Drug Delivery

ISSN: 1071-7544 (Print) 1521-0464 (Online) Journal homepage: http://www.tandfonline.com/loi/idrd20

Ophthalmic controlled release in situ gelling

systems for ciprofloxacin based on polymeric
carriers

Raida S. Al-Kassas & Mona M. El-Khatib

To cite this article: Raida S. Al-Kassas & Mona M. El-Khatib (2009) Ophthalmic controlled release
in situ gelling systems for ciprofloxacin based on polymeric carriers, Drug Delivery, 16:3, 145-152,
DOI: 10.1080/10717540802689008

To link to this article: http://dx.doi.org/10.1080/10717540802689008

Published online: 01 Apr 2009.

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Drug Delivery, 2009; 16(3): 145–152

R e s e a r c h Ar t i c l e

Ophthalmic controlled release in situ gelling systems

for ciprofloxacin based on polymeric carriers
Raida S. Al-Kassas, and Mona M. El-Khatib
Department of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia

Abstract
The objective of the present study was to design controlled release ophthalmic delivery systems for cip-
rofloxacin based on polymeric carriers that undergo sol-to-gel transition upon change in pH or in the
presence of cations in an attempt to prolong the effect of ciprofloxacin and improve its ocular bioavail-
ability. Carbopol and alginates polymers were used to confer gelation properties to the formulations.
Hydroxypropyl methylcellulose and methylcellulose were combined with carbopol to increase the viscos-
ity of the gels and to reduce the concentration of the incorporated carbopol. The release exponents (n)
for the designed systems were close to 1, indicating that the drug release occurred by zero-order kinetics.
Controlled release in situ gels consisting of carbopol and cellulose derivatives showed an increase in viscos-
ity, gelling capacity, and adhesiveness as the concentration of each polymeric component was increased.
On the other hand, these parameters possessed lowest values when alginate was used as an in situ gelling
agent. The antimicrobial efficiency of the selected formulation against gram-positive and gram-negative
organisms including Echerichia coli, staphylococcus strains and Pseudomonas aeruginosa confirmed that
the designed formulation has prolonged the antimicrobial effect of ciprofloxacin and retained its proper-
ties against bacteria.
Keywords:  Ciprofloxacin; in situ gels; pH triggered systems; ophthalmic delivery systems; controlled release gels

Introduction
Development of controlled release delivery systems for
ocular therapy is a challenging task. Conventional eye
drops results in poor bioavailability and therapeutic
response owing to drainage by gravity-induced lac-
rimal and normal tear turnover (Schoenwald, 1990).
Frequent dosing ­therefore is necessary to compensate
for the decreased pre-­corneal drug administration,
however it’s usually associated with non-patient com-
pliance. Inclusion of excess drug in the formulation in
an attempt to overcome the bioavailability problem is
potentially dangerous if the drug solution drained from
the eye is systemically absorbed from the nasolacrimal
duct (Middleton et al., 1990). It has been suggested that
incorporation of polymers into aqueous vehicle can
increase its viscosity and slow down the drug elimina-
tion from the conjuctival sac; however, limited improve-
ment in bioavailability has been obtained (Patton and
Robinson, 1975; Zaki et al., 1986). Mucoadhesive prop-
erty of the vehicle is also a critical factor that affects the
corneal contact for drugs (Greaves and Wilson, 1993).
Ophthalmic delivery systems based on in situ gel
have gained increasing interest. These systems consist of
polymers that undergo sol-to-gel phase transition upon
change in the physical conditions (pH, temperature).
The gelation can be triggered by a shift in temperature
as poloxamer (Miller and Donovan, 1982) and ethyl
(hydroxy ethyl) cellulose (Lindell and Engstrom, 1993),
a shift in pH as for cellulose acetate phthalate (Gurny,
1981) and carbopol (Srividya et al., 2001), or by presence
of cations as for deacetylated gellan gum (Carfors et al.,

