NIOSOMES

CONTENTS
 Introduction
 Composition of Niosomes
 Structure of Niosomes
 Advantages of Niosomes
 Disadvantages of Niosomes
 Comparaison with Liposomes
 Methods of Preparation
 Characterization of
Niosomes
 Applications of Niosomes.
 Marketed product
 Introduction
 Niosomes are a novel drug delivery system, in which the medication is
encapsulated in a vesicle composed of a bilayer of non-ionic surface
active agents .
 Niosomes are essentially non-ionic surfactant based multilamellar or
unilamellar vesicles in which an aqueous solution of solute (s) is
entirely enclosed by a membrane resulted from the organization of
surfactant macromolecules as bilayers”
 These are very small, and microscopic in size that lies in the nanometric
scale. Although structurally similar to liposomes, they offer several
advantages over them.
 Niosomes made-up of self assembly of hydrated nonionic surfactant
molecules (eg: tweens and spans) with or with out cholesterol and
dicetyl phosphate.
 Niosomes have recently been shown greatly increase use in transdermal
drug delivery and also in targeted drug delivery.
 Used for a variety of drugs : accommodate hydrophilic, lipophilic as
well as amphiphilic moieties.
 Most surface active agents when immersed in water yield micellar
structures, how ever some surfactants can yield bilayered vesicles
which are niosomes.
 A diverse range of materials have been used to form niosomes such as
sucrose ester surfactants and polyoxyethylene alkyl ether surfactants,
alkyl ester, alkyl amides, fatty acids and amino acid compound.
 Niosomes also called as Nonionic surfactant vesicles or
NSV s

Nios = non ionic surfactant

somes= vesicles
COMPOSITIONS OF
NIOSOMES
 The two major components used for the preparation of niosomes
are,
1. Cholesterol
2. Nonionic surfactants
3. Cholesterol: Cholesterol is used to provide rigidity
and proper shape, conformation to the niosomes
preparations.
4. Nonionic surfactants: The surfactants play a major role in
the formation of niosomes.The following non-ionic
surfactants are generally used for the preparation of
niosomes.
 E.g. Spans (span 60, 40, 20, 85, 80)
Tweens (tween 20, 40, 60, 80) and
Brijs (brij 30, 35, 52, 58, 72, 76).
 Structure Of
Niosomes
 Niosomes are microscopic lamellar structures, which are formed on the
admixture of non-ionic surfactant of the alkyl or dialkyl polyglycerol
ether class and cholesterol with subsequent hydration in aqueous
media.
 Niosomes may be unilamellar or multilamellar depending on the
method used to prepare them.
 The hydrophilic ends are exposed on the outside and inside of the
vesicle, while the hydrophobic chains face each other within the
bilayer.
 Hence, the vesicle holds hydrophilic drugs within the space enclosed in
the vesicle, while hydrophobic drugs are embedded within the bilayer
itself.
 Advantages of
niosomes
They are osmotically active and stable.
 They increase the stability of the entrapped
drug . require any special
 Handling and storage of surfactants do
 not
Can conditions
increase the oral bioavailability of drugs
 Can enhance the skin penetration of drugs
 They can be used for oral, parenteral as well topical use.
 The surfactants are biodegradable, biocompatible, and non-
immunogenic .
 Improve the therapeutic performance of the drug by protecting it from
the biological environment and restricting effects to target cells, thereby
reducing the clearance of the drug.
 In cosmetics, niosomes was first used by L’Oreal as they offered the
following advantages:
 The vesicle suspension being water based offers greater patient compliance
over oil based systems
 Since the structure of the noisome offers place to accommodate hydrophilic,
lipophilic as well as amphiphilic drug moieties, they can be used for a
variety of drugs.
 The characteristics such as size, lamellarity etc. of the vesicle can be varied
depending on the requirement.
 The vesicles can act as a depot to release the drug slowly and offer a
controlled release.
Disadvantages/problems associated
with niosomes

 As high input of energy is required in the size reduction of niosomes,

they aggregate and fuse together on prolonged storage.
 Physicochemical instability remains the major problem in
the
development of Niosomal system at industrial levels.
DISTRIBUTION OF DRUGS IN NIOSOMES
 Carriers for both lipophilic & hydrophilic drugs
 Highly hydrophilic drugs are exclusively located in the aqueous
domain
 Highly lipophilic drugs are entrapped within the lipid bilayers of
the niosomes
 Drugs with intermediary partition coefficient equilibrate
b/w lipid & aqueous domains
Types of Niosomal system on basis of
sizes
 1. Small unilamellar vesicles:
(SUV, size - 20-50 nm )

2. Multilamellar vesicles:
(MLV, size 100-1000 nm) They are formulated by hand-shaking
method. They show variations in lipid compositions.

