Journal of Photochemistry and Photobiology B: Biology 102 (2011) 232–240

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Journal of Photochemistry and Photobiology B: Biology

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

UV-prepared salep-based nanoporous hydrogel for controlled release

of tetracycline hydrochloride in colon
Ghasem Rezanejade Bardajee a,⇑, Ali Pourjavadi b,⇑, Somayeh Ghavami a, Rouhollah Soleyman b,
Farnaz Jafarpour c
a
Department of Chemistry, Payame Noor University, Qazvin Branch, P.O. Box 878, Qazvin, Iran
b
Polymer Research Laboratory, Department of Chemistry, Sharif University of Technology, P.O. Box 11365-9516, Tehran, Iran
c
School of Chemistry, University College of Science, University of Tehran, P.O. Box 14155-6455, Tehran, Iran

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

Article history: A highly swelling nanoporous hydrogel (NPH) was synthesized via UV-irradiation graft copolymerization
Received 22 July 2010 of acrylic acid (AA) onto salep backbone and its application as a carrier matrix for colonic delivery of tet-
Received in revised form 7 December 2010 racycline hydrochloride (TH) was investigated. Optimized synthesis of the hydrogel was performed by
Accepted 10 December 2010
the classic method. The swelling behavior of optimum hydrogel was measured in different media. The
Available online 30 December 2010
hydrogel formation was confirmed by Fourier transform infrared spectroscopy (FTIR) and thermo-gravi-
metric analysis (TGA/DTG/DTA). The study of the surface morphology of hydrogels using SEM showed a
Keywords:
nanoporous (average pore size: about 350 nm) structure for the sample obtained under optimized con-
Nanoporous hydrogel (NPH)
Salep
ditions. The drug delivery results demonstrated that this NPH could successfully deliver a drug to the
UV-irradiation colon without losing the drug in the stomach, and could be a good candidate as an orally administrated
Colonic delivery drug delivery system.
Ó 2010 Elsevier B.V. All rights reserved.

1. Introduction
Hydrogels are cross-linked hydrophilic polymers absorbing
large quantities of water or aqueous solutions without dissolving.
Because of softness, smartness and the capacity to store water,
hydrogels are a unique carrier matrix for drug delivery systems
[1]. Among all the routes of drug administration, oral drug delivery
has been regarded as the most convenient method of drug man-
agement. The time period of oral drug delivery has been limited
to a day because the maximum time available for drug absorption
from an orally administered formulation is determined by the total
transit time from mouth to colon, which is usually less than 24 h
[2]. Colon targeting is valuable for the topical treatment of diseases
of the colon such as Crohn’s disease, ulcerative colitis, and colorec-
tal cancer and can be useful in the treatment of nocturnal asthma,
angina, and arthritis [3]. Many pH-sensitive or bacterial degradable
polymers have been used for colon-specific drug delivery [4]. These
polymers include chitosan [5], methacrylic acid or hydroxyethyl
methacrylate [6], azo-aromatic polymers [7], methacrylated inulin
[8], cellulose derivatives such as ethyl cellulose [9], hydroxylpropyl
methylcellulose acetate succinates, cellulose acetate phthalate
⇑ Corresponding authors. Tel.: +98 21 66165311; fax: +98 21 66005718.
E-mail addresses: rezanejad@pnu.ac.ir (G.R. Bardajee), purjavad@sharif.edu
(A. Pourjavadi).
[10], cellulose acetate butyrate [11], pectin and its salts [12,13],
and chondroitin sulphate [14].
Nanoporous materials as a subset of nanostructured materials
possess unique surface, structural, and bulk properties that under-
line their important uses in various fields. Nanoporous materials
are also of scientific and technological importance because of their
vast ability to adsorb/absorb and interact with atoms, ions and
molecules on their large interior surfaces and in the nanometer
sized pore space [15]. Inorganic-based nanoporous materials as
aerogels were widely investigated by many researchers but much
less attention has been paid to organic-based ones. In the current
decade, organic-based nanoporous hydrogels (NPHs) on the basis
of natural biomaterials (such as collagen and cellulose) were syn-
thesized and investigated [16,17]. Due to their low cost, low toxic-
ity, biodegradability, and biocompatibility of the large amount of
natural biomaterials incorporated in their networks, many works
have been reported in the case of preparation of hydrogels based
on polysaccharides such as chitosan, starch, sodium alginate, and
carrageenan [18–24].
The high energy radiation (60Co gamma rays or electron beam)
cross-linking and chemical cross-linking are two main methods for
the preparation of natural based hydrogels. However, the high
investment, critical requirement for safety and irradiation protec-
tion and complicated operation and processing technology are
the deficiency of high energy radiation. Chemical methods have
low heat efficiency, toxic reagents, long cross-linking time, and

