Crosslinking of poly(vinylpyrrolidone)/acrylic acid with tragacanth gum

for hydrogels formation for use in drug delivery applications

a bs t r ac t
Tragacanth gum (TG) is generally recognized as safe by the Food and Drug Administration. The present article discusses the
design of ciprooxacin loaded TG based hydrogels for use in drug delivery especially to improve the pharmacotherapy of
diverticulitis. The polymers were characterized by SEMs, FTIR, 13C NMR, XRD, TGA, DSC, gel strength and swelling studies. The
polymer network parameters, mucoadhe- sion, gel strength, drug release mechanism and kinetic model were also determined.
The release of drug occurred through non-Fickian diffusion mechanism and best tted in the Korsmeyer-Peppas model. The pH of
the swelling medium has also exerted a strong effect on polymer network structure and mechanical strength. These hydrogels
have been observed pH responsive and mucoadhesive in nature and could be utilized for site specic drug delivery.
1. Introduction

Tragacanth gum (TG) is generally recognized as safe (GRAS) by the Food and Drug Administration (FDA), when used
in accordance with good manufacturing or feeding practice. It is also classied as GRAS at the 0.21.3% level in food
stuffs in the USA, since 1961 and included in the list of additives approved by the Scientic Commit- tee for Food of
the European Community (Anderson & Bridgeman, 1985; Eastwood, Brydon, & Anderson, 1984). TG has been used as
an adhesive or binding agent for pills and tablets. It is a bulk-forming laxative, which increases the bulk of the stools
by absorbing water which stimulates the bowel muscles to become active. In addition, it softens the stools and help to
relieve constipation which is a cause diverticulitis (pez-Franco, Higuera-Ciapara, Goycoolea, & Wang, 2009;
Golmohammadi, 2013).
Poly(vinyl pyrrolidone) (PVP) is a hydrophilic, mucoadhesive, and biocompatible polymer. Due to these properties, it
has been used for various biomedical applications (Zhao, Xu, Mitomo, & Yoshii, 2006). The hydrogels developed from
poly(acrylic acid) [PAAc] are pH responsive and bio-adhesive in nature. These drug loaded hydrogels stick to the
mucosal lining of small intestine and increase the bioavailability of drugs. It has been found that the release rate of
ketoprofen from the [PVP]/PAAc based mucoad- hesive hydrogels was mainly controlled by the dissolution rate
and diffusion rate of drug in solution of different pH (Chun, Cho, & Choi, 2002). The use of anionic mucoadhesive
hydrogels have been proposed for the prolonging gastric retention time (Mura, Cirri,. Mennini, Casella, &
Maestrelli, 2016). However, in another research report, it has been found that as carbopol proportion increased in
the hydrogels, the hardness and compressibility of the hydrogels decreased, but their bio-adhesion force and pH-
responsiveness increased (Tenreiro, Bueno, Concheiro, Labandeira, & Lorenzo, 2007).
Ciprooxacin is a broad spectrum antibiotic drug and is effec- tively used to treat diverticulitis and a variety of
bacterial infections of Gram-negative and Gram-positive pathogens (Chouhan & Bajpai, 2010). Its conventional
formulations liberate the drug contents along the intestine which results to diminish its efcacy. Hence, its
controlled release systems are required to improve its phar- macokinetic and pharmacodynamic prole ( Mostafavi,
Emami, Varshosaz, Davies, & Rezazadeh, 2011). Its mmucoadhesive for- mulations, further improve the bioavailability,
residence time in gastrointestinal tract and efcacy for the treatment of microbial infections ( Hardenia, Jain, Patel, &
Kaushal, 2011; Jain, Kare, Jain, & Singh, 2011). In general, the hydrogels can be prepared by modication of
polysaccharides through grafting and crosslinking (Athawale & Rathi, 1997). The diffusion of drug from the hydrogels
can be controlled by tailoring the crosslinked network structure (Sannino, Demitri and Madaghiele, 2009). In order to
understand the cross-linked structure of the gel, the most common approach is to study the swelling of the hydrogels
(Caykara, Birlik, & Izol, 2007; Jridi et al., 2015, 2014).
Keeping in view of the above stated facts, (i.e. pH responsive nature of PAAc, mucoadhesive nature of PVP,
antimicrobial action of ciprooxacin for diverticulitis and laxative action of TG), in the present study, ciprooxacin
loaded mucoadhesive hydrogels were prepared for the use in slow drug delivery system for diver- ticulitis. These
hydrogels were designed by using TG-PVP-PAAc. The swelling, drug release, blood-compatibility, mucoadhesive and
mechanical properties of the hydrogels were also determined. TG also reduces the intraluminal pressure in the
bowel, shortening the stool transit time which reduces the stress on the colon wall and therefore avoids the
formation of diverticuli. Overall the poten- tial of the present drug delivery system for the treatment of the
diverticulitis will enhance.