3. Large unilamellar vesicles:

(LUV, size >0.10 μm), the injections of non ionic surfactants
solubilised in an organic solvent into an aqueous buffer, can
result in spontaneous formation of LUV. But the better method of
preparation of LUV is Reverse phase evaporation, or by
Detergent solubilisation method.
Small Large
Unilamellar Multilamellar
Unilamellar Vesicle
Vesicle Vesicle
(SUV) (MLV)
(LUV)

Typical Size Ranges: SLV: 20-50 nm – MLV:100-1000 nm

COMPARISION OF NIOSOMES VS
LIPOSOMES
 In both basic unit of assembly is Amphiphiles, but they are
phospholipids in liposomes and nonionic surfactants in
niosomes.
 Niosomes behave in-vivo like liposome, prolonging the
circulation of entrapped drug And altering its organ distribution
and metabolic stability .
 Both can entrap hydrophilic and lipophilic drugs.
 Both have same physical properties but differ in their chemical
composition.
 Niosomes has higher chemical stability than liposomes.
 Niosomes ~ made of uncharged surfactant molecules
 Liposomes ~ made of neutral or charged phospholipids.
Similarity: Niosomes &
Liposomes
 Function
 Increase the bioavailability
 Decrease the clearance
 Used for targeted drug delivery
 Properties depends on both composition of bilayer and method
of preparation
Different Niosomes
Types
1. Bola-Surfactant containing Niosomes:
Niosomes made of alpha,omega-hexadecyl-bis-(1-aza-18-crown-6)
(Bola- surfactant)-Span 80-cholesterol (2:3:1 molar ratio) are named as
Bola- Surfactant containing Niosomes.
2. Proniosomes:
A dry product which may be hydrated immediately before use to yield
aqueous Niosome dispersions.These ‘proniosomes’ minimize problems of
Niosome physical stability such as aggregation, fusion and leaking, and
provide additional convenience in transportation, distribution, storage, and
dosing.
In short;
3. Carrier + Surfactants = Proniosomes
4. Proniosomes + H2O = Niosomes
 In case of Frusemide delivery in the body, it has been found that
proniosomal formulations have been found effective .
Formation of niosomes from
proniosomes
Factors Affecting NIOSOMES
Formation
 Non ionic surfactant structure

 Membrane additives

 Nature of the encapsulated material/drug

 Surfactant & lipid levels

 Temperature of hydration
 alkyl
group
Should beabove chain length :
the gelto liquid C12-C18 Span
phase transition
temperature of Hydration Non-ionic
the system
surfactants
Temperatur
surfactant with HLB values
e
nature 4 and 8

Factors
affecting
Surfactant niosomes
s and lipid Membrane
formatio additives
levels n
surfactant/
lipid Nature of Cholesterol:
ratio: 10-30 encapsulate
mM d drug Preve
Dicetyl pnt
hosphate: -
v
cha
esiclre aggregation.
6
ge
3
Nature of non-ionic
surfactant
 Type of surfactant influences encapsulation efficiency, toxicity,
and
stability of niosomes.
 The alkyl group chain length is usually from C12-C18.
 Span surfactants with HLB values between 4 and 8 were found to be
compatible with vesicle formation.
 The water soluble detergent polysorbate 20 (HLB value 16.7)
also forms niosomes with cholesterol.
 Polyglycerol monoalkyl ethers and polyoxylate analogues are the most
widely used single-chain surfactants.
Membrane additives
1.Cholesterol:
 Cholesterol, a natural steriod, is the most commonly used membrane
additive.
 Usually incorporated in 1:1 molar ratio.
 Prevent vesicle aggregation by the inclusion of molecules that stabilize
the system against the formation of aggregates by repulsive steric or
electrostatic effects.
 Leads to the transition from the gel state to liquid phase in niosomes
systems.
2. Dicetyl phosphate and Stearic
acid:
 Dicetyl phosphate provides negative charge to
 vesicles.
It is used to prevent aggregation of hexadecyl ether
diglycerol niosomes.
 Stearic acid is used in the preparation of cationic niosomes.
Surfactant and lipid
levels
 The surfactant/lipid ratio is generally 10-30 mM (1-2.5% w/w).
 If the level of surfactant/lipid ratio is high, increasing
the
surfactant/lipid level increases the total amount of drug encapsulated.