1011-1344/$ - see front matter Ó 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.jphotobiol.2010.12.008
 G.R. Bardajee et al. / Journal of Photochemistry and Photobiology B: Biology 102 (2011) 232–240
complicated processing requirement due to the critical reaction
conditions.
In recent years [18,19], we developed NPHs based on salep [20–
24] as a novel hydrogel backbone and these nanoporous materials
utilized as a nanoreactor for synthesis of hematite nanoparticles
[25]. We synthesized salep-g-poly (sodium acrylate) hydrogel
(salep-g-poly (NaAA)) via two initiation polymerization systems
(thermal classic condition and c-irradiation) [19,20]. Because of
outstanding properties of these hydrogels, we prepared them by
the UV-irradiation initiation method in the absence of toxic
cross-linking agent and initiator. The UV-irradiation technique is
a promising approach as it benefits of no disposal of solvents into
the atmosphere, low energy requirements, minimization of the
thermal abuse of the polymer compositions during processing
and a simple manufacturing procedure [26]. As there are no reports
on the UV-irradiation synthesis of hydrogels based on natural bio-
materials, in the present work, the synthesis of salep-g-poly
(sodium acrylate) NPH was carried out via applying UV-irradiation
for the first time. Neither both initiators, crosslinkers, nor waste,
relatively low operating costs and simple procedure control are
the advantages of our work. The hydrogel preparation was opti-
mized through the classic method (OFAT, one factor at a time)
and its potential as a carrier matrix for colonic release of drugs
was tested with tetracycline hydrochloride (TH).
2. Materials and methods
2.1. Materials
The palmate-tuber salep (PTS) was purchased from a supplier in
Kordestan, Iran (Mn = 1.17  106 g/mol, Mw = 1.64  106 g/mol
(high Mw), PDI = 1.39, eluent = water, flow rate = 1 mL/min, acqui-
sition interval = 0.43 s from GPC results). Acrylic acid (AA, from
Merck, Darmstadt, Germany) was used after vacuum distillation.
Tetracycline hydrochloride (TH) was purchased from Alborz Darou
Co., Tehran, Iran. All other chemicals were analytical grade. Double
distilled water was used for hydrogel and solution preparation for
release study and swelling measurements.
2.2. NPH preparation
In general, certain amount of acrylic acid (AA) was added to a
homogenized solution of salep (1.0 g) in 30 mL H2O at a three-neck
reactor equipped with a mechanical stirrer. Then the solution
transferred to a quartz tube and degassed by purging N2. Irradia-
tions (20–80 min) were carried out by a low-pressure mercury
lamp with a transmission maximum of 254 nm at room tempera-
ture. The product was neutralized with 40 mL aqueous solution
of NaOH (neutralization percentage: 20–90% based on moles of
AA monomer). Finally, the product was poured into 100 mL of eth-
anol, remained for 2 h and then cut to small pieces for further dry-
ing. In order to remove the sol fraction of mixture (uncross-linked
and not grafted poly (acrylic acid), uncross-linked salep and possi-
bly also some unreacted monomer), the dewatered hydrogel was
allowed to completely swell (4 h) in distilled water and then dewa-
ter in ethanol (200 mL, 2 h). The non-solvent ethanol was decanted
and then 100 mL fresh ethanol was added again. The chopped par-
ticles were remained for 24 h in ethanol to completely dewater.
The dewatered gel particles were filtered and dried in oven (at
50 °C) for 24 h. After grinding, the powdered superabsorbent
hydrogel was stored in the absence of moisture, heat and light.
The optimization of swelling capacity of the hydrogels was per-
formed based on the classic OFAT method. In this method, the
amount of one factor has been changed in the favorable range
without any alteration in the other factors, supposing that effective
233
factors which affecting swelling capacity of the hydrogels are
independent.
2.3. Characterization
FTIR spectra of samples in the form of KBr pellets were
recorded using an ABB Bomem MB-100 FTIR spectrophotometer.
A Shimadzu UV–visible 1650 PC spectrophotometer was used for
recording absorption spectra in solution. Photolyses were per-
formed using a low-pressure mercury lamp with a transmission
maximum at k = 254 nm (85% transmission) and transmitted light
from 254 to 579 nm (15% transmission). The dynamic weight loss
tests were conducted on a TA instrument 2050 thermo-gravimetric
analyzer (TGA). All tests were conducted in a N2 purge (25 mL/min)
using sample weights of 5–10 mg over a temperature range of
25–750 °C at a scan rate of 20 °C/min. The mass of the sample
pan was continuously recorded as a function of temperature. The
morphology of the dried samples was examined using a scanning
electron microscope (SEM) (Philips, XL30) operated at 10 kV after
coating the sol free dried samples with gold film.