Address for Correspondence: Raida Al-Kassas, Department of Pharmaceutics, College of Pharmacy, King Saud University, PO Box 17221, Riyadh 11484,
Saudi Arabia. Email: kassas_r2@yahoo.com
(Received 18 August 2008; accepted 15 December 2008)
ISSN 1071-7544 print/ISSN 1521-0464 online © 2009 Informa UK Ltd
DOI: 10.1080/10717540802689008 http://www.informapharmascience.com/drd
146   R. S. Al-Kassas and M. M. El-Khatib

1998) and alginate (Cohen et al., 1997; Liu et al., 2006). In Materials and methods
situ gel system is formulated as liquid preparation suit-
able to be instilled into eyes which upon exposure to the Materials
physiologic environment changes to gel, thus increasing
the precorneal residence time of the delivery system, Ciprofloxacin was obtained from Sigma Chemical Co
and enhances the ocular bioavailability of the drug. (USA). Carbopol 934 and Carbopol 971 were purchased
Ciprofloxacin (CPFX) (Scheme 1) is a synthetic fluo- from BF Goodrich (USA). Sodium alginate was obtained
roquinolone antibiotic with a broad spectrum antimicro- from Hopkins and Williams Ltd. (UK). Hyroxy propyl
bial activity (Goodman and Gilman, 1996). It is effective methyl cellulose, Methyl cellulose, and Tween 80 were
against external infection of the eye, such as conjuncti- obtained from Sigma Chemical Co. (USA). All other
vitis and kerato conjunctivitis (Appelbaum and Hunter, chemicals were commercially available products of ana-
2000). The solubility of ciprofloxacin is pH-dependant lytical grade.
and is ranging from 6.19 mg/ml at pH 5 to 0.15 mg/ml at
pH 7 and 37°C. The low aqueous solubility of this com- Preparation of in-situ ciprofloxacin formulations
pound at pH close to 7 is due to that in aqueous solution,
ciprofloxacin exists in its zwitterionic form due to the Table 1 shows the composition of controlled released
acid/base interaction between the basic nitrogen of the in-situ ciprofloxacin gels. Ciprofloxacin gels based on
piperazine and the carboxylic acid groups (Vilches et al., HPMC and Cabopol (CP) were prepared as follows.
2002). This interaction is the main reason that prevents Initially, HPMC was dispersed in pH 6 phosphate buffer
the design of liquid dosage forms such as ophthalmic containing 1% (w/w) Tween 80 using a mechanical stir-
and parenteral solution, because the aqueous compat- rer. Following complete hydration, CP was sprinkled
ibility of these drugs occurs at rather basic or acidic pH over the solution and allowed to hydrate over night.
(Allemandi et al., 1999). The efficacy of market fluoroqui- Ciprofloxacin was then dissolved in 0.5 N NaOH solu-
nolone products, mostly aqueous solubility, is limited by tion and incorporated manually into the HPMC/CP
poor bioavailability (Lin et al., 1996). When ciprofloxacin mixture until homogeneity was attained. Alginate gels
0.3% solution was used to treat bacterial corneal ulcers, were prepared by dispersing the required amount of
crystalline precipitates appeared in the superficial por- alginate polymer in pH 6 phosphate buffer containing
tion of the corneal defect (Thomson Micromedex, 2003). 1% w/v Tween 80. Ciprofloxacin was then added to the
Therefore, and based on what has been mentioned mixture after dissolving it in 0.1 N NaOH. The samples
above, the main objective of the present investigation were then transferred into amber ointment jars after
was to control the rate of dissolution of ciprofloxacin adjusting the pH to 6 and stored at 4°C prior to further
in an attempt to control its rate of absorption from the analysis.
cornea, improve its ocular bioavailability, and reduce its
side-effects by incorporating it into a controlled release Uniformity of drug content and pH measurements
in situ gelling system based on polymeric carriers.
Carbopol and alginate were investigated as vehicles for Drug content of ciprofloxacin formulations was deter-
formulation of ciprofloxacin eye drops which undergo mined by dissolving an accurately weighed quantity of
gelation when instilled into the cul de sac. Carbopol (CP) formulation (1 g) in 50 ml of pH 6 phosphate buffer. The
is widely used in in-situ gel forming systems, owing to solutions were then filtered through 0.45 m membrane
the property of its aqueous solution to transform into gel filter and analyzed for ciprofloxacin content by UV spec-
when the pH is raised. However, the concentration of CP trophotometer at 270 nm.
to form stiff gel results in highly acidic solutions which The pH was measured for each formulation using a
are not easily neutralized by the buffer action of the tear pH meter (Sartorius, USA) which was calibrated before
fluid. Therefore, viscosity-enhancing agents such as cel- use with buffered solution at pH 4, 7, and 10.
lulose derivatives were used in order to reduce CP con-
centration without compromising the gelling capacity Table  1.  Composition of prolonged-action in situ ciprofloxacin
and the rheological properties of the prepared systems. eye gels.
In the present study hydroxypropylmethylcellulose Formulation Concentration (%W/W)
(HPMC) and methyl cellulose have been used as viscosi- code Drug HPMC CP934 CP971 MC Tween alginate
ty-enhancing agents to carbopol, and their effects on the F1 0.3 1.5 0.5 — — 1
mechanical properties and the release characteristics of F2 0.3 1.5 0.5 — 1 —
ciprofloxacin from the gel formulations were studied. A F3 0.3 — 0.5 — 1.5 1 —
further objective of the study was to evaluate alginate as F4 0.3 — — — — 1 2
an ocular delivery system of ciprofloxacin which has the F5 0.3 1 0.4 — — 1 —
ability to gel in the presence of cations. F6 0.3 1 0.5 — — 1 —
 Ophthalmic controlled release in situ gelling systems   147

