investigation of regenerated cellulose/poly (acrylic acid

Research ArticleInvestigation of Regenerated CellulosePoly(acrylic acid)Composite Films for Potential Wound Healing ApplicationsA Preliminary Study

Manjula Bajpai S K Bajpai and Dinesh Gautam

Polymer Research Laboratory Department of Chemistry Government Model Science College Jabalpur 482001 India

Correspondence should be addressed to Manjula Bajpai mnlbpirediffmailcom

Received 20 January 2014 Revised 14 March 2014 Accepted 17 March 2014 Published 6 May 2014

Academic Editor Alejandro Rodriguez

Copyright copy 2014 Manjula Bajpai et alThis is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Regenerated cellulosepoly(acrylic acid) composite films have been synthesized for wound dressing applications The waterabsorbency of these films was studied as a function of amount of cross-linker NN1015840-methylenebisacrylamide and cellulose contentsin the feed mixture The samples having different compositions showed tensile strength and percent elongation in the range of998 times 10

5 to 1340 times 105 Nm2 and 110 to 265 respectively The water vapor transmission rate (WVTR) for various films was foundto be in the range of 203 to 718mgcm2hThese films were loaded with antibacterial drug miconazole nitrate and their release wasstudied in the physiological pH at 37∘C The release data was found to fit well the diffusion controlled Higuchi model Finally thefilms demonstrated fair antibacterial and antifungal action thus establishing their strong candidature as wound dressing materials

1 Introduction

Wound dressing films are three-dimensional cross-linkedpolymers which are permeable to gases and moisture andare used to prevent the microbial contamination in wounds[1] In the past materials such as honey pastes plant fibersand animal fats were frequently employed as wound dressingmaterials [2] A potential wound dressing film should haveproperties such as moisture and gas permeation capacitymechanical strength folding endurance (FE) capacity torelease the loaded bioactive material and antimicrobialefficacy Ultimately themajor function of the wound dressingfilm is to provide the highest rate of healing and the bestaesthetic repair of the wound [3 4] A wound dressingfilm is desirable to have the following prompt functions (a)to provide moist environment (b) to protect wound frommicrobial infection and (c) to ease application and removalavoiding dressing-related trauma [5 6] Wound dressingsloaded with antibacterial agent or drug are often used totreat wound locally such as anti-infections due to secondaryinfection or for pain control especially in chronic wounds[7 8]

Recently due emphasis has been given to various micro-bial polysaccharides towards the development of wounddressing materials [9] because of their biocompatiblenature biodegradability fair water absorbing capacity andtransparency [10] Some of these polysaccharides includehyaluronic acid cellulose alginate chitosan and kappacarrageenan [11 12]

Cellulose a natural polysaccharide and found in abun-dance belongs to the most promising class of biopolymers[13] The unique physical and mechanical properties ofcellulose as well as its purity and uniformity determineapplications that range from high quality audio membranes[14] and electronic paper [15] to fuel cells [16] and medicalmaterials [17ndash19] In recent past sincere efforts have beenmade to develop cellulose as a new skin substituent andwound dressing material [20 21] due to its hydrophilicityfair water retention capacity high permeability optimalgelatinization and high mechanical strength [22]

Hydrogels are three-dimensional polymer networks andthey possess tendency to retain water which may be help-ful in providing moist environment to the wound Theirtransparent look can be useful in monitoring the healing

Hindawi Publishing CorporationJournal of Applied ChemistryVolume 2014 Article ID 325627 9 pageshttpdxdoiorg1011552014325627

2 Journal of Applied Chemistry

Ominus

OminusOminus

ON

Cl

ClCl

Cl

N

Miconazole nitrate

Figure 1 Molecular structure of drug miconazole nitrate

process visually [23] In addition variation in the degreeof crosslinking of polymeric hydrogel may be helpful incontrolling its mechanical properties moisture permeabilityand release rate of loaded bioactive ingredient [24]Thereforecombination of cellulose and synthetic polymeric hydrogelcould result in an efficient wound dressing film with most ofthe desirable properties

Recently coinage metal nanoparticles such as silver havebeen used as antibacterial agents in fabrication of wounddressing films [25] But as per some reports silver nanoparti-cles are harmful for human osteoarthritic chondrocytes whenused in higher concentrations of around 160ndash250 120583M [26]Moreover nanoparticles present in blood are associated withthrombosis and activation of immunological reactions [27ndash29] As an alternative to metal nanoparticles we here reportdrug miconazole nitrate loaded cellulosepoly(acrylic acid)hydrogel films for antibacterial applications

2 Experimental

21 Materials Cellulose (Cell) powder was purchased fromCentral Drug House Mumbai India Its viscosity averagemolecular weight (Mn) was calculated according to equation[(p)] = 385 times 10minus2 Mn076 [25] and was found to be186 times 10

4 The monomer acrylic acid (AAc) cross-linkerNN1015840-methylenebisacrylamide (MB) and initiator potassiumper sulfate (KPS) were obtained from Hi Media MumbaiIndia The antibiotic drug miconazole nitrate was obtainedfrom Ranbaxy pharmacy as commercial product Micogel(Figure 1 shows structure of drug miconazole nitrate usedin the study) Nutrient broth nutrient agar and agar-agartype-1 were obtained from Hi Media Chemicals MumbaiIndiaThe department of biotechnology (GovernmentModelScience College Jabalpur (Madhya Pradesh) India) providedstandard culture of the organisms Double distilled water wasused throughout the investigation