2. Experimental

2.1. Materials and methods

Acrylic acid (AAc) was obtained from Merck Specialities Pri- vate Limited (Mumbai, India), polyvinyl pyrrolidone,
PVP (K 90, molecular weight 360000) was obtained from SIGMA-ALDRICH (USA), N,N-methylenebisacrylamide
(NN-MBA) and ammonium persulphate (APS) were obtained from Qualigens Fine Chemicals (Mumbai, India) and
tragacanth gum (TG) was obtained from Loba Chemie (Mumbai, India). Ciprooxacin HCl (Ranbaxy Laboratories
Limited, Himachal Pradesh, India) was procured from the market.

2.2. Synthesis of TG-cl-(PVP-co-PAAc) hydrogels

The polymerization reaction was carried out with a denite concentration of tragacanth gum (6% w/v), AAc (0.291
mol/L), PVP (2% w/v), initiator APS (0.0263 mol/L) and cross-linker NN-MBA 6300 thermal analyzer under air
atmosphere at 10 C/min heating rate. DSC scans of the powdered samples were recorded NETZSCH DSC 204,
USA. Gel strength of hydrogels was carried out by texture analyzer (TA.XT plus Stable Micro system, UK) with 5 kg
load cell.

2.4. Drug release studies

The release prole of model drug ciprooxacin HCl, from the drug loaded polymers was determined from the
standard curves prepared by using UV Visible Spectrophotometer (Cary 100 Bio, Varian Australia, ISO 9001)). The
standard curves were prepared at hmax.276 nm (in distilled water), at hmax. 277 (in pH 2.2 buffer) and at hmax.271 (in
pH 7.4 buffer solutions). All the studies were carried out in triplicate. The mechanisms of swelling of TG -cl- (PVP-co-
PAAc) hydrogels and drug release from hydrogels were determined by applying the various equations given by Ritger
 and Peppas (Ritger & Peppas, 1987a,b).
B. Singh, V. Sharma / Carbohydrate Polymers 157 (2017)
Different parameters of release kinetics of drug from the drug loaded
185195 3
hydrogels were determined. Different kinetic models (i.e. zero order, rst order, Higuchi square root law, Korsmeyer-
Peppas model and Hixson-Crowell cube root) were also applied to drug release data obtained for release of drug
from hydrogels in different medium (Singh & Sharma, 2014). Drug delivery system used for the drug release studies
was like a tablet in shape and size. Drug loading efciency of samples was
69.80 0.37%. The loading of a drug into the hydrogels was carried
out by swelling equilibrium method.

2.5. Determination of network parameters of hydrogels

The polymer network parameters such as polymer volume frac- tion in the swollen state ($), Flory-Huggins
interaction parameter (y), molecular weight of the polymer chain between two neigh-
(0.0389 mol/L), taken in the aqueous reaction system in a beaker.
The reaction contents were stirred for 3 h at 25 C, and polymer- ization reaction was carried out for 2 h at 65 C
temperature. After completion of reaction, polymer was then stirred in distilled water to remove the soluble fractions
and then it was dried in an oven at 40 C until the constant weight was obtained. The crosslinked poly- mers were
named as of TG-cl-(PVP-co-PAAc) hydrogels/polymers. The term TG-cl-(PVP-co-PAAc) polymer is used for the dry
samples and term TG-cl-(PVP-co-PAAc) hydrogel is used in case of swelled samples. The optimum reaction parameters
for synthesis of hydro- gels were evaluated, on the basis of swelling of hydrogels after 24 h, by varying [AAc] from
0.291 to 0.874 mol/L, [PVP] from 2 to 6% (w/v) and [NN-MBA] from 0.0130 to 0.0649 mol/L during polymerization
reaction.

