INTRODUCTION:

The liposome is defined as the spherical vesicle having at least one or more lipophilic bilayer membranes sandwiched between two hydrophilic layers. It is considered one of the advanced approaches in the drug delivery system which can be used for the delivery of both hydrophilic as well as lipophilic drugs into the body.^1,2 It has great importance in the pharmaceutical, biological and medical fields as it is considered the most proper carrier for the delivery of various drugs into the body such as antibiotics, anti-inflammatory, antifungal anticancer genes, and various other drugs.^3,4 Liposomes are the first enclosed microscopic nanocarrier which was discovered by Alec D Bangham in 1961 and were approved in 1965.^5,6

Liposomes are a type of nanomedicine that has found application in the treatment of patients suffering from a variety of diseases such as cardiovascular disease, neurodegenerative disease, diabetes, cancer, and inflammation.^7,8,9 In the case of drug encapsulated in liposomes the kidney excretion can be reduced causing prolonged half-life and increased activity of the drug. Liposome drug carriers can also be used in tumor targeting via the enhanced permeation and retention (EPR) phenomenon.^10,11,12

Liposome as drug carrier has various advantages such as protection of the drug from degradation in the body, control of the release of the drug, and target drug delivery to the site of disease. It can also modify the biodistribution of the drug and enhance its solubility and bioavailability.^13,14,15 On the other hand, their benefits for topical applications is to reduce serious incompatibilities and side effects that may result from excessive systemic drug absorption, as well as the significant enhancement of drug accumulation at the required site of action due to the high similarity between the liposome composition and biological membranes can also be seen.^16,17,18

The liposome can also be used as an effective vaccine adjuvant due to its ability to protect antigens (protein and peptide) and deliver them to the antigen-presenting cell and stimulate protective immune responses.^19,20,21

Figure.1: structure of the liposome

Composition of liposome:

Phospholipids are the main component of liposomes having two main categories including sphingomyelins and glycerophospholipids.²² The glycerophospholipids are the phospholipids obtained from the eukaryotic cell in which glycerol is the main component. It consists of a hydrophilic head and a hydrophobic side chain.^23,24 According to variation in the head part different glycerophospholipids are obtained such as Phosphatidylcholine, phosphatidylserine, phosphatidic acid, and cardiolipin.²⁵ Along with the head part the length of the nonpolar tail part also results in various glycerophospholipids for example dimyristoyl and stearoyl. The ratio of the hydrophobic tail can also affect the volume of the hydrophobic cavity of the liposome.²⁶

The hydrophilic head part of phospholipid can further be divided into three types according to their surface charges i.e. cationic anionic and neutral head.²⁷ The type of charge present on the lipid head also affects the activity of the liposome, for example, the cationic head helps the liposome to attach to the negatively charged cell membrane and increases the cell incorporation rate.²⁸ Other than the cationic lipid head Phosphatidylcholine and cholesterol is also used as a lipid that is neutral and nontoxic to the cell membrane.²⁹The addition of these adjutants’ lipids also helps to stabilize the liposome membrane and decreases damage to the cell membrane.^30,31

Other than these some other ingredients are also added to the liposome to increase their activity and specificity to a particular target such as a PEGylated liposome (long circulating liposome).^32,33 Long-circulation liposomes are a type of modified liposome that has their surface coated with inert polymeric molecules such as oligosaccharides, glycoproteins, polysaccharides, and synthetic polymers.^34,35Among these hydrophilic molecules, polyethylene glycol (PEG) polymer has proven to be a successful material for liposome stabilization. For example, Gambogenic acid-loaded PEGylated liposomes (GNA-PEG-LPs), were used to reduce toxicity, prolong half-life, and improve anticancer efficacy.³⁶

Method of preparation:

Most of the conventional techniques for the preparation of liposomes include the dissolution of phospholipid in an organic solvent followed by evaporation of the solvent.^37,38 The phospholipid used in the preparation of liposomes have a critical micelle concentration in the nanomolar range and the quantity of phospholipid used is much more than the critical micelle concentration which ultimately leads to the formation of liposome when comes in contact with aqueous media.^39,40

Both conventional and novel methods are used for the preparation of liposomes which is discussed below in this article.⁴¹ The conventional methods of liposome preparation are as follow:

Solvent evaporation (Thin film hydration method):

In this process, the phospholipid is dispersed in the organic solvent followed by its evaporation under a vacuum to get a fine thin film. If the drug is lipophilic it is also added to the organic solvent to form a one-phase solution.⁴² If the drug is hydrophilic then it is dissolved in an aqueous buffer and added to the phospholipid thin layer leading to the formation of multilamellar vesicles liposome. This method is much more suitable for the lipophilic drug due to its high entrapment i.e. 80 – 90% as compared to hydrophilic drugs.^43,44

Solvent dispersion method:

The technique includes dissolving the phospholipid in organic solvent and the solvent should be miscible with water such as ethanol. The lipophilic drug is dissolved in the organic solvent solution which is further added into the aqueous buffer leading to the dilution of the solvent into the water and the formation of multilamellar vesicles takes place which is in the micrometer range.^45,46 Both of these techniques i.e. solvent evaporation and solvent dispersion method produce multilamellar vesicles type of liposome which is in the micrometer range and has a great impact on the activity and pharmacokinetic activity of the drug which ultimately reduces the therapeutic efficacy of the drug in the final formulation. So, it is important to reduce the size of liposomes below the micron range (nanometer).⁴⁷There are various methods to reduce the size of liposomes such as freeze-thaw, sonication, and homogenization which leads to the increased permeation and activity of the drug-loaded liposomes.⁴⁸

Reverse phase evaporation method:

This technique is mostly used for the loading of hydrophilic drugs in liposomes. For the loading of hydrophilic drugs the technique required to load a large number of drugs in the central aqueous core of the liposomes has high entrapment efficiency.(49)(50) In this method two different solutions are prepared one is the solution of drug in aqueous media and another one is the solution of phospholipid in water-miscible organic solvent and finally, water in oil type emulsion is prepared from the above two solutions. Now the evaporation of organic solvent takes place under a vacuum to form an el phase which is further evaporated to get liposomes with high entrapment efficiency in an aqueous core.^51,52

Supercritical fluids method for liposome preparation:

The use of supercritical fluids for the preparation of liposomes is one of the novel techniques for preparation of liposomes. Supercritical fluids possess the properties of both liquid and gas and a small change in pressure leads to a change in the state of the fluid.⁵³ These supercritical fluids can be used in place of organic solvent for the preparation of liposomes because of the solubility of a large variety of species in them. Various researchers found that the drug encapsulation efficiency of liposomes is also increased by supercritical fluids preparation as compared to the conventional method of liposome preparation.⁵⁴ The size and entrapment of liposomes can also be controlled by controlling the pressure and the concentration of the organic co-solvent in the solution. Various methods have been studied for the preparation of liposomes using supercritical fluids which are discussed below.⁵⁵

Supercritical anti-solvent (SAS) is one of the methods for the preparation of liposomes. In this method firstly the phospholipid and drug are dissolved in an organic co-solvent which is further sprayed into a supercritical fluid that acts as anti solvent leading to the precipitation of the phospholipids forming liposomes.⁵⁶ The size of the liposome depends upon the size of the solvent sprayed into the fluid.⁵⁷ The method studied for the preparation of liposomes using supercritical fluid is the decompression method. In this method, a solution of a phospholipid, drug, and organic co-solvent is prepared in the compressed fluid which is further transferred into the aqueous phase through the fine nozzle which leads to the formation of liposomes in the range of 0.2–4 micrometer. Other than this method various other methods also use supercritical fluids for liposome preparation such as the supercritical reverse-phase evaporation method.^58,59,60

Microfluidic methods for liposome formation:

This method includes the flow of liquid from the cross-section channel in the range of 5 to 500 micrometers. The method includes the ability to accurately dispense nanoliter volumes.⁶¹The prepared liposome is more stable with high encapsulation efficiency, mean size, and polydispersity as compared to the conventional method.⁶² There are various processes including a microfluidic method for the preparation of liposomes such as

a. Micro hydrodynamic focusing (MHF) is the most studied method for the preparation of SUVs and LUVs. The method includes the aqueous buffer flowing between two opposite walls of a rectangular channel and the solution of phospholipid in isopropyl alcohol moves along with the axis of the channel in the aqueous phase. The interaction between the alcohol and water leads to the regions with low fraction alcohol where the phospholipid is forced to convert into bilayer leading to the formation of the liposome.^63,64

b. Thin-film hydration in microtubes: The method includes dissolving phospholipid in an organic solvent such as chloroform and was dried in microtubules having 200 to 500 micrometer in diameter resulting in the formation of a thin film which is further hydrated with an aqueous buffer via tubes.⁶⁵

c. Microfluidic droplets method (for giant vesicles): In the microfluidics method both the phases i.e. oil phase and water phase are passed through a microchannel and form small drops of uniform size. These water droplets are then frozen for the purpose to prevent coalescence and its removal from the organic phase due to precipitation. The frozen water droplet was again added into the organic phase with lipid and the solvent was evaporated to obtain a liposome (giant vesicle) having a particle size range between 2 to 50 micrometer.^66,67

Electroformation method:

This method is commonly performed for the preparation of giant unilamellar vesicles. The vesicle can only be formed when the charged phospholipid is used for the preparation of the liposome.^68,69 Giant unilamellar vesicle can be obtained by using this method in a chamber (flow chamber) and allows initial switching of the glycerol solution, which is used for rehydration of phospholipid film, into an ionic solution, and GUVs were still attached to the electrode side.⁷⁰