22 Dissolution of Cellulose in NaOHUrea Solvent SystemCellulose solution was prepared in NaOHurea solvent

system according to procedure reported by Jin et al [30] Inbrief 6 g of NaOH and 4 g of urea were dissolved in 90mL ofdistilled water and the resulting solution was filtered with aG2 sand filter Now 4 g of cellulose powderwas dispersed into100 g of above solution under constant stirring for 20min andthen stored in refrigerator (minus5 to minus10∘C) for 12 h The frozensolid was thawed and stirred extensively at room temperatureto obtain a colorless and almost transparent solution Thecellulose solution was dialyzed against distilled water using adialysis membrane with 10000 molecular weight cut-off Thedialysis was carried out for a period of 2 days The resultingsuspension was used to prepare the hydrogels as described inthe next section

23 Synthesis of Regenerated CellulosePoly(AAc) Hydro-gel Film The regenerated cellulosepoly(acrylic acid) (RCpoly(AAc)) hydrogel film was prepared by free-radical poly-merization of AAc in the presence of above dissolved cellu-lose In brief 1266mMofmonomer AAc 0194mMof cross-linker MB and 0295mM of initiator KPS were mixed into25mL of above dialyzed cellulose solution (4wt) and thetotal volume was made up to 5mL by addition of water Thereaction solution was transferred into Petri plates and keptin electric oven (Tempstar India) at 60∘C for a period of 2 hThe resulting hydrogel film was peeled off and transferredinto 200mL of ammonium sulfate solution (8wv) for aperiod of 8 h Finally RCpoly(AAc) hydrogel film was takenout and allowed to equilibrate in excess of refreshing waterfor a period of 7 days till all the remaining chemicals in thehydrogel film were leached out The compositions of varioushydrogel films prepared are given in Table 1

24 Yield of Hydrogel Film Freshly prepared RCpoly(AAc)hydrogel film was placed in refreshing distilled water tillthe external medium acquired pH almost that of distilledwater Now the swollen hydrogel was placed in 50 percent(vv) aqueous solution of methanol for a period of twodays In order to ensure the complete extraction the gel wastransferred into acetone for 12 hours followed by completedrying

The percent hydrogel yield was calculated as per thefollowing expression

Hydrogel yield =Dry weight of polymerTotal weight of reactants

times 100 (1)

Here total weight of reactants includes monomer AAcpolymer cellulose and cross-linker MB

25 Preparation of Drug-Loaded Film Thedrug-loaded filmswere prepared by adding precalculated amount of drug tothe reaction mixture before the polymerization started Thismethod enables us to incorporate a desired quantity of drugin the polymer matrix The hydrogel film samples weredenoted as HG(X) where the number in the parenthesisdenotes the amount of drug present in one gram of film

26 Water Absorption Study In the gravimetric method thepreweighed sample was placed in 500mL of distilled water

Journal of Applied Chemistry 3

Table 1 Compositions of various hydrogel films synthesizeda

Hydrogel film code Cellulose (mg) AAc (millimolar) MB (micromolar) KPS (millimolar) Glycerol (mL)HG1 100 1266 194 0295 mdashHG2 100 1266 324 0295 mdashHG3 60 1266 324 0295 mdashHG4 60 1266 194 0295 mdashHG5 20 1266 194 0295 mdashHG6 100 1266 194 0295 05HG7 100 1266 194 0295 10aTotal volume of polymerization mixture was 50mL

at 37∘C and it was taken out at different time intervalswiped superficially with tissue paper to remove extra surfacewater weighed accurately on an electronic balance (DenberGermany) and then placed back in water The swelling ratioSR obtained at different time intervals was determined usingthe following expression [31]

SR =(119872119905minus1198720)

1198720

times 100 (2)

where1198720and119872

119905are the initial mass and mass at different

time intervals respectively

27 Test forMechanical Properties Themechanical propertiesof the films were determined according to the procedurereported elsewhere [32] Film samples with the dimensions39mm times 58mm were equilibrated under the RH of 50at 23∘C for a period of 24 h and their tensile strength (TS)and percent elongation at break (E) were measured by usingan Instron Universal Testing Instrument (Model 1011) Theinitial grip separation and crosshead speed were set to 40and 200mm per min respectively All the determinationswere made in triplicate TS was calculated by dividing themaximum load (F N) on the film before failure by the initialcross-sectional area (S m2) that is TS = FS

28 Moisture Permeation Study As per ASTM E 96-93method film sample was placed in between the cup and thesilicon coated ring and held tightly using screws provided Apreweighed quantity of silica gel was placed inside the cup toproduce RH of zero percent The distance between the filmand silica gel was about 15 cm The cup was now placed at25∘C in thermostated desiccators which contained saturatedsalt solution to produce 40 RH The cup was taken out ofdesiccators at definite time intervals and mass was measuredusing an electronic balance (Denber Germany) The watertransmission rate (WVTR) was calculated [33] as

WVTR = Δ119882Δ119905 sdot 119860

mghminus1 cmminus2 (3)

where Δ119882Δ119905 is the amount of water gain per unit time ofmoisture transfer and A = area exposed to water surface incm2

29 Drug Release Study The preweighed drug-loaded filmwas placed in 25mL of release medium (ie physiologicalfluid) at 37∘C After definite time intervals film was trans-ferred into fresh release medium and the amount of drugreleased was determined spectrophotometrically at 200 nmThe quantity of drug was calculated using Lambert-Beerrsquos lawobtained for drug solutions of known concentrations

210 Antimicrobial Experimentations To test the biocidalactivity of drug-loaded Cellpoly(AAc) film in quantitativemanner [34] appropriate number of colony-forming units(CFU) of microbes (5 times 109 CFUmL of E coli) were culturedon a nutrient agar plate supplemented with drug-loadedcircular film (diameter 5mm) placed at the center of theplates The plates were examined for a possible clear zonearound the fibers after incubation at 37∘C for a period of 24 hThe plate supplemented with plain film was used as controlset

