Biomedicine & Aging Pathology 4 (2014) 59–64

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Original article

Application of chitosan and herbal nanocomposites to develop

antibacterial medical textile
S. Chandrasekar a , S. Vijayakumar a,∗ , R. Rajendran b
a
PG and Research Department of Botany and Microbiology, A. V. V. M. Sri Pushpam College (Autonomous), Poondi, Thanjavur, Tamil Nadu, India
b
Professor and Principal, PG and Research, Department of Microbiology, PSG College of Arts and Science, Coimbatore, Tamil Nadu, India

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

Article history: Aim: To enhance the efficiency of biological, chemical and physical properties like antibacterial activ-
Received 10 October 2013 ity, wash durability, air-permeability and biocompatibility, cotton fabric materials were finished with
Accepted 18 October 2013 chitosan and herbal nanocomposites.
Available online 20 November 2013
Methodology: Extracts of Senna auriculata and Achyranthes aspera with chitosan solution was bulk finished
on 40 cotton fabrics. To increase the functional properties, chitosan and herbal extract nanocomposites
Keywords: were finished on to another set of fabrics (nanocomposite finishing). Different functional properties
Chitosan
were carried out for both the sets of fabrics and comparatively analyzed. Antibacterial activity was
Coacervation
Biocompatible
carried out using standard EN ISO 20645 method and durability of finished fabrics was done using a
Nanoparticles standard AATCC–124 test method. Physical properties like tensile strength, abrasion resistance and air-
Air-permeability permeability were analyzed using the standards, ASTM D 5035-2006, AATCC 119-2004 and ASTM D
737-1996 respectively. Biocompatible properties of the finished fabric were determined by HET-CAM to
ensure that material do not cause irritation points on skin surface.
Results: Antibacterial activity of nanocomposite finished fabrics showed more inhibitory zones of 31 mm
for E. coli and 29 mm for S. aureus when compared to bulk finished fabrics. Nanocomposite finished fabrics
showed good durable properties and physical properties than bulk finished fabrics. Bulk and nanocom-
posite finished fabrics showed good biocompatible properties when analyzed using a standard HET-CAM
test.
Conclusion: The study concludes that, nanocomposites could provide better functional properties than
the bulk finished fabrics. The nano-sized particles in the composites was considered significant for its
functional applications in hospital based fabrics to prevent the transmission of nosocomial infections.
© 2013 Elsevier Masson SAS. All rights reserved.

1. Introduction infection by pathogenic microorganisms, especially bacteria such

Textiles for medical and hygienic use have become important
areas in the textile industry. Antibacterial textile production has
become increasingly prominent for hygienic and medical appli-
cations. The application of antimicrobial finishes to textiles can
prevent bacterial growth on textiles [1]. According to [2], dramatic
effects like deterioration, defacement and odours, which occur from
the microbial contamination of surfaces varied as carpeting and
medical non-woven fabrics. Purwar and Joshi [3] explained that
these surfaces can also act as a microbial “harbour”, as most offer
ideal environments for the proliferation of microorganisms that are
harmful to buildings, textiles and humans. The antibacterial mate-
rials such as fabrics, clothes are becaming important to avoid cross
∗ Corresponding author.
E-mail addresses: svijaya kumar2579@rediff.com (S. Chandrasekar),
svijaya kumar2579@rediff.com (S. Vijayakumar).
as Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae,
to control the infestation by microbes, and to arrest metabolism
in microbes in order to reduce the formation odour [1]. The ability
to make surfaces resistant to microbial contamination has advan-
tages in many applications and market segments. Krueger [4] also
confirmed that this is especially true in medical markets where
many products have contributed a degree of aseptic sophistication
beyond that required of consumer products.
As consumers have become more aware of hygiene and poten-
tially harmful effects of microbes, the demand for antimicrobial
finished clothing is increasing [5]. In the development of fab-
rics, functional aspects such as anti-bacterial and UV protection
are playing an increased important role [6]. Different chemi-
cals and heavy metal finished fabrics even though provide good
antimicrobial activity still due to factors like toxicity, and non-
biodegradability, the compounds cannot be used for medical
applications. Hence, natural polymers like chitosan and herbal
extract finished fabrics were considered significant for fulfilling