Other novel methods for the preparation of liposomes:

a. Freeze drying of double emulsion for liposome preparation: The freeze-drying of liposomes by forming lipids and water-soluble carrier materials when dissolved in tert-butyl alcohol/water as co-solvent systems form cakes of an isotropic single-phase solution. After the addition of water to the freeze-dried product, it leads to the formation of spontaneously homogeneous dispersion of MLVs. The vesicles size can further be reduced through various methods such as extrusion. ^{71, 72}

b. Hydration of phospholipids deposited on nanostructured materials: It involves the hydration of phospholipids that have been deposited on electrospun amphiphilic nanofibres made of the hydrophilic polymer that is polyvinyl pyrrolidone and soybean lecithin combination. The templating and confinement properties of nanofibres allowed for the self-assembly of Phosphatidylcholine. When the fibers were mixed with water, liposomes spontaneously formed.⁷³

Application of liposome:

Liposomes as drug carriers:

liposomes are the vesicles used for the loading of hydrophilic as well as hydrophobic drug. The hydrophilic drug is loaded into the central aqueous core of the liposome whereas the hydrophobic drug is inserted into the bilayer of lipid. The encapsulation of a drug into a liposome has various advantages such as increased solubility of the hydrophilic drug, increase tissue uptake of the drug, increase stability and also protect the drug from body enzymes and fluids.^76,77

Application in Small Molecule Therapeutics:

liposomes are also used for the delivery of drugs having small molecules. These small molecule drugs are generally divided into two categories i.e. anti-infective agents and chemotherapeutic agents.⁷⁸ Conventional cytotoxic drugs kill healthy cells along with diseased cells, causing side effects in blood cells and hair follicles, cells lining the intestinal mucosa can also be affected, and imposing practical limits on drug dose and dosing frequency.^79,80 The pathophysiological conditions of tumor tissue like leaky tumor vasculature, in conjunction with drug encapsulation within a liposome is a valuable strategy for cancer treatment.⁸¹ Classical liposomes or PEGylated liposomes use an EPR-mediated passive targeting strategy to preferentially accumulate the encapsulated drug within tumor tissue, increasing therapeutic efficacy while minimizing side effects.^82,83 Doxil®/Caelyx®, injectable PEGylated liposomes (100 nm) encapsulating doxorubicin (DOX), were the first liposomes approved in 1995 for the treatment of the various type of cancers.⁸⁴

Gene therapy and delivery of macromolecules:

After the successful delivery of small drug molecules in liposomes the ability of liposomes the delivery macromolecules is studied. The macromolecule such as protein, peptides, antisense oligonucleotides (asODNs), and small interfering RNA (siRNA) can be delivered to their target sites with the help of liposomes.^85,86 These macromolecules are nucleic acid-based materials having high molecular weight, hydrophilic in nature, and highly charged molecules that cannot cross cell membrane passively and are also susceptible to enzymatic degradation and rapid systemic clearance.^87,88 The clinical application of these molecules is further decreased due to their low selectivity for target tissue and poor cellular uptake. As a result, liposomes' ability to deliver nucleic acid-based therapeutics to target cells/tissues has been called into question.⁸⁹ For this purpose, Charge-imparting lipids like 1, 2-bis (oleoyloxy)-3 (trimethylammonium) propane (DOTAP) and 3[N, N-dimethylamino-ethane] carbomoyl] cholesterol (DC-CHOL) were incorporated into the membrane of liposomes to give them a net positive charge. These positively charged liposomes interact with the negatively charged macromolecules leading to the formation of a complex i.e. “lipoplex”.⁹⁰ These lipoplexes can easily enter the cell membrane through fusion followed by the release of endosomal membrane and internalization. Some of the other techniques are applied to increase the entrapment of macromolecules into the liposome along with a net neutral charge or anionic surface charge.

As a Vaccine carrier:

liposome can be used as a vaccine carrier and can carry several antigens into the body because some antigens can be absorbed or can bind covalently to the surface of liposomes.⁹¹ The liposome can penetrate the tissue and each to the lymphatic system where it releases antigen along with the maintenance of their integrity. The released antigen then can bind to the B cell receptor and activate the immune system of the body by major histocompatibility type I complex.⁹²^,⁹³ The antigen can be delivered in liposomes along with other immune adjutants.^94,95 Immune adjutants are the components that can enhance the immune response of the vaccine and are given along with antigen to increase the chance of contact between antigen and antigen-binding site and also increase the time of release (prolong release).⁹⁶ Virosomes is an example of liposome which are reconstituted influenza virus envelopes devoid of an inner core and genetic information.⁹⁷ In this process the influenza virus surface antigens are integrated into phospholipids bilayer liposomes leading to the formation of unilamellar Virosome in a nanometer range of 150 - 200 nanometers. Some of the clinically approved Virosome for antigen delivery are Epaxal®, Inflexal®, and Mosquirix®.⁹⁸

Application in diagnostic application:

Various diseases required early diagnosis for their better treatment such as cancer. As a result, various researches have been made for the development of new methods and improving the activity of the already present diagnostic method.⁹⁹ commonly diagnostic agents are used for the determination of the area of interest to differentiate between normal cells and abnormalities or the better activity of the diagnostic agent and its attachment to the required tissue.¹⁰⁰ It is important to transfer and accumulate the agent at the site of action. For this purpose, various carriers are used for the delivery of these agents. Among these carriers, liposome is an important carrier due to their ability to manipulate physicochemical and pharmacokinetic properties.

