Introduction

1. INTRODUCTION
Ocular diseases were widely noticed since the beginning of human race and animals.
Scientists doing research in pharmaceutical areas possess problems while formulating
ocular/opthalmic drug delivery systems (ODDS). Recently two main areas have been
discovered for attention namely:
(A) To identify and develop new more effective and safe drug molecules for ocular
infections and diseases.
(B) To modify existing ODDS for higher ocular bioavailability and sustained release
action of drug. [1-3].
Recent developments in ODDS include replacement of existing dosage form with new
ones which will offer good pharmacokinetic properties and targeted action in eye. [4,5].
Designing of new ophthalmic formulation involves hurdles like anatomy of eye,
physiology of eye and biochemistry of eye because the drug acts as a foreign material and
thus is not absorbed readily by the eye. Many efforts are taken in order to successfully
develop a new formulation but final result depends on drug’s intrinsic activity, drug
permeability through biological membrane and drug retention in eye for prolong period.
The reason behind slow development of ophthalmic formulations as compared to that of
oral, transdermal, nasal are the constraints offered by eye [6]. The formulator has
challenge to overcome the membrane barriers of eye without causing damage to tissue.
[7] Drug delivery is not asy since eye possesses efficient removal mechanisms in
precorneal region. In an ideal condition, pharmacodynamic and pharmacokinetic
parameter of drug defines delivery of drug. In case of drug delivery to eye, the absorption
of drug will occur it should remain in contact with eye for long time and must have good
permeation through corneal and non corneal ways however usually drug corneal
permeation is low. [8,9] The available dosage forms like solutions, suspensions,
ointments are conventional and inefficient. Still, eye drops are most prescribed dosage
form especially for local action of drug in eye. Repeated use of these drops (dose being
less) can cause injury (mechanical/ biochemical) leading to blepharitis or conjunctivitis.
The local and systemic toxicity can be significantly minimized by intervention with
newer ophthalmic delivery systems. Frequency of dosing being high for eye drop, can
cause high drug and preservative concentration at epithelial surface. [10] Eye diseases

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and disorders can cause uneasiness and worry state in patient and fear to loss of vision or
even facial distortion. Systemic or orally administered drug mostly do not show local
effect on eye. Hence it’s a necessity to administer the drug topically drop form for topical
action. The intention of pharamceutics is the achievement of a prerequisite drug extent in
the eye for the required length of time.
Improvement in dosing by mean of controlled and enhanced delivery, prolonged contact
time and targeting within the globe will go a long way towards achieving safe and
reliable ophthalmic delivery system. A number of drugs are available in conventional
dosage form such as eye drops, eye ointment, gels, etc. for the treatment of the eye but
from a biopharmaceutical standpoint, their use has faced objections over their
effectiveness as drug delivery systems. It is observed that bioavailability of drugs
administered by ODDS is only 1 to 10% of administered dose. This is due to to the fast
preclearance kinetics resulting from reflux tear formation and blinking, where half-life of
administered isotonic solutions is approximately 15 seconds in the humans. [11] Under
normal conditions the human tear volume is about 7 μl with 1 μl in the tear film and
about 3 μl in each marginal tear meniscus. [12] This film occupies the area of the cornea
and conjunctiva’s epithelial surface. The osmolality of the tear is mainly depended on the
number of dissolved ions and crystalloids [13]. Its pH is defined by various substances
solubilised in the aqueous layer of lachrymal secretions i.e.bicarbonate, carbon dioxide,
enzymes (lysozyme) and tear albumin. Lachrymal glands secrete fatty acids gives an acid
solution with a pH of about 6.6. Due to the frequent blinking and secretion of fresh tears,
the pH increases slowly during the waking hours of the day and varies from 6.9 to 7.5.
The tear fluid has a small buffer capacity. This is principally due to the carbon dioxide
and the carbonate solubilized in the tears [14].
1.1. Mechanisms of Ophthalmic Drug Absorption
The topical delivery into the cul-de-sac is most preferred route for ODDS. The three
types of mechanisms by which a drug is made available in the tear film is diffusion,
dissolution and erosion. [15] Absorption in the eye can be noncorneal or corneal (major).
The morphology of the cornea is complex and hence drug should possess unique biphasic
solubility. The cornea contains three major barriers for penetration that is epithelium,
stroma and endothelium. [16]