Gelling capacity
The gelling capacity of ciprofloxacin formulations was
determined by placing a drop of the prepared systems
in a vial containing 2 ml of pH 7.4 artificial tear fluid
freshly prepared and equilibrated at 37°C. The gelling
capacity was determined visually by assessing the gel
formation and noting the time taken for the gel formed
to remain intact in tear fluid before complete dissolu-
tion took place. The composition of artificial tear fluid
was 0.670 g Sodium Chloride, 0.2 g Sodium bicarbo-
nate, 0.008 g Calcium Cloride.2H2O, deionized water
q.s. 100 g (Vo’oleghem, 1993).
Viscosity measurement
A Viscosimeter (Brookfield R.V II, UK) was used to
measure the viscosities of the developed CPs/Cellulose
derivatives—ciprofloxacin formulations (pH 6). The
spindle (number 21) was rotated at 10 rpm. The viscos-
ity was also measured for ciprofloxacin formulations
after raising their pHs to 7.4 using 0.5 N NaOH solution.
The change in viscosities of the formulation based on
alginate was evaluated after mixing it with artificial tear
fluid.
Adhesion study
The adhesion study of controlled release ciprofloxacin
formulations, after gelling, was measured using the
apparatus described by Agrwal and Mishra (1990). The
apparatus consisted of two circular aluminium discs
each of 3 cm diameter. One disc was allowed to hang
on an iron stand with the help of an aluminium wire
fastened with a hook provided on the back side of the
disc. The other disc was connected to a pre-weighed
lightweight plastic bag using a hook attached on its
back. The studied gel was placed between the two discs,
which were then kept under constant pressure for 5 min
(preload time) to initiate adhesion bond. After that,
water was added to the plastic bag through an intrave-
nous infusion set at a rate of 1 drop/min until the two
adhered discs became detached from each other. The
water collected in the bag was weighed. The weights of
collected water were converted into force required for
detachment.
In vitro diffusion studies of ciprofloxacin through a
membrane
The in vitro diffusion of the drug through a mem-
brane was carried out in a system composed of a glass
tube in which a cellophane membrane (soaked over
night in artificial tear fluid, pH 7.4) was stretched and
securely fastened with a rubber band; 1 g of the 0.3%
w/w formulation was placed in the tube (phase I or the
donor phase). This was hung vertically in a beaker con-
taining 50 ml artificial tear solution, pH 7.4 (phase II or
the acceptor phase). The diffusion system was placed
in a thermostatically shaking water bath at 37°C ± 1.
At predetermined time intervals, 1 ml of the solutions
were removed from the acceptor phase and analyzed
for ciprofloxacin using a UV spectrophotometer at 270
nm and an equal volume of fresh, pre-warmed artificial
tear was replaced into the dissolution vessels. The pres-
ence of formulation excipients did not interfere with the
analysis. The amount of ciprofloxacin released at each
time was calculated from a calibration curve (2–10 µg/
ml, r > 0.99 with zero intercept).
Antimicrobial efficiency of controlled release
­ciprofloxacin gel
The antimicrobial efficiency and prolonged effect of
selected controlled release ciprofloxacin gel were deter-
mined on Echerichia coli, staphylococcus strains, and
Pseudomonas aeruginosa as a function of time. The
inhibitory effect of ciprofloxacin formulation on the
studied microorganisms was evaluated using agar dif-
fusion test. Wells were punched into the agar (Mueller
Hinton agar) previously seeded with test organisms (105
cfu/ml by Macferland method) and wells were filled with
100 µl of the samples which were collected from the in
vitro diffusion test of ciprofloxacin gel formulation. After
incubation for 24 h at 37°C the diameters of inhibition
zones were measured. The results were compared with
control (artificial tear fluid).
Results and discussion
Formulation of the gels
The pH of the buffer has a major contribution in formu-
lation of a stable and therapeutically effective ophthal-
mic delivery system. It has been reported that ocular
penetration of levofloxacin, which is a ­derivative of
ciprofloxacin, is at a maximum at pH 6.5 (Kawazu et al.,
1996). In addition, and as mentioned earlier in the intro-
duction, the aqueous solubility of ciprofloxacin occurs
at acidic or basic pHs. Therefore, the pHs of ciprofloxacin
formulations prepared in this study were adjusted to 6.
Physicochemical properties of controlled release
ciprofloxacin ophthalmic in-situ gels
Drug content and pH
All prepared ciprofloxacin formulations possessed sat-
isfactory pHs, ranging from 6–6.2. Drug content values
ranged from 98–100%.
148   R. S. Al-Kassas and M. M. El-Khatib
Mechanical properties of ciprofloxacin
ophthalmic gels
The desired outcome of a prolonged action gel must be
to preserve it’s integrity without erosion in the tear fluid
for a long period of time. However, it has been reported
that a prolonged precorneal residence of formulations
containing the in situ gelling polymer, alginic acid, was
looked for, not only based on its ability to gel in the eye
but also because of its mucoadhesive properties (Smart
et al., 1984). For these reasons, mechanical properties of