2.4. Swelling measurement
The degree of swelling was determined by routine gravimetric
method. A 100 mesh nylon screen containing an accurately dried
weighed powdered sample (0.1 ± 0.01 g) with average particle
sizes between 40 and 60 mesh (250–400 lm) was immersed en-
tirely in distilled water (400 mL) and allowed to soak for 60 min
at room temperature. The equilibrium swelling (ES) capacity was
measured twice at room temperature using the following formula:
ESðg=gÞ ¼ ðW 2  W 1 Þ=W 1 ð1Þ
where W1 and W2 are the weights of dry and swollen gel,
respectively.
For swelling kinetics, at consecutive time intervals, the water
absorption of the hydrogel was measured according to the men-
tioned method. To determine the pH-sensitivity of the hydrogel,
the swelling was measured at favored pH solutions instead of dis-
tilled water. Aqueous stock NaOH (pH 13.0) and HCl (pH 1.0) solu-
tions were diluted with distilled water to reach the desired basic
and acidic pHs, respectively.
2.5. In vitro drug delivery study
2.5.1. Drug loading
0.05 g of powdered NPH was added in 60 mL of drug (TH) solu-
tion (0.6  104 mol/L) and put in a dark place for 10 days to com-
plete drug loading. For drug loading tracing, the amount of loaded
drug was determined at favored times by UV spectroscopy (kmax)
using a calibration curve constructed from a series of drug solu-
tions with known concentrations.
2.5.2. Drug releasing
The release rate experiments were performed in acidic (pH 2.0),
and alkaline (pH 8.0) phosphate buffered solutions at 25 °C under
unstirred conditions. The hydrogel (0.05 g) containing a known
amount of drug (TH) was added to the release medium (100 mL).
At a given time interval, 1 mL filtered samples were withdrawn
and assayed for the amount of released drug as a function of time.
The amount of released drug was determined by UV spectroscopy
at characteristic kmax using a calibration curve constructed from a
series of drug solutions with known concentrations. The results
were expressed as cumulative release ratio (amount of released
drug/all amount of loaded drug). Also, method validation regarding
the assay of the drug for the determination of drug loading and
drug releasing was performed by HPLC using the SGX C18 column
234 G.R. Bardajee et al. / Journal of Photochemistry and Photobiology B: Biology 102 (2011) 232–240
in the methanol–water system and based on the HPLC calibration
curve.
3. Results and discussion
3.1. Synthesis and mechanism aspects
Cross-linking and graft copolymerization of poly (sodium acry-
late) onto backbones of salep substrates was carried out in an
aqueous medium using UV light as a radical initiator and without
using a cross-linking agent. The UV light produced hydroxyl radi-
cals that abstract hydrogen atoms from the anomeric carbon or hy-
droxyl groups of salep backbones. This redox system results in
active centers capable of radically initiating the polymerization
of AA leading to a cross-linked graft copolymer (Fig. 1).
3.2. Characterization
The grafting process was confirmed by comparing the FTIR
spectra of the salep before and after copolymerization reaction
while the sol fraction of hydrogels was removed. Fig. 2 shows the
FTIR spectra of salep, poly (sodium acrylate) and UV-prepared
salep-g-poly (sodium acrylate) hydrogel. In Fig. 2a, the broad band
at 3500 cm1 is due to stretching of OH groups in the polysaccha-
ride (salep) substrate. Another signal around 1600 cm1 was
attributed to the stretching band of C@O group in the glucomannan
Fig. 1. Proposed mechanism pathway for synthesis of the UV-prepared salep-g-poly (sodium acrylate) hydrogel.
of salep [23,24]. In Fig. 2b, broad peak around 1600–1700 cm1 are
due to stretching band of carboxyl groups of poly (sodium acrylate)
and single broad peak around 3000–3600 cm1 was attributed to
hydroxyl groups of poly (sodium acrylate). As indicated in Fig. 2c,
the salep-g-poly (sodium acrylate) showed three new characteris-
tic absorption bands at 1718, 1557 and 1409 cm1 verifying the
formation of graft copolymer product. These peaks were attributed
to carbonyl stretching of the carboxylic acid groups and symmetric
and asymmetric stretching modes of carboxylate groups of AA,
respectively.
TGA traces of salep (as an initial material), thermally-prepared
salep-g-poly (sodium acrylate) hydrogel [19] and UV-prepared
hydrogel are presented in Fig. 3A. According to this TGA curves,
values related to the salep around 280–460 °C such as T30
(296.0 °C) and T50 (352.3 °C) are lower compared to the UV-pre-
pared hydrogel (T30 = 355.6 °C and T50 = 445.8 °C). It is well-known
that the ionic structure in the network may act as heat barriers and
as a consequence, enhance the overall thermal stability of the
hydrogel. Another factor in enhancing the thermal stability of the
hydrogels is their cross-linking structures. At lower than 280 °C
