method of preparation, evaluation, applications of niosomes,aquasomes,phytosomes and electrosomes

1. PRESENTED BY:
AYUSHI JOSHI
M PHARM SEM – 2
ROLL NO: MU003
GUIDED BY:
DR.TEJAL SONI
HOD PHARMACEUTICS

2. CONTENT
 Introduction
 Structure
 Types
 Composition
 Method of preparation
 Evaluation
 Applications

3. NIOSOMES

4. INTRODUCTION
 Niosomes are a novel drug delivery system, in which the
medication is encapsulated in a vesicle. The vesicle is composed
of a bilayer non-ionic surface active agents and hence the name
niosomes.
 The niosomes are very small, and microscopic in size. Their size
lies in the nanometric scale. Although structurally similar to
liposomes, they offer several advantages over them.
 Niosomes have recently been shown to greatly increase
transdermal drug delivery and also can be used in targeted drug
delivery, and thus increased study in these structures can provide
new methods for drug delivery.

5. STRUCTURE OF NIOSOME
• Structurally, niosomes are similar to liposomes, in
that they are also made up of a bilayer.
• However, the bilayer in the case of niosome is made
up of non-ionic surface active agents rather than
phospholipids as seen in the case of liposomes.
• Most surface active agents when immersed in water
yield micellar structures however some surfactants
can yield bilayer vesicles which are niosomes.

6. •Liposomes may be
unilamellar or multimellar
depending upon method
used to prepare them.
•The niosome is made of a
surfactant bilayer with its
hydrophilic ends exposed on
the outside and inside of the
vesicle, while the
hydrophobic chains face
each other within the
bilayer.

7. Niosome Liposome
1. Less expensive 1. More expensive
2. Chemically stable 2. Chemically unstable
3. Niosomes are prepared from
uncharged single-chain surfactant.
3. Liposomes are prepared from double-
chain phospholipids.
4. They do not require special storage
and handling.
4. They require special storage, handling
& purity of natural phospholipid is
variable.
5. Non ionic drugs carriers are safer. 5. The ionic drugs carriers are relatively
toxic & unsuitable.

8. TYPES OF NIOSOMES
1) Multi lamellar vesicles (MLV)
2) Large unilamellar vesicles (LUV)
3) Small unilamellar vesicles (SUV)

9. COMPOSITION OF NIOSOME
• 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 role surfactants play a
major role in the formation of niosomes. the
following non-ionic surfactants are generally
used for the preparation of niosomes.

10. • E.g.
Spans (span 60, 40, 20, 80)
Tweens (tween 20, 40, 60, 80)
• The non ionic surfactants possess a
hydrophilic head and a hydrophobic tail.

11. METHOD OF PREPARATION
• The preparation methods should be chosen according to the
use of the niosomes since the preparation methods influence
the number of bilayers, size, size distribution, and entrapment
efficiency of the aqueous phase and the membrane
permeability of the vesicles.
A. Ether injection method:
Surfactant + cholesterol is dissolved in diethyl ether
Then injected in warm water maintained at 60°Cthrough a 14
gauze needle
Ether is vaporized to form single layered niosomes.

12. B. Hand shaking method (thin film hydration
technique):
C. Sonication method:
Surfactant + cholesterol + solvent
Remove organic solvent at room temperature
Thin layer formed on the walls of flask
Film can be rehydrated to form multilamellar niosomes
drug in buffer + surfactant/cholesterol in 10 ml of aqueous phase
Above mixture is sonicated for 3 minutes at 60°C using titanium probe
yielding niosomes

13. D. Multiple membrane extrusion method:
 Mixture of surfactant, cholesterol in chloroform
is made into thin film by evaportion.
 The film is hydrated with aqueous drug solution
and the resultant suspension extruded throungh
polycarbonate membrane. It is a good method
for controlling niosome size.

14. E. Reverse phase evaporation technique:
(REV)
Cholesterol + surfactant dissolved in ether+ chloroform
Sonicated at 50°C and again sonicated after adding PBS
Drug in aqueous phase is added to above mixture
viscous niosomes suspension is diluted with PBS
Organic phase is removed at 40°C at low pressure
Heated on a water bath for 60°c for 10 minutes to yield niosomes.

15. F. The bubble method:
 It is novel technique for the one step preparation of
liposome 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
temprature.
 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 the
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.