2210-5220/$ – see front matter © 2013 Elsevier Masson SAS. All rights reserved.
http://dx.doi.org/10.1016/j.biomag.2013.10.007
60 S. Chandrasekar et al. / Biomedicine & Aging Pathology 4 (2014) 59–64
these objectives. Chitosan derived from acetylation of chitin, the
marine polymer was a best alternative of heavy metals for finishing
medical fabrics. Chitosan is a natural, non-toxic, microbial resis-
tant biodegradable polymer. Its derivatives as antimicrobial agents
have received more attention to finish antimicrobial textiles. Simi-
lar effects from the extracts of medicinal plants like Senna auriculata
and Achyranthes aspera were also considered significant for the
functional finishing of textile materials for antibacterial, antifungal
and anti-odor properties.
Even though reactive exhaust method and microencapsula-
tion method have been used extensively in textile industries for
functional finishing of fabrics, a novel technique called nanoencap-
sulation is rapidly emerging and widely used in pharmaceutical,
chemical, cosmetics and food processing industries [7]. More recent
years, the method was also expanded to textile finishing industries
due to its significant binding properties on cellulosic substrates.
Nanoencapsulated drugs after finishing onto the textile materials
provides a slow and controlled release of the active antimicrobial
ingredient to achieve the desired delay until the right stimulus is
obtained [8]. Nanocomposites are formed by the combination of
two or more materials that have quite different properties. These
different materials work together to give the unique properties of
the composite, which is the materials’ individual properties [9].
Considering the significant characteristics of this technique, in our
present research, chitosan and herbal extract nanoparticles were
prepared and were functionally finished on the cotton fabrics. To
find out the efficiency of nanocomposite finished cotton fabrics, the
parameters like antibacterial activity, wash durability, biocompat-
ibility, and physical properties were compared with chitosan and
herbal extracts (bulk) finished cotton fabrics.
2. Materials and methods
The entire research work was carried out from February 2012
to March 2013.
2.1. Materials used in the study
Fabric material selected for the study was plain weave 40s med-
ical grade cotton with 60 ends per inch (EPI) and 56 picks per inch
(PPI).
2.2. Medicinal plants and chitosan
Two medicinal plants, Senna auriculata, Achyranthes aspera and a
natural polymer chitosan were collected from Department of Plant
Science, Tamil Nadu Agricultural University, Coimbatore, India.
Leaves of the plants were used in the study.
2.3. Test bacterial cultures
The test cultures, Escherichia coli and Staphylococcus aureus used
in the study, were the significant pathogens isolated from the
wound dressing fabric materials of a diabetic foot.
2.4. Preparation of antibacterial agents
2.4.1. Methanol extracts of medicinal plants (Thilagavathi and
Kannaian, 2008)
Fresh leaves of Senna auriculata and Achyranthes aspera was
shadow dried at 37 ◦ C. Drying was done to reduce the moisture con-
tent of leaves to less than 20%. Dried leaves were grounded to make
fine powder for the extraction of desired materials. Fine powdered
material was extracted to obtain the active substances with suitable
solvent (methanol). Ten grams of powdered leaves were extracted
in 100 mL of 80% methanol for 18 hours under shaking condition.
For every 6 hours the solution was sonicated for 20 minutes to
obtain the exact antibacterial substances of the medicinal plants
[10].
2.5. Preparation of Chitosan solution (Rajendran et al., 2012)
Chitosan (1%) solution was prepared by mixing with acetic acid
(1%) and stirred in a magnetic stirrer at 60 ◦ C till a fine homogenous
suspension was formed. The polymer solution was kept overnight
at stand still condition to remove air bubbles formed during stirring
[9].
2.6. Preparation of antibacterial nanoparticles
2.6.1. Herbal extract nanoparticles (Sumithra and Vasugi Raaja,
2012)
Herbal extract nanoparticles were prepared by coacervation
process [11] by cross-linking with glutaraldehyde. In this method,
the herbal extract was incubated with bovine serum albumin (wall
material – 2% w/v) for one hour at room temperature. Using 1 M HCl
pH was adjusted to 5.5. Ethanol was added to the solution in
the ratio of 2:1 (v/v) at the rate of 1 mL/min. Coacervate thus
formed was hardened with 25% glutaraldehyde for 2 hours to allow
cross-linking of protein. Organic solvents were removed by rotary
vacuum evaporator and resultant nanocapsules were purified by
centrifugation at 4 ◦ C at 10 000 rpm. Pellets were suspended in
0.1 M PBS (pH 7.4) and lyophilized with mannitol (2% w/v).
2.6.2. Chitosan nanoparticles (Rajendran et al., 2012)
Chitosan nanoparticles were prepared by emulsion method [9].
Briefly, 100 mL of 1.5% of tripolyphosphate solution was added into
chitosan solution (100 mL of 2 mg/mL in 1% diluted acetic acid) at
a rate of 10 mL/min under constant stirring condition till a milky
emulsion was obtained (pH −5.0). The emulsion was frozen at −4 ◦ C
followed by thawing in the atmosphere to get solid nanoparticles.
The emulsion was centrifuged at 10 000 X for 30 min. Finally, the
deposited nanoparticles were washed, vacuum dried at 60 ◦ C for
18 hours and stored at 4 ◦ C.
2.7. Bulk and nanocomposite finishing of cotton fabrics
Two sets of fabric samples were used in this study. In the first set
of samples, the prepared antibacterial agents were directly finished
(bulk finishing) at a ratio of 1:1 (herbal extract: chitosan). In the
second set of fabric samples, herbal and chitosan nanoparticles at
a ratio of 1:1 were used (nanocomposite finishing). All the samples
were padded with 8% citric acid in a padding mangle at a pressure of
3 psi with 100% wet pickup followed by drying and curing at 160 ◦ C
for 5 min.
2.8. Comparative analysis of bulk and nanocomposite finished
cotton fabrics
Fabrics finished with antibacterial agents and with antibacterial
nanoparticles were compared for different biological and physical
properties.
2.9. Antibacterial assessment of the finished fabric (EN ISO 20645
test method)
The antibacterial activity of two sets of finished fabric (bulk
and nanocomposite) was tested according to EN ISO 20645 [12]
against the test bacterial cultures, Escherichia coli and Staphylo-
coccus aureus. The finished cotton fabric with the diameter of
20 ± 1 mm was placed on the surface of Nutrient agar medium,
which was swabbed with the bacterial cultures. The plates were
 S. Chandrasekar et al. / Biomedicine & Aging Pathology 4 (2014) 59–64 61