Computerized tomography (CT) is a technique used to create a detailed image of the areas inside the body. The process required a CT contrast agent for imaging and these agents can be encapsulated in the inner aqueous core of the liposome for example encapsulation of iodinated contrast agent and iohexol in the inner aqueous compartment of ligand targeted liposome to enhance their diagnostic potential of iodinated contrast agent against atheromatous plaques in activated human coronary artery endothelial cells (HCAEC) with specific targeting.^101,102

Liposomes in cosmetics:

liposome formulations are also used in cosmetics due to their properties such as increasing skin moisture, increasing fluidity of cell membrane, and encapsulation of both oil and aqueous phase active ingredients along with deep penetration into the skin which leads to the deposition in the dermis to form a reservoir which can prolong the time of action of the active ingredient.¹⁰³^,¹⁰⁴ “Capture” was the first liposomal cosmetic launched in the market.

Theranostic Applications:

Theranostics is gaining traction as a promising therapeutic paradigm. Theranostic means the co-delivery of medicament and diagnostic agents in a single formulation; as a result, theranostic-based strategies may be useful in the selection of treatment, dosage regimen design, objective response monitoring, and follow-up therapy planning according to the specific molecular characteristics of a disease. Liposomes are currently regarded as the best platform for theranostic nanomedicine due to the high cargo capacity and flexible encapsulation capabilities of both imaging and therapeutic agents.^105,106

Theranostic nanomedicine, which uses liposomes, has recently been extensively researched in cancer.¹⁰⁷ the antibody-targeted PEGylated liposome i.e. immunoliposome are loaded with indocyanine green dye and anticancer drug doxorubicin is used to identify the tumor accumulation of this immunoliposome over time in a murine breast cancer mouse model noninvasively. The therapeutic liposome has the ability to bind with tumor-specific targeted therapy with diagnosis using multispectral optoacoustic tomography (MSOT).^108,109

Table- 2: Marketed approved drugs along with their application and drug-loaded in them:

Product name	Drug	Disease	Company	Reference
Atragen	Tretinoin	Acute promyelocytic leukemia	Aronex pharmaceuticals Inc.	¹¹⁰
Amphotec	Amphotericin B	Fungus infections leishmaniasis	Sequus pharmaceuticals Inc.	¹¹¹
Ambisome	Amphotericin B	Serious fungal infection	neXstar pharmaceuticals Inc.	¹¹¹
Abelcet	Amphotericin B	Serious fungal infection	The liposome company	¹¹¹
ALEC	Dry protein free powder of DPPCPG	Expanding lung disease in infants	Brotannia pharm, UK	¹¹²
Amphocil	Amphotericin B	Serious fungal infection	Sequus pharmaceuticals Inc.	¹¹¹
Depocyt	Cytarabine	Lymphomatous meningitis	Pacira pharmaceutical Inc.	¹¹³
Doxil	Doxorubicin	Kaposi sarcoma	Sequus pharmaceuticals Inc.	¹¹⁴
Daunoxone	Daunorubucin citrate	Kaposi sarcoma	Galen Ltd.	¹¹⁵
Depodur	Morphine	Post-surgical pain reliever	Pacira pharmaceutical Inc.	¹¹⁶
Avian retrovirus vaccine	Killed avain retrovirus	Chickenpox	Vineland lab, USA	¹¹⁷
Estrasob	Estradiol	Menopausal therapy	Novavax	¹¹⁸
Epaxal Berna vaccine	Inactivated hepatitis-A virions	Hepatitis A	Swiss serum and vaccine institute, Switzerland	¹¹⁹
Fungizone	Amphotericin B	Serious fungal infection	Bristol-myers squibb, Netherland	¹¹¹
Evacet	Doxorubicin	Metastatic breast cancer	The liposome company, USA	¹¹⁴
Mikasome	Amikacin	Bacterial infection	neXstar pharmaceutical Inc.	¹²⁰
vinaXome	Vincristine	Solid tumors	neXstar pharmaceutical Inc	¹¹⁵
Topex Br	Terbutaline sulphate	Asthma	ozone pharmaceutical Ltd.	¹²¹
Ventus	Prostaglandin- E1	Systemic inflammatory disease	The liposome company	¹²²
Nyotran	Nystatin	Systemic fungal infection	Aronex pharmaceutical Inc.	¹²³
Myocet	Doxorubicin	Breast cancer	Enzon pharmaceuticals	¹¹⁴
Onivyde	Irinitecan	Pancreatic cancer	Merrimark pharmaceuticals	¹²⁴
Merqibo	Vincristine	Acute lymphoblastic leukemia	Arbutus	¹¹⁵