16. EVALUATION OF NIOSOMES
Entrapment efficiency:
•Depend on the method of preparation.
•Niosomes prepared by ether injection method have better
entrapment efficiency than those prepared by the film or
sonication.
•Addition of cholesterol to non-ionic surfactants with single or
dialkyl-chain significantly alters the entrapment efficiency.
•Surfactants of glycerol type lead to reduction in entrapment
capacity as the amount of cholesterol increases.
•Niosomes in the form of liquid crystals possess better entrapment
efficiency than gel type vesicles.
•Entrapment efficiency (EF)=(Amount entrapped/total amount)x100
•Niosomes, similar to liposomes, assume spherical shape and so
their diameter can be determined using light microscopy, photon
correlation microscopy and freeze fracture electron microscopy.

17. EVALUATION (CONTND.)
In-vitro release:
•. A method of in-vitro release rate study includes the
use of dialysis tubing.
•A dialysis sac is washed and soaked in distilled water.
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 of
vesicles during the cycle.

18. APPLICATIONS OF NIOSOMES:
Drug targeting :
One of the most useful aspect of
niosomes is their ability to target
drugs.
Niosomes can be used to target
drugs to the reticuloendothelial
system.
It can be achieved by coating with
polymer e.g. PEG.
In diagnosis :
Niosomes have also been used as
carriers for iobitridol, a diagnostic
agent used for x-ray imaging.

19. Anti-neoplastic treatment :
Most antineoplatic 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.
Niosomes, is decreased rate of
proliferation of tumor and higher
plasma levels accompanied by
slower elimination.
Leishmaniasis :
Leishmaniasis is a disease in
which a parasite of the genus
leishmania invades the cells of
the liver and spleen.
Use of niosomes in tests
conducted showed that it was
possible to administer higher
levels of the drug without the
triggering of the side effects,
and thus allowed greater
efficacy in treatment.

20. Delivery of peptide drugs:
Oral peptide drug delivery has long been faced with a
challenge of bypassing the enzymes which would
breakdown the peptide.
Use of niosomes to successfully protect the peptides
from gastrointestinal peptide breakdown is being
investigated.
In an in vitro study conducted by oral delivery of a
vasopressin derivative entrapped in niosomes showed
that entrapment of the drug significantly increased the
stability of the peptide.
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 anaemic patients.

21. AQUASOME

22. INTRODUCTION
• Aquasomes are nanoparticulate carrier system but instead
of being simple nanoparticle these are three layered self
assembled structures.
• It comprised of a solid phase nanocrystalline core coated
with oligomeric film on which biochemically active
molecules are absorbed with or without modification.
• Aquasomes are like “bodies of water” and their water like
properties protect and preserve fragile biological
molecules, and this property of maintaining conformational
integrity as well as high degree of surface exposure is
exploited in targeting of bioactive molecules.

23. These three layered structures are self-assembled by non
covalent and ionic bonds. These carbohydrate stabilize
nanoparticles of ceramic are known as “aquasomes”.

24. STRUCTURE OF AQUASOME

25. PRINCIPLE OF SELF ASSEMBLY
• Self assembly implies that the constituent parts of some
final product assume spontaneously prescribed structural
orientations in two or three dimensional space.
• The self assembly of macromolecules in the aqueous
environment for the purpose of creating smart
nanostructure materials is governed basically by three
physicochemical processes:
1. The interactions of charged groups
2. Hydrogen bonding
3. Structural stability

26. METHOD OF PREPARATION OF AQUASOME
The method of preparation of aquasomes involves three
steps
The general procedure consists of Formation of an
inorganic core, followed by Coating of the core with
polyhydroxy oligomer, and finally loading of the drug of
choice to this assembly.

27. I- Formation of an inorganic core
 It involves the fabrication of a ceramic core, and the
procedure depends upon the materials selected.
 The two most commonly used ceramic cores are calcium
phosphate and diamond.
a) Synthesis of nanocrystalline tin oxide core ceramic - It can
be synthesized by direct current reactive magnetron
sputtering.
b) Self assembled nanocrystalline brushite (calcium
phosphate dihydrate) - These can be prepared by colloidal
precipitation and sonication by reacting solution of
disodium hydrogen phosphate and calcium chloride.
c) Nanocrystalline carbon ceramic, diamond particles -
These can also be used for the core synthesis after ultra
cleansing and sonication.

