I am a pharmacist. These slide describe biotechnology topic noisome. I hope students get good benefits about it.

1. NIOSOMES

2. 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

3. 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 .
 Nios= non ionic surfactants
 Somes = body NSV=niosomes
 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.

4.  Niosomes made-up of self assembly of hydrated
nonionic surfactant molecules alkyl or dialkyl
polyglycerol ether (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, however some surfactants can
yield bilayered vesicles which are niosomes.
 Surfactants don’t assemble into closed bilayers
spontaneously and typically required input of energy as
physical agitation or heat.

5.  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.
PROPERTIES OF NSV:
 Drug carrier
 Biodegradable, biocompatible and non immunogenic.
 Large quantity of drug can load.
 Very small in size nm
 Primarilly SAA (surface active agent) when immersed in water yield
micellar structure, how ever some surfactants( havning specific
shape) can yield bilayered vesicles which are Niosomes.

6. COMPOSITIONS OF
NIOSOMES
 The two major components used for the preparation of
niosomes are,
1. Cholesterol
2. Nonionic surfactants
1. Cholesterol: Cholesterol is used to provide rigidity and
proper shape, conformation to the niosomes preparations.
2. 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).

7. 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.

10. CRITICAL PACKING PARAMETERS:
 Important parameters is shape of surfactant
 If ratio of hydrophobic properties of surfactants to
hydrophilic properties of surfactants.
 Expressed as ratio of volume of hydrophobic chain to
product of length of hydrophobic chain(lc) and cross
section area of hydrophilic head group (ao).
CPP= V/ lc ao

11. One surfactant molecule

14. 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/ 1 micro m) exhibit increased-trapped
volume and equilibrium solute distribution, and require hand-shaking
method. They show variations in lipid compositions.
3. Large unilamellar vesicles: 100-200 nm
(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.
 Oligolamellar vesicles(OLv): 2-5 layers of surfactant 100-1000nm.

15. Small
Unilamellar
Vesicle
(SUV)
Large
Unilamellar
Vesicle
(LUV)
Multilamellar
Vesicle
(MLV)
Typical Size Ranges: SLV: 20-50 nm – MLV:100-1000 nm

16. 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.
 Niosome can also prepared for targeting like liposomes such
as pH dependent, charged, immunogenic.

19. 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

20. ADVANTAGES OF NIOSOMES OVER LIPOSOMES
o Ester bonds of phospholipids are easily hydrolyzed, this
can lead to phosphoryl migration at low PH.
o Peroxidation of unsaturated phospholipids.
o As liposomes have purified phospholipids they are to be
stored and handled at inert(N2) atmospheres where as
Niosomes are made of non ionic surfactants and are
easy to handle and store.
o Phospholipid raw materials are naturally occurring
substances and as such require extensive purification
thus making them costly.
o Niosomes has high adjuvanticity when compared to
liposomes.

21. ADVANTAGES OF NIOSOMES
They are osmotically active and stable.
They increase the stability of the entrapped drug .
Handling and storage of surfactants do not require any
special conditions
Can 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.

22. In cosmetics 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.

23. 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

25. 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;
1. Carrier + Surfactants = Proniosomes
2. Proniosomes + H2O = Niosomes
 In case of Frusemide delivery in the body, it has been found
that proniosomal formulations have been found effective .

26. Formation of niosomes from
proniosomes

28. Factors Affecting NIOSOMES
Formation
 Non ionic surfactant structure
 Membrane additives
 Nature of the encapsulated material/drug
 Temperature of hydration

29. Factors
affecting
niosomes
formation
Non-ionic
surfactant
nature
Membrane
additives
Nature of
encapsulate
d drug
Surfactants
and lipid
levels
Hydration
Temperatur
e
alkyl group
chain length
: C12-C18
Span
surfactants
with HLB
values
4 and 8
Cholesterol:
Prevent vesicle
aggregation.
Dicetyl
phosphate: -ve
charge
surfactant/lipi
d ratio: 10-30
mM
Should be
above the
gel to liquid
phase
transition
temperature
of the
system
63

30. 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.