Hydration temperature
 The hydrating temperatures used to make niosomes should usually be
above the gel to liquid phase transition temperature of the system.
METHOD OF
PREPARATION
 Ether Injection Method
 Film Method
 Sonication
 Reverse Phase Evaporation
 Micro fluidization Method
 The bubble method
Ether Injection Method
 Slow injection of an ether solution of niosomal ingredients into an
aqueous medium at high temperature.
 A mixture of surfactant and cholesterol (150 μmol) is dissolved in ether
(20 ml) and injected into an aqueous phase (4 ml) using a 14- gauge
needle syringe.
 Temperature of the system is maintained at 60oC during the process.
 Vaporization of ether leads to formation of single layered vesicles.
Depending upon the conditions used, the diameter of the vesicle range
from 50 to 1000 nm.
 Niosomes in the form of large unilamellar vesicles (LUV) are formed.
 14
Ether injection Method:
guage
needle

6
9
 Film method(Hand shaking
method):
 The mixture of surfactant and cholesterol is dissolved in an organic
solvent (e.g. diethyl ether, chloroform, etc.) in a round-bottom flask.
 Organic solvent is removed at room temperature using rotary
evaporator leaving a thin layer of solid mixture deposited on the wall of
the flask.
 Dried surfactant film hydrated with aqueous phase at 50-60°C with
gentle agitation.
 This process forms typical multilamellar niosomes Multilamellar
vesicles (MLV).
Hand shaking method:

Rotary
evaporat6o7r
 Sonication
 Aliquot of drug solution in buffer is added to the surfactant/cholesterol
mixture in a vial.
 Homogenized using a sonic probe.
 Mixture is probe sonicated at 60°C for 3 minutes using a sonicator with
a titanium probe to yield niosomes.
 The resultant vesicles are of small unilamellar (SUV) type niosomes.
 The SUV type niosomes are larger than SUV type liposomes.
 It is possible to obtain SUV niosomes by sonication of MLV
type
vesicles.
 Probe sonicator used for small volume samples.
 Bath sonicator used for larger volumes samples.
 Probe sonicator

Sonicators use probes that are

put directly into the sample to
be sonicated.
Bath
Sonicator
Sonicators produce sound waves
into a water bath, where
samples are placed.
Reverse phase evaporation
 Cholesterol and surfactant (1:1) are dissolved in chlorofom and 0.25
volume of phosphate saline buffer (PBS) is emulsified to get w/o
emulsion.
 The mixture is sonicated and subsequently chloroform is evaporated
under reduced pressure.
 The surfactant first forms a gel and then hydrates to form niosomal
vesicles.
 The vesicles formed are unilamellar and 0.5 μ in diameter.
Reverse phase evaporation
technique :
Surfactant and cholesterol is dissolved in chloroform and
0.25 volume of PBS buffer is emulsified to get a W/O
emulsion.
sonicated
chloroform is evaporated under reduced pressure.

The lipid or surfactant forms a gel first and hydrates to form

vesicles.

Free drug (unentrapped) is generally removed by dialysis.

Micro fluidization Method
 Micro fluidization 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.
 A microfludizer is used to pump the fluid at a very high pressure
(10,000 psi) through a screen.
 It is then forced along defined micro channels, which direct two
streams of fluid to collide together at right angles, thereby affecting a
very efficient transfer of energy.
 The lipids/surfactants can be introduced into the fluidizer.
 The fluid collected can be recycled until spherical vesicles are obtained.
 Uniform and small sized vesicles are obtained.
 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 are positioned in the first and
second neck and nitrogen supply through the third neck.
 Cholesterol and surfactant are dispersed together in the buffer (pH
7.4) at 70°C, the dispersion mixed for 15 secs. with high shear
homogenizer and immediately afterwards “bubbled” at 70°C using
nitrogen gas.
 Niosomes are formed.
Bubble
method:
It is novel technique for the
one step preparation of
liposomes and niosomes
without the use of organic
solvents.
RBF as bubbling unit with three necks in
water bath.
Reflux , thermometer and nitrogen supply
by three necks
Cholesterol+ Surfactant dispersed in buffer
pH 7.4 at 70°C
Above dispersion is homogenized for 15 sec
and then bubbled with nitrogen gas at 70°C
to get niosomes
71
 Drugs incorporated into niosomes
by various methods
Method of preparation Drug incorporated