28. II- Coating of the core with
polyhydroxy oligomer
• In the second step, ceramic cores are coated with
carbohydrate (polyhydroxyl oligomer).
• The coating is carried out by addition of carbohydrate into
an aqueous dispersion of the cores under sonication.
• These are then subjected to lyophilization to promote an
irreversible adsorption of carbohydrate onto the ceramic
surface.
• The unadsorbed carbohydrate is removed by
centrifugation.
• The commonly used coating materials, are cellobiose
citrate, pyridoxal-5- phosphate, and sucrose.

29. III- Loading of the drug of choice to this
assembly
• The final stage involves the loading of drug to the coated
particles by adsorption.
• For that, a solution of known concentration of drug is
prepared in suitable pH buffer, and coated particles are
dispersed into it.
• The dispersion is then either kept overnight at low
temperature for drug loading or lyophilized after some
time so as to obtain the drug-loaded formulation (i.e.,

30. APPLICATIONS
Insulin Delivery
• Aquasomes for pharmaceuticals
delivery i.e. insulin, developed because
drug activity is conformationally
specific. Bio activity preserved and
activity increased to 60% as compared
to i.v administration and toxicity not
reported.
Oral Delivery of Enzyme
• Aquasomes also used for delivery of
enzymes like DNAase and
pigments/dyes because enzymes
activity fluctuates with molecular
conformation and cosmetic properties
of pigments are sensitive to molecular
conformation.

31.  Antigen Delivery
• The adjuvants generally used to enhance the immunity to
antigens have a tendency either to alter the conformation
of the antigen through surface adsorption or to shield the
functional groups.
• The efficacy of a new organically modified ceramic
antigen delivery vehicle.
• These particles consisted of diamond substrate coated
with a glassy carbohydrate (cellobiose) film.
• These aquasomes provided conformational stabilization
as well as a high degree of surface exposure to protein
antigen.
• Diamond, being a material with high surface energy, was
the first choice for adsorption and adhesion of cellobiose.
It provided a colloidal surface capable of hydrogen
bonding to the proteinaceous antigen.

32. As oxygen Carrier
• Aquasomes as red blood cell substitutes,
haemoglobin immobilized on oligomer surface
because release of oxygen by haemoglobin is
conformationally sensitive.
• By this toxicity is reduced, haemoglobin
concentration of 80% achieved.

33. PHYTOSOMSES

34. INTRODUCTION
• The term “phyto” means plant and “some”
means cell-like.
• Phytosomes are little cell like structures.
• This is advanced form of herbal formulations
which contains the bioactive phytoconstituents
of herb extracts surround and bound by a lipid.
• Most of bioactive constituents are water soluble
compounds like flavonoids, glycosides.
• Because of their water soluble property and
lipophilic outer layer it shows better absorption
and produce better bioavailability.

35. STRUCTURE OF PHYTOSOME
•Phytosome structures contain the active
ingredients of the herb surrounded by the
phospholipids.
•The presence of a surfactant i.e. the
phospholipids in the molecule these are
shielded from water-triggered degradation
while, at the same time, allows obtaining a
higher adhesion of the product itself to the
surface it comes into contact with and a
better interaction of various molecules with
cell structure.
•Example-pc is a bifunctional compund.
Specifically the choline head (hydrophilic)
comprising the body and tail which then
envelops the choline bound material and
forms phyto- phospholipid complex.
•Molecules are anchored through chemical
bonds to the polar choline head of the pc, it
can be demonstrated by specific
spectroscopic techniques.

36. CHEMICAL PROPERTIES
• Phyosome is a complex between natural product
and natural phospholipid.
• The phytosome complex is obtained by reaction of
suitable amount of phospholipid and the substrate
in appropriate solvent such as glycerol.
• The main phospholipid-substrate interaction is due
to formation of hydrogen bonds between polar head
of phospholipid and polar functionalities of
substrate.
• When treated with water, they assume a micelle
shape, forming structures which resembles
liposomes.

37. BIOLOGICAL PROPERTIES
• Phytosomes are advanced forms of herbal
products that are better absorbed, utilized and
as a result produce better results than
conventional herbal extracts.
• Freely soluble in non-polar and aprotic solvent ;
Solvents in which the hydrophilic moiety is not
present.
• Moderately soluble in fats.
• Insoluble in water.

38. ADVANTAGES
• Enhanced absorption of herbal constituent.
• As the absorption of active constituents is improved, its dose
requirement is also reduced.
• Phosphotidylcholine acts as hepatoprotective, giving
synergistic effect.
• It shows better stability profile.
• It assures proper delivery of drugs to the respective tissues.
• Entrapment efficiency is high.
• Phyotosomes are also superior to liposomes in skin care
products.
• Better bioavailability.
• Nutritional benefit.
• Enhanced permeation of drug through skin

39. DISADVANTAGES:
• When administered orally or topically they limit
their bioavailability.
• Phytoconstituents is quickly eliminated from
phytosome.
• Stability problem.