and higher than 480 °C, the salep was found to be the most ther-
mally stable sample studied and this could be attributed to the nat-
ure of salep composition [23,24]. For the better study of the
thermal behavior of UV-prepared hydrogel, DTG and DTA curves
of the hydrogel were provided in Fig. 3B. The first derivative of
the TGA curve (DTG) shows that the maximum decomposition rate
of the hydrogel occurs in the sharp peak at 433.5 °C and according
 G.R. Bardajee et al. / Journal of Photochemistry and Photobiology B: Biology 102 (2011) 232–240
Fig. 2. FTIR spectra of (a) salep, (b) poly (sodium acrylate) and (c) optimized salep-g-poly (sodium acrylate) hydrogel.
to DTA, at this temperature an endothermic reaction cause to
decomposition of the hydrogel. Other decomposition points of
the hydrogel are at 216.6, 341.5, 443.3, 508.9 and 685.9 °C and
all are endothermic decompositions.
3.3. Surface morphology of the hydrogel
It is well-known that the pores are the regions of water perme-
ation and interaction sites of external stimuli with the hydrophilic
groups of the graft copolymers. Fig. 4 showed the scanning electron
microscopic (SEM) pictures of the optimized UV-prepared NPH in
(a) 20 and (b) 50 magnification. These pictures verify that our
hydrogel have a nanoporous and cellular structure (average pore
size: about 350 nm), where the pores might be induced into the
NPH by water evaporation resulting from heat of neutralization
reaction.
3.4. Optimization of water absorption capacity
The different variables affecting the ultimate swelling capacity,
i.e., amounts of AA, neutralization percentage of AA and UV-irradi-
ation time were systematically optimized by the OFAT method.
3.4.1. Effect of monomer amount
The effect of monomer amount on the hydrogel water absorp-
tion was investigated by varying the acrylic acid amounts from
0.5 to 3 mL, while the amount of salep was kept constant (1.0 g).
As shown in Fig. 5A, with increasing monomer amount up to
1.5 mL, the free swelling equilibrium increases then it considerably
decreases with further increase in monomer amount. The maxi-
mum absorption (790 g/g) is obtained at 1.5 mL of AA. The initial
increase in water absorption can be attributed to enhance in the
diffusion of AA molecules into the salep backbones resulting in
an increase in swelling capacity. In addition, the higher AA content
improves the hydrophilicity of the hydrogel, which in turn, causes
a higher absorption of water. The subsequent decrease after max-
imum absorption may be attributed to (a) an increase in viscosity
of the reaction medium that hinders the movement of the macro-
radicals and monomer molecules, (b) preferential homo-polymer-
235
ization over graft copolymerization, and (c) the enhanced chance
of chain transfer to monomer molecules.
3.4.2. Effect of UV-irradiation time
Fig. 5B represents the effect of UV-irradiation time on capability
of water absorption of the hydrogel. The UV light is a well-known
initiator of polymerization because of its high energy rays. As indi-
cated in Fig. 5B, the maximum ES (1320 g/g) was attained at 50 min
irradiation by UV-rays. Since UV-rays act as initiators, by the
increasing of the UV-irradiation time, the graft copolymerization
more proceed and led to the ES increasing. By the UV-irradiation
increasing more than 50 min, more active sites (macro-radicals)
created and hydrogel structure becomes rigid and cannot swell,
properly.
3.4.3. Effect of neutralization percentage of AA
In these series of experiments, after completion of the reaction,
the carboxylic acid groups of obtained hydrogel were partially con-
verted to carboxylate anions by NaOH solution. Without the neu-
tralization stage, the carboxylate anions are protonated, so the
main anion–anion repulsive forces are eliminated and conse-
quently the water absorption is decreased. According to Fig. 5C,
the best neutralization degree was found to be 65%. At high neu-
tralization percentage of the carboxylic acid groups, reduced swell-
ing is observed due to a different phenomenon that is related to a
‘charge screening effect’ of excess Na+ ions in the swelling media.
The excess cations shield the carboxylate anions and prevent effec-
tive anion–anion repulsion (screening/shielding effect). In addi-
tion, with increasing the NaOH concentration, the ionic strength
of the swollen solution is increased and the osmotic pressure be-
tween the aqueous and the gel phases is reduced and the swelling
is consequently decreased.
3.5. Swelling kinetics
Fig. 6 represents the swelling capacity of the hydrogel in dis-
tilled water at consecutive time intervals. Initially, the rate of
water uptake sharply increases and then begins to level off. The
equilibrium swelling was achieved after 30 min approximately.
236 G.R. Bardajee et al. / Journal of Photochemistry and Photobiology B: Biology 102 (2011) 232–240

