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Liposome-A Comprehensive
Approach for Researchers
Mani Sharma, Jyoti Joshi, Neeraj Kumar Chouhan,
Mamta N. Talati, Sandeep Vaidya and Abhiram Kumar
Abstract
1. Introduction
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Molecular Pharmacology
Figure 1.
Liposome molecule with lipid bilayer.
A. Phospholipids
• Phosphatidylcholine
• Phosphatidylserine
• Phosphatidylethanolamine
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Liposome-A Comprehensive Approach for Researchers
DOI: http://dx.doi.org/10.5772/intechopen.93256
2. Synthetic phospholipids:
• Disloyal phosphatidylethanolamine
• Disloyal phosphatidylcholine
B. Cholesterol
Figure 2.
Hydrophilic and lipophilic terminals of lipid.
Figure 3.
Inner and outer structure of liposome.
2. Physio-chemical properties
3. Applications of liposomes
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Molecular Pharmacology
Table 1.
Physical characterization [6].
Table 2.
Chemical characterization [6].
Table 3.
Biological characterization [6].
• Due to the encapsulation of drug, overall stability is increased and reduced the
adverse effects of encapsulated drug.
• Biodegradability
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Liposome-A Comprehensive Approach for Researchers
DOI: http://dx.doi.org/10.5772/intechopen.93256
• Biocompatibility
• Biodegradable
• Liposomes are able to provide both aqueous “milieu internee” and the lipo-
philic environment in a single system
• Study of membranes
• In gene delivery
• In multidrug resistance
• In immunology
• In cosmetology (Table 4)
In parasitic diseases After IV injection liposomes are comfortly digested by phagocytic cells in the
body and hence considered as one of the best vehicle to dispatch cargo into
macrophages
Anticancer therapy Liposomes are effective for the cells not only in tumors but also in the
gastrointestinal mucosa
Other medical These liposomes are sterically stabilized vesicles and are long circulating micro-
applications reservoirs or tumor (or site of inflammation and infection) targeting vehicles
In bioengineering Fragments of siRNA and DNA are delivered with the help of modern genetic
engineering and gene recombinant technology
In vaccination Liposomes are considerably used in proper vaccination due its fine active targeting
In agro-food industry Due to its versatile physio-chemical properties lipids are extensively manufactured
and used in large scale up sectors
Table 4.
Applications of liposomes [6].
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Molecular Pharmacology
4. Methods of preparation
See Figure 4.
This is one of the widely used methods for the preparation of liposomes. As
it has no such complicated steps involved in it. Multilamellar vesicles (MLV) are
prepared by solubilizing natural or synthesized phospholipid in chloroform, dichlo-
romethane, ethanol or in a mixture of chloroform and methanol in a ratio of 3:1 v/v;
2:1 v/v or 9:1 v/v. A homogeneous thin film forms when this mixture is revolved and
dried in a rota-evaporator under vacuum at a temperature around 45–60°C. Layes is
kept under nitrogen drying for overnight. Next, comes the hydration process where
completely dried thin film is hydrated using aqueous phase—phosphate buffer
solution of pH 7.2 for 1–2 h at 60–70°C.
This kind of procedure can be applied to almost any kind of lipid mixtures, but
has some drawbacks that majorly includes—low encapsulation space, a bit difficult
to scale up and layer formed are not always homogeneous thus shows heterogeneous
size distribution during later physio-chemical examination of liposomes through
zetasizer.
Here, the lipid mixture is dissolved in ether or diethyl ether under continuous
stirring that is later injected into a PBS or aqueous phase. Which under injection
pressure causes the removal of almost all organic solvent that ultimately forms
liposomes. This method also suffers with the heterogeneous liposomal formulation
defect.
Figure 4.
General representation for method of preparation of liposome.
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Liposome-A Comprehensive Approach for Researchers
DOI: http://dx.doi.org/10.5772/intechopen.93256
In ethanol injection method the lipid mixture is dissolved in ethanol under contin-
uous stirring that is later injected into a preheated TRIS-HCl buffer or distilled water.