CONCLUSION:

Liposomes are lipid bilayer nanocarriers used for drug delivery in various diseases like cancer tuberculosis and various other skin problems. Liposomes can protect the drug from degradation in the body such as first-pass metabolism and degradation by other body fluids therefore used as a carrier for various biodegradable drugs. Liposomes can be prepared by various methods. Novel methods such as supercritical fluids can also be used for the preparation of liposomes. In this article, we studied liposomes, a novel method for their preparation and their application in the pharmaceutical and cosmetic industry. In this, we also studied modification on liposomes like Virosome and PEGylated liposomes for specific targeting and delivery of large molecules like proteins and peptides. The various marketed product and products under trial are also mentioned in this article.

REFERENCE:

1. Gomez-Hens A, Fernández-Romero JM. The role of liposomes in analytical processes. TrAC Trends Anal Chem. 2005;24(1):9–19.

2. Has C, Sunthar P. A comprehensive review on recent preparation techniques of liposomes. J Liposome Res. 2020;30(4):336–65.

3. Manconi M, Caddeo C, Sinico C, Valenti D, Mostallino MC, Biggio G, et al. Ex vivo skin delivery of diclofenac by transcutol containing liposomes and suggested mechanism of vesicle–skin interaction. Eur J Pharm Biopharm. 2011;78(1):27–35.

4. Mukherjee S, Ray S, Thakur RS. Solid lipid nanoparticles: a modern formulation approach in drug delivery system. Indian J Pharm Sci. 2009;71(4):349.

5. Akhtar N. Vesicles: a recently developed novel carrier for enhanced topical drug delivery. Curr Drug Deliv. 2014;11(1):87–97.

6. Jindal S, Awasthi R, Singare D, Kulkarni GT. Preparation and in vitro evaluation of Tacrolimus loaded liposomal vesicles by two methods: A comparative study. J Res Pharm. 2021;25(1):34–41.

7. Chowdhury A, Kunjiappan S, Panneerselvam T, Somasundaram B, Bhattacharjee C. Nanotechnology and nanocarrier-based approaches on treatment of degenerative diseases. Int nano Lett. 2017;7(2):91–122.

8. Siddiqi KS, Husen A, Sohrab SS, Yassin MO. Recent status of nanomaterial fabrication and their potential applications in neurological disease management. Nanoscale Res Lett. 2018;13(1):1–17.

9. Yetisgin AA, Cetinel S, Zuvin M, Kosar A, Kutlu O. Therapeutic nanoparticles and their targeted delivery applications. Molecules. 2020;25(9):2193.

10. Greish K, Fang J, Inutsuka T, Nagamitsu A, Maeda H. Macromolecular therapeutics. Clin Pharmacokinet. 2003;42(13):1089–105.

11. Iyer AK, Khaled G, Fang J, Maeda H. Exploiting the enhanced permeability and retention effect for tumor targeting. Drug Discov Today. 2006;11(17–18):812–8.

12. Nakamura H, Jun F, Maeda H. Development of next-generation macromolecular drugs based on the EPR effect: challenges and pitfalls. Expert Opin Drug Deliv. 2015;12(1):53–64.

13. Çağdaş M, Sezer AD, Bucak S. Liposomes as potential drug carrier systems for drug delivery. Appl Nanotechnol drug Deliv. 2014;1:1–50.

14. Ranade V V. Drug delivery systems. 1. Site‐specific drug delivery using liposomes as carriers. J Clin Pharmacol. 1989;29(8):685–94.

15. Choudhury H, Gorain B, Pandey M, Chatterjee LA, Sengupta P, Das A, et al. Recent update on nanoemulgel as topical drug delivery system. J Pharm Sci. 2017;106(7):1736–51.

16. Rahimpour Y, Hamishehkar H. Liposomes in cosmeceutics. Expert Opin Drug Deliv. 2012;9(4):443–55.

17. Kaur IP, Kakkar S. Topical delivery of antifungal agents. Expert Opin Drug Deliv. 2010;7(11):1303–27.

18. Li W, Joshi MD, Singhania S, Ramsey KH, Murthy AK. Peptide vaccine: progress and challenges. Vaccines. 2014;2(3):515–36.

19. Gregory AE, Williamson D, Titball R. Vaccine delivery using nanoparticles. Front Cell Infect Microbiol. 2013;3:13.

20. Tandrup Schmidt S, Foged C, Smith Korsholm K, Rades T, Christensen D. Liposome-based adjuvants for subunit vaccines: formulation strategies for subunit antigens and immunostimulators. Pharmaceutics. 2016;8(1):7.

