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Carbohydrate Polymers 158 (2017) 5157

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Carbohydrate Polymers
journal homepage: www . elsevier . com/locate/carbpol

Guar gum oleate-graft-poly(methacrylic acid) hydrogel as a

colon-specific controlled drug delivery carrier

D. Sathya Seeli, M. Prabaharan
Department of Chemistry, Hindustan Institute of Technology and Science, Padur, Chennai 603 103, India
for colon-specific drug delivery. The structure
of GGO-g-PMAc hydrogel was characterized
abstra 1
by FT-IR, H NMR and X-ray diffraction
article info ct
(XRD) analysis. The swelling degree of the
GGO-g-PMAc hydrogel at pH 7.4 was found
Article history: A novel type to be higher than that at pH 1.2. The drug
Received 12 August 2016 of ethylene release studies performed in pH 7.4 and 1.2
Received in revised form 9 November 2016 Accepted 30 glycol
November 2016 buffer solutions at 37 C revealed that the
Available online 2 December 2016 (EGDMA) rate and amount of drug released from the
cross-linked GGO-g-PMAc hydrogel at pH 7.4 were higher
Keywords: guar gum than that at pH 1.2. The MTT assay revealed
Guar gum oleate-graft- that there is no notice-able cytotoxicity of
Hydrogel poly(methacryl GGO-g-PMAc hydrogel at the concentration
Colon ic acid) (GGO- range of 0100 g/ml against the mouse
Controlled release g-PMAc) mesenchymal stem cell line (C3H10T1/2).
Cytotoxicity hydrogel was These results suggested that GGO-g-PMAc
prepared as a hydrogel can be a prospective pH-sensitive
pH-responsive carrier for colon-targeted drug delivery.
release carrier 2016 Elsevier Ltd. All rights reserved.
consi gradability, The o
dered hydrophilicity and non-difficulties
as toxic nature (Mishra,involved in the o
potent Yadav, Mishra, &oral colon drug r
1. Introduction
ial Behari, 2011;delivery are g
materi Prabaharan, 2011;absorption and /
Guar gum is a naturalals for Subhraseema &the degradation 1
polymer which is obtainedbiome Usharani, 2015; Yadav,of 0
from the seeds of Cyamopsisdical, drug .
Sand, Mishra, &molecules in the 1
tetragonolobus (Deepak,phar Behari, 2010; Yadav,
upper 0
Sheweta, & Bhupendar,mace Srivastav, Verma, & 1
2014). It is a non-ionicuti-cal Behari, 2013). Due to 6
galactomannan comprisingand the vul-nerability to 1 /
of 80% galactoman-nan,enviro Corresp
onding author. j
microbial degradation E-mail address:
12% water, 5% protein, 2%nment mprabaharan@y .
in the colon (M. c
acidic insoluble ash, 0.7%al environment and Prabaharan). a
ash and 0.7% fat. Theapplic stability over a wide pH r
backbone of guar gum is aations range, guar gum based b
linear -d-(14) linkeddue to materials haveh p
mannose units having -d-their capability to be usedt o
galactopyranose unitsbioav l
for colon-targeted drugt .
connected by (16) linkagesailabili p
delivery through the: 2
(Panariello, Favaloro,ty, oral administration/ 0
Forbicioni, Caputo, &bioco (Elias, Anil, Ahmad, &/ 1
Barbucci, 2008). In recentmpati Daud, 2010; Madan etd 6
years, guar gum and itsbility, al., 2014; Sathya Seelix
derivatives are widelybiode 1
& Prabaharan, 2016). . 1
.0 colon-specific drugand practical improved
92 delivery carriers suchapplications (Yihong, Huiqun,
as time-dependent(Nazar & & Chaobo,
44 gastro
- delivery carriers, pH-Umbreen, 2014; 2007).
intesti dependent deliverySathya Seeli, In this work,
nal carriers and bacteria-Dhivya,
17 EGDMA cross-
/ (GI) dependent deliverySelvamurugan, linked GGO-g-
20 tract carriers have been
& Prabaharan, PMAc hydrogel
16 that devel-oped (George & was prepared as
El lead 2016). Due to
Abraham, 2007; a colon-targeted
to the presence of
vie Kuntal, Tejraj, & drug delivery
syste weakly acidic
r Anandrao, 2011; carrier by
mic groups, the pH-
Lt Susan, Ellen, Jennifer, grafting PMAc
d. side dependent
& Simon, 2015; onto GGO in the
All effect delivery carriers
Wakerly, Fell, Attwood, presence of
rig s and can present the
& Parkins, 1997). EGDMA as a
low limited swelling
s These carriers can cross-linking
bioav behavior in the
re deliver the drug more agent and
ailabili acidic stomach
se specifi-cally to the potassium
rv ty of fluid, which limits
colon site due to their persulfate as a
ed the the drug release
physicochemical free radical
. drug from the carriers.
properties and thereby initiator. The
in the However,
improve the GGO-g-PMAc
colon because of the
bioavailability and hydrogel is
site. increased pH in
absorption of drugs for expected to
To the large
the potential provide a pH-
overc intestine and
therapeutic effects. responsive
ome colon
However, among the character and
these environment, the
drug deliv-ery carriers, sus-tained
drawb extent of
in recent years, pH- release of the
acks, swelling and
dependent delivery loaded
variou thereby the
carriers have attracted hydrophobic
s amount of drug
increasing attention drug due to the
types release from the
because of their presence
of hydrogel will be
desirable properties
52 D.S. Seeli, M. Prabaharan / Carbohydrate Polymers 158 (2017) 5157