Hydrophobicity and hydrophilicity of a drug accounts for drug intake in a liposomal
vesicle. It has an advantage of using non-toxic and ethanol and is also easily scalable.
Here, the lipid mixture is dissolved in organic solvents ether or diethyl ether
or a mixture of diethyl ether and chloroform (1:1 v/v); a mixture of methanol-
chloroform (1:2 v/v) under continuous stirring that is later injected into a PBS
or aqueous phase comprising citric-Na2HPO4 to improve the overall efficacy of a
formulation. Which under injection pressure causes the removal of organic solvent
that ultimately leads to the formation of liposomes. This method also suffers with
the heterogeneous liposomal formulation defect. Organic solvent is then dried using
rota-vapor instrument thus forming homogeneous liposome. The major disad-
vantage of this procedure is the leftover of remaining organic solvent in the final
formulation also faces difficulty in scale up procedures.
Here acidic phospholipids are used to prepare SUV by following the thin film
hydration process followed on with the addition of calcium that causes fusion
to form MLV. Final addition of ethylenediaminetetraacetic acid (EDTA) to MLV
results in the formation of large unilamellar vesicles LUV.
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Molecular Pharmacology
Table 5.
Methods for the preparation of liposomal formulation to deliver drugs [2].
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Liposome-A Comprehensive Approach for Researchers
DOI: http://dx.doi.org/10.5772/intechopen.93256
in order to govern the ionic strength and phospholipid concentration of the final
liposomal formation. Physical disruption of lamellar structure occurs due to freeze-
thaw of liposomal formulation giving it a final ionic structure.
4.9 Microfluidization
Boltic et al. was the first to introduce such method for the preparation of lipo-
somes. Here liposomes are prepared using thin film hydration method as explained
in Section 4.1, which is then sonicated and microfluidized in order to obtain partial
homogenization. This method has its wide application in industrial formulation of
liposomes.
Supercritical fluids (SCF) were introduced to replace toxic organic solvents for
the preparation of liposomes. Supercritical carbon dioxide is the most widely used
supercritical fluid as it has many advantages over conventionally used organic sol-
vents such as—it is not flammable, can be recycled, non-toxic, can be comparatively
easily removed from the solvents, requires moderate temperature and also exclude
the product degradation in inert surroundings. Karn et al. experimented and
explained the comparative study between thin film hydration method and super-
critical fluids using method evaluating the non toxicity and better field approaches
in term of using super critical fluids for the formulation of liposomes (Table 5).
• Phospholipids shows affinity for polar molecules as well as for aqueous phase
due to a hydrophobic tail, that has 2 fatty acids which are made up of 10–24 C
atoms comprising of 0–6 double bonds in every chain [8].
6. Evaluation
The average size, size distribution, and zeta potential shall be determined by
zetasizer.
Transmission electron microscopy is used to study the shape and surface mor-
phology of a liposomal structure.
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Molecular Pharmacology
Figure 5.
Mechanism of liposome formation.
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Liposome-A Comprehensive Approach for Researchers
DOI: http://dx.doi.org/10.5772/intechopen.93256
Table 6.
Liposomal formulations present in the market [9].
Table 7.
Liposomal cosmetic formulations present in the market [10].
8. Conclusions
Acknowledgements
I thank all my coauthors who are listed, and the work was not funded by any
institute or person.
Conflict of interest
We wish to declare that there are no known conflicts of interest associated with
this publication, and there has been no significant financial support for this work
that could have influenced its outcome.
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Molecular Pharmacology
Author details
© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms
of the Creative Commons Attribution License (http://creativecommons.org/licenses/
by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,
provided the original work is properly cited.
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Liposome-A Comprehensive Approach for Researchers
DOI: http://dx.doi.org/10.5772/intechopen.93256
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