40. PREPARATION OF PHYTOSOME
Phospholipid
Dissolved in organic solvent containing drug extract
Solution of phospholipid in organic solvent with drug extract
Drying/ solvent evaporation
Formation of thin film
Hydration
Formation of phytosomal suspension

41. 1. Antisolvent precipitation technique
Drug + soya lecithin
Refluxed with20ml dichloromethane at 60 degree for 2hrs
Concentrate mixture to 5-10ml
Add hexane 20ml
Filter the ppt formed. Dry, crush and pass through #100

42. 2. Rotary evaporation technique
Drug and soya lecithin
Dissolved in 30ml of tetrahydrofuran
stirring for 3hours at a temperature not exceeding 40degree
Thin film is formed
Add n-hexane with stirring
Precipitate obtained ↓ Dry and pass through mesh
Phytosomes formed

43. 3. Solvent evaporation method
Drug and soya lecithin
Refluxed with 20ml of acetone at a temperature 50-60 degree for 2hours ↓
Concentrate mixture to 5-10ml
Obtain the precipitate
Filter and collect
phytosomes obtained

44. EVALUATION
Transition temperature:
• The transition temperature of
the vesicular lipid system can be
settled via differential scanning
calorimetry.
Entrapment efficiency:
•The entrapment efficiency of a
phytosomal preparation can be
determined by exposing the
preparation to ultracentrifugation
method.
Vesicle size and zeta potential:
•The particle size and zeta potential of
phytosomes can be confirmed by
dynamic light scattering which usages a
computerized examination system and
photon carrelation spectroscopy.
Surface tension activity
measurement:
•The surface tension activity of the
drug in aqueous solution can be
determined by the Du Nouy ring
tensiometer.

45. EVALUATION (CONTD.)
Visualization:
Visualization of phytosomes
can be accomplished using
scanning electron microscopy
(SEM) and by transmission
electron microscopy.
Vesicle stability:
The steadiness of vesicles can be
measured by calculating the size
and structure of the vesicles over
time. The mean size is calculated
by DLS and structural changes are
monitored by tem.
In vitro and in vivo
evaluations:
Models of in-vivo evaluations are
selected on the basis of the expected
therapeutic activity of biologically
active phytoconstituents present in
the phytosome.

46. APPLICATION OF PHYTOSOMES
Silymarin phytosome:
•Most of the phytosomes are focused to
silybum marianum which contains
premier liver-protectant flavanoids.
•The fruit of the milk thistle plant (s.
marianum, family steraceae) contains
flavanoids known for hepato-protective
effects.
•Silymarin has been shown to have
positive effects in treating liver diseases of
various kinds, including hepatitis,
cirrhosis, fatty infiltration of the liver
(chemical and alcohol induced fatty liver)
and inflammation of the bile duct.
Silymarin phytosome.
Phytosomes of
grape seed
•Grape seed phytosome is
composed of oligomeric
polyphenols of varying
molecular size complexed
with phospholipids.
•The main properties of
paracyanidin flavanoids of
grape seed are an increase in
total antioxidant capacity and
stimulation of physiological
defenses of plasma.

47. APPLICATION (CONTD.)
Phytosome of green tea:
•Green tea leaves is characterized by presence of a
polyphenolic compound epigallocatechin 3-O-gallate
as the key component.
•These compounds are patent modulators of several
biochemical process linked to the breakdown of
homeostasis in major chronic-degenerative diseases
such as cancer an atherosclerosis.
•Green tea also furnishes us with a number of
beneficial activities such as antioxidant,
anticarcinogenic,antimutagenic,hypocholesterolemic
, cardioprotective effects.

48. ELECTROSOME

49. INTRODUCTION
• These are the transmembrane protein generate and propagate
the electrical signals that allow us to sense our surroundings,
process, information, make decisions, and move.
• Ion channel proteins act as gates that span the lipid bilayer that
surrounds all electrochemical gradients.
• The ion flux through a channel pore can be extremely high.
• They are high resolution in function and 3D structure to
description of their molecules.
• The high resolution structure of ion channel and ion channel
associated protein are providing the substrates for sophisticated
tests of the mechanisms of channel gating and permeation.
• Ion perform two basic function open and close to control the
passage of ion across the cell membrane.

