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VESICULAR DRUG DELIVERY SYSTEM: A NOVEL APPROACH

Article · September 2014

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 VESICULAR DRUG DELIVERY SYSTEM: A NOVEL APPROACH
*1 1 1 1 1
KALPESH CHHOTALAL ASHARA , JALPA S. PAUN , M.M SONIWALA , J.R.CHAVDA , S. V. NATHAWANI ,
2,3 2
NITIN M. MORI AND VISHAL P. MENDAPARA
1Department of Pharmaceutics, B. K. Mody Govt. Pharmacy College Rajkot-360003, Gujarat, India, 2 Department of Pharmaceutical Sciences, Saurashtra

University, Rajkot-360005, Gujarat, India, 3 Torrel (Hospital Division) a member of Torrent group, Ahmedabad, Gujarat, India, E mail-kalpeshshr5@gmail.com
Received 19-06-14; Reviewed and accepted 03-07-14

ABSTRACT
A novel drug delivery system is that delivers drug at predetermined rate decided as per the requirement, pharmacological aspects, drug profile, physiological conditions of
body etc. In current conditions not a single novel drug delivery system behaves ideally those high goals with fewer side effects. A Vesicular drug delivery system (VDDS) is
the system in which encapsulation of active moieties in vesicular structure, which bridges gap between ideal and available of novel drug delivery system.Varrious types of
vesicular drug delivery system like liposome, niosome, archeosome, transferosome etc. were developed. Advances have since been made in vesicular drug delivery system.
Focus of this review is to bring about a brief of vesicular drug delivery system as novel approach.
Keywords: Vesicular Drug Delivery System, VDDS, Liposome, Niosome.

INTRODUCTION
Discussion never ends that from very ancient era discussion is
already going on newer & better alternatives & in case of drugs it
will continue till we got a drug with maximum efficacy & no side
effects. Many drugs have narrow therapeutic window so their
clinical uses are limited. Thus therapeutic effectiveness of existing
drugs is improved by formulating them in advantageous ways.[1]In
previous years, noticable work had been done to develop Novel
Drug Delivery System (NDDS), fulfills desirable characteristics
that it should deliver drug at a rate directed by need of body, over
period of treatment & should channel active entity at site of
action.Convetional dosage forms including prolonged released
dosage forms unable to fulfilled none of these desired
characteristics. At present, no available drug delivery systems
behave ideally but attempts have been made to bridge gap
between ideal & available.[2]
Definition
“Vesicles have become the vehicle of choice in drug delivery
system called Vesicular Drug Delivery System.”, e.g. liposomes,
Niosomes, Pharmacosomes etc.[1]
Advantages
Vesicular drug delivery systems have several advantages over the
conventional dosages forms as well as prolonged released
dosage forms as[1, 3]:
 Effective permeation of drugs into cells
 Prolongation of existence of drugs in systemic circulation.
 As selective uptake is taken place so reduces toxicity.
 Reduces the cost of therapy.
 Improves bioavailability.
 Hydrophilic-Lipophilic drugs can be incorporated.
 Sustained-release system function.
 Delayed elimination of rapidly metabolized drugs.
 Overcomes the problems of the drug insolubility, instability,
and rapid degradations.
Why VDDS
Conventional chemotherapy for treatment of intracellular
infections is not effective due to limited permeation of drugs into
cells. To improve bioavailability at the site of diseases, reduces
harmful side effects of conventional & controlled release drug
delivery systems, overcome problem of degradation of drug &/or
drug lose.[4]
Disadvantages
Along with numbers of advantages VDDS has some serious
disadvantages which restrict their use. Drugs passively , which
may lead to low drug loading efficiency and drug leakage in
preparation, preservation and transport in vivo. Need of intensive
sonication, lead to leakages of drug during storage. Thus the
major problem of their stability acts as a barrier and thus limiting
their use.[5, 6]
TYPES OF VDDS
There are various types of VDDS as
LIPOSOMES
Name Liposome is derived from Greek words:’Lipos’ meaning fat
& “soma’ meaning body. Liposomes are artificially prepared
vesicles made of lipid bilayer. Liposomes can be filled with drugs,
and used to deliver drugs for cancer and other diseases. Amongst
various carriers, few drug carriers reached stages of clinical trials
where phospholipid vesicles (liposome) show strong potential for
effective drug delivery to site of action. Liposomes can be
composed of naturally-derived phospholipids with mixed lipid
chains or other surfactants. Liposome has covered predominantly
medical, albeit some non-medical areas like bioreactor, catalysts,
cosmetics and ecology. However, their predominance in drug
delivery and targeting has enabled them to be used as
therapeutics tool in fields like tumor targeting, gene and antisense
therapy etc.
Liposomes were first described by British hematologist Dr Alec D
Bangham in 1961(Published 1964), at Babraham Institute, in
Cambridge. They were discovered when Bangham and R. W.
Horne were testing institute's new electron microscope by adding
negative stain to dry phospholipids[2, 7]
“Liposome is simple microscopic vesicles in which an aqueous
volume is entirely enclosed by a membrane composed of lipid
molecule.” Various amphipathic molecules have been used to
form liposome; drug molecules can either be encapsulated in
aqueous space or intercalated into lipid bilayer.
A liposome is a spherical vesicle with a membrane composed of a
phospholipid bilayer used to deliver drug or genetic material into a
cell, can be composed of naturally-derived phospholipids with
mixed lipid chain like egg phosphatidylethonalimine or of pure
components like DOPE (dioleolylphosphatidylethanolamine)[8]

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VII) Stable Plurilamella air Vesicles Method: SPLV

Based on In-Vivo applications
I. Conventional Liposomes
These can be defined as liposomes that are typically composed of
only phospholipids (neutral and/or negatively charged) and/or
cholesterol. Most early work on liposomes as a drug-carrier
system employed this type of liposomes. Conventional liposomes
are a family of vesicular structures based on lipid bilayers
surrounding aqueous compartments. Conventional liposomes are
characterized by a relatively short blood circulation time due to
rapid uptake by MPS system. They are useful for macrophage
Fig.1: Liposome targeting, as local depot and for vaccination purpose. The fast and
efficient elimination from the circulation by liver and spleen
macrophages has seriously compromised application for the
treatment of the wide range of diseases involving other tissues.
II. Long circulatory Liposomes
The advent of new formulations of liposomes that can persist for
prolonged periods of time in the bloodstream led to a revival of
interest in liposomal delivery systems at the end of the 1980s. In
fact, the long-circulating liposomes opened a realm of new
therapeutic opportunities that were up to then unrealistic because
of efficient MPS uptake of conventional liposomes. Perhaps the
most important key feature of long circulating liposomes is that
they are able to extravagate at body sites where the permeability
Fig.2: Ambiosome
of the vascular wall is increased.
Advantages
It provides selective passive targeting to tumor tissue (liposomal
doxorubicin), increased efficacy and therapeutic index of drug
(Actinomycin-D) and stability via encapsulation, biocompatible,
completely biodegradable, non-toxic, flexible and no immunogenic
for systemic and non-systemic administrations, reduce toxicity of
the encapsulated agent (Amphotericin B, Taxol) and exposure of
sensitive tissues to toxic drugs, site avoidance effect, Flexibility to
couple with site-specific ligands to achieve active targeting.
Disadvantages
High production cost, leakage and fusion of encapsulated drug /
molecules; phospholipid undergoes oxidation and hydrolysis like
reaction, Short half-life, and Low solubility.
CLASSIFICATION Fortunately, regions of increased capillary permeability include
pathological areas such as solid tumors and sites of infection and
Liposome may be produced by variety of methods. Their inflammation. It is illustrative for the importance of the long-
nomenclature also depends upon the method of preparation, circulation concept that the only two liposomal anticancer products
structural parameters or special functions assigned to them (table
that are approved for human use are based on the use of long-
1)1.Classification according to size: circulating liposomes for tumor-selective delivery of antitumor
Table 1: Classification of Liposomes according to size drugs (Doxil, DaunoXome). At present the most popular way to
produce long-circulating liposomes is to attach hydrophilic
Type Specifications polymer polyethylene glycol (PEG) covalently to the outer surface.
