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International Journal of Biological Macromolecules 98 (2017) 366378

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International Journal of Biological Macromolecules


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

Investigation on Curcumin nanocomposite for wound dressing


G. Devanand Venkatasubbu , T. Anusuya
Department of Nanotechnology, SRM University, Kattankulathur, Tamil Nadu, India

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

Article history: Curcuma longa (turmeric) has a long history of use in medicine as a treatment for inammatory condi-
Received 16 November 2016 tions. The primary active constituent of turmeric and the one responsible for its vibrant yellow color is
Received in revised form 24 January 2017 curcumin. Curcumin is used for treatment of wound and inammation. It had antimicrobial and antiox-
Accepted 1 February 2017
idant property. It has low intrinsic toxicity and magnicent properties like with comparatively lesser
Available online 3 February 2017
side-effects. Cotton cloth is one of the most successful wound dressings which utilize the intrinsic prop-
erties of cotton bers. Modern wound dressings, however, require other properties such as antibacterial
Keywords:
and moisture maintaining capabilities. In this study, conventional cotton cloth was coated with Curcumin
Curcumin
Wound dressing
composite for achieving modern wound dressing properties. Curcumin nanocomposite is characterized.
Antibacterial activity The results show that coated cotton cloth with Curcumin nanocomposite has increased drying time (74%)
Composite and water absorbency (50%). Furthermore, they show antibacterial efciency against bacterial species
Water holding time present in wounds.
2017 Elsevier B.V. All rights reserved.

1. Introduction functions including safeguard against external physical, chemical


and biological attackers. It serves as a protective barrier to external
Curcumin, 1,7-bis-(4-hydroxy-3-methoxyphenyl)-1,6-hepta- noxious agents including microorganisms. It prevents the excess
diene-2,5-dione (Fig. 1) is a yellow coloured principle phenolic pig- loss of water from the body and a role in thermoregulation. The
ment obtained from turmeric, the powdered rhizome of Curcuma composition of the skin is intricate, involving multi-faceted layers
longa Linn. (Family: Zinziberaceae). The turmeric (Curcuma longa) responsible for various functions. There are two major components
plant, a perennial herb belonging to the ginger family, is cultivated of the skin: the epidermis and the dermis. The epidermis, covered
extensively in south and southeast tropical Asia. The rhizome of with keratin, is responsible for the physical, biochemical/chemical
this plant is also referred to as the root and is the most useful and adaptive immunological barrier functions. This protective bar-
part of the plant for culinary and medicinal purposes. The most rier could be disrupted due to injuries or wounds. Injuries may
active component of turmeric is curcumin, which makes up 2 to 5% occur due to physical, chemical, thermal, microbial and immuno-
of the spice. The characteristic yellow color of turmeric is due to logical injury to the body. Wound is determined as the disruption
the curcuminoids.It has effective therapeutic properties not only of cellular and anatomical continuity of the tissue. Wound results
as an anti-inammatory drug, but also as a chemo-preventive, in the loss of continuity of epithelium in the skin with or with-
chemotherapeutic, anti-oxidant, antiamyloid, antiarthritic, anti- out the loss of underlying connective tissue. Some of these forms
HIV, antimicrobial and thrombosuppressive agent. In addition, of injuries can turn into chronic wounds, which cause long-term
curcumin is being used in the treatment of cystic brosis and agony and complications in many patients, as a result of the inabil-
Alzheimer disease. Curcumin is not toxic and safe. One of the most
important factors considered to be responsible for all the activity
of curcumin is its ability to scavenge reactive oxygen and nitrogen
free radicals [18].
Skin is one of the largest organs in human body, accounting for
about 15% of the total body weight. It carries out many essential

Corresponding Author: Department of Nanotechnology, SRM University, Kat-


tankulathur - 603 203, Kancheepuram District, Tamil Nadu, India.
E-mail address: gdevanandvenkatasubbu@gmail.com (G.D. Venkatasubbu). Fig 1. Structure of Curcumin.

http://dx.doi.org/10.1016/j.ijbiomac.2017.02.002
0141-8130/ 2017 Elsevier B.V. All rights reserved.
G.D. Venkatasubbu, T. Anusuya / International Journal of Biological Macromolecules 98 (2017) 366378 367