2.3. Characterizations

The polymers were characterized by scanning electron micro- graphs (SEMs), Fourier transform infrared
spectroscopy (FTIR), solid state 13C NMR spectroscopy, X-ray diffraction study (XRD), thermo gravimetric analysis
(TGA), differential scanning calorimet- ric (DSC), gel strength and swelling studies. SEMs were taken on FEI SEM
Quanta 256, Model D9393 (Singapore). FTIR spectra of poly- mers were recorded in KBr pellets on Nicolet
5700FTIR THERMO (USA). XRD measurements of polymers were made using PAN- analytical XPert Pro powder
diffraction system (The Netherland). The X-ray generator was operated at 40 kV and 40 mA using the Cu- ka
radiation. A denite amount of sample was scanned between 10 and 80 (20) with the increment of 0.05 in 1.0 s
at 25 C and count by using an automatic divergence slit assembly and a pro- portional detector. The solid state
13
C NMR was carried on solid state NMR spectrometer (BRUKER DSX-300). The spectrophotome- ter was operated at
a magnetic eld of 7.0 T and at a frequency of 75.4 MHz. TGA, DTA and DTG were carried out on EXSTAR TG/DTA boring
cross links (Mc), crosslink density (p) and mesh size ($) of TG-cl-(PVP-co-PAAc) hydrogels were determined (Singh &
Sharma, 2014).

2.6. Biomedical properties of hydrogels

Blood-compatibility study of polymers was carried out by the procedure reported in International Standard
Organization (ISO) (ISO10993-4, 1999). The types of blood interactions were stud- ied; thrombogenicity and
haemolytic potential. The evaluation of thrombus formation on polymer surface was carried out by the
gravimetric method. The haemolysis tests were performed as described in American Society for Testing and
Materials (2000) (ASTM F 756-00). Mucoadhesion study and gel strength of poly- mers ware determined by using
a texture analyzer (TA.XT Stable micro System, UK) equipped with 5 kg load cell and a mucoadhe- sive holder. In
case of mucoadhesion, the maximum force required to separate the probe from the goat mucosa (i.e. maximum
detach- ment force; Fmax in N) was directly recorded in the instrument and the total amount of force involved in
the probe withdrawal from the tissue (work of adhesion; W ad) was then calculated from the area under the force
versus distance curve in N mm. The materials tested with texture analyzer having thickness = 0.64 0.01 cm
and radius = 0.59 0.01 cm.

3. Results and discussion

3.1. Characterizations

3.1.1. SEM and EDAX of polymers

SEMs images of polymers showed porous surface topogra- phy (Fig. 1). The roughness and heterogeneity was
observed in SEM images (Nakason, Wohmang, Kaesaman, & Kiatkamjornwong,
2010). The completely different surface morphology and pore size of composite hydrogels has been found as
compared to raw mate- rials (Mishra, Datt, & Banthia, 2008; Thakur, Wanchoo, & Singh, 2011). The electron
dispersion X-ray analysis (EDAX) spectrum of polymers has shown the increase in the weight percentage of nitro-
gen in cross-linked polymers as compared to TG which indicates the presence of cross-linker NN-MBA and PVP in
crosslinked polymer matrix.

Fig. 1. Scanning Electron Micrographs (SEMs) of (a) dried and (b) swelled TG-cl-(PVP-co-PAAc) hydrogel.
 3.1.2. FTIR spectra of polymers
FTIR spectrum of TG-cl-(PVP-co-PAAc) polymer is given in Fig. 2a. The absorption bands at 3406.4 cm 1 (-OH
stretching vibrations), 2928.1 cm 1 (aliphatic CH2 stretching vibrations), 2361.7 cm1 (overtones and combinations
of OH bending and C O stretching vibrations), 1733.2 cm 1 (C O stretching of
COOH), 1653.8 cm 1(C O group of PVP), 1542.0 cm 1 (asym- metrically stretching vibration of carboxylate ion on
PAAc chain), 1448.0 cm 1 ( CH2 bending), 1252.5 cm1 (due C N stretching vibrations of NVP ring), and between 1040
and 1190 cm1 (C O
stretching of COOH) were observed along with the other absorp- tion. The presence of more predominant bands
at 1733.2 cm1 and 1653.8 cm1 showed the incorporation of PVP and PAAc in crosslinked polymer matrix
(Srivastava, Mishra, & Behari, 2010).

3.1.3. XRD studies of polymers

XRD of TG-cl-(PVP-co-PAAc) polymers showed a broad low intensity peak (at 20 equal to 21) due to amorphous
nature of the crosslinked polymers (Fig. 2b). Grafting of vinylic monomers onto TG polysaccharide has changed their
micro-crystalline nature to amorphous nature as intensity of diffraction peak decreased or diminished (Nguyen & Liu,
2013). Vakili and Rahneshin (Vakili & Rahneshin, 2013) have also observed that grafting of L-aspartic acid onto starch
decreased its crystallinity by decreasing peak intensity due to breaking of the inter-chain hydrogen bonding in starch.