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Noncorneal (absorption route) is not so effective and includes passage through the sclera
and conjunctiva into the intraocular tissues. The noncorneal route is applicable for the
drugs which show poor corneal permeability. Studies with timolol maleate, gentamicin,
inulin and prostaglandins entered inside the anterior chamber by penetrating through the
conjunctiva and sclera. The drug (in tear film) is absorbed by cornea/conjunctiva; part of
it is drained by rapid tear turnover, adsorbed to protein (tear) and metabolized. [17] The
transport through channels in between the cell (transcellular transport) around the
epithelium (corneal) and stroma is the major route of absorption of topically administered
ophthalmic preparations. The diffusion dependens upon various factors i.e. surface area,
diffusitivity, concentration gradient and time for which concentration gradient is
maintained.
Following are the factors which are responsible for low drug bioavailability ODDS:
blinking process and high tear turnover) [18], systemic absorption, low dwelling time in
the cul-de-sac, low permeability through corneal epithelial membrane, drainage by
gravity along with tear fluid, increased frequency of instillation, nasolacrimal drainage,
enzymatic metabolism and the lack of controlled release [19-26]. The physical and
morphological obstacles cause small portion of the instilled drug (approx.1%) accessible
for absorption [27-28]. Thus it becomes necessary to increase dosing frequency to
achieve pharmacological action. This leads to local and systemic effects which include
stomach ulcers and imbalance in GI motility [29]. This disadvantage can be overcome by
administering the drug systemically but due to blood aqueous humor barrier and retinal
blood barrier leading to high loading. Various approaches to overcome limitations of
current ODDS like vesicular drug delivery (niosomes and liposomes), particulate drug
delivery (nanoparticles), viscosity enhancement (gels), use of mucoadhesive (bioadhesive
polymers), prodrugs, and other OCDDS like ocuserts, iontophoresis, ocular insert,
contact lenses etc. are being explored [20,30-31].
The two obstacles namely static and dynamic obstacles cause hurdles in the ODDS and
affect bioavailability of drug. Static barrier includes different eye segments like sclera,
cornea, retina and blood-retinal barriers. Dynamic barriers include lymphatic clearance,
tear dilution, conjunctival and choroidal blood flow. Most widely used dosage form for
topical activity in ODDS is eye drops. The percentages (relative) of conventional dosage

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forms that is solution (62.4%), suspensions (8.7%) and ointments (17.4%) covers the
whole market [32]. These conventional dosage forms capture nearly ninety percent of
current marketed formulations [1].
1.2. Physiological and Pharmacokinetic Limitations
Topical administration is favored over systemic to prevent systemic toxicity. It has added
advantage of fast action and dose reduction. Topical administration is used to treat
anterior structures’ disorders of eye but possess drawback of low bioavailability offered
by various biological factors (Fig. 1.1), which protects the eye and hence stop the drug
entry into eye.

Fig.1.1. Factors attributing to poor bioavailability of an ophthalmic formulation [2].

1.2.1. Pre-ocular retention
Blinking and lachrymation are protective mechanisms that keep away the foreign
particles from eye and also prevent effective ocular activity [22, 33-35]. When dosage is
instilled into eye, drop volume of 25–56 µl (average 39 µl) is delivered [35-37]. After
instillation in to the eye, the human cul-de-sac (conjunctival fornix) receives 30 µl of
volume approximately. The remaining instilled solution is removed by nasolachrymal
drainage or through spillage from the conjunctival sac until original volume of tear is
regained. (7–10 µl) [35, 36]. The tear formation rate is 16% /min during day hours [38].
Upon administration in the eye, the drug gets dissolved in the tear fluid and faces small
residence time (approximately 1– 2 min) due continuous lachrymation process (0.5–2.2
µl/min) [20]. The lacrimal fluid formation rate is less (1 µl/min) and the surplus volume
of the administered fluid is removed by nasolacrimal drainage.

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1.2.2. Corneal absorption

The other reason for limited ocular drug bioavailability is the systemic drug absorption
[39,40]. It occurs directly from local blood capillaries present in conjunctival sac or
through nasolachrymal drainage [39]. Once the drug enters into nasal cavity, the drug is
directed into the blood stream [40]. This cause drug loss and possible adverse effects
[40]. Out of the total drug administered, only 5% of drug gets absorbed through cornea
and remaining is absorbed through conjunctiva/ enters into lacrimal sac through upper
and lower canaliculi [41, 42]. Drugs permeation around the epithelium (corneal) occurs
by either transcellular or paracellular pathway [20, 23, 38].
1.2.3. Permeation through cornea:
The mode and permeation rate in cornea is affected by following physicochemical drug
properties like shape and molecular size, lipophilicity, solubility, degree of ionization and
charge [23]. The cornea is barrier containing a major hydrophilic barrier (corneal
epithelium), a main lipophilic hindrance (stroma) and a minor lipophilic barrier
(endothelium); therefore, lipophilic drugs pass the epithelium (corneal) through the cells
by simple diffusion or by carrier transport across the bilayer [43,44]. In contrast,
hydrophilic drugs face the difficulty to move from film (tear) to the epithelia
(corneal/conjunctival). Thus entry of drug by moving in between the cells across epithelia
(both corneal and noncorneal) becomes most preferred route of penetration for
hydrophilic drugs [44].
1.2.4. Drug distribution:
After instillation, when the drug is in cul de sac near to the corneal surface, it partitions
into the epithelium. Lipophilic compounds remains attached to the epithelia and slowly
diffuse to the stroma (corneal) and later to the anterior chamber [34]. Later from cornea
the drug slowly diffuses into the humor (aqueous) and further to the intraocular tissues
(lens, iris-ciliary body, vitreous and choroid-retina) [20]. The drug can easily move from
aqueous humor to the ciliary body and iris and can attach to melanin [33, 34]. Melanin-
bound drugs acts as a reservoir and releases the drug gradually in the surrounding and
thus prolong action is observed [34]. The drugs from the aqueous humor are further
supplied to the intraocular tissues and eliminated through aqueous-humor into the