50. • The electrosomes, a novel surface-display system
based on the specific interaction between the
cellulosomal scaffoldin protein and a cascade of
redox enzymes that allows multiple electron-release
by fuel oxidation.
• The electrosomes is composed of two
compartment:
 (i) a hybrid anode, which consists of dockerin-
containing enzymes attached specifically to cohesin
sites in the scaffoldin to assemble an ethanol
oxidation cascade, and
 (ii) a hybrid cathode, which consists of a dockerin-
containing oxygen- reducing enzyme attached in
multiple copies to the cohesin-bearing scaffoldin.

51. STRUCTURE OF ELECTROSOME

52. METHOD OF PREPARATION
1. Strains and Constructs method.
2. Enzyme Binding to Scaffoldin.
3. Biofuel-Cell Assembly and Characterization.
4. Protein Expression.
5. Enzyme Activity Assays.
6. Construction of YSD of Chimeric Scaffoldins.
7. Cyclic Voltammetry (CV) and
Chronoamperometry (CA).

53. ENZYME BINDING TO SCAFFOLDIN
• 2.0 mL of yeast cells displaying scaffoldin, for which absorbance at
a wavelength of 600 nm was 1.0 , were incubated with bacterial
lysates containing the expressed enzymes at room temperature
for 1 h. 1.0 mL of the bacterial lysates were used for the binding,
which was performed in a final volume of 15 ml.
• As a binding buffer, 50 mM Tris buffer at pH 8.0 with 1 mM CaCl2
was used. Upon binding, the yeast cells were precipitated, and
binding was repeated using fresh lysate.
• After the second binding cycle, the yeast cells were washed four
times in the buffer to remove non-specifically bound enzymes.
• Following binding, the yeast cells were resuspended in 2.0 ml of
buffer

54. BIOFUEL-CELL ASSEMBLY AND
CHARACTERIZATION
• Air was continuously purged to the fuel-cells. A potentiostatically
controlled anode set to −0.2 V versus Ag/AgCl was used.
• In all experiments, the cells were left to stabilize overnight,
following fuel cell assembly, before characterization was
performed.
• The characterization of fuel cell performance was done by
measuring the voltage of the cells under variable external loads.
• A background current cell was used as a negative control for all
fuel cell experiments and did not contain any yeast. Graphite rods
of 5 mm diameter served as both anodes and cathodes.
• The counter electrode that served for the potentiostatically
controlled electrode was of a larger surface area, as described for
the CV and CA measurements

55. ADVANTAGES
• It perpetuates the endurance of active drug molecule in the
systemic circulation. Deferment the elimination reactions
of promptly metabolize drugs and contributes to
controlled release.
• Incorporates both hydrophilic and lipophilic drugs.
• Intensifies the stability of medicament.
• Cost of therapy is minimized by reducing the dose per unit
formulation
• Elevate bioavailability especially in water disfavouring
drugs.
• Selective uptake by tissues due to direct drug delivery.

56. DISADVANTAGES
• The production cost of electrosomes are
generally high since these come under the class
of nanotherapeutics.
• The constituent phospholipids present in lipid
vesicular structures may undergo oxidation or
hydrolysis.

57. APPLICATION
• They use enzymatic reactions to catalyze the conversion of
chemical energy to electricity in a fuel cell.
• The use of enzymatic cascades in enzymatic fuel cell
anodes resulted in very high power outputs, as the
electron density achieved was much higher when the fuel
was fully oxidized.
• Its used as a carrier in drug targeting.
• Used in the treatment of cancer.
• Used in studying immune response.
• Ear targeting
• Muscle targeting

58. REFRENCE
• N.K. Jain “Advances in Controlled and Novel Drug
Delivery” CBS Publisher and Distributors, First edition.
• Patil S, Pancli SS, Agrawal S, Agrawal GP. Surfacemodified
mesoporous ceramics as delivery vehicle for haemoglobin.
Drug Delivery 2004;1:193-9.
• Kommineni S, Ahmad S, Vengala P, Subramanyam CV. Sugar
coated ceramic nanocarriers for the oral delivery of
hydrophobic drugs: Formulation, optimization and
evaluation. Drug Dev Ind Pharm 2012;38:577-86.
• Priyanka R Kulkarni, Jaydeep D Yadav, Kumar A Vaidya.
Liposomes: A Novel Drug Delivery System. Int J Curr Pharm
Res, 2011; 3(2): 10-18.

59. THANK YOU