3 Results and Discussion

31 Percent Hydrogel Yield (PHY) The total weight of cel-lulose monomer acrylic acid and cross-linker (MB) in thefeed mixture was 1030 g whereas after extraction in differentsolvents the final dry weight was found to be 1016 g ThePHY calculated from (1) is 9814 which can be consideredas high yield It was found that for all the samples synthesizedthere was almost 96ndash99 percent yield The films synthesizedwere almost transparent as shown in Figure 2

32 Water Absorption Behavior of Hydrogel Films Thedynamic water uptake of hydrogel films HG1 HG2 and HG3is shown in Figure 3 It can be observed that samples HG1and HG2 exhibit equilibrium water uptakes of nearly 94and 56 gg respectively The observed difference in wateruptake can simply be attributed to the fact that these samplescontain almost 194 and 324 micromoles of cross-linker MBin their polymer networks respectively The hydrogel withgreater amount of cross-linker produces denser cross-linkednetwork thus allowing less quantity of water to enter into thenetwork In addition higher amount of cross produces morerigid chains due to greater number of cross-link junctions[35] As both the samples contain same quantity of cellulose(ie 100mg) the contribution of cellulose towards water


(a) (b)

Figure 2 A strip of hydrogel film put on the currency note to reveal its transparency

0123456789

10

0 100 200 300 400

Swel

ling

ratio

(gg

)

Time (min)

HG1HG2

HG3

Figure 3 Dynamic uptake of water as a function of time for thehydrogel films HG1 HG2 and HG3 at 37∘C

absorption is the same and the amount of cross-linking agentused is the main factor to govern their water absorptionbehavior

It is also noteworthy that sampleHG3 which contains 324micromoles of cross-linker MB (like sample HG2) but 60mgof cellulose exhibits a total water uptake of nearly 285 ggwhich is the minimum of all the three samples studied Thisis because the sample HG3 contains less quantity of celluloseand therefore its contribution towards total water uptake islow as compared to the sample HG2 which contains 100mgcellulose

Thewater penetrationmechanism is best described by thefollowing logarithmic equation [36]

ln119865 = ln 119896 + 119899 ln 119905 (4)

where119865 is the fractional water uptake at time 119905 119896 is gel charac-teristic constant and 119899 is swelling exponent which indicatesFickian or non-Fickian behavior of the swelling device Theslope and intercept of linear plot obtained between ln 119905 andln119865 enable us to evaluate 119899 and 119896 respectively The dynamicwater uptake data displayed in Figure 3 was used to get linearplots between ln119865 and ln 119905 (see Figure 4) and the swellingexponent ldquo119899rdquo and gel characteristic constant 119896were evaluated

The values of swelling exponent ldquo119899rdquo for the hydrogelsamplesHG1HG2 andHG3were 041 023 and 047 respec-tively In addition values of gel characteristic constants 119896were2454 times 10minus3 3706 times 10minus3 and 2798 times 10minus3 respectively Itis to be noted that for all the samples ldquo119899rdquo values are less than

minus3

minus25

minus2

minus15

minus1

minus05

0

05

1

0 1 2 3 4 5 6

HG1HG2

HG3

ln MtMinfin

lnt

Figure 4 ln 119905 versus ln119872119905119872infinplots for the evaluation of swelling

exponent 119899 and gel characteristic constant 119896 for the samples HG1HG2 and HG3

05 thus indicating diffusion-controlled swellingmechanismThis may simply be attributed to the fact that since thecellulosic chains are entangled with poly(AAc) segments andcreate a densely populated entangled network the solventdiffuses slowly into the network [37] thus indicating simplediffusion-controlled swelling mechanism

33 Mechanical Properties of Hydrogel Films Themechanicalproperties of wound dressing films play a significant rolein establishing their suitability The films must be able towithstand the appropriate stress when used on woundsMoreimportantly the film must have significant flexibility so thatit does not break during the process of exudates absorptionTheflexibility of a film is usuallymeasured in terms of percentelongation (PE) Finally elongation at break (E) is also animportant parameter which is also required to be quite fairduring the application on the affected area of wound [38]In the present study the mechanical properties namely TSand PE (percent elongation) were tested for the samples HG1and HG2 which contained 194 and 324 micromoles of cross-linker MB respectively The results are shown in Figure 5

Fairly higher percent elongations of 175 and 115 for thesamples HG1 andHG2 indicated a higher degree of flexibilityThe fairly higher PE values indicate that these films willbe beneficial in the case of wounds with higher exudatesAfter absorption of fluid the films shall maintain their


0123456789

0 100 200 300 400 500

Elon

gatio

n (c

m)

Weight (g)

HG1 HG2HG7

Figure 5 Elongation versus load profiles to determine the percentelongation at break for the samples HG1 HG2 and HG7

structural integrity The observed decrease in PE with MBcontent is attributable to the increased number of cross-linkjunction points thus imparting more rigidity to the filmsThe TS values for these samples were 099MPa and 131MParespectively The observed increase in TS is simply indicativeof increased rigidity of the films Here it is noteworthythat the two samples contain same amount of celluloseand so the contribution made by intermolecular H-bondinginteractions of ndashOH groups of cellulosic chains is almost thesame The values of TS obtained with the samples HG1 andHG2 are relatively low as compared to those reported byothers For example in a study [39] the alginate based singleand bilayer hydrocolloid films possessed TS values of 2082and 2722 respectively Similarly the wound dressing filmsprepared from Haruan and fusidic acid spray were reportedto have TS values in the range of 1509 to 2817 [40] Thedrawback of poor TS of these films may be overcome byusing these films over wounds along with external supportslike netted cotton strip which may be wrapped over the filmFinally for the sample HG7 containing 10mL of plasticizerglycerol the PE and TS values were found to be 248 and13MPa respectively Contrary to this sample HG1 whichdoes not contain glycerol has PE value of 175 This indicatesthat presence of glycerol enhances the percent elongation byreducing the intermolecular H-bonding between cellulosicndashOH groups