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Sclemm’s canal and chamber angle [20,34]. Scleroconjunctival route is more preferred
and fast route for penetration for hydrophilic than across transcorneal route.
1.3. ODDS
A multitude of ophthalmic dosage forms are available for drug delivery to the eyes.
These can be classified according to their physical forms as follows [45]
1.3.1. Liquids
a. Solutions
b. Suspension
c. Sol to Gel system
d. sprays
1.3.2. Solids
a. Ocular inserts
b. Contact lenses
c. Corneal shields
d. Artificial tear inserts
e. Filter paper strips
1.3.2. Semi-solids
a. Ointments
b. Gels
1.3.3. Miscellaneous
a. Ocular iontophoresis
b. Vesicular systems (Niosomes, liposomes, etc.)
c. Mucoadhesive dosage forms
d. Particulates
e. Use of ocular penetration enhancers
f. Viscosity imparting agents
g. Target noncorneal route
Some of the above mentioned ODDS are available in the market. Nevertheless, about
90% preparations in the market are conventional eye drops. This is because of factors
such as expensiveness, difficulty in bulk manufacture, patient compliance, efficacy and
stability

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1.4. Ophthalmic Controlled Drug Delivery Systems (OCDDS)

Various OCDDS are available for effective treatment of eye disease as well disorders.
These include ocular inserts, sol to gel systems, corneal shield, soft contact lens, vesicular
systems like liposomes niosomes and others. There are various advantages of these
systems over the conventional dosage forms (e.g. eye drops such as solutions and
suspensions, eye ointment etc).
1.4.1 Criteria for OCDDS
1. The OCDDS should be comfortable.
2. It should not interfere with the vision and oxygen permeability.
3. It should have reproducible release rate.
4. It should be sterile, stable and easy to prepare.
5. It should demonstrate long residence and contact times with eyes
6. It should deliver a number of drugs to the eye.
7. The ocular inserts should be so designed that there would be no expulsion during
insertion.
8. The ocular in situ gel should be low of viscosity and free flowing liquid during
administration.
9. The ocular sol to gel transformation should form a gel upon administration in the eye
and gel formed must be enough to endure forces (shear) in the eye.
1.5. Anatomy and Physiology of the Eye [46, 47]
The eye is the sensory organ of the sight located in the orbital cavity. The nervous system
is connected to it by the optic nerve. The eye is actually two spheres, one set in the other,
the front sphere is small and is bordered anterior by the cornea, whereas the larger
posterior sphere is an opaque fibrous shell encased by sclera. The combined weight of
both spheres has been given as 6.77-7.5 g with a volume approximately 6.5 ml. The
circumference of eye is about 7.5 cm. To design effective drug delivery system, it is first
necessary to understand the relevant anatomical and physiological constraints that impede
or modify ocular drug and vehicle disposition. The cross section of eye are shown in
Figures 1.2

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Fig 1.2. Cross section of eye

1.5.1. Anterior section of the eye [47]
It comprises the cornea, iris, cilliary body, the chambers (anterior & posterior), and the
lens. In addition, the lachrymal apparatus and the eyelids are also included in the anterior
segment. The posterior section of the eye refers to the parts of the eye which are situated
behind the lens. The cornea is the anterior most transparent membrane of the eye
continuing posterior as the sclera. The various membrane, drug and formulation factors
that affect corneal permeability are listed (Table 1.1).
Table 1.1: Factors that influence corneal permeability
Parameters Affecting Factor
Membrane Factors Area available for absorption (cornea
and/or sclera)
Thickness
Porosity
Tortousity
Lipophilicity/hydrophilicity balance
Drug Factors Concentration instilled into the eye
Solubility
Partition coefficient
Molecular weight