ity of the body to properly heal itself. Dermal reparation is an microorganisms at the injury site can disrupt the healing processes
elaborate process entailing the precise synchronization of countless in a variety of ways. Thus, preventative measures for inhibiting the
components of the skin to x damaged tissues, restore protective growth of harmful microorganisms can be an effective and straight-
barrier function, and re-establish the homeostatic equilibrium state forward course of action to help remove all the possible disruptions
of the wound area [911]. that bacteria may produce [21,22].
Wound healing is a complicated procedure involving a combina- Nanoparticles have signicantly different characteristics. They
tion of activities of different tissues and cell lineages and has been have been applied in a wide variety of medical research. Hydrox-
the subject of concentrated research for a long time. It is a highly yapatite nanoparticles are used as bone replacement materials
controlled process characterized by four distinct but overlapping [23,24]. They are also used as drug delivery systems [25]. Hydrox-
phases: hemostasis, inammation, proliferation and remodeling. yapatite nanoparticles have anticancer property [26]. They are used
Keratinocytes, broblasts, endothelial cells, macrophages, and as drug carrier in targeted drug delivery systems [27]. Metal oxide
platelets cells are among the essential cells involved in wound nanoparticle like titanium dioxide nanoparticles are proved to have
repair [12,13]. Haemostasis is characterized as the release of blood anticancer and antibacterial properties [26,28,29]. Titanium diox-
components into the injury site, allowing the platelets to come ide nanoparticles are used in drug targeting [30]. The toxicity prole
into contact with the exposed collagen and other components of of these nanoparticles shows they are non-toxic [31].
the extracellular matrix. This release then triggers the release of Silver is often utilized as an antibacterial agent, in order to pro-
clotting factors, essential growth factors, and cytokines such as vide a sanitary environment for the wound healing process. Silver
platelet-derived growth factor (PDGF) and transforming growth was considered, historically, as one of the most frequently used
factor (TGF-), from the platelets. During haemostasis, platelets antibacterial substances before the invention of antibiotics. Sil-
adhere to the wound site, where they aggregate. Aggregation is ver was used in sutures to prevent postoperative inammation
then followed by coagulation cascade activation, which results in and infections. The multiplicity of silvers bactericidal mechanisms
the formation of platelet clots in a protein mesh composed of gives it a wide range of effective applications in the inhibition of
cross-linked brin. The inammation stage involves neutrophils bacterial growth [32,33]. Utilizing broad-spectrum antimicrobial
entering the injured area and beginning phagocytosis, in order silver in wound treatment, to prevent the initial growth of any
to remove unwanted substances, bacteria, and unhealthy tissue. bacteria, may contribute to the advancement of wound treatment
Macrophages also contribute to the phagocytosis process by further systems by providing an ideal sanitary environment for maximum
releasing PDGF and TGF during inammation. The sheer number wound repair. Nanoparticles has allowed the scientic community
of growth factors and cytokines involved in the overall wound heal- to enhance the antibacterial properties of silver. The increased sur-
ing task, including broblast growth factor, vascular endothelial face area of the nanoparticles in turn induces an increased rate
growth factor, granulocyte-macrophage colony-stimulating factor, of interaction between the test subjects and the ionic silver. The
platelet-derived growth factor, connective tissue growth factor, increase in bactericidal efciency lowers the minimum inhibitory
interleukin, and tumour necrosis factor-alpha, indicates the com- concentration, meaning that a lower concentration of the mate-
plexity and fragility of the process [1416]. The cytokines and rial may be incorporated into wound healing systems to exert the
growth factors released during the haemostasis and inammation same degree of effect. A lower concentration of silver nanoparti-
phases act as chemical signals that control matrix production, cell cles will reduce the possible interference of biological and chemical
migration, differentiation, and enzyme expression. Through these wound healing processes from the AgNPs themselves. Additionally,
chemical signals, new blood supplies and connective tissue cells are the modication capability of silver nanoparticles further enhances
summoned into the damaged area. After the inammation phase, its utilitarian spectrum in skin reparation [3436].
broblasts migrate to the wound site and initiate the proliferative Wound dressing has evolved according to the pathogenesis
phase by releasing fresh extracellular matrices. The released matri- of different wounds. An ideal wound dressing has the ability to
ces are then cross-linked and reorganized during the remodeling maintain moisture, act against microorganisms, be nontoxic, non-
stage. Each step of the process must be carried out appropriately adherent, and promote wound healing. Modern wound dressing
in a timely manner, in order for the wounded area to retrieve its theory, suggests promoting dynamic equilibrium between exudate
homeostatic equilibrium. Any disruptions in the process may lead absorption and optimal surface moisture at the wound surface. In
to undesirable conditions that may present more serious problems addition, it should be able to exchange gas to provide the wound
at the wound site. Thus, it is critical for the health of the patient with sufcient oxygen tension [37,38].
that the processes involved in the healing of dermal damage are In the present study silver is added with curcumin nanopar-
not disrupted [1720]. ticles to improve the wound healing efciency. Polyvinyl alcohol
The bacterial infection of wounds is one of the major con- (PVA), a water soluble synthetic polymer, have less toxicity, possess
cerns in wound treatment. Chronic wounds, a major cause of excellent wound dressing properties. Polyvinyl alcohol has excel-
morbidity, are an example of a condition resulting from bacterial lent lm forming, emulsifying and adhesive properties. It is also
infection. The clinical spectrum of wound colonization and infec- resistant to oil, grease and solvents. It has high tensile strength and
tion describes the stages of bacterial interaction with a wound: exibility, as well as high oxygen and aroma barrier properties.
contamination, colonization, local infection/critical colonization, these properties are dependent on humidity. Higher the humidity
the spreading of invasive infection, and septicaemia. Each stage of more water is absorbed. The water, which acts as a plasticiser, will
infection produces various effects on the injury site, making it dif- then reduce its tensile strength, but increase its elongation and tear
cult to understand the interaction of bacteria with the disruption strength. [39,40]. They have an interesting application as supports
of wound healing. Biolms self-secreted extracellular polysaccha- for antimicrobial, nutritional and antioxidant substances. In this
ride matrices provide protection for bacteria, through benets such work we report a new method for preparing wound dressing by
as increased infection rates, substrate accessibility, metabolic ef- spin coating. Curcumin nanocomposite is synthesized. It is made in
ciency, and resilience towards environmental stress, along with to a composite with poly vinyl alcohol. In this study, conventional
inhibitors of agents that remove or terminate bacteria. Thus, the cotton cloth was coated with curcumin polymer nanocomposite
presence of biolms further complicates the interaction between to benet from the supreme properties of Ag, curcumin such as
dermal injury and bacteria. Chronic wounds and delayed wound prevention of embedded nanoparticles agglomerations, beside the
repair are often characterized by elongated inammation, faulty intrinsic properties of cotton bers. Some essential factors of mod-
re-epithelialization, and Poor matrix remodeling. A proliferation of ern wound dressings like water absorbency, drying time (water
368 G.D. Venkatasubbu, T. Anusuya / International Journal of Biological Macromolecules 98 (2017) 366378