3.1.4. 13C NMR spectroscopy of polymers

The solid state 13C NMR spectra of polymer samples are shown in Fig. 2c. TG-cl-(PVP-co-PAAc) polymers showed
peaks at 18.429 ( CH3), 21.233 ( CH2 ), 31.649 (C-1 of PVP), 43.268 ( CH of
polyacrylic acid and PVP), 70.911 ( C OH), 104.964 (anomeric car-
bon of polysaccharide) and 176.976 ppm ( C O group of polymer matrix). Presence of peaks at the chemical shift
between 30 and 50 ppm may be due grafting of (CH 2 CH)n units of PAAc and PVP onto polysaccharide backbone TG
(Zou et al., 2012).

3.1.5. TGA, DTA and DTG of polymers

The TGA/DTA/DTG of TG and TG-cl-(PVP-co-PAAc) polymers are shown in Fig. 2d. In case of the TG-cl-(PVP-co-PAAc)
polymers, ini- tial 9.1% weight loss occurred due to removal of bounded water of polymer between 25 and 100 C.
Initial decomposition temperature (IDT) and nal decomposition temperature (FDT) were obtained at 193 and 544 C
(residue left = 2%), respectively. Three stages decom- position mechanism was observed for the decomposition of the
crosslinked polymers. These three stages i.e rst, second and third, started at 193 C (residue left = 86.8%), 292 C
(residue left = 55.1%) and 390 C (residue left = 17.8%) respectively. The second stage was mild stage of
decomposition. The TGA of grafted polymer sam- ples showed gradual decrease in weight without any abrupt loss of
weight like tragacanth gum and PVP. Crosslinked polymer showed different behavior in term of thermal stability as
compared to tra- gacanth gum and PVP (Bianco et al., 2003). However, the most of the weight loss occurred within the
temperature range 225450 C, in which the PVP based polymers displayed the highest thermal sta- bility at the
different heating temperatures with the lowest weight loss (Alla, El-Din & El-Naggar, 2007). DTA curves of TG and TG-
cl- (PVP-co-PAAc) polymers showed endothermic peaks at 83 C and 77 C respectively due to endothermic release of
water from poly- mers. The temperature of maximum rate of weight loss (T max) for polymer matrix was found at 501 C
(496.1 g/min) in third stage of thermal decomposition. DTG of polymer matrix showed that cross-linked polymers
decomposed at lower rate as compared to tragacanth gum and PVP.

3.1.6. DSC analysis of polymers

DSC curves of TG-cl-(PVP-co-PAAc) polymers are shown in Fig. 2e. The broad endothermic peak at 88.6 C with a
heat of fusion of 449.2 J/g indicated the amorphous nature of tragacanth gum. The glass transition temperature (T g)
has not been observed. In case of the TG-cl-(PVP-co-PAAc) polymers, a broad endother- mic peak centered at 80.4 C
with heat of fusion of 295.7 J/g has been observed with glass transition state in the temperature range of 185.3
200.5 C (6Cp = 0.669 J/gK). The lower-temperature endotherm between 40 C and 120 C is attributed to the evolu-
tion of unbound water from the polymer matrix. Presence of glass transition state in DSC curve of the TG- cl-(PVP-co-
PAAc) polymers showed the modication of tragacanth gum. This increase in glass transition temperature (T g) of
polymer matrix, as compared to PVP, may be attributed to the presence of intermolecular hydro- gen bonding
between PAAc ( COOH) and PVP ( C O). Single T g peak for crosslinked polymer showed good miscibility between
tragacanth gum, PVP and poly(AAc). Broad endothermic peak above 200 C may be due to de-polymerization of
backbone chains (Soares, de Castro, Cury, & Evangelista, 2013). In case of the grafted samples of another
polysaccharide, i.e. guar gum-g-AAc/PVA, sim- ilar type of new broad endothermic peak have also been found at
higher temperature (>200 C). This may be due to enhanced inter- action between carbonyl groups of the grafted
copolymer and the hydroxyl groups of guar gum which conrming the grafting of AAc onto guar gum (Sullad,
Manjeshwar, & Aminabhavi, 2010).

Fig. 2. (a) FTIR, (b) XRD, (c) C13-NMR, (d) TGA/DTA/DTG and (e) DSC analysis of TG-cl-(PVP-co-PAAc) hydrogel.