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pKa
Tonicity
Adjuvant effect (e.g. Surfactant)
Release rate (If semisolid or insert)
Formulation Factors Retention time at absorption site
pH
Tonicity
Viscosity
Adjuvant effect (e.g. Surfactant)
Release rate (If semisolid or insert)
The inner surface of eyelid is covered by mucous membrane called conjunctiva and the
visible parts of the sclera. The surfaces (conjunctival and corneal) are covered by tear
film, fluid secreted by glands (conjunctival and lachrymal). A large number of fine ducts
supply the tears (secretion of lachrymal gland) into conjunctival fornix (Figure 1.1). The
movement of eye helps for tear spread over the conjunctival surface. Tears are drained
from the lachrymal lake by two small tubes, the lachrymal cancaliculi, which lead into
superior part of the nasolachrymal duct. Blinking action exerts force and help in
removing tears from lachrymal lake and direct the tears into the nasal cavity. Lacrimation
is caused reflexly by nerve ending stimulation of cornea and conjunctiva. The
composition of the human tears is depicted in Table 1.2
Table 1.2: Physical properties and chemical composition of human tears
Properties Composition Approximate value
Physical Osmotic pressure 0.9% NaCl (300 mOsm/l)
Properties pH 7.4 (7.3 to 7.7)
Refractive Index 1.357
Volume 0.50 to 0.67 g/16h (walking)
Electrolytes Bicarbonate 26 m Eq/l
Chloride 120 to 135 m Eq/l
Potassium 15 to 29 m Eq/l
Sodium 142 m Eq/l
Calcium 2.29 mg/100 ml

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Nitrogenous Total protein 0.669 to 0.800 g/100 ml

Substances Albumin 0.394 g/100 ml
Globulin
Ammonia
Urea
Total Nitrogen
Non protein nitrogen
(*m Eq/l = Milliequivalent per liter)
The eyelid consists four main layers such as skin, muscle, fibrous tissue layer or tarsus
and conjunctiva. The tear fluid has a nutritional function as well as an antibacterial
function. The tear fluid also lubricates the eye and washes away debris. The anterior uvea
comprises cilliary body and the iris, and between the lens, cilliary processes and the iris
there is a small cavity called the posterior chamber. Anterior chamber is a cavity present
in between the iris, chamber angle and cornea. The two chambers, posterior and anterior,
communicate through the pupil. The cilliary body comprises the cilliary muscles and the
cilliary processes. The iris is composed of the pigmented epithelial cell layer, the irradial
sphincter and dilator muscles, and the stroma. The meshwork (trabecular) is the structure
between the cornea, the sclera and the iris in the chamber angle. The crystalline lens is
located in ahead of vitreous and behind iris.
1.5.2. Posterior section of the eye [47]
It comprises of the retina with the optic nerve, the vitreous the choroids, and sclera. The
coat in the interior of the eye is retina. It is a transparent tissue that divides the two-third
posterior of the eyeball. The vitreous is a hydrogel which fills the cavity between the
retina and the lens. The highly vascularized tissue between retina and sclera is choroid.
1.6. Pathology of Eye [48]
The eye consists of a number of individual anatomic and functional elements. In keeping
with this, pathology of the eye covers the conjunctiva, cornea, uvea, lens, retina, vitreous
and optic nerve. Systemic diseases affect the above mentioned elements causing disorders
like glaucoma, cataract and macular degeneration which are called as blinding diseases.
There are some inflammatory disorders of eye, in which steroidal drug is use as primary

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drug of choice. Different acute and chronic allergic and inflammatory responses affecting
eye are as follows:
1.6.1 Allergic conjunctivitis:
Allergy cause conjunctival inflammation which is referred allergic conjunctivitis. Hay
fever is the source of allergic conjunctivitis. Oedema of conjunctiva, redness, scratching
and augmented lacrimation is the symptoms of allergic conjunctivitis. Allergic
rhinoconjunctivitis is combo of rhinitis and conjunctivitis. Histamine and other
substances are released by mast cells irritate nerve endings, cause blood vessels dilation
and increase tear formation. Pathophysiological condition mainly shows: The allergic
response in eye is a sequence of events that is handled by mast cells. Production and mast
cells activation is done by eotaxin and MIP which are types of chemokines. In presence
of particular allergen, sensitization takes place and signals the antigen specific response.
The antigen specific immunoglobulin E (IgE) production is promoted by cytokines
released from TH2 differentiated T cells. IgE receptors are situated on the mast cells’
surface to which IgE binds. Histamine is secreted by mast cells which further leads to the
release of prostaglandins, cytokines and platelet-activating factor. Histamine (secreted
from mast cells) combines with H1 receptors and cause itching. Receptors H1 and H2 on
conjunctival vasculature binded with histamine cause vasodilatation. Chemokine and
interleukin IL-8 are type of cytokines recruit neutrophils. IL-5 (TH2 cytokine) recruit
eosinophil and TH2 cytokine (IL-13, IL-6, and IL-4 ) promote increased sensitivity.
Molecular cascade lead to immediate symptoms. Advancement of acute inflammation
results to a state of chronic allergic inflammation.
1.6.2 Keratitis: Keratitis is a situation where cornea is inflamed. Moderate- intense pain
occurs and also involves impairment of eye. Itching occurs which may lead to scratching
of eye feeling on blinking action.
1.6.3 Herpes zoster ophthalmicus: Herpes Zoster Ophthalmicus (HZO) or shingles is a
viral disease which contains a painful skin rash. It occurs in dermatome distributions (1
or more) of the cranial nerve (5th) which the eye shares with orbit.
1.6.4 Iritis and iridocyclitis or Uveitis: Uveitis is, broadly, inflammation of the uvea. The
uvea comprises of pigmented, middle organelles of the eye like ciliary body,iris and

