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International Journal of Pharmaceutical and Clinical Research 2013; 5(2): 64-71
ABSTRACT
Ocular drug delivery is one of the most interesting and challenging endeavors facing the pharmaceutical scientist for past
10‐20 years. As an isolated organ, eye is very difficult to study from a drug delivery point of view. Despite these limitations,
improvements have been made with the objective of maintaining the drug for an extended period. Recently, controlled and
sustained drug delivery has become the standard in modern Pharmaceutical design and an intensive research have been
undertaken in achieving much better drug product effectiveness, reliability and safety. The formation of ocular in-situ gels
depends on factors like temperature modulation, pH change, presence of ions and ultra violet irradiation, from which the
drug gets released in a sustained and controlled manner. Various biodegradable polymers that are used for the formulation
of in situ gels include gellan gum, alginic acid, xyloglucan, pectin, chitosan, poly(DLlactic acid), poly(DL-lactide-co-
glycolide) and poly-caprolactone. The in situ gel forming polymeric formulations offer several advantages like sustained
and prolonged action in comparison to conventional drug delivery systems. From a manufacturing point of view, the
production of such devices is less complex and thus lowers the investment and manufacturing cost.
Keywords: In-Situ, Hydrogels, biodegradable polymers, controlled release, Novel ocular drug delivery system.
and strategies for this drug delivery including at pH 4 form hydrogel at neutral pH condition14.
physiological factors, physiochemical factors and In Situ Formation Based on Physical Mechanism
Swelling: In situ formation may also occur when material Fluoroquinolones. The fluoroquinolones represent an
absorbs water from surrounding environment and expand expanding class of broad-spectrum antibacterials which
to occur desired space15. One such substance is myverol cover a host of Gram-negative and anaerobic species
18-99 (glycerol mono-oleate), which is polar lipid that responsible for ocular infections. These antibacterials have
swells in water to form lyotropic liquid crystalline phase gained popularity in the ophthalmology field since they
structures.It has some Bioadhesive properties and can be have been shown to be equivalent to combination therapy
degraded invivo by enzymatic action16. in the treatment of many ocular infections.
Diffusion: This method involves the diffusion of solvent Fluoroquinolones are also effective against a variety of
from polymer solution into surrounding tissue and results Gram-positive organisms, including Streptococcal and
in precipitation or solidification of polymer matrix. N- Staphylococcal species; however, resistance is emerging
methyl pyrrolidone (NMP) has been shown to be useful among some of these organisms. 19-21The classification
solvent for such system17. (Table.2) and mechanism of action of fluoroquinolones are
In Situ Formation Based on Chemical Reactions: given below.
Chemical reactions that results in situ gelation may involve Classifications of ocular polymeric systems:
precipitation of inorganic solids from supersaturated ionic Pectin: Pectins are a family of polysaccharides, in which
solutions, enzymatic processes, and photo-initiated the polymer backbone mainly comprises α-(1-4)-
processes. Dgalacturonic acid residues. Low methoxypectins (degree
Ionic Crosslinking: Polymers may undergo phase of esterification <50%) readily form gels in aqueous
transition in presence of various ions. Some of the solution in the presence of free calcium ions, which
polysaccharides fall into the class of ion-sensitive ones18. crosslink the galacturonic acid chains in a manner
While k-carrageenan forms rigid, brittle gels in reply of described by egg-box model . Although the gelation of
small amount of K+, i-carrageenan forms elastic gels pectin will occur in the presence of H + ions, a source of
mainly in the presence of Ca2+. Gellan gum commercially divalent ions, generally calcium ions is required to produce
available as Gelrite is an anionic polysaccharide that the gels that are suitable as vehicles for drug delivery22.
undergoes in situ gelling in the presence of mono- and Xyloglucan: Xyloglucan is a polysaccharide derived from
divalent cations, including Ca2+, Mg2+, K+ and Na+. tamarind seeds and is composed of a (1-4)-β-D-glucan
Enzymatic Cross-Linking: In situ formation catalysed by backbone chain, which has (1-6)-α-D xylose branches that
natural enzymes has not been investigated widely but are partially substituted by (1-2)-β-D-galactoxylose23.
seems to have some advantages over chemical and When xyloglucan is partially degraded by β- galactosidase,
photochemical approaches. For example, an enzymatic the resultant product exhibits thermally reversible gelation
process operates efficiently under physiologic conditions by the lateral stacking of the rod like chains . The sol-gel
without need for potentially harmful chemicals such as transition temperature varies with the degree of galactose
monomers and initiators. elimination. It forms thermally reversible gels on warming
Drugs Used Under Ocular Preparations: Common topical to body temperature.
antibacterials used in the treatment of ocular infectious Gellangum: Gellan gum (commercially available as
diseases include sulfonamides, aminoglycosides, Gelrite TM or Kelcogel TM ) is an anionic deacetylated
polymyxin-based combinations, exocellular polysaccharide secreted by Pseudomonas
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Fig.2 Various problems encountered in poor bioavailability of the eye installed drugs
polysaccharide consisting of β-D-mannuronic acid and α- Xanthum gum: Xanthan gum is a high molecular weight
L-glucuronic acid residues joined by 1,4-glycosidic extra cellular polysaccharide produced by the fermentation
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linkages . The proportion of each block and the of the gram-negative bacterium Xanthomonas campestris.