3.2. Swelling studies

In order to study the effect of synthetic reaction parameters (i.e. AAc, PVP and crosslinker concentration) on
the structure of the polymer network hydrogels, swelling studies of hydrogels werecarried out at 37 C in distilled
water. Fig. 3 indicate the effect of (a) [AAc],
B. Singh, V. Sharma / Carbohydrate Polymers 157 (2017)
(b) [PVP], (c) [NN-MBA], (d) pH, (e) salt concentration and (f) temperature
185195 5
of swelling medium on swelling of TG-cl-(PVP-co- PAAc) hydrogels as a function of time and reaction parameter. Insert
gures show the effect of (a) [AAc], (b) [PVP], and (c) [NN-MBA] on the swelling of TG-cl-(PVP-co-PAAc) hydrogels after
24 h. Swelling of hydrogels decreased (from 5.178 0.137 to 1.687 0.043 g/g of gel) with increase in feed [AAc]
(0.2910.874 mol/L) during polymerization reaction (Fig. 3a). This may be due to increase in crosslink density of the
hydrogels with increase in feed monomer concentration. Gel content inuences the crosslink density of the polymer
network and it depends on the composition of the reac- tion mixture. The increase in total monomer concentration in
the pregel reaction mixture increases the chain interpenetration and efciency of the cross-linking reaction in the
resulting hydrogels (Baker, Hong, Blanch, Prausnitz 1994). The values of diffusion expo- nent n and gel characteristics
constant k have been evaluated and are results are presented in Table 1. The results show that the swelling of the
polymers prepared with different monomer con- centration occurred through a non-Fickian diffusion mechanism.

Fig. 3. Effect of (a) [AAc], (b) [PVP], (c) [NN-MBA], (d) pH, (e) salt concentration and (f) temperature of swelling medium on swelling of TG-cl-
(PVP-co-PAAc) hydrogels as a function of time and reaction parameter. Insert show the effect of (a) [AAc], (b) [PVP], and (c) [NN-MBA] on the
swelling of TG-cl-(PVP-co-PAAc) hydrogels after 24 h.
 19 B. Singh, V. Sharma / Carbohydrate Polymers 157 (2017)
0 185195

Table 1
Results of diffusion exponent n, gel characteristic constant k and various diffusion coefcients for the swelling kinetics of TG-cl-(PVP-co-PAAc)

hydrogels.

In this mechanism during polymer swelling the rate of solvent diffusion is comparable to the rate of polymer chain
relaxation. The solvent enhances the mobility of polymer chains by convert- ing the glassy matrix to a swollen
rubbery material (Rebenfeld, Makarewicz, Weigmann, & Wilkes, 1976). The values of diffusion coefcients are given in
Table 1.
To study the effect of feed [PVP] on network structure of TG-cl- (PVP-co-PAAc) hydrogels, the PVP concentration was
varied from 2 to 6% (w/v) during polymerization reaction and swelling of hydro- gels was determined ( Fig. 3b). The
swelling of hydrogels increased with increase in [PVP]. This may be due to increase in hydrophilicity of hydrogels due
to incorporation of more PVP in the copolymers (Lanthong, Nuisin, & Kiatkamjornwong, 2006). Swelling of hydro- gels
decreased with increase in cross-linker concentration from 0.0130 to 0.0649 mol/L (Fig. 3c). As the cross-linker
concentra- tion increases in the reaction system, the probability of forming cross-links in the hydrogels increases. The
exibility of macro- molecular chains decreases which makes the network chains stiffer and results in poor chain
relaxation during swelling. These fac- tors may be responsible for decrease in swelling of the hydrogels ( Pourjavadi,
Ghasemzadeh, & Hosseinzadeh 2004).
The results of swelling studies of the TG-cl-(PVP-co-PAAc) hydrogels in pH 2.2 buffer and pH 7.4 buffer are
shown in Fig. 3d. The higher swelling of hydrogels was observed in pH
7.4 buffer (7.581 0.231 g/g of gel) as compared to pH 2.2 buffer (2.746 0.043 g/g of gel). This may be due to
opening of the pores due to ionic repulsion of the constituted ions ( COO ) formed
after partial hydrolysis/ionization of COOH in solution of pH 7.4. Presence of ionic moieties in hydrogels make them pH
responsive. Alla and coworkers (Alla et al., 2007) have reported the swelling of copolymer network hydrogels of
PVP/AAc prepared by gamma irradiation in aqueous solutions. They have shown pH responsive swelling behavior of
PVP/AAc hydrogels. The equilibrium degree of swelling of these hydrogels has been mainly inuenced by the charge of
the ionic monomer, crosslinking density and structure of the polymer backbone. The water uptake by the hydrogels in
0.9% NaCl solution (2.977 0.061 g/g of gel) has been observed less as compared to the distilled water (4.161
0.078 g/g of gel) (Fig. 3e) due to the screening effect of the ions. The swelling of hydro- gels increased with increase
in temperature of swelling medium between 27 and 47 C with the increment of 10 C (Fig. 3f). The swelling of
hydrogels in different medium has followed non-Fickian diffusion mechanism. The values of diffusion coefcients are
pre- sented in Table 1.