incubated at 37 ◦ C for 24 hours to measure the zone of inhibition in Table 1

Antibacterial assessment of the finished fabric by EN ISO 20645.
millimeters formed around the fabric.
Test culture Zone of inhibition (mm)
2.10. Wash durability of finished fabric (AATCC–124 test method) Escherichia colib Staphylococcus
aureusb
Bulk and nanocomposite finished cotton fabrics were analyzed Unfinished fabric 0 0
for their wash durability by subjecting the sample to repeated Chitosan finished 21.3 22.9
washing and antibacterial testing using the standard AATCC-124 Herbal finished fabric 21.6 21.3
[13] and AATCC–147 [14] test methods (Parallel streak method). Chitosan+herbal (bulk) 23.3 22.6
finished fabrica
All the samples were washed and its antibacterial activity was ana-
Nanocomposite finished 31 29
lyzed after 1st, 5th and 10th wash. fabrica
a
Values in mm was measured including the diameter size of the fabric (20 mm).
2.11. Physical properties of finished cotton fabrics b
Mean values were tabulated after performing three times for each test culture.

Difference in the physical properties of two sets of finished

appearance of vessel lysis, coagulation time = time (in seconds) of
fabrics was analyzed with untreated control cotton samples. Four
first appearance of protein coagulation.
significant parameters, which were considered for comfort prop-
erties of cotton fabric were selected for the study. Tensile strength
3. Results and discussion
is the measure of the resistance of the fabric tensile load or stress
in either warp or weft direction. Abrasion test was determined the
In the present study, bulk finished and nanocomposite fin-
ability of a fabric to withstand damage by friction. Air permeabil-
ished cotton fabrics were analyzed to determine their efficiency
ity of a fabric is the volume of air measured in cubic cm passed
of biological and physical properties. Antibacterial activity and the
per second through 1 sq.cm for the fabric at a pressure of one cm.
durable properties of the nanocomposite finished cotton fabrics
head of water. Also the difference in the weight of the finished
were analyzed along with physical properties like air permeability
fabrics was measured to determine the presence of antibacterial
and abrasion resistance. Biocompatible nature of the antibacterial
agents. Tensile strength, abrasion resistance and air-permeability
agents was also analyzed to determine the regular use of the fin-
were analyzed using the standards, ASTM D 5035-2006, AATCC
ished fabric for hospital workers and patients.
119-2004 and ASTM D 737-1996 respectively.