holding time) and the amount of vertical wicking were determined 2.5. Estimation of curcuminoids by HPLC
and were compared with untreated samples.
2.5.1. Sample preparation
2. Experimental procedure Weigh accurately, about 100 mg of sample into a 100 ml vol-
umetric ask. Add about 20 ml of Acetonitrile and sonicate for
2.1. Synthesis of curcumin nanocomposites 10 min. Cool to room temperature. Make up the volume with
Acetonitrile and mix well. Pipette 5 ml solution into clean 50 ml
30 ml of aqueous solution of AgNO3 is taken. Equal volume of volumetric ask. Makeup with mobile phase up to the mark and
Curcumin Solution in DMSO was added in drops under constant mix well.
stirring at the ratio of 5:1 (W/V) (curcumin: silver). Stirring is
continued for 5 h. A clear orange coloured solution was obtained. 2.5.2. Method
Solution was centrifuged at 2500 rpm for 15 min. Residue was Inject 20 l of mobile phase into chromatograph as blank run.
washed repeatedly with cold water to remove DMSO. The sample Discard any peak due to the solvent in the sample and standard
is freeze dried. chromatograms. Inject 20 l standard solution in duplicate into the
Curcumin nanocomposite is characterized with UV spec- chromatograph. Relative standard deviation for replicate injection
troscopy, FTIR, DLS and HRSEM. The amount of silver present in of standard preparation should not be more than 1.0%. Inject 20 l
the complex is analyzed by ICP-OES. The concentration of cur- sample in duplicate in to the chromatograph. Record the response
cumin in the complex is analyzed by HPLC analysis. The functional of standard and sample at 424 nm.
groups present in pure curcumin and Curcumin nanocomposite the Curcumin:[A]
sample was analyzed by FTIR (FTIR, Perkin Elmer Spectrum One). Curcumin area of sample X Standard wt in g X 5 100
The FTIR spectra were collected in mid-IR range 4000400 cm-1 X 50 X std purity
at 4 cm-1 resolutions, averaging 256 scans. Perkin Elmer Optima . . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . . = % (1)
5300 DV is used to analyse the amount of silver in the sample. Curcuminoids area of standard X 100 50 X Spl wt in g X 5
Dynamic light scattering measurements were taken by using a Demethoxycurcumin:[B]
Malvern Instruments Zetasizer functioned in backscatter (173 ) DemethoxyCurcumin area of sample X Standard wt in g
mode. The sample slurry containing solid content of 1% by volume X 5 100 50 X std purity
was diluted with methanol. Ultra sonication was done for 10 min
. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . . = % (2)
in order to make sample dispersion. This dispersed sample was
then shifted to square cuvettes with a path length of 10 mm for Curcuminoids area of standard X 100 50 X Spl wt in g X 5
measurements. BisDemethoxycurcumin:[C]
BisDemethoxyCurcumin area of sample X Standard wt in g
2.2. Synthesis of curcumin polymer nanocomposite X 5 100 50 X std purity
. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . . = % (3)
Poly vinyl alcohol (5% W/V) solution was prepared. On continu-
Curcuminoids area of standard X 100 50 X Spl wt in g X 5
ous stirring 0.5% (W/V) of Curcumin nanocomposite is added slowly
Curcuminoids = A +B + C
to the PVA solution. Stirring is continued for 2 h.
2.6. Estimation of curcuminoids in the coated sample
2.3. Coating of curcumin nanocomposite on cotton cloth
Initial weight of Cloth sample is measured. The coated cloth is
Cotton cloth is pretreated before coating with water and cut in to 1 1 cm size. Curcumin are extracted from the coated