Fig. 3. (A) TGA traces of salep, thermally-prepared salep-g-poly (sodium acrylate) and UV-prepared salep-g-poly (sodium acrylate) and (B) TG/DTG/DTA curves of optimized
salep-g-poly (sodium acrylate) hydrogel.

A power-law behavior is obvious from Fig. 6. The data may be well the particle size of the comparing samples are the same or, at least,
fitted with a Voigt-based following equation [27]: in the same range.

St ¼ Se ð1  et=s Þ
where St (g/g) is the swelling at time t, Se is equilibrium swelling
(power parameter, g/g), t is time (min) for swelling, and s (min)
stands for the ‘‘rate parameter’’. For calculating the ‘‘rate parame-
ter’’, by using the above formula (after a little modification), one
can plot Ln(1  St/Se) versus time (t). The slope of the fitted straight
line (slope = 1/s) gives the rate parameter. According to Fig. 6 and
using Eq. (2), the ‘‘rate parameter’’ for swelling of the hydrogel in
water is 1.54 min. This appropriate ‘‘rate parameter’’ of this hydro-
gel put it in the fast-swelling hydrogel category. Since the s value is
a measure of swelling rate (i.e., the lower the s value, the higher
the rate of swelling), it can be used for comparative evaluation of
the rate of water absorption of hydrogels on the condition that
ð2Þ
3.6. pH-sensitivity of the hydrogel
Ionic hydrogels exhibit swelling changes at a wide range of pHs.
Therefore, in these series of experiments, the equilibrium swelling
for the synthesized hydrogels was measured in different pH solu-
tions ranging from 1.0 to 13.0 (Fig. 7A). Since the swelling capacity
of all anionic hydrogels is appreciably decreased by addition of
counter-ions (cations) to the swelling medium, no buffer solutions
were used. Therefore, aqueous stock NaOH (pH 13.0) and HCl (pH
1.0) solutions were diluted with distilled water to reach the de-
sired basic and acidic pHs, respectively. Maximum swelling
(2710 g/g) was obtained at pH 7.0. Under acidic (pH 2.0), most of
the carboxylate anions are protonated, so the main anion–anion
 G.R. Bardajee et al. / Journal of Photochemistry and Photobiology B: Biology 102 (2011) 232–240 237

Fig. 4. SEM photographs of optimized UV-prepared NPH at (A) 20 and (B) 50
magnification.

repulsive forces are eliminated and consequently swelling values

are decreased. Furthermore, some additional attractive interac-
tions, like AOH hydrogen bonding, lead to more decreased absorp-
tions. At pH 7.0, some of the carboxylate groups are ionized and the
electrostatic repulsion between ACOO groups causes an enhance-
ment of the swelling capacity. The reason of the swelling loss for
the highly basic solutions (pH > 7.0) is a ‘‘charge screening effect’’
of excess Na+ in the swelling media, which shields the carboxylate
anions and prevents effective anion–anion repulsion. Similar
swelling-pH dependencies have been reported for other hydrogel
systems [28–31]. The optimally prepared pH-sensitive hydrogel,
showed a reproducible On–Off switching behavior when the envi-
ronmental pH of the sample was alternatively changed between
8.0 and 2.0 (Fig. 7B). This responsiveness behavior may be of signif-
icant importance in colonic controlled delivery of drugs having
alkaline medium.
3.7. Drug delivery studies
Tetracycline hydrochloride (Fig. 8) is a well-known broad-spec-
trum antibiotic with low toxicity (LD50 = 6443 mg/kg) and efficacy
against gram-negative bacteria (MIC 90% = 8 lg/mL) which exhib-
its anti-collagenase activity, inhibition of bone resorption, anti-
inflammatory action, and have ability to promote attachment of
fibroblasts and connective tissue to root surfaces [32–34].
Fig. 5. Effect of (A) monomer amounts (B) UV-irradiation time (C) neutralization
percentage of AA on the water absorption capacity of the hydrogel.
3.7.1. Measurement of the extinction coefficient of the TH drug
The TH drug was dissolved in water at four different concentra-
tions. The UV absorption spectra of these solutions were measured
in a 1.00 cm UV cell and plotted versus concentrations. The plot
gave straight line that obey to the Beer–Lambert law (A = ecl, where
A is the absorbance of the solution, e is the extinction coefficient, l
is the cell length and c is the drug concentration). According to this
results, the extinction coefficient at kmax = 356 is 1.44  104
M1 cm1.
3.7.2. Determination of the drug loading in the hydrogel matrix
In order to determine the loading and releasing of the TH drug
in the entitled hydrogel, UV measurements were performed on sol-
free hydrogel samples. An accurately weighed powdery sample
(0.2 g) with average particle sizes between 40 and 60 mesh
238 G.R. Bardajee et al. / Journal of Photochemistry and Photobiology B: Biology 102 (2011) 232–240

3000 3.0

2500 2.5

UV Absorption
2000 2.0 4 Hour (36% Loading)
ES (g/g)

1500 1.5
2 Day (58% Loading)

1000 1.0

500 0.5
6 Day (74% Loading)
0.0
0
250 300 350 400
0 10 20 30 40 50 60
Time (min) Wavelength (nm)

Fig. 6. Swelling kinetics of the optimized NPH in distilled water. Fig. 9. The loading extent of the TH at different times.

(250–400 lm) was immersed entirely in a solution of TH drug

(0.6  104 mol/L) and allowed to soak at room temperature. Using
3000 the extinction coefficient of the TH in water, we calculated the
(A) amount of medicine incorporated into the polymer matrix during
2500 the loading of the drug. The results showed that the absorption
of drug depends on the concentration of drug in the solution,
2000 amount of hydrogel and the time of swelling. With increasing these
ES (g/g)