3.1. Antibacterial assessment of the finished fabric by EN

2.12. Biocompatible properties of finished fabrics
ISO 20645

2.12.1. HET-CAM Test [NIH Publication No. 06-4515 (2006)]

Two sets of finished fabric were assessed for their antibacterial
HET-CAM (Hen’s egg test-chorio allantoic membrane) method
activity by EN ISO 20645 against test bacterial cultures. The zone of
uses the vascular fetal membrane of chicken embryos. It is assumed
inhibition for the first set of bulk-finished fabric was 23.3 mm and
that acute effects induced by a test substance and the small blood
22.6 mm for E.coli and S. aureus respectively (Table 1). Second set of
vessels and proteins of this soft tissue membrane are similar to
fabric treated with nanocomposites showed more inhibitory zones
effects induced by the same test substance in the skin of a treated
than the first set of finished fabric against same test bacterial cul-
rabbit. The membrane was evaluated for the development of irri-
tures. Nanocomposite finished fabric inhibited the organisms with
tant endpoints (vascular lysis, hemorrhage and coagulation) and
the zones of 31 mm for E. coli and 29 mm for S. aureus (Fig. 1). The
qualitative assessments of the irritation potential of test substances
measured zone of inhibition thus indicated that nanocomposites
are made. All test materials, finished and untreated fabric samples
not only prevented the growth under the fabric also it constantly
were evaluated undiluted. A 0.9% NaCl negative control should be
leached out from the material by restricting the growth of orga-
included in each experiment in order to provide a baseline for the
nisms to a greater extent than the first set of finished fabric.
assay endpoints and to ensure that the assay conditions do not in
appropriately result in an irritant response and 0.1 N NaOH was
3.2. Wash durability of finished fabrics (AATCC–124 test method)
used as positive control. Preparation and treatment of CAM with
test materials was done using the method described by Valdes
Difference in the durable properties between two sets of
et al. (2001) [15]. Measuring the irritation scores with the standard
finished fabric was analyzed by repeated industrial washings.
formula was presented in the catalogue of NIH Publication No. 06-
4515 (2006) [16].