arrangement of blocks along the molecule vary depending The primary structure of this naturally produced cellulose
BASF Corporation) are the series of commercially crosslinking of individual polymer chains. They resemble
available difunctional triblock copolymers of non-ionic natural living tissue more than any other class of synthetic
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nature. They comprise of a central block of relatively biomaterials due to their high water content; furthermore,
hydrophobic polypropylene oxide surrounded on both
the high water content of the materials contributes to their physiological temperature, was determined by the time
biocompatibility10. (seconds), the apparatus took to sink 5cm down through
B. Smart hydrogels: Smart hydrogels, or stimuli-sensitive the prepared gel33.
hydrogels, are very different from inert hydrogels in that Gelling capacity: The gelling capacity of the prepared
they can sense changes in environmental properties such formulation is determined by placing a drop of the
as pH and temperature and respond by increasing or formulation in a vial containing 2.0 ml of freshly prepared
decreasing their degree of swelling. The volume-changing simulated tear fluid and visually observed. The time taken
behavior of smart hydrogels is particularly useful in drug for its gelling is noted34,35.
delivery applications as drug release can be triggered upon Rheological studies: The viscosity measurements can be
environmental changes. calculated using Brookfield viscometer, Cone and Plate
Evaluations of ocular drug delivery system viscometer. The insitu gel formulations were placed in the
The prepared ocular formulations were evaluated for sampler tube. From the literature it was evident that, the
clarity, pH measurement, gelling capacity, drug content, formulation before gelling should have a viscosity of 5 to
rheological study, in vitro diffusion study, isotonicity, 1000 mPas. And after ion gel activation by the eye, will
antibacterial activity, in vivo ocular testing in rabbits and have a viscosity of from about 50‐ 50,000 mPas10, 13. The
accelerated stability studies. The pH of insitu gel solution samples are analyzed both at room temperature at 25°C
was found to be 7.4 for all the formulations. The and thermostated at 37°C ± 0.5°C by a circulating bath
formulation should have an optimum viscosity that will connected to the viscometer adaptor prior to each
allow for easy instillation into the eye as a liquid (drops), measurement. The angular velocity of the spindle was
which would undergo a rapid sol‐to‐gel transition increased 20, 30, 50, 60, 100, 200 and the viscosity of the
(triggered by pH, temperature or ion exchange). Marketed formulation is measured. All the formulations exhibited
products of ocular insitu formulations are also Newtonian and pseudoplastic flow characteristics before
included(table.3). and after gelling in the simulated tear fluid
Physical Parameters: The formulated insitu gel solution is respectively36,37.
tested for clarity, pH, gelling capacity, and drug content In vitro drug release studies: In vitro release study of insitu
estimation. gel solution was carried out by using Franz diffusion cell.
Appearance and Determination of Ph: The appearance of The formulation placed in donor compartment and freshly
the formulation was observed which included clarity, color prepared simulated tear fluid in receptor compartment.
of solution visually and pH was measured using pH meter. Between donor and receptor compartment dialysis
Drug Content: The drug content was determined by taking membrane is placed (0.22μm pore size). The whole
1ml of the formulation and diluting it to 100 ml with assembly was placed on the thermostatically controlled
distilled water. Aliquot of 5 ml was withdrawn and further magnetic stirrer. The temperature of the medium was
diluted to 25 ml with distilled water. Drug concentration maintained at 37°C ± 0.5°C. 1ml of sample is withdrawn
was determined at by using UV is spectrophotometer form at predetermined time interval of 1hr for 6 hrs and same
the standard graph of r2=0.99. volume of fresh medium is replaced. The withdrawn
Spreadability: For the determination of Spreadability, samples are diluted to 10ml in a volumetric flask with
excess of sample was applied in between two glass slides respective solvent and analyzed by UV spectrophotometer
and was compressed to uniform thickness by placing at respective nm using reagent blank. The drug content
1000g weight for 5 min. weight (50 g) was added to the calculated using the equation generated from standard
pan. The time in which the upper glass slide moves over to calibration curve. The % cumulative drug release (%CDR)
the lower plate was taken as measure of Spreadability. calculated. The data obtained is further subjected to curve
S= ML/T fitting for drug release data. The best fit model is checked
Where, M = weight tide to upper slide, L = length moved for Krosmeyers Peppas and Fickinian diffusion
on the glass slide, T = time taken. mechanism for their kinetics34.
Sterility test: All ophthalmic preparations should be sterile, Texture analysis: The consistency, firmness and
therefore the test for sterility is very important evaluation cohesiveness of insitu gel are assessed by using texture
parameter. 2 ml of the formulation from test container was profile analyzer which mainly indicated gel strength and
removed with a sterile pipette or with a sterile syringe or a easiness in administration in vivo. Higher values of
needle. The test formulation was aseptically transferred to adhesiveness of gels are needed to maintain an intimate
fluid thioglycolate medium (20 ml) and soya bean-casein contact with mucus surface38.
digest medium (20 ml) separately. The Isotonicity evaluation: Isotonicity is important
inoculated media were incubated for not less than 14 days characteristic of the ophthalmic preparations. Isotonicity
at 30°C to 35°C in the case of fluid thioglycolate medium has to be maintained to prevent tissue damage or irritation
and 20°C to 25°C in the case of soya bean-casein digest of eye. All ophthalmic preparations are subjected to
medium. isotonicity testing, since they exhibited good release
Measurement of Gel Strength: A sample of 50 gm of gel characteristics and gelling capacity and the requisite
was placed in a 100 ml graduated cylinder and gelled in a viscosity. Formulations are mixed with few drops of blood
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thermostat at 370c. The apparatus for measuring gel and observed under microscope at 45X magnification and
strength was allowed to penetrate in ocular gel. The gels compared with standard marketed ophthalmic
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