri plate. A swab stick
was taken and dipped in the nutrient that contained
bacterial cells. Soon after dipping , it was swabbed in
the petri plate that contained the nutrient. The
nanofibrous matrix was placed in the prepared petri
plate and the antibacterial activity was analyzed from
the inhibition zone.

2.2.5 In-vivo Wound Healing
The wound healing characteristic of the calcium
alginate/PVA blended nanofibre was evaluated using
two rat models. Two partial thickness wounds of 1cm
length were prepared towards the upper side of the
organ separately in each rat model. Nanofibre
dressing was compared with control wound which
was left to the atmosphere to heal. All the
experiments with the rat model were preliminary and
it was performed with the approval of Institutes
Animal ethics committee. Female Wistar rats
weighing approximately 180 g were anesthetized by
intramuscular injection of Ketamine and Xylaxin, at a
dose of 40 mg/kg and 5 mg/kg. The hairs on the skin
of the animal were removed and disinfected using
70% ethanol. The wound size was photographed and
the length was measured using a scale. The wounds
were grossly examined for a period of 5 and 8 days.

3 Results and Discussion
3.1 Electrospinning of Blended Solution of 4% Calcium
Alginate and 7% PVA
PVA was used for blending with calcium alginate
solution to provide spinnability of blended solution
and to get better mechanical properties of the fibre.
Figure 1 shows SEM photographs of nanofibre web
that was obtained from the blended solution of
calcium alginate and PVA. At ratio of 70:30
(PVA/calcium alginate), fibres are seen along with
beads [Fig. 1(c)]. The average diameter of the fibre is
found to be 98.1 nm. The viscosity of the blended
solution is one of the perceived parameters that
affects the structure and diameter of the fibre. At ratio
of 80:20 (PVA/calcium alginate), fibres along with
beads are also seen [Fig. 1(b)]. But the beads are
reduced, when it is compared with the nanofibres
spun from the ratio of 70:30 (PVA/calcium alginate).
The average diameter of the fibre is found to be
113.6 nm. At ratio of 90:10 (PVA/calcium alginate),


Fig.1 SEM images of electrospun nanofibres prepared from
solutions of 4% calcium alginate and 7% PVA at different volume
ratios of PVA: calcium alginate (a) 90:10, (b) 80:20, and (c) 70:30
INDIAN J. FIBRE TEXT. RES., JUNE 2012


130
uniform fibres are seen [Fig 1(a)], but beads are seen
only in few places. The average diameter of the
nanofibres is 191.5 nm. It is inferred that the diameter
of the nanofibres decreases as the calcium alginate
proportion in the blend increases.

3.2 FTIR Spectral Study
Fourier transform infrared was used to observe
peak shift caused due to the interactions between two
blended polymers such as hydrogen bonding or any
complex structure formation. The FTIR spectra show
spectral features similar to those for individual
polymers, but some bands shifts from their original
positions. Hydrogen bonding is formed between the
proton donor and proton acceptor molecules. The
intensity of the hydrogen bonding depends upon the
acidity of the hydrogen in proton-donor, alkalinity of
the proton-acceptor, and distance between the groups.
Due to the hydrogen bond formation, the covalent
bonds in donor and acceptor become weaker. Figure 2
shows FTIR spectra of PVA and calcium alginate.
Figure 3 shows FTIR spectra of calcium alginate
and PVA blended nanofibres in the wavelength ranges
4000 500 cm
-1
. Figure shows the characteristic
bands of PVA and calcium alginate at 3418 cm
-1
and
3347 cm
-1
which are observed to be OH stretching
vibration brands. It is evident that the bands appeared
at the region 3400 cm
-1
belong to all types of
hydrogen bond. It is observed from Figs 3(a)-(c) that -
OH stretching has shifted to lower frequencies
compared to pure calcium alginate [Fig. 2(a)].
Hydroxyl stretching frequency becomes slightly lower
with the increase in calcium alginate content. It is
evident that OH stretching has a favorable extent in
changing the molecular spectra.

3.3 Rate of Moisture Transmission and Porosity Measurement
An ideal wound dressing should control the water
loss from a wound at an optimal rate. It is found that
the loss of water increases linearly with time. MVTR
of the nanofibre web that is produced from three
different proportions is calculated as the gradient of
the weight loss versus time plot. For the first two
hours, the weight loss in 90:10, 80:20 and 70:30
(PVA/Calcium alginate) blend is found to be 306.6,
259.4 and 224.5 g/m
2
/2h respectively. Subsequently,
the weight loss in the three samples after 24 h is found
to be approximately 3312.5, 3019.5 and
2725.8 g/m
2
/2h respectively. It is concluded that
MVTR decreases as the calcium alginate content in
the nanofibre increases.
Figure 4 shows the SEM image of the blended
nanofibre matrix that was water dipped. It is found
that calcium alginate is not degraded because of the
water insoluble nature but PVA is degraded because
of its solubility in the water. Calcium alginate appears
like beads due to its insoluble nature in the water.
Table 1 shows that pore diameter decreases negligibly
as the average fibre diameter decreases.