2.12.2. Irritation Score (IS) calculation – IS [B] analysis method

The time taken for the development of each endpoints,
hyperemia, hemorrhage and coagulation was substituted in the
standard formula as per IS [B] analysis method. The time values
assigned/obtained to each endpoint were totalled to give an over-
all IS value for the test substance. An IS score could be calculated
using the following general formula:
 301 − Hemorrhage time    301 − Lysis time  
×5 + ×7
300 300
 301 − Coagulation time  
+ ×9
300
where, hemorrhage time = time (in seconds) of the first appearance Fig. 1. Antibacterial assessment of the nanocomposite finished fabric. Assessment
of blood hemorrhages, lysis time = time (in seconds) of the first was made by EN ISO 20645 against two test bacterial cultures E. coli and S. aureus.
62 S. Chandrasekar et al. / Biomedicine & Aging Pathology 4 (2014) 59–64
Table 2
Wash durability of finished fabric by AATCC–124 test method.
Type of fabric Zone of inhibition (mm) after washes (in numbers)
Escherichia coli#
1st 5th
Unfinished fabric 25# 25#
Chitosan finished 26.3 26.3
Herbal finished 26.9 26.3
Chitosan + herbal finished 27.3 25.3
Nanocomposite finished 27.9 26.9
#
No inhibitory zone was observed (value given is the actual diameter of the fabric).
Durability was tested based on their antibacterial activity using
standard Parallel streak method (AATCC–147 test method). The
set of bulk-finished fabric showed inhibitory zones of 27.3 mm,
25.3 mm and 25.3 mm for E. coli and 27.3 mm, 26.3 mm and
26.3 mm for S. aureus after 1st, 5th and 10th wash respec-
tively (Table 2). Nanocomposite finished samples provided more
inhibitory zones than the bulk finished fabric. This was evi-
dent after 10 washes, reported that 26.6 mm and 26.3 mm of
inhibitory zones were observed for E. coli and S. aureus respec-
tively (Fig. 2) Nanocomposite fabric thus provided evidence for long
term durability by retaining their antibacterial activity even after
10 industrial washes.
Nano-sized particles of chitosan and herbal extracts played a
vital role in providing the durability in finished fabrics. Nanosized
particles due to their low concentration also considered signifi-
cant in reducing the colour in finished fabrics. The colour of plant
extracts was a widely-met problem in the applications of textile
because treated cotton often change to green colour since higher
concentration of crude extracts were used for finishing the fabrics.
In the present study, nanocomposites containing low concentration
of antibacterial agents were finished which provided more antibac-
terial activity with greater durable properties. Durability of fabric
treated with nanoparticles was mainly due to their size of nature
in which they are present in the cotton fabric. The nanosized par-
ticles embedded easily within the cellulose moieties of cotton, so
that it remains constant and released at low concentrations, which
ultimately required for inhibiting the growth of organisms.
3.3. Physical properties of finished cotton fabrics
The physical properties of the nanocomposite finished fabric
and bulk finished fabric was compared with untreated control
cotton fabric in Table 3. The tensile strength of nanocomposite fin-
ished cotton fabrics was found to be similar to untreated control
Fig. 2. Wash durability of nanocomposite fabric. Durable properties was deter-
mined after antibacterial assessment against E. coli and S. aureus. Assessment was
made using AATCC–147 parallel streak method. 5th wash fabric was presented in
the figure.
Staphylococcus aureus#
10th 1st 5th 10th
25# 25# 25# 25#
25# 27.3 26.3 25.3
25# 26.3 26.3 25#
25.3 27.3 26.3 26.3
26.6 28.3 26.9 26.3
cotton fabrics. Due to nanosized particles the nanocomposite fin-
ished fabrics does not vary with the tensile property of untreated
control cotton fabrics. Whereas tensile strength of bulk finished
fabric showed slightly higher tensile strength to the tune of 3.65% as
compared to that of untreated cotton fabrics. This was mainly due to
the larger size and concentration of the particles that was finished
in the fabrics. No change in the abrasion resistance was detected
for both the finished fabrics when compared to the untreated
cotton after 5 hours at 18 000 rpm. Similarly, no significant differ-
ence in the weight between the nanocomposite finished fabric and
untreated control cotton fabric was detected. After finishing with
either bulk particles or nanocomposites, addition of antibacterial
agents on each fabric samples was calculated. The weight of the
samples was measured before and after finishing individually with
bulk particles and nanocomposites. Bulk finished samples showed
more fabric weight than the nanocomposite finished fabric sam-
ples. In this regard, the bulk finished samples showed 3.08 and
2.27% more weight than unfinished and nanocomposite finished
samples respectively. Whereas, nanocomposite finished samples
showed only 0.82% more weight than the unfinished samples. The
results indicated that nanocomposite finished fabric may strongly
influence the comfort properties of the wearer.
Air-permeability, which ultimately tested to decide the com-
fort properties showed interesting phenomenon of differences
between bulk and nanocomposite finishes. Bulk finished fabrics
showed relatively low air-permeability than the nanocompos-
ite finished fabrics. Air-permeability of bulk finished fabric was
102 cm3 /cm2 /s which was 6.27% less than that of untreated control
cotton (95.6 cm3 /cm2 /s). Whereas the nanocomposite finished fab-
ric was measured as 96.6 cm3 /cm2 /s which was only 1.03% less than
the untreated control samples, ensuring more air-permeability and
also influencing the comfort property of the fabric. This may be due
to the particle size which was considered to be in nano size; that
may not block the pores of fabrics. But the pores in bulk finished
fabric may get blocked due to the size of chitosan and herbal parti-
cles, which in turn leads to low-permeability of air thus decreasing
the comfort properties of the fabric.
3.4. Biocompatible properties of finished fabrics
In the present research the initial irritation scores of the CAM
implanted test materials was calculated with the reference of
positive (0.1 N NaOH) and negative controls (0.9% NaCl). The devel-
opment of endpoints (haemorrhage, hyperemia and coagulation)
as demonstrated by Luepke (1985) [16] was analysed for the CAM
samples of test material, negative and positive control samples. A
variety of endpoints were evaluated in the HET-CAM test method
protocols reviewed after the original protocol described by [16].
Budai and Varnagy [17] observed other endpoints that were eval-
uated to assess the irritancy potential of test substances, which
included injection (mild haemorrhage), vasoconstriction (narrow-
ing of the vessels), dilation (expansion of the vessels), and lysis
(disintegration of the vessels).
 S. Chandrasekar et al. / Biomedicine & Aging Pathology 4 (2014) 59–64 63

Table 3
Physical properties of finished cotton fabrics.