ethanol. It is dried at 80 C. Curcumin polymer nanocomposite is sample. Weight of solids extracted is calculated. Purity of Total
coated on cloth substrates using spin-coating technique. A clear Curcumin is estimated by UV Visible spectroscopy. From this the
homogeneous solution of Curcumin polymer nanocomposite was Content of Curcumin is estimated.
spin coated on cloth substrates at 3000 rpm for 20 s. After each spin
coating, the substrates were dried in a furnace at 100 C for 20 min 2.7. Static immersion test
for evaporating the solvent. The entire process was repeated 25
times. The water absorption test was carried out according to BS 34491
(1990) standard called as Static immersion test for fabrics (like
2.4. In vitro evaluation of antibacterial activity of curcumin bandages, gauzes and, cotton for medical applications) with high
nanocomposite capability of water absorption. Weighted dry samples were soaked
in a lled beaker with distilled water for a minute then hanged for
The antibacterial activity was done against Escherichia coli, Bacil- two minutes and weighted. The water absorption percentage (W)
lus subtilis, Staphylococcus aureus, Proteus vulgaris, Enterococcusi was obtained by Eq. (1). Each test repeated ve times and the mean
faecalis, Staphylococcus epidermis, Klebsiella pneumoniae, Enterobac- values for sample were reported.
ter aerogenes, Pseudomonas mendocina, Coliform using agar well
W = (M2 M1 )/M1 (4)
diffusion method [4144]. Different concentrations of Samples
(250 g, 500 g, 750 g and 1000 g/well) were used in this study. M2 = weight of wet sample
Nutrient Agar (NA) plates were inoculated with test organisms. M1 = weight of dry sample.
The plates were evenly spread out. Then wells were prepared in
the plates with a cork borer. Each well was loaded with 0.1 ml 2.8. Water holding time
of corresponding concentration of sample and 10 g of Tetracy-
cline dissolved in 1 ml of DMSO was used as a Positive control for Drying time of samples was determined based on T-PACC
antibacterial activity. The plates were incubated for 24 h at 37 C. method. First circular cuts (3.5 inch diameter) of samples were
The development of inhibition zone around the well was measured prepared and weighed. Then gauzes were wetted (sprayed) with
and recorded. 1CC distilled water and weighted. The reduced weight of samples
G.D. Venkatasubbu, T. Anusuya / International Journal of Biological Macromolecules 98 (2017) 366378 369

due to water evaporation was measured every 15 min until the period of 600 min. 5 ml samples were withdrawn by a pipette and
weight measuring differences between three successive measure- replaced immediately with 5 ml of fresh PBS medium, which was
ments reached to a non signicance level. Each test was repeated accounted for when calculating the amount released. The released
ve times, and the mean value was reported. amount of curcumin was determined at different time intervals by
recording the absorbance of release medium by UVvis spectropho-
tometer. Drug release kinetics were analyzed by using the
2.9. Vertical wicking test
The percentage of cumulative release
To determine the wicking rate, samples were cut (in ribbon Mt /M0 100 (5)
shape) with dimensions of 170 mm by 25 mm and placed straight
in a beaker containing distilled water. The height of wicked water (where Mt is the amount of drug released at time t and Mo is the
in different times (1, 5, and 10 min) recorded as wicking ability fac- initial loaded drug amount), the amount of curcumin released in
tor. Each test was repeated three times, and the mean value was pH 7.4 buffer was measured spectrophotometrically at 492.2 nm.
reported. Mean, standard deviation and one way analysis of variance (Anova)
is carried out.
2.10. In vitro drug release
3. Results and discussion
The in vitro release studies was carried out by placing the coated
cloth of size 1 1 cm, known weight of curcumin coated cloth The curcumin nanocomposite when suspended in water, the
sample in denite volume (50 ml) of releasing medium (7.4 pH lyophilized powder formed a very ne dispersion and appeared to
phosphate buffer) at 37 C. The drug release study was done for a be soluble (Fig. 2a (i)), unlike curcumin, which is completely insol-