factors, the absorption of drug increased. For example, Fig. 9 shows

1500 that with increasing the time of soaking from 4 h to 6 day, the load-
ing of drug increases from 36% to 74%. The same effect was ob-
1000 served with increasing the amount of swelling hydrogel and the
concentration of drug solution which was used for loading experi-
500 ment. The higher drug solution concentration, the higher loading
will result. The maximum of TH absorption (98%) was achieved
0 for 0.2 g hydrogel in a 0.6  104 mol/L of drug solution after
0 2 4 6 8 10 12 14 10 days.
pH
3.7.3. Study of the TH release from the hydrogel matrix
Fig. 10A shows the curves of UV–visible spectra of TH release
2000
(B) from optimum NPH at various time intervals in pH 8.0. Fig. 10B
pH= 8 represents successful slow delivery of TH after 24 h with cumula-
pH= 2 tive release of 94%. There is a burst release initially for the 3 h in
1500 basic medium, followed by an almost constant release of TH from
ES (g/g)

the hydrogel for the remaining period of 24 h. The initial burst re-
1000 lease may be attributed to the release of TH molecules loaded near
the surfaces of the hydrogel. The in vitro TH release was also tested
in the conditions chosen to simulate the pH and time interval likely
500 to be encountered during transit from stomach to colon. First, the
TH loaded optimum NPHs were put in pH 2.0 buffer solution for
1 h, then in pH 6.5 buffered solutions for 3 h, and finally in pH
0 8.0 buffer solution for about 20 h. The main amount of TH (almost
0 10 20 30 40 50 60 50%) was released in the pH attributed to the colon (pH = 8.0;
Time (min) Fig. 11). This can be a good result in comparison with some earlier
reported cases [35] which without using enzyme in release med-
Fig. 7. (A) Swelling dependency of optimized hydrogel on pH and (B) On–Off ium, the cumulative release of drug has not exceeded from 30%.
switching behavior of the optimized hydrogel in buffered solutions with pH 8.0 and As one can see in Fig. 11, the amount and percentage of TH release
pH 2.0.
is much higher in basic media in comparison with acidic solutions.
The drug in the hydrogel could be released as a result of the hydro-
gel volume change and interaction between the polymer network
and TH. The fractional release is directly proportional to the swell-
ing ratio of the hydrogels, that is, pH 8.0 > pH 6.5 > pH 2.0 (Fig. 11).
This result indicates that the higher swelling ratios of the hydrogel
create larger surface areas to diffuse the drug. With the changing of
the pH of the solution, the charges of TH are also varied. In basic
solutions (pH 8.0), the electrostatic repulsion between dimethyl-
amine and phenoxide groups of TH and the carboxylate anions of
Fig. 8. Chemical structure of TH. grafted poly (sodium acrylate) on the hydrogel, accelerates the
 G.R. Bardajee et al. / Journal of Photochemistry and Photobiology B: Biology 102 (2011) 232–240 239

0.6 Despite the all advantages of enteric drug delivery systems,

(A) however, it should be remembered that enteric-coated formula-
tions are not suitable in some situations. Enteric-coated tablets
9 Hour (55% Releasing) contain indigestible solids and are often of considerable size. Thus,
UV Absorption

0.4 15 Hour (68% Releasing) seriously ill patients, who may have gastric hypomotility or pyloric
channel narrowing, are probably not good candidates for therapy
with large enteric-coated dosage forms. A further disadvantage of
this approach is the uncertainty of the location in which the enteric
0.2
coating starts to dissolve. Large inter and intra-patient variations
exist in such parameters as gastrointestinal (GI) motility and pH
3 Hour (34% Releasing) profile. Thus, there is a lack of control associated with the system,
0.0 so that depending on the prevailing GI motility pattern, pH profile
300 400 within the GI tract and fed-state, drug release may occur within the
small intestine or deep within the colon [36]. The released data im-
Wavelength (nm)
ply that this polymer network has the desired protective effect for
oral delivery of the drug, because a significant fraction of the drug
100 remains in the hydrogel pass through the low-pH environment of
(B) the stomach. When the hydrogels are transferred to the pH = 8
80 solution, a great deal of TH is released from the polymer network
system. Thus, in a more practical point of view, these biocompati-
Release (%)

ble hydrogel systems can bypass the acidity of gastric fluid without
60 liberating substantial amounts of the loaded drug. After 24 h, the
cumulative TH released cannot be delivered completely. It can be
40 attributed to some entrapped TH molecules within the hydrogel
network, and these cannot be released unless the polymer ma-
trixes are degraded.
20

4. Conclusions
0
0 5 10 15 20 25 In the present work, we prepared a novel nanoporous hydrogel
Time (h) by UV-irradiation cross-linking graft copolymerization of AA onto
salep (as a composite polysaccharide) backbone in the absence of
Fig. 10. The releasing extent of the TH at different times.
toxic cross-linking agent and initiator. The optimum reaction con-
ditions (obtained by the OFAT method) to achieve maximum water
absorption (2710 g/g) were found to be: salep = 1.0 g, AA = 1.5 mL,
pH= 6.5
neutralization percentage = 65% and UV-irradiation time 50 min.
pH= 2 pH= 8
The SEM images proved the porosity of the hydrogel in nanoscale
100 (average pore size: about 350 nm). The drug delivery results re-
Cumulative Release (%)