MLV Multilamellar large vesicles- >0.5μm
OLV Oligolamellar vesicles- 0.1-1μm III. Immunoliposomes
UV Unilamellar vesicles (all size range) Immunoliposomes have specific antibodies or antibody fragments
SUV Small Unilamellar vesicles- 20-100nm (like Fab9 or single chain-antibodies) on their surface to enhance
MUV Medium sized Unilamellar vesicles target site binding. They are useful for site specific targeting.
LUV Large Unilamellar vesicles- >100
GUV Giant Unilamellar vesicles- >1μm
MV Multivesicular vesicles- 1μm
Classifications according to method of preparations
I) Extraction method: VET (Vesicles prepared by Extraction
Technique)
II) French Pressure Cell method
III) Fusion method
IV) Reverse Phase Evaporation method: SUVs, MLVs & OLVs are
made by reverse phase evaporation (REV) Method
V) Frozen & Thawed Multilayered Vesicles:
VI) Dehydration & Rehydration method: DRV

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Ashara et. al.
A. Temperature-sensitive Immunoliposomes
The heat induced drug release concept is based on the large
increase in the permeability of liposomal bilayers around their
phase transition temperature. Local heating of tumor tissue up to
this phase transition temperature will enhance drug release from
liposomes present in the heated area. Both the degree of
extravasation and the rate of drug release increases in this case.
Example of polymer that is used for preparation of temperature
sensitive immunoliposomes is N-isopropyl acrylamide.
B. pH-Sensitive Immunoliposomes
pH sensitive ImmunoLiposomes targeted to internalizing
receptors will end up in endosomes, where acidification will
trigger liposome destabilization and possible fusion with
endosomal membrane. They have been successfully applied in
vitro for the delivery of antitumor drugs into cytoplasm of tumor
cells. Example of polymer that is used for preparation of
temperature sensitive immunoliposomes is propylacrylic acid.
IV. Cationic Liposomes
These delivery systems are under development for improving the
delivery of genetic material. Their cationic lipid components
interact with, and neutralize, the negatively-charged DNA, thereby
condensing the DNA into a more compact structure. The resulting
lipid–DNA complexes, rather than DNA encapsulated within
liposomes, provide protection and promote cellular internalization
and expression of the condensed Plasmid.
V. Fusogenic Liposomes
Here reconstituted Sandal Virus is enveloped.
General steps involved in the preparations of Liposomes
various general steps that are evolved in the preparations of
liposomes are:
1. Preparation of Lipids for Hydration
2. Hydration of lipid film/cake
3. Sizing of lipid suspension
i. Sonication
ii. Extraction
Methods of Liposomes Preparations[9]
All the method of preparing liposomes involves four basic stages:
1. Drying down lipids from organic solvent.
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2. Dispersion of lipid in aqueous media.
3. Purification of resultant liposome.
4. Analysis of final product.
PASSIVE LOADING TECHNIQUE
Loading of entrapped agents before or during the manufacturing
process. Used for drugs which are aqueous soluble but lipid
insoluble. It is further divided into
1. Mechanical dispersion Method
Lipid hydration Method
Fig.3: Multilamelar Vesicles (MLVs) formed either Hand
shaking technique or Rotatory flash Evaporator
In this method, lipid mixtures are dissolved in solvent mixture of
chloroform: methanol (2:1) in rotary evaporator flask and dried thin
film of lipid is made using rotary evaporator under reduced
pressure (60 rpm, 30ºC, and about 15 min). Flask is flushed with
nitrogen and Hydration of lipid is done by adding 5ml of saline
phosphate buffer containing drug/solute to be encapsulated and
again use of rotary evaporator for making homogeneous milky
white suspension. It is allowed to stand for 2 hr at RT/above Tc for
complete swelling process. This will give MLVs.
Sonication
At high energy level, preformed MLVs are sonicated using either
probe or bath ultrasonic disintegrator.
Fig.4: Preparations of Small Unilemilar Vesicles (SUVs) by
Bath/Probe Sonication Process from MLVs
Using Probe: Used for suspensions which require high energy in
a small volume. And contamination of preparation with metal can
lead to degradation of lipid.
Using Bath: Used for large volume of dilute lipids where may not
necessary to reach the vesicle size limit. Finally, they are purified
into the SUVs by ultracentrifugation and collected from supernant
of centrifuge tube. Size of liposome is influenced by temperature,
composition, and concentration, sonication time & power, volume
of product.
Microfludization/Micro-emulsification Method
In this method, Micro fluidizer pumps the fluid at very high
pressure through a 5μm screen. Then, it is forced along defined
micro channels which direct two streams of fluid to collide together
at right angles at a very high velocity, thereby affecting a very
efficient transfer of energy. The lipid can be introduced into the
fluidizer, either as a suspension of large MLVs, or as slurry of
anhydrate lipid in an organic medium. The fluid collected can be
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Ashara et. al.
recycled through the pump and interaction chamber until vesicles
of the spherical dimension are obtained.
Fig.5: Representation of use of micro-fluidizer to prepare
Small Unilamellar Vesicles (SUVs) from MLVs
Advantages
Excellent size reduction up to 0.2mm, High rate of production, for
encapsulation of water soluble materials due to high proportion of
lipid.
French Pressure Cell
In this method, liquid sample of preformed MLVs are introduced
into the sample cavity, then the position of piston and pressure is
set up to fill sample up to the outlet hole. Then power is switched
on. At high pressure (2000 psi) and at 40ºC, MLVs are extruded
through small orifice, which is collected in suitable container. This
technique yields uni- or oligo lamellar liposome of intermediate
size. More stable than they obtained by sonication method and
also leakage of the content from the liposomes are lesser.
Fig. 6: French pressure cell & parts used for pre-parations of
Uni or oligo Lamellar Vesicles
Drawback
High cost of the pressure cell.
Membrane Extraction Method
Size of prepared liposomes is reduced by gently passing them
Fig.7: Liposomes Preparations using extractions techniques
based on polycarbonate filters
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Through membrane filter of defined pore size and this can be
achieved at much lower pressure.
In this process, the vesicles content are extruded with the
dispersion medium during breaking and resealing of phospholipids
as they pass through the polycarbonate membrane in order to
achieve high entrapment. The liposomes produced by this method
have been termed as LUVETs and 30% encapsulation can be
obtained using high lipid concentration.
Dried Reconstituted Vesicles
Fig.8: Preparations of Dried-Reconstituted Vesicles (DRVs)
membrane r restructures enclosing a proportion of solutes,
which was originally present in Extra-Liposomal Medium
It starts with freeze drying of a dispersion of empty SUVs and
rehydrating it with the aqueous fluid containing the materials to be
entrapped. This leads to dispersion of solid lipids in finely
subdivided form. Freeze drying is used to freeze and lyophilize the
preformed SUVs dispersion rather than to dry the lipids from an
organic solution. This leads to organized membrane structure
which on addition of water can rehydrate, fuse and reseal to form
vesicle with high capture capacity. It is used for manufacturing of
uni - or olio lamellar of the order of 1.0μm or less in diameter.
Advantages
High entrapment of water soluble content and use of mild
condition for preparation & loading of bioactive.
Freeze Thawed Sonication
This method is based on freezing of unilamellar dispersion and
thawing (melting) by standing at RT for 15 min. and finally
subjected to a sonication cycle. This process ruptures and refuses
SUVs during which the solute equilibrates between inside and
outside, and liposomes themselves fuse and markedly increase in
size. The second step of the sonication considerably reduces the
permeability of the liposome membrane, by accelerating the rate
at which the packing defects are eliminated. For producing giant
vesicles of diameter having 10 – 50 μm, the sonication step is
replaced by the dialysis against hypo-osmolar buffer. In this case,
SUVs are mixed with salt solution followed by freeze thawing.
During this dialysis, the large vesicles formed by freeze thawing
swell and rupture as a result of the osmotic lysis, where the fuse
and prepare as giant vesicles.
Disadvantage
- Lesser encapsulation efficiency,
- Presence of charge particle for the formation of ice crystal to aid
in the rupture or fusion process, so neutral liposomes cannot be
resulted.
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Ashara et. al.
Advantage
- Simple, rapid, result in proportion of large unilamellar vesicles
formation.