Fig. 2. (a) Solubility of (i) curcumin (ii) Curcumin nanocomposite. (b) UV absorption of (i) curcumin (ii) Curcumin nanocomposite.
370 G.D. Venkatasubbu, T. Anusuya / International Journal of Biological Macromolecules 98 (2017) 366378

uble in water (Fig. 2a (ii)), with undissolved akes clearly visible pure curcumin and this peak gets shifted in curcumin nanocom-
in the suspension. It is shown in Fig. 2(a). It was observed that this posite. The peaks at 3380 cm1 corresponding to O H stretch,
sample dissolved to clear solution very quickly and easily, with no 1625 cm1 corresponding to N H bend, 1219 cm1 correspond-
noticeable curcumin precipitation. Stabilizers or surfactants were ing to C N stretch, 810 cm1 corresponding to N H wagging from
not used, and the nished product entirely consisted of curcumin curcumin. The peak at 3490 cm1 is assigned to the stretching
in the form of nanoparticles. Reduction in the particle size of active vibrations of the OH group in raw curcumin. The appearance of
ingredients to nanoparticle size has shown improvement in its ef- a peak at 1624 cm1 to predominantly mixed C C and C O bonds.
cacy, solubility, and bioavailability [45,46]. Another peak at 1600 cm1 is attributed to symmetric aromatic
The UV-Spectroscopy image of pure curcumin and curcumin ring stretching vibrations of C C. In addition, the vibration of C O
nanocomposite are given in Fig. 2(b). The UVvis absorption spec- appeared at 1506 cm1 , whereas enol peaks for C O and C O C
trum of the curcumin (Fig. 2b (i)) displays band at 420 nm. On the appeared at 1272 and 1110 cm1 respectively. In both Cur-NPs
other hand, the spectrum of the curcumin nanocomposite showed and curcumin nanocomposite, the OH peak of curcumin is shifted
band at 438 for 1:5 ratio (Fig. 2b (ii)). The observation that the to 3384 cm cm1 and appeared broader. The peaks assigned for
band of curcumin at 420 (420 nm, -* transition) have shifted to C O C at 1110 cm1 were also obviously detected in both Cur-NP
higher wavelength, is an indicative of involvement of the carbonyl and curcumin nanocomposite, thus indicating the presence of cur-
group of curcumin in metal complexation [47]. The shoulders are cumin nanocomposite. The shifting of the peak is due to formation
attributed to a curcumin metal (M2+ ) charge transfer, specic of co-ordination bond between the silver atom and the electron
complex formed. We believe that the variation of the absorption rich groups (oxygen/nitrogen) present in curcumin [49].
peak of curcumin and shoulders apparition in complex depend on The particle size analysis and distribution of the nanoparticles
the nature of metal ion [48]. was performed by DLS analysis. The results for curcumin nanocom-
The FTIR image of curcumin and curcumin nanocomposite is posite are given in Fig. 3(b). DLS of an aqueous dispersion of nano
shown in Fig. 3(a). From the FTIR results it can be conrmed that curcumin revealed the formation of nanoparticles with an average
there is interaction between the curcumin and curcumin nanocom- hydrodynamic diameter of 300 400 nm (Fig. 3b (i)). For curcumin
posite. The peak of C O stretching is observed at 1614.47 of nanocomposite it is 290350 nm (Fig. 3b (ii)).

Fig. 3. (a) FTIR image of curcumin and Curcumin nanocomposite. (b) DLS image of (i) curcumin (ii) Curcumin nanocomposite.
G.D. Venkatasubbu, T. Anusuya / International Journal of Biological Macromolecules 98 (2017) 366378 371

Fig. 4. (a) SEM image of (i) curcumin (ii) Curcumin nanocomposite. Circle shows the presence of silver nanoparticles. (b) HPLC spectrum of curcumin in Curcumin nanocom-
posite.

SEM image of curcumin nanocomposite are given in Fig. 4 (a). curcumin nanocomposite samples. The amount of silver present
The image shows that the particles are in nanometer range. The in 1 g of Curcumin nanocomposite is 0.1868 mg. The HPLC spec-
SEM image of pure curcumin (Fig. 4a (i)) curcumin nanocomposite trum of curcumin nanocomposite is given in Fig. 4(b). Curcumin
(Fig. 4a (ii)) are in nanometer size. SEM images reveals the presence nanocomposite shows a Curcuminoids content of 93.6% by HPLC
of axes shaped nanoparticles. The enhanced aqueous solubility of analysis.
nano-sized curcumin particles could be attributed to their larger The anti-microbial ability of curcumin nanocomposite was
surface area, which promotes dissolution. Encircled area in Fig. 4a investigated by both agar diffusion method the antimicrobial prop-
(ii) shows the silver nano particles. erty of curcumin nanocomposite is given in Table 1. Antimicrobial
The curcumin nanocomposite samples were analyzed by ICP- studies as done in E. coli, B. subtilis, S. aureus, E. faecalis, S. epi-
OES spectroscopy to nd out the amount of silver present in the dermis, K. pneumonia, E. aerogenes, P. vulgaris, P. mendocina,
372 G.D. Venkatasubbu, T. Anusuya / International Journal of Biological Macromolecules 98 (2017) 366378

Table 1
Zone of inhibition (mm) of silver curcumin nanoparticles.