3.3. Network parameters of hydrogels

3.3.1. Effect of [AAc], [PVP] and [NN-MBA] on network parameters of hydrogels

The results of the network parameters of the TG-cl-(PVP- co-PAAc) hydrogels as a function of feed AAc, PVP and
NN-MBA concentrations are presented in Table 2. The values
Mc and ($) of network parameters of hydrogels decreased with

increase in feed [AAc]. Maximum value of Mc (84326.902 g/mol) and $ (41.252 nm) was evaluated for the hydrogel
prepared with [AAc] = 0.291 mol/L. The crosslink density increased from
1.50725 to 6.78578 105 mol/cm3 while polymer volume fraction
increased from 0.12483 to 0.29532 with increase in [AAc]. These results are corresponding to swelling trends of
hydrogels. The y val- ues increased with increase in feed [AAc]. It means that interaction between polymer-water
decreases and polymerpolymer chains increases, which leads to the decrease in pore size and swelling (Li, Wu,
Wang, & Duan, 2006).

Table 2
The network parameters of TG-cl-(PVP-co-PAAc) hydrogels as a function of different variations.
 7
With increase in [PVP] from 2 to 6% w/v, the mesh size ( $) increased from 23.761 to 34.035 nm and molecular weight
of the
polymer chain between two neighboring cross links Mc increased (from 33341.160 to 52466.569 g/mol), except in case of
polymers prepared with 4% PVP. These trends are corresponding to swelling trends of hydrogels. However, decrease in p and $
values of polymer network parameters have been observed with increase in [PVP]. This may be due to increase in hydrophilic
PVP units in the polymer matrix and hydrogels expand to greater extent which leads to less
p values and more Mc values of hydrogels (Katime & de Apodaca, 2000).
The crosslink density (p) of polymer network of hydrogels was increased from 2.06984 to 4.71823 105 mol/cm3 with
increase in [NN-MBA] from 0.0130 to 0.0649 mol/L. The values of $ regularly
decreased (from 33.274 to 21.022 nm) and Mc (from 62054.165 to 27698.309 g/mol) with increase in [NN-MBA] (Table
2). These trends may be due to the increase in number of crosslinks in the gel network which results in a reduction in pore
size of the voids avail- able between the network chains (Lu & Anseth 1999). Similar trends
have also been found for M c , p and $ values on increasing the NN- MBA concentration of PVP/AAc and sodium alginate-g-
poly(sodium acrylate)/PVP hydrogels (Sohail, Khan, Shahzad, Hussain, & Ranjha, 2014; Wang & Wang, 2010). The y values
have been observed more than 0.5 for [AAc], [PVP] and [NN-MBA] variations.

3.3.2. Effect of nature of swelling medium on network parameters of hydrogels

The effect of pH on network parameters of TG-cl-(PVP-co-PAAc) hydrogels have been studies by taking swelling in pH 2.2
buffer, distilled water and pH 7.4 buffer and results are presented in
Table 2. The increase in mesh size $ (from 16.974 to 36.616 nm)
and Mc values (from 21281.126 to 55994.033 g/mol) while decrease in p (from 5.79546 to 2.20263 mol/cm3) and $
(from 0.22722 to 0.096603) values have been observed with change in pH from
2.2 to 7.4 (Table 2). This may be due to more swelling of hydro- gels in pH 7.4 buffer because of ionic repulsion of COO
ions in
basic medium. Gudmann and Peppas (Gudeman & Peppas, 1995) have also observed increase in mesh size of PVP/AAc
polymers on increasing pH of swelling medium.
The $ and Mc values decreased in 0.9% NaCl solution as com- pared to distilled water. The mesh size decreased from 25.899
nm to 21.390 nm in 0.9% NaCl solution. The p and $ values have been increased in salt solution. These results may be due
to screen- ing effect of additional ions (Soppimath, Kulkarni, & Aminabhavi, 2001).
With increase in temperature of swelling medium, $ and p val-

ues of hydrogels decreased while corresponding $ and Mc values of polymer network increased. At xed cross-linker
concentration, increase in temperature of swelling medium leads to decrease in crosslink density, polymer-solvent interaction
parameter y and increase in water content of hydrogels (Xue, Huglin & Liao, 2007). Effect of temperature on the network
parameters such as mesh size, volume fraction of polymer in swollen state, solventsolute inter- action parameter etc. is due to
increase in swelling ratio of polymer with increasing temperature. When the temperature increased, the disentanglement
between polymer chains happened and the poly- meric network expanded ( Li et al., 2006). This increase in swelling ratio with
increasing temperature is due to increase in the rate of diffusion of water molecules into the polymeric networks at high
temperature (Baselga, Hernandez-Fuentes, Masegosa, & Llorente, 1989). More swelling ratio means more swelling of
polymer which means there is an increase in the chain length between two cross -links which results in increase in Mc and
$ and decrease in crosslink density of polymeric network. The y values are more than 0.5 for all cases for different
variation in swelling medium (Table 2).