80
60
40
20
0
0 5 10
Time (h)
Fig. 11. Release profile of TH from NPH under different conditions: 0–1 h, pH = 2.0;
1–4 h, pH = 6.5; 4–24 h, pH = 7.5.
release of TH from the hydrogel. These repulsion forces compete
with a large number of H-bonds between the polar groups in TH
(amide and hydroxyl groups) and the hydroxyl, neutralized and
un-neutralized carboxylic acid groups in the polymer network
which serve to hinder the TH release from the hydrogel. As a result
of the two effects, the release of TH from the NPH at pH 8.0 solution
is good and may be suggested as a successful device for drug re-
lease in colon. Due to the instability of tetracycline in acidic media,
this system which protects the drug crossing over the stomach will
increase the efficiency of the medicine.
vealed that the release profiles of TH from the NPH were slow in
simulated gastric fluid (pH 2.0) but almost of 50% of drug content
was released in simulated colonic fluid (pH 8) within 20 h. Overall,
the results demonstrated that this NPH could successfully deliver
the drug to the colon without losing the drug in the stomach,
and could be a good candidate as an orally administrated drug
delivery system for drugs such as TH.
References
[1] G. Gerlach, K.F. Arndt, Hydrogel Sensors and Actuators: Engineering and
Technology, Springer, London, 2009.
15 20 25 [2] G.S. Kwon, Polymeric Drug Delivery Systems, Taylor and Francis, New York,
2005.
[3] B. Wang, T. Siahaan, R. soltero, Drug Delivery: Principles and Applications, John
Wiley and Sons, New Jersey, 2005.
[4] R. Hejazi, M. Amiji, Chitosan-based gastrointestinal delivery systems, J.
Control. Release 89 (2003) 151–165.
[5] H. Tozaki, J. Komoike, C. Tada, T. Maruyama, A. Terabe, T. Suzuki, A. Yamamoto,
S. Muranishi, Chitosan capsules for colon-specific drug delivery: improvement
of insulin absorption from the rat colon, J. Pharm. Sci. 86 (1997) 1016–1021.
[6] S. Davaran, J. Hanaee, A. Khosravi, Release of 5-amino salicylic acid from acrylic
type polymeric prodrugs designed for colon-specific drug delivery, J. Control.
Release 58 (1999) 279–287.
[7] C.L. Cheng, S.H. Gehrke, W.A. Ritschel, Development of an azopolymer based
colonic release capsule for delivering proteins/macromolecules, Methods Find.
Exp. Clin. Pharmacol. 16 (1994) 271–278.
[8] L. Vervoort, I. Vinckier, P. Moldenaers, G.V.D. Mooter, P. Augustijns, R. Kinget,
Inulin hydrogels as carriers for colonic drug targeting: rheological
characterization of the hydrogel formation and the hydrogel network, J.
Pharm. Sci. 88 (1999) 209–214.
[9] S.Y. Lin, J.W. Ayres, Calcium alginate beads as core carriers of 5-aminosalicylic
acid, Pharm. Res. 9 (1992) 1128–1131.
240 G.R. Bardajee et al. / Journal of Photochemistry and Photobiology B: Biology 102 (2011) 232–240
[10] M. Marvola, P. Nykanen, S. Rautio, N. Isonen, A. Autere, Enteric polymers as
binders and coating materials in multiple-unit site-specific drug delivery
systems, Eur. J. Pharm. Sci. 7 (1999) 259–267.
[11] M. Rodriguez, J.L. Vila Jato, D. Torres, Design of a new multiparticulate system
for potential site-specific and controlled drug delivery to the colonic region, J.
Control. Release 55 (1998) 67–77.
[12] M. Ashford, J. Fell, D. Attwood, H. Sharma, P. Woodhead, An evaluation of
pectin as a carrier for drug targeting to the colon, J. Control. Release 26 (1993)
213–220.
[13] M. Ashford, J. Fell, D. Attwood, H. Sharma, P. Woodhead, Studies on pectin
formulations for colonic drug delivery, J. Control. Release 30 (1994) 225–
232.
[14] A. Rubinstein, D. Nakar, A. Sintov, Colonic drug delivery: enhanced release of
indomethacin from cross-linked chondroitin matrix in rat cecal content,
Pharm. Res. 9 (1992) 276–278.
[15] G.Q. Lu, X.S. Zhao, Nanoporous Materials: Science and Engineering, Imperial
College Press, London, 2004.
[16] L.I. Mikhalovska, V.M. Gun’ko, W. Turov, V.I. Zarko, S.L. Jamesa, P. Vadgama,
P.E. Tomlins, S.V. Mikhalovsky, Characterisation of the nanoporous structure of
collagen-glycosaminoglycan hydrogels by freezing-out of bulk and bound
water, Biomaterials 27 (2006) 3599–3607.