Organism Positive control 250 g/ml 500 g/ml 750 g/ml 1000 g/ml

E. coli 19 7 10 19
B. subtilis 18 8 11 18
S. aureus 41 6 12 14
E. faecalis 19 6 14 19
S. epidermis 15 7 13 15
K. pneumoniae 20 7 10 20
E. aerogenes 28 8 12 28
P. vulgaris 22 7 13 22
P. mendocina 16 8 10 16
Coliform 19 8 11 19

Coliform. These microorganisms are commonly found in wounds. could be one of the reasons for the observed antibacterial prop-
The analysis was done for 250, 500, 750 and 1000 g/ml of cur- erty, nanoparticles also modulate the phosphotyrosine prole of
cumin nanocomposite. The antibacterial activity of nanocurcumin putative bacterial peptides, which could thus affect bacterial signal
against E. coli, B. subtilis, S. aureus, E. faecalis, transduction and inhibit the growth of the organisms. The released
S. epidermis, K. pneumonia, E. aerogenes, P. vulgaris, P. men- silver nanoparticles from curcumin nanocomposite can interact
docina, Coliform showed that it exhibits a broad spectrum with sulfur containing intracellular proteins in bacteria and kill
inhibitory effect against all microorganisms. The results showed them [5456]. Silver nanoparticles are reported to show better
that curcumin nanocomposite showed a concentration dependent wound healing capacity, better cosmetic appearance and scar less
bactericidal activity. curcumin nanocomposite at high concentra- healing when tested using an animal model. For biomedical appli-
tion of 1000 g/ml showed high antibacterial activity against all cations, use of biomaterials with anti-microbial property will be an
the test bacterium. Fig. 5 shows the antibacterial analysis of cur- added advantage. Curcumin nanocomposite having such antimi-
cumin nanocomposite. The formation of inhibition zone clearly crobial activity will prevent infection, sepsis and can be a good
indicates that the mechanism of antibacterial action is due to the material for preparing wound dressing.
activity of curcumin and leached Ag+ ions. The results revealed that Antioxidant activity is very important in wound healing.
curcumin nanocomposite are more effective against gram positive Curcumin has antioxidant activity. This is mediated through
bacteria and gram negative bacterium. The variation in the zone antioxidant enzymes such as superoxide dismutase, catalase, and
of inhibition among various bacterium could be due to differences glutathione peroxidase. Curcumin has been shown to serve as a
in their cell membrane constituents and structure. It is known that Michael acceptor, reacting with glutathione and thioredoxin. Reac-
Gram-positive bacteria contain an outer peptidoglycan layer, while tion of curcumin with these agents reduces intracellular GSH in the
Gram-negative bacteria contain an outer phospholipidic mem- cells. The suppression of LPO by curcumin could lead to the suppres-
brane, both of which undergo different types of interaction when sion of inammation. In curcumin, the phenolic and the methoxy
encountered by curcumin. It is shown that curcumin can be sol- group on the phenyl ring and the 1,3-diketone system seem to be
ubilized in water when it is in the nano form and that it is as important structural features that can contribute to these effects
much or even more effective than curcumin. The rationale behind [5759].
the stronger activity of nano curcumin is related to the particle In Fig. 6, FTIR spectrum of pure PVA sample is showed. It clearly
size. Once curcumin nanoparticles are formed, the size reduces, reveals the major peaks associated with poly (vinyl alcohol). A
which is much less than the size of curcumin particles, which is strong hydroxyl bands for free alcohol (non-bonded OH stretch-
responsible for better penetration and higher uptake by the cells. ing band at = 36003650 cm1 ), and hydrogen bonded band
The in vitro biological assays clearly demonstrated that transfor- ( = 32003570 cm1 ). Intramolecular and intermolecular hydro-
mation to the nano form greatly improves the water solubility and gen bondings are expected to occur among PVA chains due to high
efcacy of curcumin as an antimicrobial agent. Curcumin nanocom- hydrophilic forces. An important absorption peak was veried at
posite will broke the peptidoglycan layer and penetrated inside the 1142 cm1 . This band has been used as an assessment tool
cell, thereby causing disruption of the structure of cell organelles of poly (vinyl alcohol) structure because it is a semicrystalline
and killing the cell through lysis. The mechanism of antibacterial synthetic polymer. The peaks at 28302695 cm1 (C H from alde-
activity of curcumin involves perturbing the GTPase activity of FtsZ hyde), 28403000 cm1 (C H from alkyl groups), 1414 cm1 for the
protolaments, which are known to play a critical role in bacte- C O group and 11501085 cm1 (C O C) corresponds to PVA.
rial cytokinesis. This perturbation becomes lethal to the bacteria The FTIR spectrum of Curcumin polymer nanocomposite clearly
and inhibits bacterial cell proliferation by inhibiting the assem- shows the typical signatures of both PVA and curcumin. Signa-
bly dynamics of FtsZ in the Z ring. Curcumin nanoparticles inhibits tures of curcumin are free O H group (3681 cm1 ), C O and C C
bacterial surface protein sortase A and prevents cell adhesion to (enol) (14501630 cm1 ), C H (methyl) (2845 cm1 ), C H (aryl)
bronectin, thereby acting as an antibacterial agent [5053]. The (3015 cm1 ) and C O C (10001300 cm1 ) typically attributed
major mechanism through which silver nanoparticles manifested to symmetric and asymmetric congurations of C O C chains.
antibacterial properties was by anchoring to and penetrating the Curcumin molecule may have attached to the PVA molecule by
bacterial cell wall, and modulating cellular signaling by dephos- intermolecular hydrogen bonding to enolic hydroxyl group which
phorylating putative key peptide substrates on tyrosine residues. results in shift of O H stretching from 3653 to 3681 cm1 .
Silver will attach to the surface of the cell membrane and drastically In this research, we coated textile substrates with silver
disturb its proper function, like permeability and respiration. They nanoparticles by a spin coating method. The images of the uncoated
are able to penetrate inside the bacteria and cause further dam- and Curcumin polymer nanocomposite coated cotton cloth are
age by possibly interacting with sulfur- and phosphorus-containing presented in Fig. 7a and 7b respectively. The SEM image of Cur-
compounds such as DNA. Curcumin nanocomposite release sil- cumin nanocomposite coated sample is shown in Fig. 8a and Fig. 8b.
ver ions, which have an additional contribution to the bactericidal The SEM images conrms the coating of Curcumin nanocomposite
effect of the silver nanoparticles. Although bacterial cell lyses in the cotton cloth substrate. The presence of PVA acts as a binder
G.D. Venkatasubbu, T. Anusuya / International Journal of Biological Macromolecules 98 (2017) 366378 373