Fig. 4. Release prole of ciprooxacin HCl from drug loaded TG-cl-(PVP-co-PAAc) hydrogels in different medium at 37 C.

3.4. Gel strength of TG-cl-(PVP-co-PAAc) hydrogels

Fig. 5. Plot of ln(Mt/Moo) versus ln t for the evaluation of diffusion exponent n and gel characteristics constant k for the release of ciprooxacin HCl from
drug loaded TG-cl-(PVP-co-PAAc) hydrogels in different medium at 37 C.

The values of the gel strength of TG-cl-(PVP-co-PAAc) hydro- gels have been recorded on texture analyzer as (78.991 11.4),
(55.085 3.2) and (47.798 3.1) g cm, respectively in pH 2.2 buffer, distilled water and pH 7.4 buffer. The higher swelling
of hydro- gels in pH 7.4 buffer results in decrease in degree of crosslinking and gel strength. The degree of crosslinking in the
hydrogels is an important factor to determining their mechanical properties, which are further dependent upon chemical
composition of hydrogels and nature of external swelling medium. In general the grafting of synthetic polymers onto
polysaccharides enhances the mechan- ical strength of polysaccharide based hydrogels (Caykara, Demirci, Eroglu, & Guven,
2006).

3.5. Drug release studies of TG-cl-(PVP-co-PAAc) hydrogels

The release prole of ciprooxacin from the drug loaded TG-cl-

(PVP-co-PAAc) hydrogels is shown in Fig. 4. Drug released in pH 7.4 buffer was found higher than the pH 2.2 buffer solution.
Here the swelling of the hydrogels is more dominating factor as compared to the solubility of the drug for the release of drug
from the drug loaded hydrogels. The drug release followed a non-Fickian diffu- sion mechanism in which both diffusion and
8
relaxation processes contribute (Fig. 5
B. Singh, V. Sharma / Carbohydrate Polymers 157 (2017)
, Table 3) (Das & Subuddhi, 2013). The values of various diffusion coefcients are
185195
presented in Table 3. The kinetic parameters were evaluated for the release ciprooxacin in differ- ent releasing medium (Fig. 6,
Table 3). The maximum amount of drug (C max) was released in pH 7.4 buffer (255.10 mg/L) solution at highest initial release
rate (ro) as compared to pH 2.2 buffer (117.51 mg/L). Different kinetic models were applied for the release of loaded drug and it
has been found that drug release from hydro- gel has obeyed all kinetic models (R 2 > 0.98) and followed best by Korsmeyer-
Peppas model with highest value of regression coef- cient (R 2) in all release medium (Table 4). Sullad and coworkers (Sullad et
al., 2010) have prepared the pH-sensitive hydrogel blend of poly(vinyl alcohol) with acrylic acid-graft-guar gum and investi-
gated the controlled release of an anti-tuberculosis drug, isoniazid from the drug loaded microspheres. In vitro release
experiments were performed under both gastric (pH 1.2) and intestinal (pH 7.4) conditions. In both dissolution media,
however, drug release was extended up to 8 h. So the pH sensitive hydrogels are effec- tive drug carriers for site specic
release of ciprooxacin where drug release can be controlled by changing the pH of the envi- ronment and have potential
applications in the biomedical eld (Demirci, Celebioglu, Aytac, & Uyar, 2014). Broad spectrum antibi- otics like amoxicillin
and ciprooxacin are generally proposed as a therapy for the treatment of bacterial intestinal infection like crohns
disease, acute ulcerative colitis and diverticulitis (Beckham & Whitlow, 2009; Khan et al., 2011; Mantzaris et al., 1997).

Fig. 6. Release rate curves for release of ciprooxacin HCl from drug loaded TG-cl-
(PVP-co-PAAc) hydrogels in different medium at 37 C.

Table 3
Results of diffusion exponent n, gel characteristic constant k, various diffusion coefcients and release kinetic parameters for the release of ciprooxacin HCl
from drug loaded TG-cl-(PVP-co-PAAc) hydrogels.

Table 4
Kinetic interpretation of drug release by using different models for release of ciprooxacin HCl from TG-cl-(PVP-co-PAAc) hydrogels.