of PMAc and amphiphilic GGO,2.3. Preparation of GGO-

respectively. In addition, due to the cross-g-PMAc hydrogel GE% = [(weight of pure
graft copolymer
linked polymer networks, this hydrogel
would have the improved stability and GGO-g-PMAc hydrogel
swelling behavior in the aqueous media,was prepared by reacting 1 weight of
which control the drug release2.22 g (25.8 mmol) of GGS)/weight of
mechanisms from the hydrogel (Todd &methacrylic acid with 0.5 g monomer] 100
Daniel, 2008). Therefore, the GGO-g-(0.82 mmol) of GGO in
PMAc hydrogel could be usefull topresence of 0.02 g (0.1
release the drugs more specifically to themmol) of EGDMA using
colon in a controlled manner by avoiding0.05 g (0.185 mmol) of 2.4. Characterization
the premature release of drugs in thepotassium persulfate as a
stomach when travelling through the GIfree radical initiator in 100 The FT-IR spectra of
tract. In this study, the prepared GGO-g- ml of distilled water. The the products in the range of
PMAc hydrogel was analyzed with FT-IR, reaction was performed 1
4000400 cm were
1 under constant stir-ring recorded on a double-
H NMR and XRD techniques. The
swelling character of the hydrogel was and nitrogen environment beam Perkin-Elmer 1600
investigated in pH 1.2 and 7.4 buffer at 70 C. After 3 h, the FT-IR spec-trometer. H