Fig. 5. Antibacterial activity of Curcumin nanocomposite.

and forms a layer on the substrate. The presence of silver is seen as with high surface coverage were formed on the cotton cloth sub-
white dots on the nanoparticles. This is also seen clearly in the SEM strate. The NPs were agglomerated. The images demonstrates that
image of the Curcumin nanocomposite in Fig. 4. The SEM image of a rough surface with a hierarchical structure was formed by coat-
the coated sample shows the presence of dened nanoparticles in ing of Curcumin nanocomposite on cotton cloth. It is difcult to
the lm coated. Curcumin nanocomposite in the coated sample are distinguish individual bres in the coated sample. The difference
essential for antimicrobial applications. Curcumin nanocomposite in clarity is most likely be a result of the coating of Nps with PVA
374 G.D. Venkatasubbu, T. Anusuya / International Journal of Biological Macromolecules 98 (2017) 366378

Table 2
Quantitative estimation of curcumin in coated cloth sample.

Tests Weight

Initial Cloth weight 0.0135 g


Weight of Cloth after Extraction and Drying 0.0124 g
Weight of solids extracted 0.0011 g
Assay by UV: 73.90%
Purity of Total Curcumin by UV Visible
spectroscopy
Content of Curcumin cloth 0.81 mg

Table 3
Water absorption (static immersion test) of samples.

S.No Sample Mean Percentage


of water
absorptiona (%)

1 Untreated cloth 280 3


2 Coated cloth 420 4
a
Each test was repeated ve times and the mean reported.
Fig. 6. FTIR spectrum of Curcumin polymer nanocomposite.