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choroid. Immediate treatment is required in uveitis along with proper diagnosis by

optometrist.
1.6.5 Chorioretinitis: Chorioretinitis involves inflamed choroid inflammation and retina.
Posterior uveitis can be called as chorioretinitis. Choroiditis is the condition where
choroid is inflamed. Floating black spots, red eyes and pain in the eye blurred vision,
sensitivity to light, lachrymation is the symptoms.
1.6.6 Optic neuritis:
Optic nerve inflammation is called as optic neuritis. The patient suffereing from it may
become half or completely blind.
1.6.7 Sympathetic ophthalmia: Sympathetic ophthalmia (SO) is a type of uveitis. It
affects both eyes followed by shock to one eye. Patient if not treated may become
completely blind. Sympathetic ophthalmia is an autoimmune disease against ocular
antigens. After a traumatic injury, the immune system is introduced to ocular antigens.
When exposed, these antigens are sensed as foreign and are attacked. The onset period
can vary from days to years.
1.7. Interest in New ODDS
Disadvantages of eye drops can be overcome by various ODDS such as microparticles
and nanoparticles, liposomes, collagen shields, etc. have been investigated. An optimum
ODDS can be claimed as one which can be administered in drop; no blurring of vision;
frequency of administration is 1-2 times per day Patient compliance is improved since
administration frequency is reduced with low toxicity and side effects.
Augmentation of dwelling time in cornea/ conjunctival sac and slow drug release
resulting into diminished drug loss were the two criteria for which formulation efforts
were taken. This was accomplished by using viscosity modifying agents which retard the
drainage of drug into systemic circulation through nasolachrymal drainage. Inserts
deliver controlled rate of administration accompanied with no blurred vision. However,
the disadvantage is difficulty in administration. Colloidal systems offer easy
administration and efficiency over long residences time [50]. Vesicular DDS like
liposomes and niosomes render enhanced movement across cornea and thus improves the
penetrability through ocular barriers. It also accomplishes the necessity for an ideal
dosage form that cause drop formation during administration for accurate dosing [51].

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But liposomes possess stability issues hence niosomes are developed. They offer
advantages like they are stable (chemically), low toxicity (non-ionic surfactant are safe).
[49]
Using niosomal suspension in the eye would not overcome the problem of drug loss. The
purpose behind development of several in situ gel was to raise the ocular bioavailability
by prolonging the precorneal residence time. Employment of one polymer system cause
formation of stiff gel upon administration for which higher amount of carbopol is
required. Increase in carbopol amount increases the acidic nature of formulation which
may irritate the eye. [52] Therefore total polymer content is reduced without altering the
gelation properties by using combination of polymers. Here in situ gel was synthesized
which can change its physical state in contact with physiological state. But the drug
diffusion from these systems was not found to be sustained release type. The two systems
were combined together to overcome the barriers associated with conventional system
and get advantage of both systems with controlled drug release. This was achieved by
incorporation of niosomes in hydrogel. [53] In the following paragraphs the niosomes
and hydrogels are discussed in more details.
Bioadhesive polymers (bind to the mucin surface) are beneficial in drug delivery for the
purposes of
(a) Retaining a drug in eye and
(b) Increasing drug contact time with the biological membrane.
Acrylic acid derivatives have bioadhesion qualities and employed with a sustained-
release delivery system for improved ODDS to eye. Acrylic acid derivatives show the
excellent binding to the ocular mucin. Due to that binding the shows increase in
absorption window.
Use of bioadhesive polymer is a remedy where polymer adheres to epithelium (corneal)
surface by ionic bonds or hydrogen bonds [54]. Literature survey [55, 56] highlighted
two points for bioadhesion regarding study of polymer’s binding quality to mucin
epithelial membrane. Firstly, the water insoluble polyanionic polymer showcase more
benefits over polycationic neutral drug delivery system. Secondly, Carboxylic group is a
key thing in bioadhesion [55]. Acrylic acid polymers are water insoluble but swell in
water which can hold large quantity of water [56]. Following properties should be