solutions at 37 C. To assess the NMR spectra of the
product formed was cooled
products were measured
suitability of the hydrogel as a carrier for to room temperature and on a Bruker AV3 HD
colon-targeted drug delivery, the washed repeatedly with
hydrophobic model drug, ibuprofen, was 60/40% (v/v) ethanol-water spectrometer at 25 C. The
encapsulated in the hydrogel and the XRD patterns of the sam-
mixture in order to ples were tested by a
amount of drug released was determined eliminate the
Bruckers D-8 advanced
in pH 1.2 and 7.4 buffer media at 37 C. homopolymers and wide-angle X-ray
In addition, the cytotoxicity of the GGO-g- unreacted materials. diffractometer using Ni filter
PMAc hydrogel against the C3H10T1/2 Finally, the product was Cu K radiation source.
cell line was evaluated using 3-(4,5- Absorbance
dried at 55 C in the measurements were
Dimethylthiazole-2-yl)-2,5-diphenyl tetra-
zolium bromide (MTT) assay. vacuum oven for 8 h and conducted on a Shimadzu
stored in the des-iccators UVvis spectropho-tometer
for further use. The grafting (UV-2450) at 222 nm.
parameters such as
2. Experimental grafting percentage (G%) 2.5. Swelling studies
and grafting efficiency
2.1. Materials percentage (GE%) of the The swelling characters
hydrogel were calculated of the GGO-g-PMAc
Guar gum was purchased from Sigmausing the following hydrogel were determined
in pH 1.2 and 7.4 buffer
Chemical Company and cleaned byformulas (Mishra, &
refluxing with diethyl ether prior to use.Goutam, 2011). solutions at 37 C. The dry
hydrogel was precisely
Oleic acid, 4-dimethylaminopyridine
weighed (W0 ) and
(DMAP), methacrylic acid, EGDMA, submerged in buffer
G% = [(weight of pure
potassium persulfate and ibuprofen were graft copolymer solutions. At prearranged
purchased from SigmaAldrich. The time periods the swollen
mouse mesenchymal stem cells weight of hydrogel was weighed
(C3H10T1/2 cell line) were obtained from GGS)/weight of GGS] (Wt ) after the removal of
100 excess water by the tissue
National Centre for Cell Sciences paper. The degree of
(NCCS), Pune, India. All other chemicals swelling of the hydrogel at
used were of analytical reagent grade. time t was determined
using the following formula.
Degreeofswelling, % = [(W
2.2. Synthesis of GGO t W 0 )/W 0 ] 100

GGO with different degree of where Wt and W0 are the

substitution (DS) was synthesized by weights of the hydrogel at
time t and in the dry state,
reacting 100 ml of 1% (w/v) guar gum respectively.
aqueous solution with the requisite
amounts of oleic acid in the presence of 2.6. Drug loading and
DMAP as shown in Table 1. The reaction release studies
was performed at room temperature
under constant stirring. After 24 h of The model drug,
reaction period, the reaction mixture was
ibuprofen, was
poured into ethanol solution (200 ml)
encapsulated into the
under stirring in order to precipitate the
hydro-gel by dipping
product. Thereafter, the precipitate was
accurately weighed amount
filtered, washed many times with ethanol
of hydrogel in 10 ml of
and vacuum dried at 55 C for 8 h. The ethanol-drug solution (50
DS of the oleic acid group in GGO was mg/ml, pH 7) in a small
determined by acid-base titration (Sarkar glass beaker at room
& Singhal, 2011).
temperature under stirring.
After 48 h, the hydrogel-drug solutionPilla, Steeber, & Gong, experiments were
was filtered out using a Whatman filter 2009). A defi-nite amount conducted in triplicate.
paper. Then, the concentration of drug in (50 mg) of drug loaded
the filtrate was determined by using ahydrogel was immersed in
UVvis spectrophotometer at 222 nm.the buffer medium and kept 2.7. Cytotoxicity studies
in a laboratory shaking
The drug loading percent by the hydrogel
water bath retaining the
(A) was calculated using the following The cytotoxicity of

formula. constant temperature (37 GGO-g-PMAc hydrogel
C) and stirring (100 rpm). against C3H10T1/2 cell
A = [V (C 0 C 1 )]/W 100 Thereafter, 2 ml of drug
solutions were removed at line was assessed using
where V is the volume of ibuprofen MTT assay (Khaled et al.,
regular intervals and the
solution (ml), C0 is the ini-tialvolume of each sample 2016). In brief, the
concentration of ibuprofen (mg/ml), C 1 iswas restored by the same C3H10T1/2 cells, which
the concentration of ibuprofen solution volume of fresh buffer
after adsorption by the hydrogel (mg/ml),solution. The amount of were maintained under
and W is the weight of the hydrogel (g). standard con-ditions (5%
released ibuprofen was
The in vitro drug release studies were
evalu-ated on a CO2 , 37 C, 10% FBS
carried out in a glass beaker holding 100
spectrophotometer at 222
ml of buffer solutions (pH 1.2, 6.8 and containing DMEM), were
nm. All the release
7.4) at 37 C ( Prabaharan, Grailer,