21. Khalaj‐Hedayati A, Chua CLL, Smooker P, Lee KW. Nanoparticles in influenza subunit vaccine development: Immunogenicity enhancement. Influenza Other Respi Viruses. 2020;14(1):92–101.

22. Li J, Wang X, Zhang T, Wang C, Huang Z, Luo X, et al. A review on phospholipids and their main applications in drug delivery systems. Asian J Pharm Sci. 2015;10(2):81–98.

23. Singh RP, Gangadharappa H V, Mruthunjaya K. Phospholipids: Unique carriers for drug delivery systems. J Drug Deliv Sci Technol. 2017;39:166–79.

24. Paltauf F, Hermetter A. Phospholipids—Natural, semisynthetic, synthetic. In: Phospholipids. Springer; 1990. p. 1–12.

25. Ahmed KS, Hussein SA, Ali AH, Korma SA, Lipeng Q, Jinghua C. Liposome: Composition, characterisation, preparation, and recent innovation in clinical applications. J Drug Target. 2019;27(7):742–61.

26. Silvius JR. Role of cholesterol in lipid raft formation: lessons from lipid model systems. Biochim Biophys Acta (BBA)-Biomembranes. 2003;1610(2):174–83.

27. Harayama T, Riezman H. Understanding the diversity of membrane lipid composition. Nat Rev Mol cell Biol. 2018;19(5):281–96.

28. Zhukovsky MA, Filograna A, Luini A, Corda D, Valente C. Phosphatidic acid in membrane rearrangements. FEBS Lett. 2019;593(17):2428–51.

29. Iscaro A, Howard NF, Muthana M. Nanoparticles: properties and applications in cancer immunotherapy. Curr Pharm Des. 2019;25(17):1962–79.

30. P Samy R, Gopalakrishnakone P, G Stiles B, S Girish K, N Swamy S, Hemshekhar M, et al. Snake venom phospholipases A2: a novel tool against bacterial diseases. Curr Med Chem. 2012;19(36):6150–62.

31. Jindal S, Awasthi R, Singhare D, Kulkarni GT. Topical delivery of Tacrolimus using liposome containing gel: An emerging and synergistic approach in management of psoriasis. Med Hypotheses. 2020;142:109838.

32. Medina OP, Zhu Y, Kairemo K. Targeted liposomal drug delivery in cancer. Curr Pharm Des. 2004;10(24):2981–9.

33. Panahi Y, Farshbaf M, Mohammadhosseini M, Mirahadi M, Khalilov R, Saghfi S, et al. Recent advances on liposomal nanoparticles: synthesis, characterization and biomedical applications. Artif cells, nanomedicine, Biotechnol. 2017;45(4):788–99.

34. Daraee H, Etemadi A, Kouhi M, Alimirzalu S, Akbarzadeh A. Application of liposomes in medicine and drug delivery. Artif cells, nanomedicine, Biotechnol. 2016;44(1):381–91.

35. Jindal S, Kumar A, Goyal K, Awasthi R, Kulkarni GT. Lipid Nanocarriers for Dermal Delivery of Lutein. In: Nanomedicine for Bioactives. Springer; 2020. p. 341–66.

36. Mura S, Nicolas J, Couvreur P. Stimuli-responsive nanocarriers for drug delivery. Nat Mater. 2013;12(11):991–1003.

37. Huang Z, Li X, Zhang T, Song Y, She Z, Li J, et al. Progress involving new techniques for liposome preparation. asian J Pharm Sci. 2014;9(4):176–82.

38. Mozafari MR. Liposomes: an overview of manufacturing techniques. Cell Mol Biol Lett. 2005;10(4):711.

39. Paternostre MT, Roux M, Rigaud JL. Mechanisms of membrane protein insertion into liposomes during reconstitution procedures involving the use of detergents. 1. Solubilization of large unilamellar liposomes (prepared by reverse-phase evaporation) by triton X-100, octyl glucoside, and sodium. Biochemistry. 1988;27(8):2668–77.

40. Edwards K, Johnsson M, Karlsson G, Silvander M. Effect of polyethyleneglycol-phospholipids on aggregate structure in preparations of small unilamellar liposomes. Biophys J. 1997;73(1):258–66.

41. Kumar S, Kaur N, Mithu VS. Amphiphilic ionic liquid induced fusion of phospholipid liposomes. Phys Chem Chem Phys. 2020;22(43):25255–63.

42. Zhang H. Thin-film hydration followed by extrusion method for liposome preparation. In: Liposomes. Springer; 2017. p. 17–22.

43. Fernández-García R, Lalatsa A, Statts L, Bolás-Fernández F, Ballesteros MP, Serrano DR. Transferosomes as nanocarriers for drugs across the skin: Quality by design from lab to industrial scale. Int J Pharm. 2020;573:118817.