2. Solvent Dispersion Method
Ethanol Injection method
Ethanol is used to dissolve the lipids and solution is rapidly
injected through a fine needle into an excess of buffer solution.
SUVs form spontaneously. Method is restricted to the production
of relatively dilute SUVs suspension. Removal of residual ethanol
is also present a problem. This can be done by ultrafiltration or
vacuum distillation
Ether Injection Method
In this method, solution of lipids in diethyl ether or ether: methanol
mixture is slowly injected to aqueous solution of materials to be
encapsulated at 55 - 65ºC. Subsequent removal of ether under
vacuum leads to the formation of liposomes.
Drawbacks
Heterogeneous size (70 - 190μm), exposure of compounds to
organic solvents or high temperature.
Fig.9: Principles of Vesicles formations by solvent dispersion
methods in which two phases (Aqueous & organic) are
miscible with each other & forms different types of vesicles
Double Emulsion Vesicles
When organic solution which already contain water droplet, is
introduced into excess aqueous phase followed by mechanical
dispersion, multi compartment vesicles are obtained. The ordered
dispersion so obtained is desirable as a w/o/w system. The
vesicles with aqueous core are suspended in aqueous medium.
So two aqueous compartments being separated from each other
by pair of phospholipids monolayer whose hydrophobic surface
face each other across a thin film of organic solvent. Removal of
this solvent clearly results in intermediate sized unilamellar
vesicle. The theoretical entrapment may reach up to 90%.
Reversed- phase Evaporation of Vesicles
The essential feature of this method is the removal of solvent from
emulsion by evaporation. In this method, lipids dissolved in
organic solvents are sonicated by bath sonication which forms
emulsion (w/o) and then emulsion is dried down to a semi solid
gel using rotary evaporator under reduced pressure. The next
step is to Bing about the collapse of a certain proportion of water
droplets by vigorous mechanical shaking with a vortex mixer. This
will give LUVs. Encapsulation percentage: up to 50%.
Stable plurilamellar vesicles
In this method, w/o dispersion is prepared as described in REV
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Method with excess lipid, but drying process is accompanied by
continued bath sonication with a stream of nitrogen. The
redistribution and equilibration of aqueous solvent and solute
occur during this time in between the various bilayers in each
plurilamellar vesicle. Entrapment percentage: 30%.
Fig.10:Formation of different liposomes using reverse-phase
Evaporation method.MLVs are formed in presence of excess
of phospholipids whereas LUV-REVs are formed in absence
of extra lipids.
3. Detergent Removal Method
In this method, the phospholipids are brought into intimate contact
with the aqueous phase via the intermediary of detergents, which
associate with phospholipid molecules and serve to screen the
hydrophobic portions of the molecule from water. Detergent
depletion is achieved by four following approaches:
Dialysis
The dialysis can be performed in dialysis bags immersed in large
detergent free buffers (equilibrium dialysis) or by using continuous
flow cells, diafiltration and cross filtration.
Gel filtration
In this method the detergent is depleted by size exclusive
chromatography. Sephadex G-50, Sephadex G-100, Sepharose
2B-6B and Sephacryl S200-S1000 can be used for gel filtration.
The liposomes do not penetrate into the pores of the beads
packed in a column.
Adsorption using bio beads
Detergent adsorption is achieved by shaking of mixed micelle
solution with beaded organic polystyrene absorbers such as XAD-
2 beads and Bio-beads SM2. The great advantage of the using
detergent absorbers is that they can remove detergents with a
very low critical micelle concentration (CMC) which are not
completely depleted by dialysis or gel filtration methods.
Dilution
Upon dilution of aqueous mixed micellar solution of detergent and
phospholipids with buffer the micellar size and the polydispersity
increases dramatically, and, as the system is diluted beyond the
mixed micellar phase boundary, a spontaneous transition from
polydisperse micelles to monodisperse vesicles occurs.
ACTIVE (REMOTE) LOADING TECHNIQUE
Certain types of drugs with ionisable groups and those with both
lipid and water solubility can be introduced into liposomes after
the formation of the intact vesicles. Drug is loaded into the
preformed liposomes using pH gradient and potential difference
across liposomal membrane.
5
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Approach for remote loading fluid. There are several factors which directly affect the phase
transition temperature including hydrocarbon length, unsaturation,
Vesicles are prepared in low pH solution, thus generating low pH charge, and head group species. When developing a new
within liposome interior followed by addition of the base to product, procedure, or method, controlling the transition
external medium of liposomes. Basic compounds with amino temperature of the lipid could be useful. Using a high transition
group are relatively lipophilic at high pH and hydrophilic at low pH. lipid when filtration is necessary could present some technical
Unprotonated form of basic drug can diffuse through the bilayer. problems.
At the low pH side, the molecules are predominantly protonated,
which lower the concentration of the drug in the unprotonated Stability
form.
The long term stability or shelf-life of a drug product containing
Structural components of Liposomes lipids can be dramatically affected by the lipid species used in the
formulation. Generally, the more unsaturated a compound, the
Various lipids and amphiphiles are available as liposome raw easier the product is oxidized, and thus the shorter the shelf life of
materials or additives that are required for the formation of lipid the product. Lipids from biological sources (e.g., egg, bovine, or
bilayers. Phospholipids, Synthetic Phospholipids, Glycerolipids, soybean) typically contain significant levels of polyunsaturated
Sphingolipids, Glycosphingolipids, Steroids, Polymeric material, fatty acids and therefore are inherently less stable than their
Charge-inducing lipids synthetic counterparts. While saturated lipids offer the greatest
Phospholipids stability in terms of oxidation; they also have much higher
transition temperatures and thus present other difficulties in
Natural Phospholipids formulation.
Phosphotidylcholine, Charge
Phosphotidylserine, The charge is generally imparted by the presence of anionic
phospholipid species in the membrane. The major naturally
Phosphotidylethanolamine occurring anionic phospholipids are phosphatidylserine,
Phosphatidylinositol phosphatidylinositol, phosphatidic acid, and cardiolipin. The
charge may provide a special function for the membrane. Several
Synthetic Phospholipids steps of the blood coagulation cascade require a lipid membrane.
The assembling of protein aggregates on the surface of platelets
1, 2-Dilauroyl-sn-Glycero-3-Phosphocholine (DLPC), 1, 2- requires a negatively charged surface.
Dioleoyl-Sn-Glycero-3-[Phospho-L-Serine] (Sodium Salt) (DOPS),
Dipalmitoylphosphotidylcholine (DPPC), Lipid mixtures
Distearoylphosphotidylcholine (DSPC),
Dipalmitoylphosphotidylserine (DPPS), In many cases, a single lipid species does not yield the exact
Dipalmitoylphosphotidylglycerol (DPPG), 1, 2-Dilauroyl-sn- physical properties needed for a particular system, or does not
Glycero-3-Phosphocholine (DLPC) adequately mimic the natural system for which it is intended to
replace or reproduce. For these issues, consider a complex lipid
Unsaturated mixture composed of two or more individual lipid species, the
composition designed to create or reproduce a particular charge
1-Stearoyl-2-Linoleoyl-Sn-Glycero-3-[Phospho-L-Serine] (Sodium ratio, unsaturation ratio, phase transition temperature, or
Salt),
biological function.
Dioleaylphosphotidylcholine (DOPC), Dioleayphosphotidylglycerol Cholesterol
(DOPG),
Dioleayphosphotidyl ethanolamine (DOPE) Cholesterol is a membrane constituent widely found in biological
systems which serves a unique purpose of modulating membrane
Sphingolipids: Sphingomyelin, Glycosphingolipids: fluidity, elasticity, and permeability. It literally fills in the gaps
Gangliosides, Steroids: Cholesterol, Polymeric material: Lipids created by imperfect packing of other lipid species when proteins
conjugated to dine, methacrylate, & thiol group, Charge-inducing are embedded in the membrane. Unfortunately, cholesterol
lipids: Dioctadecyldimethyl ammonium bromide/chloride presents certain problems when used in human pharmaceuticals.