D.S. Seeli, M. Prabaharan / Carbohydrate Polymers 158 (2017) 5157

Table 1
Reaction conditions for the preparation of GGO with different DS values.

Types of GGO Guar gum (mmol) Oleic acid (mmol) DMAP (mmol) Guar gum/oleic acid (mmol) DS Yield (%)
GGO1 2.056 2.056 2.056 1:1 0.23 74
GGO2 2.056 10.28 10.28 1:5 0.33 53
GGO3 2.056 20.56 20.56 1:10 0.5 59
was efficiently utilized
4 2 by the available free
into a concentration of 3 10 /cm in 24 wellradical sites on GGO
plates and incu-bated for a period of 12 produce the grafted
Thereafter, the cells were treated with GGO-g- product.
PMAc hydrogel at different concentrations (0, 10,
30, 50, 100 and 150 g/ml) for 24 h. The control
cells were left untreated and the positive control
cells were treated with 0.1% Triton-X 100. 3.2. Characterization
Thereafter, after washing with phosphate bufferof GGO and GGO-g-
solution (PBS), the cells were treated with 5 mg/ml PMAc hydrogel

of MTT solution at 37 C for 4 h. Then, the culture
medium was removed and 0.2 ml of DMSO was spectrum of guar gum
incorporated with the precipitates. The absorbance presents a broad OH
value of result-ing solution was measured at 570stretch absorption
band in between 3600
nm using a spectrophotometer. Using SPSS 1
software, statistical analysis was performed by and 3000 cm and
the aliphatic C H
one-way ANOVA followed by StudentNewman stretch in between
Keuls test. Inconsistencies with a P value < 0.05 2930 and 2890 cm1
were regarded as statistically significant. as revealed in Fig. 1A
(Mishra & Goutam, Fig. 1. FT-IR spectra of (A)
2011). The other major guar gum, (B) GGO, and
3. Results and discussion absorption bands at (C) GGO-g-PMAc hydrogel.
1015 and 877 cm
3.1. Synthesis of GGO-g-PMAc hydrogel correspond to C O C
stretching vibrations of
glycosidic linkages. trum of GGO (Fig. 1B),
To prepare GGO-g-PMAc hydrogel, first GGOThe absorption bands in addition to the
with different DS values were prepared by reactingin between 1225 and characteristic peaks of
1 guar gum, the new
guar gum aqueous solution with the required817 cm correspond
amounts of oleic acid in the presence of DMAP as to the highly coupled peaks at 1735, 2921,
C C O, C OH and C O 2853, 1435 and 1383
listed in Table 1. The reaction route for the C stretching modes of
synthesis of GGO is shown in Scheme 1. Toguar gum backbone. cm were observed.
determine the DS of the oleic acid group in GGO,In the IR spec- A peak at 1735 cm