ciprofloxacin ophthalmic formulations, including the gel-
ling capacity, viscosity at pH 7.4, and the mucoadhesive
properties, were evaluated and the results are presented
in Table 2. The gelling capacity was expressed as the time
it takes the gel to remain intact in tear fluid before erosion
takes place. It should be noted that all formulations under-
went immediate transition into gel phase in contact with
the tear fluid (pH 7.4). The type of in situ gelling polymer,
its concentration, and the type of cellulose derivative had
a significant effect on the gelling capacity of ciprofloxacin
formulations. Minimum gelling capacity was exhibited
by formulations containing alginate. On the other hand,
formulations based on Carbopol and methylcellulose
exhibited the maximum gelling capacity.
Table 2 shows the viscosity of ciprofloxacin formula-
tions consisting of CPs/cellulose derivatives when their
pHs were raised to 7.4 and alginate formulation after
it was mixed with artificial tear fluid. In formulations
based on HPMC and CP, increasing the concentration of
each polymeric component significantly increased the
formulation viscosity. Carbopol and HPMC are water-
swellable polymers. The extent of swelling of these poly-
mers was dependent on the amount of free water in the
formulations. Hence, in formulations containing high
concentrations of HPMC or CP, their extent of swelling
was decreased. Furthermore, in formulations containing
high concentrations of HPMC or formulations containing
high concentrations of CP, a greater proportion of theses
polymers was present as unswollen suspended solid com-
pared to formulations containing lower concentrations of
these polymers. It is proposed that the physical states of
these polymers, i.e. swollen or unswollen, were directly
responsible for the mechanical properties, including the
Table  2.  Physical properties of prolonged action in in situ
ciprofloxacin eye gels.
Formulation Gelling capacity Viscosity (CPs) Adhesiveness
code (min ± SD) (10 rpm) (N mm)
F1 11.66 ± 0.58 4296.3 0.0903
F2 3.67 ± 0.58 2870.97 0.0717
F3 > 37 ± 0.00 1600 0.2761
F4 2.33 ± 1.16 262 0.0643
F5 2.68 ± 0.58 1797 0.0863
F6 7.67 ± 1.15 2176 0.0893
viscosity of the prescribed formulations. Ferrari et al.
(1994) reported that gel strength of HPMC gels increased
as the polymer concentration was increased. Jones et al.
(2000) revealed that increasing the concentration of poly-
mer components of the gels resulted in an increase in the
hardness, adhesiveness, compressibility, and syringe-
ability of the formulations. The effect of HPMC and CP
on the viscosity of in situ prolonged action gels under
examination may also be explained by their effect on the
product viscoelasticity following dispersion of swollen or
unswollen solids. Increasing the product viscoelasticity,
particularly elasticity, produced an increased resistance
to product deformation during viscosity measurements.
The lowest viscosity value was observed from formula-
tion 4, containing alginate, indicating complete swelling
of the polymer in the buffer system. On the other hand,
the formulation consisting of MC and CP (F3) showed
lower viscosity data than formulations containing HPMC
and CP, which may be attributed to the improved swell-
ing property of methylcellulose in the vehicles used for
preparation of ciprofloxacin formulations.
One important characteristic of the prolonged action
eye gel is its ability to exhibit retention in the eye. The
polymers used in this study have been described as
mucoadhesives. Therefore, they would be expected to
show retention through formation of adhesive bonds
with ocular tissues.
The adhesiveness of ciprofloxacin formulations,
after they were gelled, are presented in Table 2. In the
method used to evaluate the mucoadhesive properties
of the ciprofloxacin gels, the work required to overcome
the attractive force developed between the surface of
the sample and the surfaces of the discs was expressed
as adhesiveness. The bioadhesive force is known to be
dependent on the nature and the concentration of the
bioadhesive polymer. In formulations consisting of
HPMC and CP, the adhesiveness of the gel increases as
the concentration of each polymer increases. This may
be attributed to the increased ability of these polymers
to interact with the surface of the discs, but may also
be a function of the increased tack of each formulation.
Furthermore, the physical state of the polymeric compo-
nent was observed to significantly affect the adhesive-
ness of the formulations. Hence, in formulations where
CP or HPMC existed as suspended unswollen solids, the
adhesiveness was greater than in formulations in which
these polymers exhibited a greater degree of swelling.
The increased mass of unswollen particles in formula-
tions containing CP 0.5%(w/w) compared to those con-
taining CP 0.4% (w/w) or those containing HPMC 1.5%
(w/w) compared to those containing HPMC 1% (w/w)
has a direct effect on the adhesive strength. These results
are in agreement with Choi et al. (1998), who reported
an increase of bioadhesive forces of the gels as the car-
bopol concentration was increased. Table 2 also reveals
 Ophthalmic controlled release in situ gelling systems   149