31. 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.
 As the result, niosomes become less leaky.

32.  2.Dicetyl phosphate and Stearic acid:
 Dicetyl phosphate provides negative charge to
vesicles.
 It is used to prevent aggregation of hexadecyl
diglycerol ether (C16G2) niosomes.
 Stearic acid is used in the preparation of cationic
niosomes.

33. Hydration temperature:
• The hydrating temperatures used to make niosomes
should usually be above the gel to liquid phase
transition temperature of the system.

34. METHOD OF PREPARATION
 Materials:
1.Non ionic surfactant ( hydrophilic group surfactant)
Glycerol, ethylene oxide , crown ether, polyhydroxy
head group, sugar head as galactose, mannose,
lactose, glucose)
2.Charge inducers( to prevent aggregation they
charged to repel each other)
3.Cholesterol
Hydrophobic moiety: 1or 2 alkyl gr or perflouroalkyl gr
or sometime single steroidal gr.

35. METHOD OF PREPARATION
1.Physical dispersion:
a) Hand shaking Film Method
b) Micronization
c) Ultra Sonication
d) Freez thaw sonication
e) Membrane extrusion
2. Solvent dispersion
a) Ether Injection Method
b) Reverse Phase Evaporation
c) Micro fluidization Method
 The bubble method
 Proniosomes

36. 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.

37. Ether injection Method: 14 guage
needle
69

38. 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).

39. Hand shaking method:
Rotary
evaporator
67

40. 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.

41. Probe sonicator
Sonicators use probes that are
put directly into the sample to
be sonicated.

42. Bath Sonicator
Sonicators produce sound
waves into a water bath,
where samples are placed.

43. 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.

44. 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.
Reverse phase evaporation technique :

45. 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 microfluidizer 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.

46. Bubble method
 It is novel technique for the one step preparation of
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.

47. Bubble method:
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
It is novel technique for
the one step
preparation of
liposomes and niosomes
without the use of
organic solvents.
71

48. Method of preparation Drug incorporated
Ether Injection Sodium stibogluconate
Doxorubicin
Hand Shaking Methotrexate
Doxorubicin
Sonication
Reverse phase evaporation
9-desglycinamide
8-arginine
Vasopressin
Oestradiol
Diclofenac sodium
Drugs incorporated into niosomes
by various methods

49. 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.

50. Post-Preparation Processes
 Size reduction of niosomes
 Separation of unentrapped material
Post-Preparation
Processes
Size reduction
of niosomes
Separation of
unentrapped
material

51. 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

52. Sonication
MLVs hazy transparent
5-10 min solution
centrifugation 30 min
clear SUV
Dispersion.
SONICATIO
N

53. Extrusion method
 The size of niosomes
is reduced by gently
passing them through
polycarbonate
membrane filter of
defined pore size at
lower pressure.

54. 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.
Constant stirring at 100rpm on a magnetic stirrer at 37 oC.
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

55. 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

57. 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.

58. EVALUATION PARAMETERS
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(
surfactant) or 2.5% Na lauryl sulfate and
analysing the resultant solution by appropriate
assay method for the drug.
Entrapment efficiency (EF)% = (Amount of drug
entrapped/ total amount of drug) x100
2. ENCAPSULATION VOLUME/ TRAPPED

59. 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.

60. 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.

61. In-vitro release :
A method of in-vitro release rate study includes the use
of dialysis 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.

63. 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.

64.  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

65. 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.

66. APPLICATIONS OF NIOSOMES
Applicatio
ns
Transderm
al
Parenteral Peroral
Antineopla
stic
Ophthalmi
c Drug
delivery

67. 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.

68. 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.

69. 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.
.

70. 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

71. 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.

72. 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.

73. 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.
A research article was published that Acyclovir entrapped
niosomes were prepared by Hand shaking and Ether
injection methods increases the oral bioavailability.