all the ester linkages present in GGO were could be assigned to

saponified by 0.1 N NaOH solu-tion and the the carbonyl group
amount of remained NaOH was determined by stretching vibration
titrating against 0.1 N HCl solution. It was found due to the presence of
that the DS value of the GGO progressively ester linkage between
guar gum and oleic
increased from 0.23 to 0.5 with the increase in
acid. The appearance
guar gum/oleic acid feed ratio from 1:1 to 1:10 as of strong absorption
shown in Table 1. Next, EGDMA cross-linked peaks at 2921 and
GGO-g-PMAc hydrogel was prepared by reacting 1
2853 cm was
GGO (DS = 0.5) with methacrylic acid and corresponding to C H
EGDMA in the presence of potassium persulfate stretching vibration of
under the nitrogen environ-ment. During the free the long chain
radical polymerization reaction, the hydroxyl aliphatic group present
groups of GGO were grafted with EGDMA cross- in GGO. Moreover, the
linked PMAc as shown in Scheme 1. Due to the absorption bands
presence of GGO and the EGDMA cross-linked observed at 1435 and
PMAc network, the resultant GGO-g-PMAc 1383 cm could be
hydrogel will have the improved stability and consigned to CH2
swelling behavior in the bio-logical fluid in addition
and CH3 stretching
to the capability to release the encapsulated
vibra-tion of the oleic
hydrophobic drugs to the targeted site in a more acid present in the
controlled man-ner. Moreover, because of the GGO, respectively.
presence of carboxyl groups on the backbone of These observations
PMAc, the GGO-g-PMAc hydrogel can show the confirmed the
pH-dependent swelling and drug release formation of GGO.
behaviors that are particularly important for the The IR spectrum of
colon-specific drug delivery. The grafting param-
hydrogel shows the
eters such as G% and GE% of the GGO-g-PMAc
hydrogel were found to be 400 and 90, absorption bands of
respectively. The higher values of grafting parame- both GGO and PMAc
ters indicate that most of the acrylic acid monomer (Fig. 1C). A strong
peak at 1695 cm can be assigned to the characteristic peaks of characteristic peaks of
stretching vibration of carbonyl groups present in guar gum were guar gum (Zhongjian,
1 observed at 3.64.0 Tianpeng, Baojian, &
PMAc. An absorption band at 1560 cm , which
ppm, which Xingwang, 2016). In
was not observed in the spectrum of GGO,
correspond to the addition, the
represents the asymmetric deformation of carboxyl
group that reveals that acid groups present inprotons in sugar units appearance of a peak
PMAc are involved in the hydrogen bonding of guar gum (Dodi, at 4.2 ppm indicated
between the polymer chains (Chen, Liu, & Zhuo,Hritcu, & Popa, 2011). the presence of oleate
2005). A new absorption band at 1480 cm wasThe spectrum of GGO
1 moiety in the grafted
(Fig. 2B) presents the product. In the
due to CH3 stretching in the grafted sample. Thenew peaks at 0.9, 1.3, spectrum of GGO-g-
reduced intensity of OH absorption band in1.65, 2.3, and 5.2 ppm PMAc (Fig. 2C), new
between 3600 and 3000 cm indicates that the due to the presence of proton signals
OH groups of GGO were involved in the grafting methyl, long chain appeared in the range
reaction with PMAc. proton, - proton, of 0.71.3 and 1.82.2
The formation of GGO and GGO-g-PMAc -proton and methine ppm because of the
hydrogel was fur-ther confirmed by H NMR asproton of oleic acid presence of methyl
shown in Fig. 2. As shown in Fig. 2A, therespectively in and methylene groups
addition to the of PMAc
54 D.S. Seeli, M. Prabaharan / Carbohydrate Polymers 158 (2017) 5157

Scheme 1. Synthesis of GGO-g-PMAc hydrogel.

( Giordanengo, Viel, Hidalgo, Allard-Breton, Thvand, & Charles,

2009). These results demonstrated the formation of GGO-g-PMAc

Fig. 3 shows the XRD pattern of guar gum, GGO, and GGO-g-
PMAc hydrogel. As shown in Fig. 3A, guar gum shows the

characteristic diffraction peaks at 17.47 and 20.12 due to its semi-
crystalline in nature (Deepak, Sheweta, & Khatkar, 2012). In the case
of GGO (Fig. 3B), in addition to the characteristic diffraction peaks of