(DODAB/C), Dioleoyl trimethylammonium propane (DOTAP), Purity sources and Stability problem for lipid based drug products
Other Substances: Stearylamine & Dicetylphosphates,
Polyglycerol & polyethoxylated mono & dialkyl amphiphiles. Source

Issues to Consider while Selecting Lipids
Phase transition temperature
The phase transition temperature is defined as the temperature
required to induce a change in the lipid physical state from the
ordered gel phase, where the hydrocarbon chains are fully
extended and closely packed, to the disordered liquid crystalline
phase, where the hydrocarbon chains are randomly oriented and
There are two basic sources of phospholipids: synthetic and
tissue-derived. Tissue-derived lipids are generally either egg-
derived or bovine-derived. For clinical applications, either of these
sources is not suitable due to stability problems and the possibility
of viral or protein contamination. The U.S. Food and Drug
Administration issued a letter restricting the source of bovine
tissue used to isolate pharmaceutical products to countries and
animals certified to be free of bovine spongiform encephalopathy
(BSE).

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Ashara et. al.
Egg sources are not currently restricted; however, additional
testing for viral contamination may be required for pharmaceutical
products.
CHARACTERIZATION OF LIPOSOMES
PHYSICAL CHARACTERIZATION
Size & size distribution
Most precise method to determine size of the liposome is electron
microscopy, since it allows to view each individual liposome and
to obtain exact information about the profile of liposome
population over the whole range of sizes.Unfortunately it is very
time consuming and requires equipments that may not always be
immediately available to hand. In contrast, laser light scattering
(quasi elastic laser light scattering) method is very simple and
rapid to perform. Above methods require very costly equipments.If
only approximate idea of size range is required then gel exclusion
chromatographic techniques are recommended, since only
expense incurred is that of buffers and gel materials.
Surface Charge
Vesicle surface charge can be estimated by measurement of
particle electrophoretic mobility & is expressed as the zeta
potential which can be calculated using the Henry equation:
ζ = µE4πη / Σ
Where, ζ = zeta potential, µE = electrophoretic mobility, η =
viscosity of the medium, Σ = dielectric constant.
Lameliarity
The average number of bilayers present in a liposome can be
found by freeze electron microscopy and by NMR.
In the latter technique, the signals are recorded before and after
the addition of broadening agent such as manganese ions which
interact with the outer leaflet of the outermost bilayers. Thus, a
50% reduction in NMR signal means that the liposome
preparation is unilamellar and a 25% reduction in the intensity of
the original NMR signal means that there are 2 bilayers in the
liposome.
Nowadays, freeze fracturing electron microscopy has become a
very popular method to study structural details of aqueous lipid
dispersions.
Encapsulation Capacity
Amount of Encapsulated drug X 100/Total amount of drug
Encapsulation/Entrapped drug capacity is measured by two
methods:
-Mini column centrifugation
-Protamine aggregation
Mini column centrifugation: This method serves for both- isolation
of liposomes and analysis of entrapped material.
 A sephadex or sepharose column is used which is
prostrated with dispersion medium.
 Sample is applied to the column and column is centrifuged at
2000 rpm for 3 min.
 Concentration of free or entrapped material is then found out.
 Entrapped material can be assayed after disrupting the
liposomes by ethanol(2ml ethanol for 10 µl of liposomes)
Protamine aggregation method
Concentration of free or entrapped material is then found out.
Entrapped material can be assayed after disrupting the liposomes
by ethanol (2ml ethanol for 10 µl of liposomes).
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Entrapped or internal Volume
“To describe entrapment of water soluble drugs in the aqueous
compartments of Liposomes.”
The best way to measure internal volume is to measure the
quantity of water directly, and this may be done very conveniently
by replacing the external medium with a spectroscopically inert
fluid, (e.g. D2O) and then
Measuring the water signal by NMR
Trapped volumes are also determined by dispersing lipid in an
aqueous medium containing non-permeable radioactive solute
such as [22Na] and [14C]. The proportion of solute is determined by
removing external radioactivity by centrifugation or dialysis.
Drug release
The mechanism of drug release from the liposomes can be
assessed by the use of a well calibrated in vitro diffusion cell.
The liposome based formulations can be assisted by employing in
vitro assays to predict pharmacokinetics and bioavailability of the
drug before employing costly and
Time consuming in vivo studies
Another assay which determined intracellular drug release
induced by liposomes degradation in the presence of mouse liver
lysosome lysate was used to assess the bioavailability of the drug.
CHEMICAL PROPERTIES
Determination of Phospholipids
The phospholipids are measured by
Bartlett assay
 The problem is that the test is easily upset by trace
contamination with inorganic phosphate. Therefore,
precaution is to be taken
– Using a set of borosilicate glass tubes which are washed
well and not used for any other purpose.
– Use of double distilled water for making up solutions.
 The sensitivity of the Bartlett assay to inorganic phosphate
creates problem with measurement of phospholipid
liposomes suspended in physiological buffers, which usually
contain phosphate ions. This can be overcome by employing
a more specific method which is unaffected by inorganic
phosphate.
Stewart assay
In Stewart assay, the phospholipid forms a complex with
ammonium ferrothiocyanate in organic solution.Samples are
measured spectrophotometrically.
7
Ashara et. al.
Phospholipid Hydrolysis
The major product of lecithin hydrolysis is lysolecithin where one
fatty acid chain is lost by de esterification. Ideally, estimation of
phospholipid hydrolysis by quantitation of lysolecithin could be
carried out by HPLC
Phospholipid Oxidation
Oxidation products of phospholipids (such a phospholipids
hydroperoxides) are analyzed using HPLC or GLC.
Cholesterol analysis
Cholesterol is qualitatively analyzed using
Chromatography.Whereas it is quantitatively estimated by
measuring the absorbance of purple complex produced with iron
upon reaction with a combined reagent containing ferric
perchlorate, ethyl acetate and sulphuric acid at 610nm.
BIOLOGICAL CHARACTERIZATION
Liposomes for parenteral use should be sterile and pyrogen free.
APPLICATION OF LIPOSOMES
Twenty five year of research into the use of liposome in drug
delivery. Liposomes are one of the unique drug delivery system,
which can be of potential use in controlling and targeting drug
delivery. Liposomes are administrated orally, parenteral and
topically as well as used in cosmetic and hair technologies,
sustained release formulations, diagnostic purpose and as good
carriers in gene delivery various drugs with liposomal delivery
systems have been approved. Nowadays liposomes are used as
versatile carriers for targeted delivery of drug.
Liposomes also have applications in other disciplines like
mathematics, physics, biophysics, chemistry, biochemistry, etc.
As of 2008, 11 drugs with liposomal delivery systems have been
approved and six additional liposomal drugs were in clinical trials.
List of clinically-approved liposomal drugs (Table 2).
Therapeutic Application of Liposome [10]
1. Liposome as drug/protein delivery vehicles, Controlled and
sustained drug release, Enhanced drug solubilization, altered
pharmacokinetics and biodistribution, Enzyme replacement
therapy and biodistribution, Enzyme replacement therapy and
lysosomal storage disorders
2. Liposome in antimicrobial, antifungal and antiviral therapy,
Liposomal drugs, Liposomal biological response modifiers
3. Liposome in tumor therapy: Carrier of small cytotoxic molecules
Vehicle for macromolecules as cytokines or genes
4. Liposome in gene delivery: Gene and antisense therapy,
Genetic (DNA) vaccination
5. Liposome in immunology: Immunoadjuvant
Immunomodulation, Immunodiagnostics
6. Liposome as artificial blood surrogates
7. Liposome as radiopharmaceutical and radio diagnostic carriers
8. Liposome in cosmetics and dermatology
9. Liposome in enzyme immobilization and bioreactor technology.
Table 2: List of Clinically-Approved Liposomal Drugs
Name Trade Name
Liposomal Amphotericin B Abelcet
Ambisome
Liposomal Citarabine Depocyte
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NIOSOMES
Controlled release formulations are often prepared to permit the
establishment and maintenance of any concentration at target site
for longer period of time. One such technique of drug targeting is
niosomes. Niosomes are microscopic lamellar structures formed
on admixture of a nonionic surfactant, cholesterol and diethyl
ether with subsequent hydration in aqueous media. Non-ionic
surfactant vesicles (or niosomes) are now widely studied as
alternates to liposomes.[11] Structure of Niosomes: Nonionic
surfactant vesicles (NSVs or niosomes) result from the self
assembly of hydrated surfactant monomers. They are similar in
physical structure and form to the more widely studied
phospholipid vesicles (liposomes) [12], consisting of single or
multiple surfactant bilayers (lamellae) enclosing an aqueous core.