Table 4
Average drying times (water holding ability) of wet samples.
polymer. The image indicates that there is less free space due to
the presence of NPs polymer composite coated. This tightly packed S.No Sample Mean Time of
dryinga (min)
network makes it difcult to perceive depth in the image. It was
also observed that the cotton cloth was physically stiffer and brit- 1 Untreated cloth 42.45 2.45
2 Coated cloth 75.23 3.27
tle when coated compared to an uncoated cotton cloth. From SEM
a
images the formed coating appears homogeneously on the cotton Each test was repeated ve times and the mean reported.
cloth surface
Surface chemical elements of the fabrics were determined by
EDX spectroscopy. Fig. 9a shows the EDX spectrum and Fig. 9b for water, wound exudates, and liquid drug absorption of a wound
shows the SEM image in which the EDX is taken. The EDX exhib- dressing [60]. The micro and porous brous 3-D network of formed
ited signicant peaks for carbon and oxygen as expected. Carbon PVA on cotton gauze can be the most important factor of water
and oxygen are the key elements of the polymer chains that make absorption capability increasing. The presence of micro brils in
up cotton. As expected, the energy dispersive X-ray (EDX) anal- PVA layer causes a great specic area that plays a signicance role
ysis conrmed the presence of Ag nanoparticles on the sample in water absorbency. This ability is very crucial for a wound dress in
surface. This analysis veried the formation of curcumin polymer which water, wound exudates, and liquid drugs absorption abilities
nanocomposite. The result agreed with the SEM as well. are highlighted.
The amount of curcumin loaded on the sample is calculated. The Moisture retaining of wound surface (preserving wet condi-
results are given in Table 2. The purity of curcumin shows as 73.90%. tion during wound care) accounts for a very important criterion
The curcumin content in the coated cloth is 0.81 mg. in modern active wound dressing. Table 4 shows average drying
The water absorption percentages of samples are shown in time for each sample. For this, the differences between untreated
Table 3. The difference between untreated cotton gauze and treated cotton gauze and the treated ones were measured. The data set
one is statistically signicant (P = 0). Comparing data shows that from ANOVA shows that the P = 0, so the null hypothesis is refused
capability in water absorption of cotton gauze had increased about and there is a statistically signicant difference between untreated
50 percent. When the spaces of cotton cloth between wraps and cotton gauze and treated one is at level of 95%. The average water
wefts are lled with PVA, the number of capillaries of cotton cloth holding ability of treated gauze is 74 percent higher than untreated
increased in both weft and wrap directions. Therefore, vertical ones. Holding capability of liquid especially water is a fundamental
wicking and water absorption increased. This ability is highlighted factor to alleviate the pain and heal the wound [61].

Fig. 7. Image of (a) Cotton cloth (b) Curcumin polymer nanocomposites coated Cotton cloth.
G.D. Venkatasubbu, T. Anusuya / International Journal of Biological Macromolecules 98 (2017) 366378 375

Fig. 8. SEM image of coated cloth (a) lower magnication (b) higher magnication.

The vertical wicking ability of treated sample with curcumin Table 5


Vertical wicking ability of untreated and treated cloth in different times.
polymer nanocomposite and untreated one were measured three
different times. Each time, the test was repeated ve times and the S.No Time (min) Untreated cloth Coated clotha
mean value reported in Table 5. The results show that the wicking 1 1 1.65 2.57 0.35
ability of treated gauze and untreated gauze both increased though 2 5 3.32 5.26 0.25
time. The results of ANOVA show P = 0.01which is smaller than 3 10 4.87 6.12 0.18
the signicant 0.5 level. So difference between treated cotton gauze a
Each test was repeated ve times and for every time the mean was reported.
and untreated one is statistically signicant (95%). So by covering
the cotton cloth with curcumin polymer nanocomposite, the rate
of therapeutic liquid transport to the wound increases and the rate Curcumin widely known for its anti-tumor, antioxidant, anti-
of exudate absorption decreases. inammatory, anti-Alzheimers, anti-cystic brosis, and wound-
376 G.D. Venkatasubbu, T. Anusuya / International Journal of Biological Macromolecules 98 (2017) 366378

Fig. 9. EDX spectrum of Curcumin polymer nanocomposites coated cotton cloth. (a) EDX spectrum (b) SEM image where EDX is taken.

healing, properties. The percentage of cumulative release of The release rate depends on the diffusion of drug from the polymer
curcumin from the lms was calculated using the following equa- pores. Diffusivity of drug is indirectly proportional to its molecu-
tion. lar weight. The drug release was highly affected by the diffusion
The percentage of cumulative release process, along with the coating degradation. The drug release was
dominated by the diffusion of drug through the coating layer.
Mt /M0 100

Where Mt is the amount of drug released at time t and M0 is the


initial loaded drug amount. Fig. 10 depicts the percentage of cumu-
lative releases of curcumin from the coated cloth sample in 7.4 pH 4. Conclusion
buffer solution at 37 C. The percentage of drug released in 600 min
from the coated cotton cloth was found to be 100%. The drug is
released gradually over a period of time. This shows that the drug In this work we successfully obtained Curcumin nanocompos-
is released in a sustained manner. The initial burst of curcumin from ite. The Curcumin nanocomposite was characterized by FTIR, UV,
the coated cotton cloth is well controlled. Error bars are the stan- SEM, ICP-OES. The present work demonstrates a simple method
dard error on the basis of triplicate determinations (mean S.E., in producing novel curcumin polymer nanoparticle nanocompos-
n = 3). In the initial stages the release rate from coated cloth is high ite lms coated on cotton cloth for wound healing applications.
for some time, then it declines and remains constant for a long The Curcumin nanocomposite have good antibacterial activity
time period. One way analysis of variance was P < 0.001. This initial against bacterial species present in wounds. These agents may nd
burst of curcumin may be occurring due to the shrinkage of PVA. potential applications in wound dressing/wound burns. The coated
G.D. Venkatasubbu, T. Anusuya / International Journal of Biological Macromolecules 98 (2017) 366378 377

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Kumar, Surface modication and Paclitaxel drug delivery of folic acid
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