guar gum, a new peak was also observed at 14.54 . Moreover, the

intensity of diffraction peaks at 17.47 and 20.12 was considerably
increased. This observation indicates that the crystallinity of guar gum
was enhanced to some extent after the chemical modifica-tion with
oleic acid due to the ordered arrangement of the grafted polymer
chains. In the diffraction pattern of GGO-g-PMAc hydrogel (Fig. 3C),
the crystalline peaks of GGO were disappeared and new characteristic

amorphous halo peaks of PMAc observed at 16 and 32 (Guangping,
Siqi, Jingquan, Zhen, & Qinglin, 2014). This obser-vation might be due
to the destruction of original semi-crystalline structure of GGO when
grafting with PMAc. This result indicated that the PMAc has been
grafted effectively onto GGO back bone.
Fig. 2. H NMR spectra of (A) guar gum, (B) GGO, and (C) GGO-g-PMAc hydrogel.
D.S. Seeli, M. Prabaharan / Carbohydrate Polymers 158 (2017) 5157 55

Fig. 3. XRD pattern of (A) guar gum, (B)

GGO, and (C) GGO-g-PMAc hydrogel.

Fig. 5. Drug release profile of GGO-

g-PMAc hydrogel at 37 C.
swelling degree of bonding the swelling degree of that the rate of drug
hydrogel at pH 7.4 between thethe hydrogel was release from the
were found to be polymer decreased at pH 1.2. Fig. ibuprofen-loaded
higher than that at chains, which4 revealed that the GGO- hydrogel was quite rapid
pH 1.2. In addition, reduces theg-PMAc hydrogel had an at the initial period of
the swelling of free volume inimproved stability for a time in both pH 1.2 and
hydro-gel at pH 7.4 the hydrogelperiod of 24 h. This 7.4 media. The presence
reached a constant matrix. observation might be due of unbound free drug
equilibrium more Therefore, to the presence of strong molecules at the surface
slowly than that at EGDMA cross-linking of the hydrogel may be
pH 1.2. The between the polymer the reason for this
increased swelling chains and the observation. After the
character of hydrophobic interactions initial abrupt release,
hydrogel at pH 7.4 between the oleate GGO-g-PMAc hydrogel
might be because moieties of the hydro-gel. showed a slow and
of the presence These results suggested controlled release of the
drug into the buffer
PMAc containing that EGDMA cross-linked
media. The slow release
many numbers of GGO-g-PMAc hydrogel
of the drug from the
carboxyl groups could be a promising
Fig. 4. Swelling behavior of GGO-g-PMAc hydro-gel might be due to

hydrogel at 37 C (Inset: photographic image of (pKa = 4.55) in carrier for colon-targeted
the hydrophobic
hydrogel after being immersed in (A) pH 1.2 the hydrogel (Glen, drug deliv-ery with an
and (B) pH 7.4 buffer medium for 24 h). interaction between the
Daniel, Qi, & improved stability in the
drug molecules and long
Jennifer, 2004). At GI tract and sensitivity to
chain oleic acid present
pH 7.4, the pH of the surrounding
in the GGO-g-PMAc
3.3. Swelling behavior of GGO-g- carboxylic acid medium.
PMAc hydrogel groups present in hydrogel ( Prabaharan,
the hydrogels are 3.4. Drug release Reis, & Mano, 2007). In
It is well known that the swelling ionized which leads behavior of GGO-g-
addition, the exis-tence
PMAc hydrogel
property of hydrogels greatlyto a strong ionic of hydrogen bonding and
influences their drug releaserepulsion between electrostatic interactions
The release of the
mechanisms (Prabaharan & Gong,the polymer chains. between the drug
hydrophobic model drug,
2008). Therefore, in this study, theThis strong ionic molecules and hydrogel
ibuprofen, from the
swelling behavior of the GGO-g-PMAc repulsion improves network could also be
GGO-g-PMAc hydrogel
hydrogel was analyzed in pH 1.2 andthe water uptake by reason for the controlled
was performed in pH 1.2
7.4 buffer solution at 37 C. As shownenhancing the free delivery of the drug from
and 7.4 buffer media at
in Fig. 4, the swelling behavior of the volume in the hydrogel. Fig. 6 reveals