A schematic diagram of a non-ionic surfactant vesicle is shown in
Fig. 4 Preliminary X-ray scattering data on unilamellar sorbitan
monostearate (C18- sorbitan monoester)-cholesterol niosomes
suggest a bilayer spacing of 15 nm and a bilayer thickness of 3.3-
3.4 nm [13], the latter similar to the figure obtained for the bilayer
thickness of phospholipid vesicles (3.4-3.9 nm) [14]. Although
terminology suggests that distinctions exist between liposomes
and niosomes, of which the basic unit of assembly is the
amphiphile, their properties are largely similar and the differences
between liposomes (phospholipid vesicles) and non-ionic
surfactant vesicles are sometimes blurred as liposomes are often
prepared incorporating a non-ionic surfactant component [15, 16],
while non-ionic surfactant vesicles may also be formulated with
various ionic amphiphiles such as stearylamine and
dicetylphosphate to achieve greater protection against
flocculation in vesicle suspensions. The association of nonionic
surfactant monomers into vesicles on hydration is a result of the
fact that there exists a high interfacial tension between water and
the hydrocarbon portion (or any other hydrophobic group) of the
amphiphile which causes these groups to associate.
Simultaneously, the steric, hydrophilic and/or ionic repulsion
between the head groups ensures that these groups are in
contact with water. These two opposing forces result in a
supramolecular assembly.
Fig.11: A Schematic diagramme of Non-Ionic Surfactant
Method of preparation of niosomes
Various methods are reported for the preparation of niosomes
such as
1. Ether injection method
2. Hand shaking method (Thin film hydration technique)
3. Sonication method
4. Reverse phase evaporation technique (REV)
5. Micro fluidization
6. Multiple membrane extrusion method
Company Indication
Enzon
Fungal & Protozoal Infections
Gilead Sciences
Pacira (formerly Sky Pharma) Malignant lymphomatous
meningitis
8
Ashara et. al.
7. Trans membrane pH gradient (inside acidic) drug uptake
process (remote loading)
8. Bubble method
9. Formation of niosomes from proniosomes
Ether injection method
This method provides a means of making niosomes by slowly
introducing a solution of surfactant dissolved in diethyl ether
(volatile organic solvent) into warm water maintained at 60°C. The
surfactant mixture in ether is injected through 14- gauge needle
into an aqueous solution of material. Vaporization of ether (volatile
organic solvent) leads to formation of single layered vesicles.
Depending upon the conditions used the diameter of the vesicle
range from 50 to 1000 nm [17, 18].
Hand shaking method (Thin film hydration technique)
The mixture of vesicles forming ingredients like surfactant and
cholesterol are dissolved in a volatile organic solvent (diethyl
ether, chloroform or methanol) in a round bottom flask. The
organic solvent is removed at room temperature (20°C) using
rotary evaporator leaving a thin layer of solid mixture deposited on
the wall of the flask. The dried surfactant film can be rehydrated
with aqueous phase at 0-60°C with gentle agitation. This process
forms typical multilamellar niosomes [18].
Sonication
In this method an aliquot of drug solution in buffer is added to the
surfactant/cholesterol mixture in a 10-ml glass vial. The mixture is
probe sonicated at 60°C for 3 minutes using a sonicator with a
titanium probe to yield niosomes.[18]
Reverse phase evaporation technique (REV)
Cholesterol and surfactant (1:1) are dissolved in a mixture of ether
and chloroform. An aqueous phase containing drug is added to
this and the resulting two phases are sonicated at 4- 5°C. The
clear gel formed is further sonicated after the addition of a small
amount of phosphate buffered saline (PBS). The organic phase is
removed at 40°C under low pressure. The resulting viscous
niosome suspension is diluted with PBS and heated on a water
bath at 60°C for 10 min to yield niosomes [19].
Micro fluidization
It is a recent technique used to prepare unilamellar vesicles of
defined size distribution. This method is based on submerged jet
principle in which two fluidized streams Interact at ultra high
velocities, in precisely defined micro channels within the
interaction chamber. The impingement of thin liquid sheet along a
common front is arranged such that the energy supplied to the
system remains within the area of niosomes formation [20]. The
result is a smaller size, greater uniformity and better
reproducibility of niosomes formed.
Multiple membrane extrusion method
Mixture of surfactant, cholesterol and dicetyl phosphate in
chloroform is made into thin film by evaporation. The film is
Difference between Liposomes & Niosomes:
Characteristics Liposomes
Availability Phospholipids are comparatively hard in availability
Cost Expensive
Storage Required special handling & storage conditions
Stability Instability problem
Chemical composition Not Précised
Double chained phospholipids(Neutral/Charged)
Preparations
Properties Dependence bilayer Composition & Method of preparations
Entrapped efficiency Depended on Nature & amount of phospholipids
Technical point of View: Non-promising drug carrier
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hydrated with aqueous drug polycarbonate membranes
and the resultant suspension extruded through which are placed
in series for up to 8 passages. Multiple membrane extrusion
method is better for the controlling of niosome size [20].
Trans membrane pH gradient (inside acidic) drug uptake
process (remote loading)
Surfactant and cholesterol are dissolved in chloroform. The
solvent is then evaporated under reduced pressure to get a thin
film on the wall of the round bottom flask. The film is hydrated with
300 mm citric acid (pH 4.0) by vortex mixing. The multilamellar
vesicles are frozen and thawed 3 times and later sonicated. To
this niosomal suspension, aqueous solution containing 10 mg/ml
of drug is added and vortexes. The pH of the sample is then
raised to 7.0-7.2 with 1M disodium phosphate. This mixture is
later heated at 60°C for 10 minutes to give niosomes [21].
Bubble method
It is novel technique for the one step preparation of liposomes and
niosomes without the use of organic solvents. The bubbling unit
consists of round-bottomed flask with three necks positioned in
water bath to control the temperature. Water-cooled reflux and
thermometer is positioned in the first and second neck and
nitrogen supply through the third neck .Cholesterol and surfactant
are dispersed together in this buffer (pH 7.4) at 70°C, the
dispersion mixed for 15 seconds with high shear homogenizer and
immediately afterwards “bubbled” at 70°C using nitrogen gas.
Formation of niosomes from proniosomes
Another method of producing niosomes is to coat a water-soluble
carrier such as sorbitol with surfactant. The result of the coating
process is a dry formulation [22]. In which each water-soluble
particle is covered with a thin film of dry surfactant. This
preparation is termed “Proniosomes”.
4. RECENT ADVANCES IN VDDS:
Fig. 12: Different Pharmaceutical Carriers[23]
Niosomes
Synthetic non-ionic surfactants are readily available
Cheaper
Don’t Required special handling & storage conditions
Chemically stable
Précised
uncharged single chain surfactant and cholesterol
Surfactant, Entrapped drug molecules, Targeted site
Depended on concentration & Lipophilicity of surfactant.