Fig. 2 FTIR spectra of (a) calcium alginate, and (b) polyvinyl
alcohol


Fig. 3 FTIR spectra of calcium alginate and PVA blended
nanofibres (a) 90:10, (b) 80:20, and (c) 70:30 PVA/calcium
alginate
TARUN & GOBI:

CALCIUM ALGINATE/PVA BLENDED NANO FIBRE MATRIX FOR WOUND DRESSING


131
3.4 Antibacterial Test
The microbial suspension was spread evenly over
the face of the sterile agar plate. The nanofibre matrix
along with the control sample was applied on the agar
plate and incubated. After 24 h time, zone of
inhibition appears around the test product. The zone
of inhibition was measured from the area on the agar
plate that remains free from microbial growth.
Johnson and Johson bandage was used as control
wound. Nanofibre matrix prepared from 4 % calcium
alginate and 7% polyvinyl alcohol at 90:10, 80:20 and
70:30 (PVA/Calcium alginate) proportions were used
to determine the antibacterial activity in
Staphylococcus aureus. Figure 5 shows that the area
of zone formed in control wound is 10 mm; at 90:10
proportions, the area of the zone formed is 10mm. In
80:20, the area of the zone formed is 12 mm and in
70:30, the area of the zone formed is 20 mm. It is
found that the increase in calcium alginate content
shows higher zone of inhibition value and that the
zone of inhibition value for 70:30 and 80:20 is higher
as compared to the control wound.

3.5 Gross Examination
One centimeter length wound was incised on the
female Wistar rats. Grossly each wound was observed
within 5 and 8 days. Wound was covered with the
blended nanofibre matrix of 70:30 proportion and the
dimension of the web was 1cm length and 2 cm wide.
At the 5 days, epithelial cells appeared on both the
control wound and test wound. The difference was the
healing rate. Figure 6 (c) shows that the control
wound remains open and the length is reduced to
0.7 cm. The test wound is partially covered with the
gel and the length of the wound is found to be 0.5 cm
[Fig. 6(d)]. The 8 days examination of both control and
test wound is shown in Fig. 6(e) and 6(d). Figure 6(e)


Fig. 4 SEM image of blended nanofibre matrix after treatment
in water
Table 1 Determined pore size of the blended electrospun
nanofibre mats

Solution ratio
(PVA: calcium
alginate)
Average fibre
diameter, nm
Areal
density
g/m
2

Mean pore
diameter
m

90:10 191.5 10 10.41
80:20 113.6 20 10.39
70:30 98.1 30 10.38


Fig. 6 Examination of nanofibre matrix on wound healing (a),
(c) and (e) control wounds, and (b), (d) and (f) test wounds


Fig. 5 Assessment of antibacterial activity in nanofibre matrix
prepared from blended solution of 4% calcium alginate and 7%
polyvinyl alcohol
INDIAN J. FIBRE TEXT. RES., JUNE 2012


132
shows that control wound is found to be open and
epithelial cells are grown well as compared to the
5 days. But, in case of test wound [Fig. 6(f)],
epithelial cells have grown well and it has covered the
wound. Hairs are also grown in both control as well as
test rat.

4 Conclusion
PVA/calcium alginate blend in different
proportions has been used to produce electrospun web
for wound dressing and it is found that the fibre
diameter varies from 191.5nm to 98.1 nm. The
increase in concentration of calcium alginate reduces
the size of nanofibre and increases the number of
beads per unit area. The calcium alginate and PVA
crosslinking has been analyzed using FTIR studies
and the results reveal that there is change in the
intensity of transmission at OH groups with respect to
the proportions. The mean pore diameter of the
electrospun web has also been calculated and the
values are found to be similar in all cases. The
increase in concentration of calcium alginate reduces
the MVTR due to the absorption of water by the
calcium alginate. The antibacterial activity against
staphylococcus aureus gradually increases while
increasing the concentration of calcium alginate.
In-vivo results show that the nanofibre matrix has the
capacity to heal the wound. The experiment suggests
that the blended polymer nanofibre bandages can be
produced by electro spinning technique
commercially.
Acknowledgement
The authors acknowledge UGC-DRS-Phase I
program for providing the instrumental facilities to
carrying out this work.

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