44. Varona S, Martin A, Cocero MJ. Liposomal incorporation of lavandin essential oil by a thin-film hydration method and by particles from gas-saturated solutions. Ind Eng Chem Res. 2011;50(4):2088–97.

45. Marsden HR, Gabrielli L, Kros A. Rapid preparation of polymersomes by a water addition/solvent evaporation method. Polym Chem. 2010;1(9):1512–8.

46. Zhang X, Coleman AC, Katsonis N, Browne WR, Van Wees BJ, Feringa BL. Dispersion of graphene in ethanol using a simple solvent exchange method. Chem Commun. 2010;46(40):7539–41.

47. Kim SW, Kim T, Kim YS, Choi HS, Lim HJ, Yang SJ, et al. Surface modifications for the effective dispersion of carbon nanotubes in solvents and polymers. Carbon N Y. 2012;50(1):3–33.

48. Ming-Jie LI, ZHANG H-Y, Xiao-Zhe LIU, Chun-Yan CUI, Zhi-Hong SHI. Progress of extraction solvent dispersion strategies for dispersive liquid-liquid microextraction. Chinese J Anal Chem. 2015;43(8):1231–40.

49. Chen J, Brooks III CL, Khandogin J. Recent advances in implicit solvent-based methods for biomolecular simulations. Curr Opin Struct Biol. 2008;18(2):140–8.

50. Szoka F, Papahadjopoulos D. Procedure for preparation of liposomes with large internal aqueous space and high capture by reverse-phase evaporation. Proc Natl Acad Sci. 1978;75(9):4194–8.

51. Mertins O, Sebben M, Pohlmann AR, da Silveira NP. Production of soybean phosphatidylcholine–chitosan nanovesicles by reverse phase evaporation: a step by step study. Chem Phys Lipids. 2005;138(1–2):29–37.

52. Taylor KMG, Taylor G, Kellaway IW, Stevens J. Drug entrapment and release from multilamellar and reverse-phase evaporation liposomes. Int J Pharm. 1990;58(1):49–55.

53. Trucillo P, Campardelli R, Reverchon E. Liposomes: From bangham to supercritical fluids. Processes. 2020;8(9):1022.

54. Jung J, Perrut M. Particle design using supercritical fluids: literature and patent survey. J Supercrit Fluids. 2001;20(3):179–219.

55. Naik S, Patel D, Surti N, Misra A. Preparation of PEGylated liposomes of docetaxel using supercritical fluid technology. J Supercrit Fluids. 2010;54(1):110–9.

56. Xia F, Hu D, Jin H, Zhao Y, Liang J. Preparation of lutein proliposomes by supercritical anti-solvent technique. Food Hydrocoll. 2012;26(2):456–63.

57. Booysen E. Characterization of a novel antibiotic isolated from Xenorhabdus khoisanae and encapsulation of vancomycin in nanoparticles. Stellenbosch: Stellenbosch University; 2018.

58. Lesoin L, Crampon C, Boutin O, Badens E. Preparation of liposomes using the supercritical anti-solvent (SAS) process and comparison with a conventional method. J Supercrit Fluids. 2011;57(2):162–74.

59. Kompella UB, Koushik K. Preparation of drug delivery systems using supercritical fluid technology. Crit Rev Ther Drug Carr Syst. 2001;18(2).

60. Amoabediny G, Haghiralsadat F, Naderinezhad S, Helder MN, Akhoundi Kharanaghi E, Mohammadnejad Arough J, et al. Overview of preparation methods of polymeric and lipid-based (niosome, solid lipid, liposome) nanoparticles: A comprehensive review. Int J Polym Mater Polym Biomater. 2018;67(6):383–400.

61. van Swaay D, DeMello A. Microfluidic methods for forming liposomes. Lab Chip. 2013;13(5):752–67.

62. Yu B, Lee RJ, Lee LJ. Microfluidic methods for production of liposomes. Methods Enzymol. 2009;465:129–41.

63. Amrani S. Microfluidic platform for the fabrication and loading of nanoscale liposomes by 2D hydrodynamic flow focusing. McGill University (Canada); 2018.

64. Cheung C. Preparation of Multifunctional Nanoparticles Using Microfluidics. Queen’s University Belfast; 2020.

65. Shum HC, Thiele J, Kim S-H. Microfluidic fabrication of vesicles. Adv Transp Phenom 2011. 2014;1–28.

66. Hu PC, Li S, Malmstadt N. Microfluidic fabrication of asymmetric giant lipid vesicles. ACS Appl Mater Interfaces. 2011;3(5):1434–40.

67. Vayssieres L, Keis K, Hagfeldt A, Lindquist S-E. Three-dimensional array of highly oriented crystalline ZnO microtubes. Chem Mater. 2001;13(12):4395–8.

68. Ding X, Zeng D, Zhang S, Xie C. C-doped WO3 microtubes assembled by nanoparticles with ultrahigh sensitivity to toluene at low operating temperature. Sensors Actuators B Chem. 2011;155(1):86–92.