Physical properties Bulk finished fabric Nano- composite finished fabric Untreated control cotton

Tensile strength 35.6 kgf 34.6 kgf 34.3 kgf

Resistance to abrasion No breakdown of the No breakdown of the specimen up No breakdown of the
specimen up to 18 000 rpm to 18 000 rpm for 5 hours specimen up to 18 000 rpm
for 5 hours for 5 hours
Fabric weight g/sq/m (GSM) 74.6 g/2 m 72.9 g/2 m 72.3 g/m2
Air permeability 102 cm3 /cm2 /s 96.6 cm3 /cm2 /s 95.6 cm3 /cm2 /s

All the tests were tested in triplicates, mean values were tabulated.

This protocol was used to evaluate the anti-inflammatory and Table 4

Comparative evaluation of irritation scores for test materials, negative control and
anti-angiogenic properties of various test substances. Therefore,
positive control by HET-CAM test.
the endpoint selected for this test method protocol was likely
unique for these effects. In the present study, no such endpoints Materials on CAM Mean value of Irritation
endpoint development scoreb
were detected on the CAM of any of the test materials. This obser-
vation was confirmed from the endpoint obtained for the CAM Hma Hya Cga
samples of negative control (Fig. 3a) and control sample (untreated Sample-1 (negative control) 0 0 0 0
fabric) (Fig. 3b). Since no endpoints were detected, the initial Sample-2 (untreated fabric) 0 0 0 0
irritation score was considered to be 0 for all the test materi- Sample-3 (treated fabric-1)d 0 0 0 0
als and negative control samples. Direct examination of the CAM Sample-4 (treated fabric-2)d 0 0 0 0
Sample-5 (positive control) 5.9 6.7 6.3 18.9c
presented in Fig. 3c and d for bulk finished and nanocomposite
finished fabrics showed no specified irritant endpoints like hyper- Hm- hemorrhage, Hy-hyperemia, Cg-coagulation.
a
Mean values of time until development of identified endpoint.
emia, hemorrhage, and coagulation. The positive control scores b
Irritation score calculated as described by IS [B] analysis
were calculated using the time taken for the development of iden- c
Irritation category – severe irritation.
tified endpoints. For the positive control samples, the mean value d
Treated fabric-1: normal sized particle finished fabric, treated fabric-2: nano-
sized particle finished fabric.

of time for the development of hemorrhage, hyperemia and coag-

ulation were identified as 5.9, 6.7 and 6.3 respectively (Fig. 3e). The
irritation score of positive control samples were thus calculated as
18.9. The IS of test materials, positive and negative controls was
presented in Table 4. From the obtained values, it was proved that
bulk and nanocomposite finished fabrics were found to be good
biocompatible in nature.

4. Conclusion

In the present study, the advantages of functionally finished

nanoparticles on the medical cotton were well determined based
on the biological, chemical and physical properties. The nano size of
chitosan and herbal extracts increases the durability and antibac-
terial activity of finished fabric to a greater extend. Also physical
properties of nanoparticle-finished cotton could also influence its
wide applications in the hospitals for the workers and patients in
providing suitable comfort properties.

Disclosure of interest

The authors declare that they have no conflicts of interest con-

cerning this article.

Acknowledgement

The authors are thankful to the Management of A.V.V.M. Sri

Fig. 3. HET-CAM Test [NIH Publication No. 06-4515 (2006)]. a: negative control; b:
untreated fabric. Well-developed blood vessels without irritation points. c: finished
fabric-1; d: finished fabric-2. No irritation points observed for the finished fabric.
Biocompatible properties assessed using HET-CAM test compared to positive control
no irritation endpoints observed for finished fabrics. e: positive control (0.1 N NaOH).
All three endpoints observed.
pushpam College (Autonomous), Poondi, for providing them nec-
essary facilities and support to carry out this work and RndBio,
Biosolution Company at Coimbatore for the preparation of chitosan
and herbal extract nanocomposites and carrying out biocompati-
bility tests.
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