that the impact of HPMC on the bioadhesive force of the
gels was lessened by the presence of CP971. Keeping the
concentration of HPMC constant, the formulation con-
taining 0.5% (w/w) of CP 971 possessed lower adhesive
force than formulation containing 0.5% (w/w) of CP 943.
On the other hand, methylcellulose has enhanced the
gel adhesiveness more efficiently than the other systems
examined. This could be due to the strong adhesive
nature of methylcellulose when it is combined with CP
in a formulation. Of all the systems prepared, the formu-
lation containing 2% of alginate possessed the poorest
adhesive properties, and would therefore be susceptible
to easy removal from the cornea after administration.
Thus, these results suggest that addition of viscosity-
enhancing agent to alginate gel may improve its adhe-
siveness and other mechanical properties.
In vitro diffusion study
The in vitro diffusion results of the drug release obtained
with the ciprofloxacin formulations in comparison with
O F3
F 4
6 3
7
1
HN N N
Scheme  1.  Chemical structure of ciprofloxacin.
between the zwitterionic species of ciprofloxacin and the
carboxylic groups of carbopol and alginates. During the
diffusion studies, Na+, K+, HCO3−, and Cl− ions present
in the acceptor phase (artificial tear fluid) diffused into
the gel and Na+ and K+ promoted the exchange of the
cationic and zwitterionic species attached to carbopol
and alginate polymers. The dissociation of ionic pairs
has contributed in facilitating the drug release from gel
matrices. Vilches et al. (2002) reported that ionic pair-
ing between fluoroquinolones and carbopol is the main
interaction that determines the enhanced aqueous com-
patibility and releasing properties of the drug. Moreover,
loading carbopol with fluoroquinolone together with an
appropriate amount Na+ yields physically stable disper-
sion that behaves as a molecular matrix of fluoroqui-
nolones, able to deliver the drug at a constant rate (zero
order) in contact with a biological fluid-like solution.
Table  3.  The time required for the release of 20% of the original mass
of ciprofloxacin from gel formulation.
Formulation code T20% (min)
F1 135
F2 108
113
COOH F4 110
F5 105
F6 123
a)
H H