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exhibited by the bioadhesive polymer: a) safe and nontoxic b) should not enter into
systemic circulation by absorption c) bioadhesion by non covalent bond d) fast adhesion
e) easy drug incorporation without affecting its release f) economical.
1.8. Niosomes
Niosomes are formed by hydration of nonionic surfactants with/without cholesterol
incorporation. These systems are similar to liposomes, used as vectors for drugs (both
hydrophilic and lipophilic). Nonionic surfactant vesicles (niosomes) offer greater stability
and overcome the drawbacks associated with liposomes like storage condition, high cost
and degradation of phospholipids by oxidation.
It acts as medium for poorly absorbable drug. It improves the drug bioavailability by
permeating through biological membranes. Niosomes acts as target specific drug delivery
system in topical route of administration. The niosomes still requires to grow on larger
scale with better results.
1.8.1. Advantages of Niosomes
i. Hydrophilic, lipophilic and amphiphilic drug moieties can be incorporated in to
noisome. Hence its use is applicable for wide variety of drugs.
ii. Niosomes are not rigid in their structure and can be modified according to the need.
iii. It acts as a vehicle for drug delivery, safe and enhance the drug therapeutic value by
targeting its action to specific cells and prtects drug from degradation through different
enzymes.
iv. Niosomes can diffuse slowly the drug through depot and thus exhibit controlled
release.
v. They can augment the drugs bioavailability.
vi. They are stable and osmotically active.
vii. They offer improved stability of the entrapped drug.
viii. They enhance the drug penetration through skin.
ix. They act as targeted drug delivery where it can attain the action site by oral, topical
and parenteral routes.
x. The surfactants degrade organically, biocompatible, and non-immunogenic.
xi.The vesicle suspension contains water as an vehicle showcase patient comfort over
oily dosage forms.

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1.8.2. Disadvantages of niosomes

i. Vesicle aggregation and vesicle fusion
Niosomes are stable as compared to that of liposomes, but still sometimes they may
possess physical instability problems like fusion, aggregation, seepage of encapsulated
drugs, or entrapped drug hydrolysis which limits the durability of product. This occurs
due absence of charge between successive bilayer, small amount or total absence
cholesterol.
ii. Vesicle leaking
Due to low cholesterol amount, vesicle leaking may occur.
1.8.4. Types of niosomes
i. Small Unilamellar Vesicles (Size = 0.025-0.05μm)
ii. Multilamellar Vesicles (Size = > 0.05 μm)
iii. Large Unilamellar Vesicles (Size = > 0.10 μm).
1.8.5. Components of niosomal system
The major formulation components of niosomes are
i. Drug
Drug moieties (Hydrophilic/ lipophilic/ampiphilic) can be incorporated into niosomes.
ii. Cholesterol [57]
Cholesterol is present in cell membrane and is waxy steroid metabolite by nature. It is
added to nonionic surfactants for rigidity to bilayer vesicle. It cause gel to liquid
conversion of vesicular systems leading to less leaky niosomes. Cholesterol in high
amount cause aggregate formation in niosomes affecting the membrane stiffness and
resluting to random dispersement along the bilayer. Increase in vesicle size and
entrapment efficiency and decrease in release of entrapped substance is associated with
cholesterol addition.
iii. Non-ionic surfactant [58]
Nonionic surfactants are used widely surface active agent in preparing vesicles as they
impart higher benefits with respect to compatibility, stability and toxicity as compared to
other counterparts. Nonionic surfactants are less hemolytic, less toxic and cause less
irritation to cellular surface and helps in maintaining pH in solution. Surfactants perform
many functions as wetting agents, emulsifiers, solubilizers, and permeability enhancers.