Ether Injection Sodium stibogluconate

Doxorubicin

Hand Shaking Methotrexate

Doxorubicin

Sonication 9-desglycinamide
8-arginine
Vasopressin
Oestradiol

Reverse phase evaporation Diclofenac

sodium
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 undergo process of freezing and thawing
3 times and later sonicated.
 To this niosomal suspension, aqueous solution containing 10 mg/ml of
drug is added and vortexed.
 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.
 Post-Preparation Processes
 Size reduction of niosomes
 Separation of unentrapped
material
Post-Preparation Processes

Size reduction of Separation of

niosomes unentrapped material
Size reduction methods
 Probe sonication 100 -140nm

 Extrusion method in the range of

140nm

 Sonication & filtration in the range of

200nm

 Microfluidizer <50nm

 High pressure homogenization <100nm

 Sonication
Sonication
ML hazy
Vs transparent 5-10 min
solution
centrifugation 30
min

clear
SUV
Disper
sion.
Extrusion method
 The size of niosomes is
reduced by gently passing
them
through
polycarbonate membrane
filter of defined pore size
at lower pressure.
Separation of Unentrapped
Drug
1. Dialysis:
The aqueous niosomal dispersion is dialyzed in a dialysis
tubing against phosphate buffer or normal saline or glucose
solution.
2. Gel Filtration:
The unentrapped drug is removed by gel filtration of
niosomal dispersion through a Sephadex-G-50 column and
elution with phosphate buffered saline or normal saline.

Gel
Filtration
3.
Centrifugation:
 The niosomal suspension is centrifuged and the supernatant is
separated. The pellet is washed and then resuspended to obtain a
niosomal suspension free from unentrapped drug.

Centrifuser
Characterization of Niosomes

Size, Shape and Morphology:

• Scanning electron microscopy (SEM): Particle size
analysis was done by scanning electron microscopy (SEM)
 Freeze Fracture Electron Microscopy: Visualize
the vesicular structure of surfactant based vesicles.
 Photon Correlation spectroscopy : Determine
mean diameter of the vesicles.
 Electron Microscopy : Morphological studies of vesicles.
Entrapment
efficiency
 After :preparing niosomal dispersion, unentrapped drug is
separated by dialysis, centrifugation, or gel filtration.
 The drug remained entrapped in niosomes is determined by
complete vesicle disruption using 50% n-propanol or 0.1% Triton X-
100 and analysing the resultant solution by appropriate assay method
for the drug.
Entrapment efficiency (EF)% = (Amount of drug
entrapped/ total amount of drug) x100
 Bilayer formation :Assembly of non-ionic surfactants to
form bilayer vesicle is characterized by light polarization
microscopy.
Number of lamellae :It is determined by using NMR
spectroscopy, small angle X-ray scattering and electron
microscopy .
Membrane rigidity :
 The biodistribution and biodegradation of niosomes
are influenced by rigidity of the bilayer.
 Membrane rigidity can be measured by means of NMR,
differential scanning calorimetry (DSC) and fourier
transform- infra red spectroscopy (FTIR) techniques.
Vesicle Surface Charge:
 The vesicle surface charge can play an important role in the behavior of
niosomes in vitro and in vivo.
 In general, charged niosomes are more stable against aggregation and
fusion than uncharged vesicles.
 In order to obtain an estimate of the surface potential, the zeta potential
of individual niosomes can be measured by microelectrophoresis.
 In-vitro
 release : in-vitro release rate study includes the use of dialysis
A method of
tubing.
 The vesicle suspension is pipetted into a bag made up of the tubing and
sealed.
 The bag containing the vesicles is placed in 200 ml of buffer solution in
a 250 ml beaker with constant shaking at 25°C or 37°C.
 At various time intervals, the buffer is analyzed for the drug content by
an appropriate assay method.
STABILITY OF
NIOSOMES
 Vesicles are stabilized based upon formation of following forces:
 Van der Waals forces among surfactant molecules.
 Electrostatic repulsive forces are formed among vesicles upon
addition of charged surfactants to the double layer, enhancing the
stability of the system.
 Niosomes in the form of liquid crystal and gel can remain stable at both
room temperature and 4oC for 2 months.
 Recommended temperature of storage 4oC.
 Ideally niosomes should be stored dry for
reconstitution.
 The factors which affect the stability of niosomes:
 Type of surfactant
 Nature of encapsulated drug
 Storage temperature
 Detergents
 Inclusion of charged molecule
 Stability study of
 Niosomes:
All niosomal formulations were subjected to stability studies by
storering at 4°C, 25°C and 37°C in thermostatic oven for the period of
three months.
 After one month, drug content of all the formulations were checked by
method discussed previously in entrapped efficiency parameter. In-vitro
release studies of selected formulations were also carried out.
APPLICATIONS OF
NIOSOMES