40 C C
35
H C
30
% Drug diffused

25 HO O n
20 F1
15 F2
F3 b) a COO−
10 F4 O
OH O
OH
F5 COO−
5 F6 OH OH OH OH
HO HO
Pure drug
0
0 0.5 1 1.5 2 2.5 3 3.5 β-D-mannuronate (M) α-L-guluronate (G)
Time (hours) b
−OOC OH −OOC HO −OOC
O O O HO O
Figure  1.  In vitro diffusion of ciprofloxacin from controlled release OH
O
HO O OH
O
gel formulations. O −OOC HO O
OH
O O
the ciprofloxacin solution are shown in Figure 1. A slow OH −OOC OH
diffusion rate was observed from ciprofloxacin solution G G M M G
which could be attributed to the low aqueous solubility c
of ciprofloxacin at pH close to 7. Surprisingly, incorpora- M MMMGMGGGGGMGMGGGGGGGGMMGMGMGGM
tion of ciprofloxacin into gels caused an increase in its
release and permeation rates. In an attempt to explain M-block G-block G-block MG-block
the reasons for these findings, the chemical structures
Figure  2.  (a) General structure of Carbopol polymers. (b) schematic
of alginates and carbopol, the polymers used in gels diagram of the structure of alginate showing (a) the constituent sug-
formulations, were analyzed (Figures 2(a) and (b)). The ars, (b) their relevant linkages, and (c) possible intramolecular pat-
chemical structures suggest formation of ionic pairs terns of the different sugars.
150   R. S. Al-Kassas and M. M. El-Khatib
Table 3 shows the time required for 20% of the cip-
rofloxacin to be released (T20%) from the various gel
formulations. Increasing the concentration of HPMC
from 1 to 1.5% and the CP from 0.4 to 0.5% has mark-
edly increased T20%. The maximum T20% value was 135
min, and was associated with F1 (formulation contain-
ing highest concentration of HPMC and CP). On the
other hand, the minimum T20% value was associated
with the formulation containing lowest CP and HPMC
concentrations (F5), and was 105 min. F3 (formula-
tion containing methylcellulose) and F4 (formulation
containing alginate) exhibited T20% values between the
two. It is thought that these effects are the function of
interaction of two parameters: the polymer type and its
concentration. As the polymer concentration increases,
the diffusion of ciprofloxacin through the formulation
reduces due to the more entangled nature of the poly-
meric network. In addition, the ingress of water into the
formulation containing high concentration of polymer
was reduced, thus lowering the rates of both dissolution/
erosion. Also, the density of the chain structure of the
gels increases at higher polymeric concentrations, and
this limits the active substance movement area (Wang et
al., 2001). Finally, the degree of swelling increases as the
concentration of the suspended solids increases, thus
decreasing the diffusion of ciprofloxacin from the swol-
len matrix. The swelling phenomenon may be directly
responsible for the effects of the type of polymer on the
drug release from the formulation.
The data generated from these release studies were
fitted to the general release equation (Korsmeyer et al.,)
using the logarithmic transformation and least-square
regression analysis, as described below, in an attempt to
investigate the mechanism of ciprofloxacin release.
log Mt/M∞ = log k + n log t
where Mt is the amount of drug released at time t; M∞
is the total drug content; k is a constant incorporating
structural and geometrical characteristics of the device;
and n is the release exponent which may indicate the
mechanism of drug release.
The kinetic parameters n and k were calculated and tab-
ulated (Table 4). For the formulations under investigation,
the release exponents (n) were close to 1, suggesting that
ciprofloxacin was delivered from gels by zero order kinetics.
Similarly, Vilches et al. (2002) reported zero-order kinetics
Table  4.  Kinetic parameters obtained from the release equation.
Formulation code k N
F1 0.1215 1.022
F2 0.1383 1.051
F3 0.1246 1.029
F4 0.1425 1.032
F5 0.2111 0.975