Table 5
Effect of contact force and contact time on mucoadhesion of TG-cl-(PVP-co-PAAc) hydrogels with intestinal mucosa using texture analyzer. Blood-
compatibility studies of TG-cl-(PVP-co-PAAc) hydrogels

Thrombogenicity of TG-cl-(PVP-co-PAAc) hydrogels was deter- mined and weight of clot formed and thrombose
percentage for polymers was found (0.216 0.010 g/2 mL of citrated blood) and (47.26 2.54)%, respectively. It has been
observed that clot formation was lower in case of hydrogels than in the positive control (0.434 0.005 g) and the hydrogels are
classied as non- thrombogenic (Imai & Nose, 1972). The haemolytic index (HI%) for hydrogels is found to be 0.75 0.04%.
From the results it was observed that TG-cl-(PVP-co-PAAc) hydrogels have a haemolytic percentage less than 2% and are found
to be non-haemolytic in nature (Nguyen & Liu, 2013).

3.6. Mucoadhesion studies of TG-cl-(PVP-co-PAAc) hydrogels

The effect of contact force and contact time on mucoadhesion of TG-cl-(PVP-co-PAAc) hydrogels with intestinal mucus
membrane was studied by determining Fmax and Wad by using texture analyzer (Table 5). The Fmax value generally increased
with increase in con- tact force (hold for 60 s). Similar trends have been obtained for work of adhesion. With increase in contact
time (from 60 to 300 s) for a xed contact force (i.e. 0.5 N) both F max and Wad values increased. Longer the contact time with
mucus membrane, higher will be the interaction of polymeric chains with biological membrane and higher will be the force of
adhesion (Wong, Yuen, & Peh, 1999). Sim- ilar trends have been observed for the adhesion of polymeric lms of PVP/PAAc with
buccal mucosa where both, Fmax and Wad values increased with increase in contact time at a contact force of 0.5N (Chary,
Vani, & Rao, 1999). Mucoadhesion of pluronic-PAAc based hydrogels has been found to dependent on pH of the buffer solu- tion.
The adhesive force is highest at pH values of 45 with values of 7580 mN/cm 2, has moderate values at lower pH, and
decreased sharply with increasing pH (Cleary, Bromberg, & Magner, 2004). Highly hydrophilic nature of PVP has limited its
adhesive properties and its blending with polysaccharides has improved its mucoadhe- sion nature ( Karavas, Georgarakis, &
Bikiaris, 2006). PAAc grafted onto the polysaccharide backbone of dextran has showed lesser oral mucoadhesion as compared
to pure PAAc but showed higher mucoadhesion than dextran at higher PAAc concentration in poly- mer (Nerkar & Gattani, 2013;
Thermes et al., 1992). Crosslinked hydroxypropyl cellulose (HPC)-AAc based hydrogels showed the contribution of the
macromolecules diffusion and chain entangle- ment into mucoadhesion with buccal mucosa while mucoadhesion of pure PAAc
is mostly related to its hydrogen bonding with the mucus layer, and the diffusion/entanglement mechanism plays a less
important role (Dubolazov, Nurkeeva, Mun, & Khutoryanskiy, 2006). Overall the combination of PVP and AAc, has developed
a pH responsive hydrogels with better blood compatibility and mucoadhesion. Hence, these hydrogels can be used as site
specic controlled drug delivery systems for GIT. Further all the compo- nents (TG, PVP and PAAc) have been reported to be non
toxic in nature. Gum tragacanth has been used in the food industry which include salad dressings, sauces, bakery emulsions,
llings and top- pings, confectionery, soft drinks, jellies, desserts and ice creams ( Weiping, 2000). Higa, Rogero, Machado,
Mathor and Lugo, (1999) have done in vitro study of biocompatibility for PVP hydrogel mem- branes and have observed no
evidence of cell toxicity and none- hemolytic nature of PVP based hydrogel dressing (Higa et al., 1999).

Ameye et al. (2005) have performed bio-compatibility and mucosal irritation test on polymeric blends of starch/polyacrylic
acid and have reported, these polymers as safe bio-adhesive carrier with no irritation with mucocal membrane

4. Conclusions

From the foregone discussion it is concluded that the compo- sition of the polymers has inuenced the network structure
of the hydrogels. The release of drugs from the drug loaded hydro- gels occurred slowly without any initial burst release and
followed non-Fickian diffusion mechanism. The pH of the swelling medium has also exerted a strong effect on polymer
 9
network structure and mechanical strength. These hydrogels have been observed pH responsive and mucoadhesive in nature
and could be utilize for site specic drug delivery.