[17] M.L. Deng, Q. Zhou, A.K. Du, J.V. Kasteren, Y.Z. Wang, Preparation of
nanoporous cellulose foams from cellulose-ionic liquid solutions, Mater. Lett.
63 (2009) 1851–1854.
[18] G.R. Bardajee, A. Pourjavadi, R. Soleyman, Novel highly swelling nanoporous
hydrogel based on polysaccharide/protein hybrid backbone, J. Polym. Res. 17
(2010) In Press.
[19] A. Pourjavadi, R. Soleyman, G.R. Bardajee, Novel nanoporous superabsorbent
hydrogel based on poly(acrylic acid) grafted onto salep: synthesis and swelling
behavior, Starch/Starke 60 (2008) 467–475.
[20] G.R. Bardajee, A. Pourjavadi, R. Soleyman, N. Sheikh, Gamma irradiation
mediated synthesis of a new superabsorbent hydrogel network based on poly
(acrylic acid) grafted onto salep, JICS 7 (2010) 652–662.
[21] G.R. Bardajee, A. Pourjavadi, R. Soleyman, N. Sheikh, Irradiation mediated
synthesis of a superabsorbent hydrogel network based on polyacrylamide
grafted onto salep, Nucl. Instrum. Methods Phys. Res. Sect. B 266 (2008) 3932–
3938.
[22] A. Pourjavadi, G.R. Bardajee, R. Soleyman, Synthesis and swelling behavior of a
new superabsorbent hydrogel network based on polyacrylamide grafted onto
salep, J. Appl. Polym. Sci. 112 (2009) 2625–2633.
[23] R. Farhoosh, A. Riazi, A compositional study on two current types of salep in
Iran and their rheological properties as a function of concentration and
temperature, Food Hydrocolloid. 20 (2006) 660–666.
[24] K.K. Tekinsen, A. Guner, Chemical composition and physicochemical
properties of tubera salep produced from some Orchidaceae species, Food
Chem. 121 (2010) 468–471.
[25] H. Saeidian, F. Matloubi Moghaddam, A. Pourjavadi, S. Barzegar, R. Soleyman,
Superabsorbent polymer as nanoreactors for preparation of hematite
nanoparticles and application of the prepared nanocatalyst for the friedel-
crafts acylation, J. Brazil Chem. Soc. 20 (2009) 466–471.
[26] I.S. Kim, S.H. Kim, C.S. Cho, Drug release from pH-sensitive interpenetrating
polymer network hydrogel based on poly (ethylene glycol) macromer and poly
(acrylic acid) prepared by UV cured method, Arch. Pharm. Res. 19 (1996) 18–
22.
[27] H. Omidian, S.A. Hashemi, P.G. Sammes, I.G. Meldrum, A model for the swelling
of superabsorbent polymers, Polymer 39 (1998) 6697–6704.
[28] W.F. Lee, W.Y. Yuan, Thermoreversible hydrogels X: synthesis and swelling
behavior of the (N-isopropylacrylamideco-sodium 2-acrylamido-2-methyl-
propyl sulfonate) copolymeric hydrogels, J. Appl. Polym. Sci. 77 (2000) 1760–
1768.
[29] C.K. Nisha, D. Dhara, P.R. Chatterji, Superabsorbency and volume phase
transition in crosslinked poly[[3-(methacryloylamino)propyl]-trimethyl-
ammonium chloride] hydrogels, J. Macromol. Sci. Part A: Pure Appl. Chem.
37 (2000) 1447–1460.
[30] K. Burugapalli, D. Bhatia, V. Koul, V. Choudhary, Interpenetrating polymer
networks based on poly(acrylic acid) and gelatin. I: Swelling and thermal
behavior, J. Appl. Polym. Sci. 82 (2001) 217–227.
[31] S. Lu, M. Duan, S. Lin, Synthesis of superabsorbent starch-graft-poly
(potassium acrylate-co-acrylamide) and its properties, J. Appl. Polym. Sci. 8
(2003) 1536–1542.
[32] S.S. Biju, S. Saisivam, N.S.M.G. Rajan, P.R. Mishra, Dual coated erodible
microcapsules for modified release of diclofenac sodium, Eur. J. Pharm. Sci.
Biopharm. 58 (2004) 61–67.
[33] P. Andrade-Vivero, E. Fernandez-Gabriel, C. Alvarez-Lorenzo, A. Concheiro,
Improving the loading and release of NSAIDs from pHEMA hydrogels by
copolymerization with functionalized monomers, J. Pharm. Sci. 96 (2007) 802–
813.
[34] K. Schwach-Abdellaoui, A. Monti, J. Barr, J. Heller, R. Gurny, Optimization of a
novel bioerodible device based on auto-catalyzed poly(ortho esters) for
controlled delivery of tetracycline to periodontal pocket, Biomaterials 22
(2001) 1659–1666.
[35] L.G. Chen, Z.L. Liu, R.X. Zhuo, synthesis and properties of degradable hydrogels
of konjac glucomannan grafted acrylic acid for colon-specific drug delivery,
Polymer 46 (2005) 6274–6281.
[36] A.M. Hillery, A.W. Lloyd, J. Swarbrick, Drug Delivery and Targeting: for
Pharmacists and Pharmaceutical Scientists, Taylor & Francis, London, 2009.