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They act as P-glycoprotein inhibitors and thus improves drug absorption and used for
specific tissue targetting like anticancer drugs (daunorubicin), steroids (hydrocortisone)
and HIV protease inhibitor(ritonavir), cardiovascular drugs (digoxin) and beta-blockers
(acebutolol).
Nonionic surfactant contains polar and non polar segments with high interfacial activity.
Bilayer vesicles formation or micelle formation will depend on Surfactant HLB value,
structure of chemical components and CPP. CPP governs the vesicle type formed. CPP is
determined from hydrophobic group volume, head group area (hydrophilic) and
lipophilic alkyl chain length which also affect the entrapment efficiency. Higher the chain
length more is the entrapment eg stearyl (C18) chain surfactants exhibit more drug
entrapment than lauryl (C12) chain ones. Tween series of surfactants show highest
entrapment for hydrophilic drug since they havelong alkyl chain and large hydrophilic
head. The HLB value governs entrapment of drug in the vesicle. HLB value 14–17 do
not form vesicles; HLB value 8.6 forms ideal niosomes with the highest encapsulation.
HLB value decrease 8.6 to 1.7 cause reduction in entrapment eficiency. For HLB > 6,
cholesterol is required for bilayered vesicle formation as it improves the stability.
Cholesterol addition enables introduction of more hydrophobic moiety to form vesicles,
surfactant tendency to form aggregates is suppressed, and gel liquid transition
temperature promoted thus providing stability to the bilayer. Tc affect entrapment
efficiency eg span 60 with high Tc shows high entrapment efficiency..
iv. Solvents
Mostly volatile solvents are used. The choice of solvent relies upon drug, method of
preparation, etc.
E.g. Ether, chloroform, methanol, ethanol, etc.
v. Hydration medium
Phosphate buffer (different pH) is the mostly used for niosomes formation. Drug
solubility being incorporated decides the pH of medium.
1.9. Polymeric Hydrogels [59]
The ophthalmic semi-solid hydrogels prove their usefulness by increasing ocular
residence time through greater viscosity and mucoadhesive properties. Hydrogels are
hydrophilic, 3D network of polymer which can incorporate water or fluids in large

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 Introduction

amount. The networks containing homopolymers/ copolymer are not soluble either due to
the chemical crosslinks (tie-points, junctions), or due to the physical crosslinks
(entanglements /crystallites). Hydrogels (thermodynamically compatible with water)
swell in aqueous media. Hydrogels are constituted of water (high contents) and their
consistency is soft which is identical to that of natural tissues and hence they are
biocompatible. The most common approach for drug retention at corneal surface is use of
polymers. Hydrogels are polymers which have water swellable property or aqueous
solvents and cause a liquid-gel transformation. Polymeric gels are grouped into two
Preformed gels
In-situ forming gels
The gels help in the bioavailability improvement and reduce the side effects associated
with systemic absorption of ophthalmic drugs. Preformed hydrogels are highly viscous
solutions and they maintain their physical state even after instillation. In-situ forming gels
are liquid formulations (solutions, sols, or suspensions) which undergo gelation after
administration due to physiological changes in to the eye.
1.9.1. In-situ Gel forming Hydrogels [60]
The major disadvantage of preformed hydrogels is non accurate and non reproducible
administration of drugs accompanied with crust formation at eyelid, blurred vision and
tear formation after instillation. In situ gel is a novel discovery where advantages of
solution for accurate dosing are achieved and advantage of gel for prolonged residence
time is also attained. Polymer chains crosslinking cause gelation process. It can occur by
chemical cross linking (covalent bond formation) or physical cross linking (non covalent
bond formation). In-situ gel systems can be explained as liquids (low viscosity solutions)
that undergo phase transition when come in contact with the ocular cul-de-sac to form
gels (viscoelastic) due to change in arrangement of polymers with respect to
environment. Before strong gel formation, in between drug administration, formation of
weak gels occurs due to fluid mechanism in eye.
Most successful strategies for prolonged ocular drug release is the viscosity rise of
administered solutions using gels. Polymers from natural, synthetic or semisynthetic
source are being used for gel formation over last decade. However, large increase in
viscosity may cause lured vision, discomfort or foreign body sensation. Thus, optimal

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 Introduction

viscosity range with suitable rheological behavior is required for better efficacy and
tolerance to the preparation. In fact, first 20-50 s after drug administration the retention of
product occurs with slight increase in viscosity ( <100mPas). Before gelling, the viscosity
is 5-1000 mPas which rise to 50-50,000 mPas after gelling. Rise in reflex action cause
significant rise in viscosity of formulations leading to elimination of administered drug
[22, 68-70]. After administration of ophthalmic products, precorneal tear film’s
pseudoplastic character is not disturbed. 0.03 s-1 is the shear rate of eye during inter
blinking periods and 4250-28,500 is during blinking. Thus, viscoelastic fluids also called
as pseudoplastic fluids, defined as fluids which possess high viscosity under low shear
rates and vice versa are often preferred.
1.9.2. Mechanisms of in-situ hydrogels
There are different mechanisms by which sol to gel phase transition takes place by
altering environmental conditions. The physical stimuli that give various responses
include temperature change, light, pressure, electric fields, sound and magnetic fields.
The chemical stimuli include pH change and change in ion activation; biochemical or
biological stimuli include glucose level change. Widely used methods among these are
pH change, temperature change and electrolyte composition change for ODDS.
1.9.1.1 Temperature induced gelation
They are liquid at 20 to 25ºC (RT) and undergo gelation at 35 to 37ºC (temperature of
body fluids) due to temperature rise. Examples of different temperature dependent gels
include poloxamer, cellulose derivatives and xyloglucan.
1.9.1.2 pH induced gelation
Gel formation of the solution occurs by pH change. At pH 7.4 the free flowing
formulation (at pH 4.4) undergoes coagulation due to tear fluid to form gel. The pH
changes by about 2.8 units after administration cause spontaneous transformation of the
fluid solution into viscous gel.
1.9.1.3. Osmotically induced gelation
In this method, gelation of the administered solution occurs by ionic strength change.
1.10. Niosomal In Situ Gel (NIG):
Vesicular DDS includes niosomes as one way through which localized drug action can be
achieved as the size and penetrability through epithelium tissue is low and gives localized