From the above discussion it is clear that hydrogel filmsHG1 and HG2 prepared without using glycerol have fairlyhigh percent elongation PE and therefore addition of glycerolis not at all necessary in the present hydrogel films Howevertheir TS needs to be enhanced or alternatively an externalsupport is required for applying these films on wounds

34 Moisture Permeation Analysis Permeation of moisturethrough a wound dressing film is a significant parameter Incase of excessive water loss via permeation through the filmthe wound may get dried and this will retard the healingprocess The wound dressing film must be permeable to themoisture to an extent that neither there should be excessive

0

002

004

006

008

01

012

014

016

0 5 10 15 20 25 30Time (h)

HG1HG4HG5

HG6HG7

Moi

sture

gai

n (m

g hminus1

cmminus2)

Figure 6 Moisture gain versus time profiles for the hydrogel filmsHG1 HG2 HG4 HG5 HG6 and HG7 under relative humidityatmosphere of 40

loss of moisture to result in a dry wound nor there should besuch a low permeability which may cause leakage of exudates[41] Therefore the water-uptake capacity and WVTR areimportant factors in determining the suitability of film for aparticular wound Normally theWVTR of the normal skin isabout 085mgcm2h whereas those of the injured skin canrange from 116 to 2141mgcm2h The WVTR of a wounddressing film should be such that the film may prevent thewound from dehydration Therefore a wound dressing witha WVTR ranging from 833 to 1042mgcm2h is generallyrecommended [42]

TheWVTRof the hydrogel film samplesHG1HG4HG5HG6 and HG7 were determined at the RH of 40 percent andthe results of moisture gain versus time profiles are shown inFigure 6

WVTR of films were found to be 591 678 718 186and 203mgcm2h respectively These values did not fallwithin the desired range of 833 to 1042mgcm2h Howeversamples HG1 HG4 and HG5 have WVTR values quite closeto the desired range The observed increase in WVTR ofsamples HG1 HG2 and HG3 can be attributed to the factthat these samples contain 100 60 and 20mg of celluloseThe decrease in cellulose content results in formation of lessdense network thus permitting more moisture permeationMoreover decreased cellulose content lowers the H-bondinginteractions among the cellulosic chains thus reducing thenumber of pseudo crosslinks It is reported that glycerol withproper size and three hydroxyl groups easily enters betweenthe film forming polymers and weakens the intermolecularforces between the polymeric chains [43] thus enhancingthe extensibility and moisture permeation through filmHowever in the current work the WVTR of samples HG6and HG7 was observed to decrease probably due to H-bonding between ndashOH groups of glycerol and polar hydroxylgroups of cellulose chains Such type of phenomenon has alsobeen reported elsewhere [44 45]


0

1000

2000

3000

4000

5000

6000

0 100 200 300 400

Dru

g re

leas

e (m

Lm

g)

Time (min)

HG1

HG2

Figure 7The drug release versus time profiles for the hydrogel filmsHG1 (54) and HG2 (54) in the physiological fluid at 37∘C

35 Drug Release Study In wound healing managementstudies nature of exudates is very importantThe compositionof wound fluid depends upon the nature of the wound Forexample Trengrove et al [46] reported that wound fluidcollected from leg ulcers contained 06 to 59mML glucoseand 25ndash51 gL protein Similarly Bonnema et al [47] analyzedserum fluid formed after auxiliary dissection and reportedthat on the first operative day the drainage fluid containedblood and high concentration of creatine phosphokinasewhile after day 1 it changed to lymph like fluid that containeddifferent cells and more proteins In this work we carried outin vitro release study in physiological fluid (PL) as suggestedby British pharmacopeiasThe PF contained 142mM of NaCland 25mM of CaCl

2

The effect of degree of cross-linking on the drug releaseprofiles for the samples HG1 (54) and HG2 (54) is shownin Figure 7 It is clear that sample HG1 (54) exhibits higherrelease as compared to the other sample HG2 (54)

This is due to the fact that as the sample HG2 (54)contains more quantity of cross-linker MB it shows lessswelling and therefore slower release is observed Sincesamples HG1 and HG2 show diffusion-controlled swelling(119899 = 041 and 023 resp) therefore diffusion-controlledmodel given by Higuchi [48] was used to interpret drugrelease data According to this model

119876119905=119872119905

119872infin

= 11987011986711990512 (5)

where 119876119905is the fractional release of drug at time 119905 and 119870

119867

is the Higuchi constant The dynamic release data of samplesHG1 (54) andHG2 (54) were applied on (4) and curves wereplotted between 119876

119905and 119905minus12 as shown in Figure 8

The plots obtained were fairly linear with an excellentregression value of 09723 and 09768 respectively thussupporting Higuchi diffusion-controlled release model Thevalues of Higuchi constants were found to be 331 times 10minus3 and261 times 10minus3 hminus12 respectively

0

01

02

03

04

05

06

0 5 10 15 20

HG1HG2

MtM

infin

tminus12

(min)12

Figure 8 Higuchi plots for the release data observed for the samplesHG1 (54) and HG2 (54)

36 Antibacterial Tests The results of antibacterial experi-ments against model bacteria E coli are shown in Figure 9

It can be seen that the Petri plate containing the plainfilm in Figure 9(a) shows a dense population of bacterialcolonies whereas the Petri plates supplemented with thehydrogel films HG1 (54) and HG2 (54) show clear zones ofinhibition with their respective areas 622 cm2 (Figure 9(c))and 414 cm2 (Figure 9(b)) respectively This may simply beattributed to the fact that more cross-linked film that isHG2 (54) shows slower release from the film and hencedemonstrates poorer inhibition efficiency as compared to theother sampleHG1 (54) which is less cross-linked and exhibitsfaster release as observed in release experiments