Promising drug carriers
9
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Table 3: Therapeutic Application of Drugs after incorporation

Different types of pharmaceutical carriers are present. (Fig with Transferosomes
No.:4)They are particulate, polymeric, macromolecular, & cellular
carrier. Particulate carrier also knows as colloidal carrier system Effects after
includes lipid particles (Low & High density lipoprotein).The Drug incorporation of drugs Applications
vesicular systems have been advanced as[8]: in transferosomes
Delivery of
Ethosome
Insulin Transferulin[28] insulin by
Ethosomes has initiated a new area in vesicular research for Dermal Route
transdermal drug delivery which can provide better skin Increases the level of IgA
Zudovudin
permeation and stability than liposomes. Application of ethosomes Higher AUC Transdermal
provides the advantages such as improved entrapment and NSAIDS Immunization
physical stability. Ketoprofen Increases Bioavailability [29, 30]
Transferosomes
Ethosomes are lipid based elastic vesicles. Phospholipids, alcohol
(In high concentration) & water. Size: Nanometers-Microns. High Pharmacosomes
concentration ethanol (20-50%).Lipid membrane packed less
tightly than conventional vesicles hence improved drug distribution Pharmacosomes are amphiphilic lipid vesicular system
through stratum corneum. Increase fluidity of cell membrane, possessing phospholipid complexes of drugs. Pharmacon means
increases cell permeability, Ulters solubility properties of stratum drugs & Soma means Carrier thus Pharmacosomes means drug
corneum & Increase solubility of drugs, e.g. Levonorgesterol, carriers. System formed by linking drugs’ o the carrier. Colloidal
hydrocortisone, 5-flurouracil(TDDS). dispersion of drugs co-valant bond to lipids. Composed of
amphiphilic prodrugs, so high drug loading amount & very low
Ethosomes are lipid “Soft malleablevesicles”embodying a
drug leakages can be achieved easily. Increases interfacial
permeation enhancer & composed of phospholipid, ethanol and
tensions so increases contact area & finely increases
water. Ethosomes are used mainly as Targeted delivery to deep
bioavailability.
skin layer. [24]
Advantages
Transferosomes
Drug targeting, Controlled released, High entrapment efficacy, No
Transferosomes was introduced for the effective transdermal
need of removal of entrapment drugs from formulations as
delivery of number of low and high molecular weight drugs.
required in Liposomes, Improves bioavailability of purely soluble
Transfersomes can penetrate the intact stratum corneum
drugs, Reduces cost of therapy[31]
spontaneously along two routes in the intracellular lipid that differ
in their bilayers properties [25]. It consist of both hydrophilic and Disadvantages
hydrophobic properties, high deformability gives better penetration
of intact vesicles [26].These vesicular transfersomes are several Co-valent bond is required to protect leakages of drugs,
orders of magnitudes more elastic than the standard liposomes Amphiphilic nature is responsible for the synthesis of the
and thus well suited for the skin penetration. Transfersomes compounds, On storages undergoes fusion, aggression &
overcome the skin penetration difficulty by squeezing themselves Chemical hydrolysis.[32]
along the intracellular sealing lipid of the stratum corneum. There
Preparation Methods
is provision for this, because of the high vesicle deformability,
which permits the entry due to the mechanical stress of Generally pharmacosomes can be prepared by two methods
surrounding, in a self-adapting manner. Flexibility of which are as follows[33]:
transfersomes membrane is achieved by mixing suitable surface-
active components in the proper ratios. Transferosome based Hand shaking method
formulations of local anesthetics- lidocaine and tetracaine showed
In this method a mixture of drug and lipid are dissolved in a
permeation equivalent to subcutaneous injections. Anti cancer
volatile organic solvent such as dichloromethane in a round
drugs like methotrexate were tried for transdermal delivery using
bottom flask. The organic solvent is removed at room temperature
transferosome technology. This provided a new approach for
using a rotary evaporator, which leaves a thin film of solid mixture
treatment especially of skin cancer.[4]
deposited on the walls of flask. The dried film can then be
Advantage hydrated with aqueous medium and readily gives a vesicular
suspension.
 Transferosomes possess an infrastructure consisting of
hydrophobic and hydrophilic moieties together and as a Ether injection method
result can accommodate drug molecules with wide range of In this method organic solution of drug-lipid complex is injected
solubility. slowly into the hot aqueous medium wherein the vesicles are
 Transferosomes can deform and pass through narrow formed readily.
constriction (from 5 to 10 times less APPLICATIONS OF PHARMACOSOMES
 Than their own diameter) without measurable loss. As Novel Drug Delivery System
 Poses high entrapment efficiency, in case of lipophilic drug In the development of novel ophthalmic dosage forms
near to 90%. pharmacosomes are the amphiphilic lipid vesicular system. Any
drug bearing a free carboxyl group or active hydrogen atom can
 Used for both systemic as well as topical delivery of drug.
be esterified to the hydroxyl group of a lipid molecule thus forming
Limitation an amphiphilic prodrug. These are converted to pharmacosome
on dilution with tear and enhance transport across cornea and
 Transferosomes are chemically unstable because of their facilitate control release profile. Pharmacosome elicit greater shelf
predisposition to oxidative degradation. stability. [L.M.Raikhman et al.[34] discussed the pharmacosomes
as building particles characterized by high degree of selectivity
 Purity of natural phospholipids is another criteria militating
(acting on target cells). These are capable of delivering various
against adoption of transfersomes as drug delivery vehicles.
biologically active substances including biopolymers (nucleic acid
 and proteins). A. Semalty et al. optimized development and
Transferosomes formulations are expensive.[27]
evaluation of pharmacosomes of aceclofenac and found the drug
content

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Ashara et. al.
Was 91.88% (w/w) for aceclofenac phospholipid complex (1:1)
and 89.03% (w/w) for aceclofenac phospholipid complex (2:1)?
Solubility of aceclofenac pharmacosome was found to be higher
than the aceclofenac. In this study the free aceclofenac showed a
total of only 68.69% drug release at the end of 4hr dissolution
study while aceclofenac pharmacosome (1:1) showed 79.78%
and aceclofenac pharmacosome (2:1) showed 76.17% at the end
of 4hr dissolution study. A. Semalty et al. studied development of
diclofenac pharmacosome and physicochemical evaluation for
drug solubility, in vitro dissolution study, drug content, surface
morphology, crystallinity and phase transition behavior. Water
solubility of diclofenac pharmacosome was found to be 22.1μg/ml
as compared to 10.5 μg/ml of diclofenac. Drug release of
diclofenac pharmacosome was 87.8% as compared to 60.4% of
diclofenac at the end of 10hr dissolution study and the drug
content of diclofenac pharmacosome was found to be 96.2+/-1.1
%. In SEM, pharmacosomes of diclofenac were found to be disc
shaped. XRPD analysis and DSC thermo grams proved the
formation of phospholipid complex. M. Han et al. optimized
preparation and evaluation of 20(S) protopanaxadiol
pharmacosome and showed that the encapsulation efficiency of
protopanaxadiol pharmacosome was (80.84+/-0.53) with the
diameter of 100.1 nm and (72.76+/-0.63) with the diameter of
117.3 nm. Thus the selected formulation and technology are
simple and preparation properties are more stable. AI PING ET
al.prepared didanosine pharmacosomes by using tetra hydro
furan injection method and investigate the in vivo behavior of
pharmacosomes in rats by determining the drug in plasma and
tissues with HPLC. From the result it can be concluded that
pharmacosomes elicit liver targeting and sustained release effect
in target tissues. Z R Zhang, J X Wang successfully optimized the
preparation of 3',5'-dioctanoyl-5-fluoro-2'-deoxyuridine
pharmacosomes [35] by using central composite design and
concluded that 3',5'- dioctanoyl-5-fluoro-2'-deoxyuridine
pharmacosomes showed a good targeting efficiency in vivo and
can improve the ability of drug to cross the blood brain barrier. JIN
Yi-Guang et al. prepared acyclovir pharmacosomes by tetrahydro
furan injection and investigate the following properties) the
negatively charged pharmacosome were nanometer vesicles
based on analysis of trans-mission electron scanning calorimetry.
ii) The effects of centrifugation and heating on stability of
pharmacosomes were weak.
iii) Freezing and lyophilisation disrupted pharmacosomal structure.
iv) The amphiphilic pharmacosomes would insert into rabbit
erythrocyte membranes and led to hemolysis. Plasma proteins in
blood absorbed pharmacosomes or interfered the interaction with
erythrocytes to reduce hemolytic reaction. A. Semalty et al.
investigate development and characterization of aspirin –
phospholipid complex (in 1:1 molar ratio) for improved drug
delivery and found that the drug content was 95.6% for aspirin-
phospholipid complex. The free aspirin showed a total of only
69.42% drug release at the end of 10 hr dissolution study while
aspirin pharmacosome showed a total of only 90.93% drug
release at the end of 10 hr dissolution study in pH 1.2 acid buffers.
Thus it can be concluded that aspirin pharmacosomes enhance
the bioavailability of aspirin. The GI toxicity is also reduced in case
of aspirin-phospholipid complex. Peng-Fei Yue et al. prepared and
investigate the characteristics of geniposide pharmacosome and
optimize the process by response surface design. The
phospholipid to drug ratio, reaction temperature and drug
concentration were determined as 3, 50⁰C, 5.5mg/ml respectively.