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 Introduction

drug action at the administration site. It results in enhancement of activity and also
reduces its systemic effects. Vesicular DDS causes prolonged and controlled drug effect
at the surface (corneal) and prevents drug loss by drug metabolism initiated by enzymes
present in lachrymal secretion and epithelial (corneal) surface. However, disadvantage of
drug wastage through nasolachrymal drainage and precorneal dilution by tear fluid is
associated with niosomes. Hence there is necessity for incorporation of these niosomes
into in situ gel [71]. These systems are in liquid form at administration time, and get
converted into gel phase once it is inserted into cul de sac of eye. The in situ gel do not
offer sustained drug release. Hence the drug release from these systems was not found to
be sustained release type. So, to overcome the individual system’s disadvantages both the
systems were combined together to overcomethe barriers related to conventional ODDS.
1.11. Prednisolone sodium phosphate (PSP)
It is sodium phosphate salt of prednisolone (synthetic glucocorticoid) with anti-
inflammatory properties and immunodulating activities. PSP binds to specific
glucocorticoid receptors intracellularly. The ligand- receptor complex is moves inside the
nucleus and initiates the transcription process of glucocorticoid-responsive genes like
cytokines and lipocortins. Lipocortins inhibit phospholipase A2, which block the
arachidonic acid release from phospholipids (membrane) and inhibit the synthesis of
leukotrienes and prostaglandins. PSP is used to treat eye inflammation or injury.
Prednisolone acts by decreasing symptoms like redness, swelling and itching,
• Scratch Wound on Cornea
• Uvea inflammation
• Corneal ulcers of the eye
• Damage to Cornea of Eye and lesions
• Allergic Conjunctivitis
• Herpes zoster causing infection to cornea
• Blood Vessel dilation of the Eye
Amongst the various available anti bacterials, PSP is still the most popular drug for the
treatment of various bacterial induced infections, inflammations, conjunctivitis,
blepharitis, iritis, and corneal ulcer. [57]. Frequent use of the solutions in concentrated

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 Introduction

form may cause damage to the ocular surface. However in case of normal eye drop
solutions, bioavailability after ocular eye drop administration is less than 5%.
Niosomes cause increase in the penetration (corneal) and in turn bioavailability which is
a limitation for conventional ophthalmic preparations. Ocular niosomes provide sustained
and controlled at ocular surface and protect the drug from metabolizing enzymes present
in tear fluid or at corneal surface. Drug entrapped in the surfactant vesicles cause
movement of drug by partitioning and transport mechanism through cornea [72].
Therefore, in order to remove the hurdles related with conventional ocular therapy like
drug loss, short residence time, permeation barrier due to structure of corneal epithelium,
increased frequency of administration, it was necessary to find an alternative dosage from
for controlled release and minimize the drug loss at the site that is cornea.
1.12. Terbinafine Hydrochloride (THCl)
Terbinafine, brand name Lamisil is an antifungal drug used to treat fungal skin and nail
infections. It is available in oral dosage form or topical dosage form. Mode of action of
terbinafine is it prevents ergosterol synthesis by inhibiting ezyme squalene epoxidase,
which converts squalene to lanosterol. In fungi, lanosterol is the source of ergosterol
whereas in humans lanosterol is converted to cholesterol. There is large genetic
difference in human and fungal squalene epoxidase. Hence on fungal infection,
specifically terbinafine obstructs fungal squalene epoxidase without affecting human
cholesterol production. This may cause change in cell membrane permeability leading to
fungal cell lysis. Fungal keratitis (keratomycosis) is a serious corneal fungal infection
usually caused by Curvularia, Aspergillus, Candida, and Fusarium in tropical climates
and wearing contact lenses in developed countries. THCl belongs to allylamines class and
inhibits ergosterol synthesis causing lysis of fungal cell membrane. Topical THCl
(0.25%, w/v) drops are effective against filamentous fungal keratitis for treatment.
However, the treatment period with THCl was longer. The dosing of THCl drops is once
per 2h at night time and once per hour in day time [73].

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