Applications

Ophthalmi
Transdermal Parenteral Peror Antineoplastic c Drug
al delivery
Anti-neoplastic treatment
 Most antineoplastic drugs cause severe side effects. Niosomes can alter
the metabolism, prolong circulation and half life of the drug, thus
decreasing the side effects of the drugs.
 Niosomal entrapment of Doxorubicin and Methotrexate (in two
separate studies) showed beneficial effects over the unentrapped drugs,
such as decreased rate of proliferation of the tumor and higher plasma
levels accompanied by slower elimination.
 Niosomes for the treatment of
 Leishmaniasis:
Leishmaniasis is a disease in which a parasite of the
genus Leishmania invades the cells of the liver and spleen.
 Niosomes are being used for the delivery of stibogluconate
an
antileishmaniasis agent for its delivery to visceral organs.
Delivery of peptide drugs:
 Oral peptide drug delivery has long been faced with a challenge
of by passing the enzymes which would breakdown the peptide.
 Use of niosomes to successfully protect the peptides
from gastrointestinal peptide breakdown is being investigated.
 Oral delivery of 9-desglycinamide, 8-arginine and
vasopressin
entrapped in niosomes increase stability of peptide significantly.
 Use in studying immune
 response:
Due to their immunological selectivity, low toxicity and greater
stability; niosomes are being used to study the nature of the immune
response provoked by antigens.
Niosomes as carriers for haemoglobin:
 Niosomes can be used as carriers for haemoglobin within the blood.
The niosomal vesicle is permeable to oxygen and hence can act as a
carrier for haemoglobin in anemic patients.
Parenteral Applications
 Niosomes in sub-micron size are used for parenteral administration
 Niosomal vesicles up to 10 μm are administered via I.V. or I.M.

.
Transdermal drug delivery systems utilizing
niosomes:
 One of the most useful aspects of niosomes is that they greatly enhance
the uptake of drugs through the skin.
 Transdermal drug delivery utilizing niosomal technology is widely used
in cosmetics, in fact, it was one of the first uses of the niosomes.
 Topical use of niosome entrapped antibiotics to treat acne is done. The
penetration of the drugs through the skin is greatly increased as
compared to un-entrapped drug. Example Oestradiol
Ophthalmic Drug Delivery:
 Niosomes> 10 μm are suitable for drug administration to eye.
 Example: Cyclopentolate (Polysorbate 20 and cholesterol were used
for niosomes formulation).
Radiopharmaceuticals:
 First application of niosomes as radiopharmaceuticals demonstrated by
Erdogan et al. in 1996.
MARKETED
PRODUCT:

 Lancôme has come out with a variety of anti-ageing

products which are based on noisome formulations.
 L’Oreal is also conducting research on anti-ageing cosmetic
products.
 RECENT ADVANCES IN
NIOSOMES
Combination of PEG and glucose conjugates on the surface of
niosomes significantly improved tumor targeting of an encapsulated
paramagnetic agent assessed with MR imaging in a human carcinoma.

 Phase I and phase II studies were conducted for Niosomal

methotrexate gel in the treatment of localized psoriasis. These studies
suggest that niosomal methotrexate gel is more efficacious than placebo
and marketed methotrexate gel.
References:
 Niosomes: A Novel Drug Delivery System. V. Pola Chandu et al. 2277
– 2782. IJNTPS.
 Modern Pharmaceutics, 5th edition Volume 2 : Applications and
advances by Alexander T. Florence, page no. 357-358