37 Antifungal Tests The results of antifungal experimentsagainst fungus Aspergillus niger are shown in Figure 10 Itis clear that the Petri plate containing the plain film inFigure 10(a) shows growth of five colonies whereas the Petriplates supplemented with the hydrogel films (Figure 10(b))HG1 (54) show clear zone of inhibition of fungusAspergillusniger

This indicates that the drug miconazole nitrate is alsoeffective in antifungal activities too Similarly Najafi et al [49]have recently reported antifungal action of a polymeric filmcomposed of hydroxypropyl cellulose and chitosan loadedwith miconazole nitrate The highest loading of drug into thefilms was 96mg

4 Conclusion

From the above study it may be concluded that cellu-losepoly(AAc) hydrogel films are highly flexible andhave fairmechanical strength They allow low permeation of moistureand show diffusion-controlled release of miconazole nitrateThese films show fair antibacterial and antifungal propertiesTheir cytotoxic studies are in progress and will be reportedsoon


(a) (b) (c)

Figure 9 ldquoZones of inhibitionrdquo obtained for Petri plates supplemented with (a) plain film (b) HG1 (54) film and (c) HG2 (54) film

(a) (b)

Figure 10 ldquoZones of inhibitionrdquo obtained for Petri plates supplemented with film loaded with (a) plain film and (b) miconazole nitrate

Conflict of Interests

Theauthors hereby declare that there is no conflict of interestsregarding the publication of this paper

References

[1] K Peh T Khan and H S Chrsquong ldquoMechanical bioadhesivestrength and biological evaluations of chitosan films for wounddressingrdquo Journal of Pharmacy amp Pharmaceutical Sciences vol3 no 3 pp 303ndash311 2000

[2] G Majno The Healing Hand Man and Wound in the AncientWorld Harvard University Cambridge UK 1975

[3] S Thomas Wound Management and Dressing PharmaceuticalPress London UK 1990

[4] L G Ovington ldquoAdvances in wound dressingsrdquo Clinics inDermatology vol 25 no 1 pp 33ndash38 2007

[5] T Abdelrahman and H Newton ldquoWound dressings principlesand practicerdquo Surgery vol 29 no 10 pp 491ndash495 2011

[6] A J Singer and A B Dagum ldquoCurrent management of acutecutaneous woundsrdquo The New England Journal of Medicine vol359 no 10 pp 1037ndash1046 2008

[7] MM G Fouda RWittke D Knittel and E Schollmeyer ldquoUseof chitosanpolyamine biopolymers based cotton as a modelsystem to prepare antimicrobial wound dressingrdquo InternationalJournal of Diabetes Mellitus vol 1 no 1 pp 61ndash64 2009

[8] B Steffansen and S P K Herping ldquoNovel wound modelsfor characterizing ibuprofen release from foam dressingsrdquoInternational Journal of Pharmaceutics vol 364 no 1 pp 150ndash155 2008

[9] H Xu LMa H Shi C Gao andCHan ldquoChitosan-Hyaluronicacid hybrid film as a novel wound dressing in vitro and in vivostudiesrdquo Polymers for Advanced Technologies vol 18 no 11 pp869ndash875 2007

[10] B Singh and L Pal ldquoSterculia crosslinked PVA and PVA-poly(AAm) hydrogel wound dressings for slow drug deliverymechanical mucoadhesive biocompatible and permeabilitypropertiesrdquo Journal of the Mechanical Behavior of BiomedicalMaterials vol 9 pp 9ndash21 2012


[11] M Pandima Devi M Sekar M Chamundeshwari et al ldquoAnovel wound dressing material-Fibrin-chitosan-sodium algi-nate composite sheetrdquo Bulletin of Materials Science vol 35 no7 pp 1157ndash1163 2012

[12] H V Pawar J Tetteh and J S Boateng ldquoPolyox and car-rageenan based composite films dressings containing antimi-crobial and anti-inflammatory drugs for effective wound heal-ingrdquo International Journal of Pharmaceutics vol 441 no 1-2 pp181ndash191 2013

[13] S Bielecki A Krystynowicz M Turkiewicz and H Kali-nowska ldquoBacterial celluloserdquo in Biopolymers PolysaccharidesI Munster A Steinbuchel Ed vol 5 pp 37ndash90 Wiley-VCHGmbH Weinheim Gremany 2002

[14] Y Nishi M Uryu S Yamanaka et al ldquoThe structureand mechanical properties of sheets prepared from bacterialcellulosemdashpart 2 improvement of the mechanical properties ofsheets and their applicability to diaphragms of electroacoustictransducersrdquo Journal of Materials Science vol 25 no 6 pp2997ndash3001 1990

[15] J Shah and R M Brown Jr ldquoTowards electronic paper displaysmade from microbial celluloserdquo Applied Microbiology andBiotechnology vol 66 no 4 pp 352ndash355 2005

[16] B R Evans H M OrsquoNeill V P Malyvanh I Lee and JWoodward ldquoPalladium-bacterial cellulose membranes for fuelcellsrdquo Biosensors and Bioelectronics vol 18 no 7 pp 917ndash9232003

[17] J D Fontana A M de Souza C K Fontana et al ldquoAcetobactercellulose pellicle as a temporary skin substituterdquo Applied Bio-chemistry and Biotechnology vol 24-25 pp 253ndash264 1990

[18] O M Alvarez M Patel J Booker and L Markowitz ldquoEffec-tiveness of a biocellulose wound dressing for the treatment ofchronic venous leg ulcers results of a single center randomizedstudy involving 24 patientsrdquoWounds vol 16 no 7 pp 224ndash2332004

[19] W Czaja M Kawecki A Krystynowicz K Wysota S Sakieland P Wroblewski ldquoApplication of bacterial cellulose in treat-ment of second and third degree burnsrdquo in Proceedings of the227th ACS National Meeting Anaheim Calif USA April 2004