Thus pharmacosomes can improve absorption and permeation of
biologically active constituents. V.E.Ivan et al. studied the effect of
temperature on cascade system of pharmacosome fusion and
demonstrated that a combination of cell-specific drug vehicles
(pharmacosomes) containing cascade fusion system, at
appropriate temp will have a prominent effect on drug delivery to
appropriate sites within an organism by using both heating and
cooling of tissues.
COLLOIDOSOME
Hollow shell microcapsules consits of coagulated or fused
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Particles at interphase of emulsion droplets. Control of size allows
flexibility in applications. Colloidosome membrane offers great
potential in controlling the permeability of entrapped spices.
Allows selective & timed release.
Colloidosomes are the hollow shell microcapsules consisting of
coagulated or fused particles at interface of emulsion droplets.
Colloidosomes have exciting potential applications in controlled
release of drugs, proteins, vitamins as well as in cosmetics and
food supplements.
Colloidosomes have a great encapsulation efficacy with a wide
control over size, permeability, mechanical strength and
compatibility. Colloidosomes is a novel class of microcapsules
whose shell consists of coagulated or fused colloid particles at
interface of emulsion droplets. The particles self assemble on the
surface of droplets in order to minimize the total interfacial energy
forming colloidosomes. Such structures were produced for first
time by templating latex particles adsorbed on the surface of
octanolin- water emulsion drops and subsequent removal of oil
after fusing the particles monolayers. Similar structures have also
been obtained by templating water-in-oil emulsions and template
solid nanoparticles on the surface of solid sacrificial micro
particles based on electrostatic attraction and layer by layer
assembly of multilayer shells consisting of alternating positively
and negatively charged nanoparticles or polyelectrolytes. The final
hollow shells are obtained by removal of central, sacrificial
colloidal particles. Colloidosomes assemble polymer latex
colloidal particles into shells around water-in-oil emulsion drops
followed by partial fusion of shell and centrifugal transfer into
water to yield stable capsules in which the shell permeability can
be controlled by adjustment of partial fusion conditions. Hairy
colloidosomes, whose shell consists of micro rod particles, are
designed and fabricated novel colloidosome capsules that consist
of aqueous gel core and shells of polymeric micro rods. This has
been achieved by templating water-in-oil emulsions stabilized by
rod like particles followed by gelling of the aqueous phase,
dissolution of oil phase in ethanol and redispersion of obtained
colloidosome microcapsules in water.
Advantages: Control of the size allows flexibility in applications
and choice of encapsulated materials, Colloidosome membrane
offer great potential in controlling the permeability of the
entrapped species and allow the selective and time release,
Control of the mechanical strength allows the yield stress to be
adjusted to withstand, varying of mechanical loads and to enable
release by defined shear rates[5].
HERBOSOMES
The term "herbo" means plant, while "some" means cell like. Over
the past century, phytochemical and phyto‐ pharmacological
sciences established the compositions, biological activities and
health promoting benefits of numerous botanical products. Most of
the biologically active constituents of plants are polar or water
soluble molecules. However, water soluble phytoconstituents (like
flavonoids, tannins, glycosidic aglycones etc) are poorly absorbed
either due to their large molecular size which cannot absorbed by
passive diffusion, or due to their poor lipid solubility, severely
limiting their ability to pass across the lipid rich biological
membranes, resulting poor bioavailability. Phytomedicines,
complex chemical mixtures prepared from plants, have been used
for health maintenance since ancient times. But many
phytomedicines are limited in their effectiveness because they are
poorly absorbed when taken by mouth. Herbosomes are also
often known as phytosomes. Herbosomes exhibit better
pharmacokinetic and pharmacodynamics profile than conventional
herbal extracts. Molecular layer consisting of PC and other
phospholipids provides a continuous matrix into which the proteins
insert.
Advantages
It enhances the absorption of lipid insoluble polar
phytoconstituents through oral as well as topical route showing
better bioavailability, hence significantly greater therapeutic
benefit, As the absorption of active constituent(s) is improved, its
11
Ashara et. al.
dose requirement is also reduced, Phosphatidylcholine used in
preparation of herbosomes, besides acting as a carrier also acts
as a hepatoprotective, hence giving the synergistic effect when
hepatoprotective substances are employed, Herbosome
permeates the non-lipophilic botanical extract to be better
absorbed in intestinal lumen, Unlike liposome, chemical bonds is
formed between phosphatidylcholine molecule and
phytoconstituent, so the Herbosomes show better stability profile.
SPHINOSOMES
Liposome stability problems are of course much more severe so it
is very important task to improve the liposomal stability. Liposomal
phospholipid can undergo chemical degradation such as oxidation
and hydrolysis either as a result of these changes or otherwise
liposome maintained in aqueous suspension may aggregate, fuse,
or leak their content. Hydrolysis of ester linkage will slow at pH
value close to neutral. The hydrolysis may be avoided altogether
by use of lipid which contains ether or amide linkage instead of
ester linkage (such are found in sphingolipid) or phospholipid
derivatives with the 2- ester linkage replaced by carbomoyloxy
function. Thus sphingolipid are been nowadays used for the
preparation of stable liposomes known as sphingosomes.
Sphingosome may be defined as “concentric, bilayer vesicle in
which an aqueous volume is entirely enclosed by a membranous
lipid bilayer mainly composed of natural or synthetic sphingolipid.
Sphingosomes are administered in many ways these include
parenteral route of administration such as intravenous,
intramuscular, subcutaneous, and intra-arterial. Generally it will be
administered intravenous or some cases by inhalation. Often it will
be administered into a large central vein, such as the superior
vena cava and inferior vena cava to allow highly concentrated
solution to be administered into large volume and flow vessels.
Sphingosomes may be administered orally or transdermal. In
simple way we can say sphingosome is liposome which is
composed of sphingolipid.
Advantages
Provide selective passive targeting to tumor tissue, Increase
efficacy and therapeutic index, Increase stability via
encapsulation., Reduction in toxicity of the encapsulated agent,
Improve pharmacokinetic effect (increase circulation time),
Flexibility to couple with site specific ligands to achieve active
targeting.[5]
LAYEROSOMES
Layerosomes are conventional liposomes coated with one or
more multiple layers of biocompatible polyelectrolytes in order to
stabilize their structures. The layer-by-layer coating concept is one
of the strategies used for the preparation or the stabilization of
nanosystems[36]. The layersomes are conventional liposomes
coated with one or multiple layers of biocompatible
polyelectrolytes in order to stabilize their structure. The
formulation strategy is based on an alternative coating procedure
of positive poly (lysine) (pLL) and negative poly (glutamic acid)
(PGA) polypeptides on initially charged small unilamellar
liposomes. The major drawback of liposomes is their instability
during storage or in biological media which is related to surface
properties. This surface modification stabilized the structure of the
liposomes and led to stable drug delivery systems. Oral
administration or their incorporation in biomaterials are among
potential fields of application[37]. Thus the concept of
layerosomes has brought forward the stable nanosystem.
UFOSOMES
The formations of fatty acid vesicles are named "ufasomes,"
ufosomes are unsaturated fatty acid liposomes. Fatty acid
vesicles are colloidal suspensions of closed lipid bilayers that are
composed of fatty acids and their ionized species (soap). They
are observed in a small region within the fatty acid-soap-water
ternary phase diagram above the chain melting temperature (Tm)
of the corresponding fatty acid-soap mixture[38]. Fatty acid
vesicles always contain two types of amphiphiles, the nonionized
neutral form and the ionized form (the negatively charged soap).
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The ratio of nonionized neutral form and the ionized form is critical
for the vesicle stability. Fatty acid vesicles are actually mixed "fatty
acid/soap vesicles". Ufasome membranes are much more
stabilized in comparison to liposomes[39].
STRATEGIES TO IMPROVE VDDS
To improve VDDS mainly two strategies are
Pro-Vesicular Drug Delivery:
Pro vesicular drug delivery developed to overcome the stability
problems associated with vesicular drug delivery systems
composed of dry products or liquid crystalline gel that can be
hydrated immediately before use, e.g. Proliposomes,
Proniosomes.[40]
Characterization
Morphology, Angle of Repose, Rate of hydration, Entrapment
efficiency, Degree of deformity & permeability measurements,
Size & Size distribution etc.