GGO-g-PMAc hydrogel was found to hydrogel. Since the 37 C. Fig. 5 shows the that the rate and amount
be greatly controlled by the pH of theionization of drug release behaviors of of drug released from the
swelling medium and swollencarboxylic acid GGO-g-PMAc hydrogel hydrogel are pH-
hydrogels were maintained their shape groups is limited at loaded with 14% (w/w) of dependent and occurs at
without undergoing disintegration evenpH 1.2, there can ibuprofen as a function of a higher rate in pH 7.4
after 24 h. The rate and amount ofbe strong hydrogen time. It was observed medium than that in pH
1.2 medium. Within 8 h, 50 and 15% of of the hydrogel that because of itsfaster rate. out in pH 1.2, 7.4 and 6.8
the drug release were observed from limits the diffusion longer To understand the
buffer media at 37 C as
the hydrogel in pH 7.4 and 1.2 of drug molecules intermolecular drug release behavior of
medium, respectively. Thisat a quicker rate as distances these pHs are
the GGO-g-PMAc
phenomenon can be explainedit hap-pens at pH might be encountered the
hydrogel in the real GI
considering the rate of drug dif-fusion 7.4. In the alkaline responsible for stomach, small
microenvironment, the
from the swollen hydrogel in the buffer pH, the improved the release of
media. As mentioned earlier, in the swelling degree of drug sequential drug release
acidic pH, there is a restricted swellingthe hydrogel molecules at aexperiments were carried
56 D.S. Seeli, M. Prabaharan / Carbohydrate Polymers 158 (2017) 5157
Fickian diffusion plays a
significant role in release of
the drug. The value of n in
pH 7.4 medium was
determined as 0.52. The
higher value of n suggests
that the drug release from
hydrogel pursues a non-
Fickian mechanism in the
basic medium, where the
drug release is mainly
influenced by the diffusion
and relaxation of
hydrogels. The values of
kinetic constant K
determined at pH 1.2 and
7.4 were found to be 0.08 Fig. 7. Viability of C3H10T1/2
2 2 cells in the presence of GGO-g-
(R = 0.9513) and 0.14 (R
PMAc hydrogel for 24 h. #
= 0.9363), respectively. Indicates significant decrease
These results revealed that compared to control (p < 0.05).
GGO-g-PMAc hydrogel
can be an idle pH-sensitive crystals. This metabolic
Fig. 6. Sequential drug release profile of carrier for colon-targeted activity of cells is an