[20] M Ul-Islam T Khan W A Khattak and J K Park ldquoBacterialcellulose-MMTNano reinforced composite films novel wounddressing material with antibacterial propertiesrdquo Cellulose vol20 no 2 pp 589ndash596 2013

[21] I Siro and D Plackett ldquoMicrofibrillated cellulose and newnanocomposite materials a reviewrdquo Cellulose vol 17 no 3 pp459ndash494 2010

[22] W Czaja A Krystynowicz S Bielecki and R M BrownJr ldquoMicrobial cellulosemdashthe natural power to heal woundsrdquoBiomaterials vol 27 no 2 pp 145ndash151 2006

[23] J V Cartmell and W R Sturtevant ldquoTransparent hydrogelwound dressingrdquo U S Patent 5106629 A 1992

[24] M-R Hwang J O Kim J H Lee et al ldquoGentamicin-loadedwound dressing with polyvinyl alcoholdextran hydrogel gelcharacterization and in vivo healing evaluationrdquo AAPS Pharm-SciTech vol 11 no 3 pp 1092ndash1103 2010

[25] M Bajpai S K Bajpai and D J Goutam ldquoAtom transfer radicalpolymerization of glycidyl methacrylate (GMA) in emulsionrdquoJournal of Macromolecular Science A Pure and Applied Chem-istry vol 50 pp 120ndash127 2013

[26] N A Pascarelli E Moretti G Terzuoli et al ldquoEffects ofgold and silver nanoparticles in cultured human osteoarthriticchondrocytesrdquo Journal of Applied Toxicology vol 33 no 12 pp1506ndash1513 2013

[27] X Q Wang H E Chang and R J Francis ldquoSilver deposits incutaneous burn scar tissue is a common phenomenon followingapplication of a silver dressingrdquo Journal of Cutaneous Pathologyvol 36 no 7 pp 788ndash792 2009

[28] X Gan T Liu J Zhong X Liu and G Li ldquoEffect of silvernanoparticles on the electron transfer reactivity and the cat-alytic activity of myoglobinrdquo ChemBioChem vol 5 no 12 pp1686ndash1691 2004

[29] L Braydich-Stolle S Hussain J J Schlager and M-C Hof-mann ldquoIn vitro cytotoxicity of nanoparticles in mammaliangermline stem cellsrdquo Toxicological Sciences vol 88 no 2 pp412ndash419 2005

[30] H Jin C Zha and L Gu ldquoDirect dissolution of cellu-lose in NaOHthioureaurea aqueous solutionrdquo CarbohydrateResearch vol 342 no 6 pp 851ndash858 2007

[31] J Zhou C Chang R Zhang and L Zhang ldquoHydrogelsprepared from unsubstituted cellulose in NaOHurea aqueoussolutionrdquoMacromolecular Bioscience vol 7 no 6 pp 804ndash8092007

[32] H Xu L Ma H Shi C Gao and C Han ldquoChitosan-hyaluronicacid hybrid film as a novel wound dressing in vitro and in vivostudiesrdquo Polymers for Advanced Technologies vol 18 no 11 pp869ndash875 2007

[33] X L Shen J M Wu Y Chen and G Zhao ldquoAntimicrobial andphysical properties of sweet potato starch films incorporatedwith potassium sorbate or chitosanrdquo FoodHydrocolloids vol 24no 4 pp 285ndash290 2010

[34] ldquoASTM standard test method for water vapour transmission ofmaterialsrdquo Designation E vol 701 pp 93ndash96 1993

[35] S Park P S K Murthy S Park Y M Mohan and W-G Koh ldquoPreparation of silver nanoparticle-containing semi-interpenetrating network hydrogels composed of pluronic andpoly(acrylamide) with antibacterial propertyrdquo Journal of Indus-trial and Engineering Chemistry vol 17 no 2 pp 293ndash297 2011

[36] H V Chavda and C N Patel ldquoEffect of crosslinker concentra-tion on characteristics of superporous hydrogelsrdquo InternationalJournal of Pharmaceutical Investigation vol 1 no 1 pp 17ndash212011

[37] A R Khare and N A Peppas ldquoSwellingdeswelling of anioniccopolymer gelsrdquo Biomaterials vol 16 no 7 pp 559ndash567 1995

[38] C Ozeroglu and A Birdal ldquoSwelling properties of acrylamide-NN1015840-methylene bis(acrylamide) hydrogels synthesized byusing meso-23-dimercaptosuccinic acid-cerium(IV) redoxcouplerdquo Express Polymer Letters vol 3 no 3 pp 168ndash176 2009

[39] V Rattanaruengsrikul N Pimpha and P Supaphol ldquoDevelop-ment of gelatin hydrogel pads as antibacterial wound dressingsrdquoMacromolecular Bioscience vol 9 no 10 pp 1004ndash1015 2009

[40] H Thu M H Zulfakar and S Ng ldquoAlginate based bilayerhydrocolloid films as potential slow release modern wounddressingrdquo International Journal of Pharmaceutics vol 434 no1-2 pp 375ndash383 2012

[41] F Febriyenti A M Noor and S B B Baie ldquoMechanicalproperties and water vapour permeability of film from Haruan(Channa striatus) and Fusidic acid spray for wound dressingand wound healingrdquo Pakistan Journal of Pharmaceutical Sci-ences vol 23 no 2 pp 155ndash159 2010



[43] Y-B Wu S-H Yu F-L Mi et al ldquoPreparation and char-acterization on mechanical and antibacterial properties ofchitsoancellulose blendsrdquo Carbohydrate Polymers vol 57 no4 pp 435ndash440 2004

[44] D Jia Y Fang and K Yao ldquoWater vapor barrier andmechanicalproperties of konjac glucomannan-chitosan-soy protein isolateedible filmsrdquo Food and Bioproducts Processing vol 87 no 1 pp7ndash10 2009