Types of pro vesicular drug delivery systems
 Proliposomes
 Proniosomes
Proliposomes
In proliposomes, lipid and drug are coated onto a soluble carrier to
form free-flowing granular material which on hydration forms an
isotonic liposomal suspension. The proliposome approach may
provide an opportunity for cost-effective large scale manufacture
of liposomes containing particularly lipophilic drugs. Proliposomes:
Comparatively high stability, less required storage & handling
conditions. High encapsulation & drug release profile than
Liposomes. Types: Dry granular Proliposomes and Mixed
mecellar Proliposomes.
Comparison between liposomes and proliposomes
Liposomes-unilamellar or multilamellar spheroid structures
composed of lipid molecules, often phospholipids. They show
controlled release and increased solubility. But have tendency to
aggregate or fuse, susceptible to hydrolysis or oxidation.
Proliposomes-an alternative forms to conventional liposomal
formulation Composed of water soluble porous powder as a
carrier, phospholipids and drugs dissolved in organic solvent.
Lipid and drug are coated on to a soluble carrier to form free-
flowing granular material. Show controlled release, better stability,
ease of handling and increased solubility.
Proniosomes
Versatile Drug Delivery System, Ease of transfer, Feasibility,
distribution, storage, and high drug loading capacity, less side
effects, high efficacy, Type & concentration of surfactant affect
encapsulation efficiency & drug release rate from Proniosomes.
Method of Preparation of Proniosome: To create proniosomes, a
water soluble carrier such as sorbitol is first coated with the
surfactant.
 The coating is done by preparing a solution of the surfactant
with cholesterol in a volatile organic solvent, which is sprayed
onto the powder of sorbitol kept in a rotary evaporator.
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Ashara et. al.
 The evaporation of the organic solvent yields a thin coat on
the sorbitol particles. The resulting coating is a dry
formulation in which a water soluble particle is coated with a
thin film of dry surfactant. This preparation is termed
Proniosome. Other methods of preparations are:
 Slurry method
 Co-acervation phase separation
 Slow spray-coating method
Higher the Lipophilicity grater will be the encapsulation
efficiency.e.g. Proniosomal TDS of Losartan potassium, Alprenolol
HCl, Valsartan proniosome.
Comparison between niosomes and Proniosomes: Niosomes-are
non-ionic surfactant based multilamellar or unilamellar vesicles,
aqueous solution of solute is entirely enclosed by a membrane of
surfactant macro-molecules as bilayers. They are cheap and
chemically stable but posses’ problems related to physical stability
such as fusion, aggregation, sedimentation and leakage on
storage.
Proniosomes-approach minimizes the problems associated with
niosomes as it is a dry and free flowing product which is more
stable during sterilization and storage. Ease of transfer,
distribution, measuring and storage make it a versatile delivery
system. Proniosomes are water-soluble carrier particles that are
coated with surfactant.
Improve permeability:
Physical means
Iontophoresis
Effective method of drug transport in deeper layer of the bladder,
e.g. Mitomycin C, Bethanecol.Electroporation (high voltage than
Ionotophoresis).Increases permeability of tissues electric field.
Helpful for delivery of drug in Bladder carcinoma
treatment.Electroporation-Sonophoresis (Low density ultrasound
waves), decreases tissue damage.
Chemical means
Perior instillation of DMSO enhances absorption of
chemotherapeutic drugs, e.g. Paclitaxal, Pirarubicin.Intravesicle
instillation of Saponin before administration of anticancer drugs,
e.g. 4-0-Tetrahydropyranaldoxorubicin (THP).Increases
concentrations of THP in bladder tissues. Topical administration of
chitosan & cyclodextrin-disturbs intracellular tight junction.
Increases paracellular transport.
FUTURE PROSPECTIVES IN VDDS
Aquasomes
Three layered self assembly compositions with ceramics carbon
nanocrystalline particulate core coated with, glassy cellobiose
specific targeting and molecular shielding[41]
Cryptosmes
Lipid vesicles with a surface coat composed of pc and of suitable
polyoxoyethylene derivative of phosphotidyl ethanolamine.
Capable of Ligand mediated drug targeting.
Discomes: Niosomes solubilized with non ionic surfactant
solutions (polyoxyethylene cetyl ether class). Show ligand
mediated drug targeting.
Table 4: Non-ionic surfactants and coating carriers used for the preparation of proniosomes
Non-ionic Coating materials Non-ionic Coating materials
Surfactants Investigated surfactants Investigated
Span20 Sucrose stearate Tween60 Lactose monohydrateSpan80
Span40 Sorbitol Tween80 Spay-dried lactose
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Emulsomes: Nanosize Lipid particles (bioadhesives
nanoemulsion) consisted of microscopic lipid assembly with
apolar core used parenteral delivery of poor water soluble drugs.
Enzymosomes: Liposomal constructs engineered to provide a
mini bioenvironmental in which enzymes are covalently
immobilized or coupled to the surface of liposomes. Targeted
delivery to tumor cell.
Genosomes: Artificial macromolecular complexes for functional
gene transfer .Cationic lipids are most suitable because they
possess high biodegradability and stability in the blood stream.
Cell specific gene transfer.
Photosomes: Photolysase encapsulated in liposomes, which
release the content photo triggered charges in membrane
permeability characteristics.
Virosomes: Liposomes spiked with virus glycoprotein,
incorporated into the liposomal bilayers based on retro viruses’
derived lipids.
Vesosomes: Nested bilayer compartment in vitro via the inter
digested bilayer phase formed by adding ethanol to a variety of
saturated phospholipids. Multiple compartments of the vesosomes
give better protection to the interior contents in serum.
Proteosomes: High molecular weight multi-submit enzyme
complexes with catalytic activity, which is specifically due to the
assembly pattern of enzymes. Better catalytic activity turnover
than non associated enzymes.
Emulsomes: Hb containing liposome engineered by immobilizing
Hb with polymerisable phospholipids.
Erythrosomes: Liposomal system in which chemically cross
linked human erythrocytes used as support to which lipid bilayer is
coated.
Enzymosomes: Enzymes are co-valently immobilized or coupled
to the surface of liposomes.
Archaeosome: made from natural archaeal membrane lipids
and/or synthetic lipid analogues have been extensively studied for
potential applications in drug and vaccine delivery over the past
decade only. Archaeal-type lipids consist of archaeol (diether)
and/or caldarchaeol (tetraether) core structures wherein regularly
branched and usually fully saturated phytanyl chains (20-40
carbons in lengths), are attached via ether bonds to the sn-2, 3
carbons of the glycerol backbone. Archaeosomes constitute a
novel generation of liposomes that exhibit high stabilities to low or
high temperatures, acidic or alkaline pH, oxidative conditions, high
pressure, action of phospholipases, bile salts and serum proteins.
These properties associated with a good safety profile are
beneficial for nanotechnological applications in drug and gene
delivery. Additionally, archaeosome formulations could be used as
efficient carriers of antigens and/or adjuvants promoting antigen-
specific, humoral and cell-mediated immune responses, in
addition to antigen-specific mucosal immune responses in the
vaccinated hosts. The immune responses are well sustained over
time, and are subject to strong memory responses. Nanodelivery-
based vaccinations using archaeosomes could then represent a
promising approach for treating and preventing infections,
allergies, and neoplastic or cancer diseases. In this review, the
Non-ionic Coating materials Non-ionic Coating materials
surfactants Investigated Surfactants Investigated
Maltodextrin M700 Span60 Maltodextrin M500
Tween20 Glucose monohydrate
13
 Ashara et. al.
few recent US, World and European patents developing
archaeosomes for these biotechnological applications in Health
are discussed.[42]
CONCLUSION
Utilization and solving of critical issues of pharmaceutics field by
outstanding examples of vesicular drug delivery system such as
liposome, niosome etc. have been proved themselves as novel
approach and very much useful as drug delivery system in current
field and have remarkable place in pharmaceutical dosage forms
over and above to the conventional drug delivery system.
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