GGO-g-PMAc hydrogel at 37 C. controlled drug delivery. appropriate tech-nique for
estimating the number of
viable cells since dead or
intestine and colon of the GI tract3.5. Cytotoxicity studies injured cells do not show
respectively and the results are shown in any mitochondrial
Fig. 6. Initially, 12% of initial drug content To assess the suitability
dehydrogenase activ-ity
was released from the hydrogel within 3 of GGO-g-PMAc hydrogel
(Prabaharan et al.,
h period of time in pH 1.2 medium (sim- as a bio-material for colon- 2009a,b). Fig. 7 shows the
ulated gastric fluid). Thereafter, the targeted drug delivery, in C3H10T1/2 cell viability in
amount of drug released from the this study, the cytotoxicity the presence of GGO-g-
hydrogel was found to be improved with of the drug free-GGO-g- PMAc hydrogel at the
the increase in time in both pH 7.4 and PMAc hydrogel against the concen-tration range of
6.8 media. Within 4 h time period, thecul-tured C3H10T1/2 cells 10150 g/ml for 24 h. It
amount of drug released from the was estimated by MTT could be seen that at 10,
hydrogel was found to be 38% in pHassay, which is based on 30, 50 and 100 g/ml of
7.4 (simulated intestinal fluid). In pH 6.8the capability of a
GGO-g-PMAc hydrogel, no
medium (simulated colonic fluid), the mitochondrial
significant dif-ference
remaining 50% of the loaded drug wasdehydrogenation enzyme occurred in the cell viability
released from the hydrogel within 21 h. in viable cells to convert of the cultured C3H10T1/2
In order to study the drug releasethe tetrazolium rings of cells with respect to the
mechanism of GGO-g-PMAc hydrogel,MTT into formazan control. However, the cell
the first 60% of the drug release data viability at 150 g/ml of the
from Fig. 6 was fitted with the Ritger hydrogel was found to be
Peppas model as shown in Eq. (1) significantly decreased into
(Korsmeyer & <85% when compared to
Peppas, 1983). the control. The reason for
this observation may be
Mt /M = K t the possible presence of
where Mt /M is the fractional drug release residual monomer in the
at time t, K is a kinetic constant, and n is hydrogel even after
the diffusional exponent that can be exhaustive washings with
related to the drug release mechanism. distilled water. Based on
For a thin hydrogel, when n 0.5, the these results, it can be said
drug release mechanism is Fickian that GGO-g-PMAc
diffusion. When n = 1, Case II (polymer hydrogel has no adverse
relaxation) transport occurs, leading to effects against the cultured
zero-order release. When the value of n C3H10T1/2 cells and
is between 0.5 and 1, both swelling and hence it could be a safe
diffusion con-trolled drug release carrier to deliver the drugs
mechanism (anomalous transport) is in the colon target.
observed (Yihong, Huiqun, & Chaobo,
2007). It is found that the value of n for
the GGO-g-PMAc hydrogel in pH 1.2 4. Conclusions
medium was 0.37, which suggest that
A novel type of EGDMA cross-linkedamount of drug released g/ml concentrations against
GGO-g-PMAc hydrogel was prepared asfrom the GGO-g-PMAc the cultured C3H10T1/2
a carrier for colon-targeted controlledhydrogel in pH 7.4 medium cell line. These results
drug delivery. Using FT-IR, H NMR andare higher than that in pH indicated that GGO-g-
1.2 medium. Therefore, in PMAc hydrogel can
XRD analysis, formation of the product
the stomach due to its potentially serve as an
was confirmed. Due to the presence of
acidic conditions, the orally administrated colon-
GGO moieties, the GGO-g-PMAc
unwanted premature targeted drug carrier for
hydrogel was found to have an improved
release of drug from the hydrophobic drugs.
drug loading efficiency and controlled
hydrogel could be reduced.
drug release ability in the environment of
However, the maximum
GI tract. Moreover, because of the
release could be achieved Acknowledgements
presence of a large num-ber of carboxyl
in the colon-site due to its
groups, the GGO-g-PMAc hydrogel
increased pH value when The authors thank Dr.
showed the pH-dependant swelling as
the hydrogel passes N. Selvamurugan and Ms.
well as drug release behavior in the
through the GI tract. The S. Dhivya, Department of
buffer solutions. The swelling degree of
MTT studies revealed that
the hydrogel in the basic medium was Biotechnology, SRM
there is no noticeable
about 5000% while it was about 800% in University, Chennai for
cytotoxicity of GGO-g-
the acidic medium. The in vitro drug their
PMAc hydrogel at 10100
release study revealed that the rate and
D.S. Seeli, M. Prabaharan / Carbohydrate Polymers 158 (2017) 5157 57

Drug release from Panariello, G., Favaloro, R.,

Forbicioni, M., Caputo, E., &
pronounced help and support in glassy poly(HEMA
conducting the cytotoxicity stud-ies. Barbucci, R. (2008).
-co-NVP) Synthesis of a new
copolymers. hydrogel, based on guar
Journal of gum: for controlled drug
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