[45] Q-P Zhong andW-S Xia ldquoPhysicochemical properties of edi-ble and preservative films from chitosancassava starchgelatinblend plasticized with glycerolrdquo Food Technology and Biotech-nology vol 46 no 3 pp 262ndash269 2008

[46] N J Trengrove S R Langton and M C Stacy ldquoBiochemicalanalysis of wound fluid from nonhealing and healing chronicleg ulcersrdquo Wound Repair and Regeneration vol 4 no 2 pp234ndash239 1996

[47] J Bonnema D A Ligtenstein T Wiggers and A N vanGeel ldquoThe composition of serous fluid after axillary dissectionrdquoEuropean Journal of Surgery vol 165 no 1 pp 9ndash13 1999

[48] T Higuchi ldquoMechanism of sustained-action medicationTheo-retical analysis of rate of release of solid drugs dispersed in solidmatricesrdquo Journal of Pharmaceutical Sciences vol 52 no 12 pp1145ndash1149 1963

[49] R B Najafi Z Maghrouri and M Peikanpour ldquoPreparationand pharmaceutical evaluation of miconazole nitrate mucoad-hesive films for vaginal candidiasisrdquo Journal of Isfahan MedicalSchool vol 30 no 216 pp 2103ndash2112 2013

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy


Carbohydrate Chemistry

International Journal of


Journal of

Chemistry


Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of


Chromatography Research International


Applied ChemistryJournal of



Theoretical ChemistryJournal of


Journal of

Spectroscopy

Analytical ChemistryInternational Journal of


Journal of


Quantum Chemistry


Organic Chemistry International

ElectrochemistryInternational Journal of



CatalystsJournal of


Ominus

OminusOminus

ON

Cl

ClCl

Cl

N

Miconazole nitrate

Figure 1 Molecular structure of drug miconazole nitrate

process visually [23] In addition variation in the degreeof crosslinking of polymeric hydrogel may be helpful incontrolling its mechanical properties moisture permeabilityand release rate of loaded bioactive ingredient [24]Thereforecombination of cellulose and synthetic polymeric hydrogelcould result in an efficient wound dressing film with most ofthe desirable properties

Recently coinage metal nanoparticles such as silver havebeen used as antibacterial agents in fabrication of wounddressing films [25] But as per some reports silver nanoparti-cles are harmful for human osteoarthritic chondrocytes whenused in higher concentrations of around 160ndash250 120583M [26]Moreover nanoparticles present in blood are associated withthrombosis and activation of immunological reactions [27ndash29] As an alternative to metal nanoparticles we here reportdrug miconazole nitrate loaded cellulosepoly(acrylic acid)hydrogel films for antibacterial applications

2 Experimental

21 Materials Cellulose (Cell) powder was purchased fromCentral Drug House Mumbai India Its viscosity averagemolecular weight (Mn) was calculated according to equation[(p)] = 385 times 10minus2 Mn076 [25] and was found to be186 times 10

4 The monomer acrylic acid (AAc) cross-linkerNN1015840-methylenebisacrylamide (MB) and initiator potassiumper sulfate (KPS) were obtained from Hi Media MumbaiIndia The antibiotic drug miconazole nitrate was obtainedfrom Ranbaxy pharmacy as commercial product Micogel(Figure 1 shows structure of drug miconazole nitrate usedin the study) Nutrient broth nutrient agar and agar-agartype-1 were obtained from Hi Media Chemicals MumbaiIndiaThe department of biotechnology (GovernmentModelScience College Jabalpur (Madhya Pradesh) India) providedstandard culture of the organisms Double distilled water wasused throughout the investigation

22 Dissolution of Cellulose in NaOHUrea Solvent SystemCellulose solution was prepared in NaOHurea solvent

system according to procedure reported by Jin et al [30] Inbrief 6 g of NaOH and 4 g of urea were dissolved in 90mL ofdistilled water and the resulting solution was filtered with aG2 sand filter Now 4 g of cellulose powderwas dispersed into100 g of above solution under constant stirring for 20min andthen stored in refrigerator (minus5 to minus10∘C) for 12 h The frozensolid was thawed and stirred extensively at room temperatureto obtain a colorless and almost transparent solution Thecellulose solution was dialyzed against distilled water using adialysis membrane with 10000 molecular weight cut-off Thedialysis was carried out for a period of 2 days The resultingsuspension was used to prepare the hydrogels as described inthe next section

23 Synthesis of Regenerated CellulosePoly(AAc) Hydro-gel Film The regenerated cellulosepoly(acrylic acid) (RCpoly(AAc)) hydrogel film was prepared by free-radical poly-merization of AAc in the presence of above dissolved cellu-lose In brief 1266mMofmonomer AAc 0194mMof cross-linker MB and 0295mM of initiator KPS were mixed into25mL of above dialyzed cellulose solution (4wt) and thetotal volume was made up to 5mL by addition of water Thereaction solution was transferred into Petri plates and keptin electric oven (Tempstar India) at 60∘C for a period of 2 hThe resulting hydrogel film was peeled off and transferredinto 200mL of ammonium sulfate solution (8wv) for aperiod of 8 h Finally RCpoly(AAc) hydrogel film was takenout and allowed to equilibrate in excess of refreshing waterfor a period of 7 days till all the remaining chemicals in thehydrogel film were leached out The compositions of varioushydrogel films prepared are given in Table 1

24 Yield of Hydrogel Film Freshly prepared RCpoly(AAc)hydrogel film was placed in refreshing distilled water tillthe external medium acquired pH almost that of distilledwater Now the swollen hydrogel was placed in 50 percent(vv) aqueous solution of methanol for a period of twodays In order to ensure the complete extraction the gel wastransferred into acetone for 12 hours followed by completedrying