F6 0.1698 0.973
for the release of ciprofloxacin from carbomer hydrogels.
The slight deviation of the n-value that was observed in
some cases could be attributed to the swelling properties
of these formulations following the ingress of water.
It has been reported that as the k-value becomes
higher, the release rate becomes faster (Choi et al., 1998).
Analysis of k-value of the various formulations revealed
that the release rate of ciprofloxacin was decreased as
the concentration of each polymeric component was
increased. These results confirm our earlier finding
based on T20% data. Moreover, replacing HPMC with MC
has slightly accelerated drug release rate, as indicated by
k-values, 0.1215 and 0.1246 for formulations containing
HPMC and MC, respectively. Based on the mechanical
properties and release characteristics of the investigated
formulations, formulation F3 containing Carbopol 934
and methyl cellulose in addition to ciprofloxacin was
selected for the microbiological studies.
Antimicrobial efficiency studies
The antimicrobial efficiency of the selected controlled
release ciprofloxacin formulation was evaluated against
gram-positive and gram-negative organisms including
Echerichia coli, staphylococcus strains, and Pseudomonas
aeruginosa. The inhibition zones produced by cipro-
floxacin gel was measured in bacteriological medium
and the results are presented in Figures 3(a–c). The inhi-
bition zones were evaluated every 30 min and reduction
in the growth of microorganisms was clearly observed.
Comparison of the diameters of the zone of growth
inhibition (Figure 4) reveals that controlled release cip-
rofloxacin formulation was most effective against E. coli.
The growth inhibition was observed in the first 30 min of
the experiment and increased as the amount of cipro-
floxacin diffused from gel was increased. Ciprofloxacin
gel has also shown activity against staphylococcus stains
but to a lesser extent than E. coli; however its antimicro-
bial activity was sustained for 4 h. On the other hand,
the result of ciprofloxacin formulation against P. aerugi-
nosa showed that the microorganism has resisted cipro-
floxacin in the first 2 h, but rapid increase in the rate of
growth inhibition was observed afterwards.
In conclusion, various controlled release in situ gelling
systems for ciprofloxacin were designed and character-
ized in terms of ciprofloxacin release, mechanical, and
mucoadhesive properties. Based on the results obtained
from the in vitro characterization techniques, a candi-
date formulation containing carbopol, methyl cellulose
in addition to ciprofloxacin was selected for microbio-
logical evaluation. The selected formulation offered
compromise between optimal ciprofloxacin release and
adhesiveness and rheological properties, and showed
a prolonged antimicrobial effect against gram-positive
and gram-negative organisms. The designed controlled
 Ophthalmic controlled release in situ gelling systems   151

a) 50
S.aureus
45

Diameter of inhibition zone (mm)

E.coli
P.aregonosa
40
35
30
25
20
15
10
5
0
0.5 1 1.5 2 2.5 3 3.5 4
Time (hours)

Figure  4.  Comparison of the diameters of the inhibition zones of E.

coli, S. aureus, and P. aeruginosa.

b) Acknowledgement

The authors would like to thank Miss Fadheela Al-Anzy

for her valuable help in the microbiological studies.

Declaration of interest: The authors report no conflicts

of interest. The authors alone are responsible for the
content and writing of the paper.

c)
Figure  3.  Evaluation of the antimicrobial efficiency of ciprofloxacin
gel on (a) E. coli, (b) S. aureus, and (c) P. aeruginosa as a function of
time.
release in situ gelling system is a viable alternative to
conventional eye drops as a result of its ability to enhance
bioavailability through its prolonged drug release.
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