The percent hydrogel yield was calculated as per thefollowing expression

Hydrogel yield =Dry weight of polymerTotal weight of reactants

times 100 (1)

Here total weight of reactants includes monomer AAcpolymer cellulose and cross-linker MB

25 Preparation of Drug-Loaded Film Thedrug-loaded filmswere prepared by adding precalculated amount of drug tothe reaction mixture before the polymerization started Thismethod enables us to incorporate a desired quantity of drugin the polymer matrix The hydrogel film samples weredenoted as HG(X) where the number in the parenthesisdenotes the amount of drug present in one gram of film

26 Water Absorption Study In the gravimetric method thepreweighed sample was placed in 500mL of distilled water





SR =(119872119905minus1198720)

1198720

times 100 (2)

where1198720and119872





WVTR = Δ119882Δ119905 sdot 119860









(a) (b)


0123456789

10

0 100 200 300 400

Swel

ling

ratio

(gg

)

Time (min)

HG1HG2

HG3





ln119865 = ln 119896 + 119899 ln 119905 (4)



minus3

minus25

minus2

minus15

minus1

minus05

0

05

1

0 1 2 3 4 5 6

HG1HG2

HG3

ln MtMinfin

lnt







0123456789

0 100 200 300 400 500

Elon

gatio

n (c

m)

Weight (g)

HG1 HG2HG7





0

002

004

006

008

01

012

014

016

0 5 10 15 20 25 30Time (h)

HG1HG4HG5

HG6HG7

Moi

sture

gai

n (m

g hminus1

cmminus2)






0

1000

2000

3000

4000

5000

6000

0 100 200 300 400

Dru

g re

leas

e (m

Lm

g)

Time (min)

HG1

HG2



2



119876119905=119872119905

119872infin

= 11987011986711990512 (5)


119867




0

01

02

03

04

05

06

0 5 10 15 20

HG1HG2

MtM

infin

tminus12

(min)12






4 Conclusion



(a) (b) (c)


(a) (b)




References





























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of





SR =(119872119905minus1198720)

1198720

times 100 (2)

where1198720and119872





WVTR = Δ119882Δ119905 sdot 119860









(a) (b)


0123456789

10

0 100 200 300 400

Swel

ling

ratio

(gg

)

Time (min)

HG1HG2

HG3





ln119865 = ln 119896 + 119899 ln 119905 (4)



minus3

minus25

minus2

minus15

minus1

minus05

0

05

1

0 1 2 3 4 5 6

HG1HG2

HG3

ln MtMinfin

lnt







0123456789

0 100 200 300 400 500

Elon

gatio

n (c

m)

Weight (g)

HG1 HG2HG7





0

002

004

006

008

01

012

014

016

0 5 10 15 20 25 30Time (h)

HG1HG4HG5

HG6HG7

Moi

sture

gai

n (m

g hminus1

cmminus2)






0

1000

2000

3000

4000

5000

6000

0 100 200 300 400

Dru

g re

leas

e (m

Lm

g)

Time (min)

HG1

HG2



2



119876119905=119872119905

119872infin

= 11987011986711990512 (5)


119867




0

01

02

03

04

05

06

0 5 10 15 20

HG1HG2

MtM

infin

tminus12

(min)12






4 Conclusion



(a) (b) (c)


(a) (b)




References





























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


(a) (b)


0123456789

10

0 100 200 300 400

Swel

ling

ratio

(gg

)

Time (min)

HG1HG2

HG3





ln119865 = ln 119896 + 119899 ln 119905 (4)



minus3

minus25

minus2

minus15

minus1

minus05

0

05

1

0 1 2 3 4 5 6

HG1HG2

HG3

ln MtMinfin

lnt







0123456789

0 100 200 300 400 500

Elon

gatio

n (c

m)

Weight (g)

HG1 HG2HG7





0

002

004

006

008

01

012

014

016

0 5 10 15 20 25 30Time (h)

HG1HG4HG5

HG6HG7

Moi

sture

gai

n (m

g hminus1

cmminus2)






0

1000

2000

3000

4000

5000

6000

0 100 200 300 400

Dru

g re

leas

e (m

Lm

g)

Time (min)

HG1

HG2



2



119876119905=119872119905

119872infin

= 11987011986711990512 (5)


119867




0

01

02

03

04

05

06

0 5 10 15 20

HG1HG2

MtM

infin

tminus12

(min)12






4 Conclusion



(a) (b) (c)


(a) (b)




References





























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


0123456789

0 100 200 300 400 500

Elon

gatio

n (c

m)

Weight (g)

HG1 HG2HG7





0

002

004

006

008

01

012

014

016

0 5 10 15 20 25 30Time (h)

HG1HG4HG5

HG6HG7

Moi

sture

gai

n (m

g hminus1

cmminus2)






0

1000

2000

3000

4000

5000

6000

0 100 200 300 400

Dru

g re

leas

e (m

Lm

g)

Time (min)

HG1

HG2



2



119876119905=119872119905

119872infin

= 11987011986711990512 (5)


119867




0

01

02

03

04

05

06

0 5 10 15 20

HG1HG2

MtM

infin

tminus12

(min)12






4 Conclusion



(a) (b) (c)


(a) (b)




References





























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


0

1000

2000

3000

4000

5000

6000

0 100 200 300 400

Dru

g re

leas

e (m

Lm

g)

Time (min)

HG1

HG2



2



119876119905=119872119905

119872infin

= 11987011986711990512 (5)


119867




0

01

02

03

04

05

06

0 5 10 15 20

HG1HG2

MtM

infin

tminus12

(min)12






4 Conclusion



(a) (b) (c)


(a) (b)




References





























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


(a) (b) (c)


(a) (b)




References





























































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of



















































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

investigation of regenerated cellulose/poly (acrylic acid

Documents