69. Li Q, Wang X, Ma S, Zhang Y, Han X. Electroformation of giant unilamellar vesicles in saline solution. Colloids Surfaces B Biointerfaces. 2016;147:368–75.

70. Stein H, Spindler S, Bonakdar N, Wang C, Sandoghdar V. Production of isolated giant unilamellar vesicles under high salt concentrations. Front Physiol. 2017;8:63.

71. Kuribayashi K, Tresset G, Coquet P, Fujita H, Takeuchi S. Electroformation of giant liposomes in microfluidic channels. Meas Sci Technol. 2006;17(12):3121.

72. Kanha N, Regenstein JM, Surawang S, Pitchakarn P, Laokuldilok T. Properties and kinetics of the in vitro release of anthocyanin-rich microcapsules produced through spray and freeze-drying complex coacervated double emulsions. Food Chem. 2021;340:127950.

73. Kim TH, Park TG. Critical effect of freezing/freeze-drying on sustained release of FITC-dextran encapsulated within PLGA microspheres. Int J Pharm. 2004;271(1–2):207–14.

74. Supramaniam P, Ces O, Salehi-Reyhani A. Microfluidics for artificial life: techniques for bottom-up synthetic biology. Micromachines. 2019;10(5):299.

75. Monteiro N, Martins A, Reis RL, Neves NM. Liposomes in tissue engineering and regenerative medicine. J R Soc Interface. 2014;11(101):20140459.

76. Xue J, Wu T, Dai Y, Xia Y. Electrospinning and electrospun nanofibers: Methods, materials, and applications. Chem Rev. 2019;119(8):5298–415.

77. Mohanraj VJ, Chen Y. Nanoparticles-a review. Trop J Pharm Res. 2006;5(1):561–73.

78. Jadhav SM, Morey P, Karpe M, Kadam V. Novel vesicular system: an overview. J Appl Pharm Sci. 2012;2(1):193–202.

79. Narvekar M, Xue HY, Eoh JY, Wong HL. Nanocarrier for poorly water-soluble anticancer drugs—barriers of translation and solutions. Aaps Pharmscitech. 2014;15(4):822–33.

80. Alam A, Farooq U, Singh R, Dubey VP, Kumar S, Kumari R, et al. Chemotherapy treatment and strategy schemes: A review. Open Access J Toxicol. 2018;2(5):555600.

81. Pérez-Herrero E, Fernández-Medarde A. Advanced targeted therapies in cancer: Drug nanocarriers, the future of chemotherapy. Eur J Pharm Biopharm. 2015;93:52–79.

82. Overchuk M, Zheng G. Overcoming obstacles in the tumor microenvironment: Recent advancements in nanoparticle delivery for cancer theranostics. Biomaterials. 2018;156:217–37.

83. Lila ASA, Ishida T. Liposomal delivery systems: design optimization and current applications. Biol Pharm Bull. 2017;40(1):1–10.

84. Sharma G, Anabousi S, Ehrhardt C, Ravi Kumar MN V. Liposomes as targeted drug delivery systems in the treatment of breast cancer. J Drug Target. 2006;14(5):301–10.

85. Singh N, Sondhi S, Sharma S, Singh D, Koundal V, Goyal K, et al. Treatment of Skin Cancer by Topical Drug Delivery of Nanoparticles: A Review. Res J Pharm Technol. 2021;14(10):5589–98.

86. Tamura A, Nagasaki Y. Smart siRNA delivery systems based on polymeric nanoassemblies and nanoparticles. Nanomedicine. 2010;5(7):1089–102.

87. Mohammadinejad R, Dehshahri A, Madamsetty VS, Zahmatkeshan M, Tavakol S, Makvandi P, et al. In vivo gene delivery mediated by non-viral vectors for cancer therapy. J Control Release. 2020;325:249–75.

88. Jindal S, Awasthi R, Singare D, Kulkarni GT. POTENTIAL OF HERBAL NANOCARRIER FORMULATION FOR THE TREATMENT OF PSORIASIS. Int J Pharm Life Sci. 2019;10(6).

89. Goldberg M, Langer R, Jia X. Nanostructured materials for applications in drug delivery and tissue engineering. J Biomater Sci Polym Ed. 2007;18(3):241–68.

90. Zhou J, Rossi JJ. Cell-specific aptamer-mediated targeted drug delivery. Oligonucleotides. 2011;21(1):1–10.

91. Saad M, Garbuzenko OB, Minko T. Co-delivery of siRNA and an anticancer drug for treatment of multidrug-resistant cancer. 2008;

92. Frezard F. Liposomes: from biophysics to the design of peptide vaccines. Brazilian J Med Biol Res. 1999;32(2).

93. Nam G, Choi Y, Kim GB, Kim S, Kim SA, Kim I. Emerging prospects of exosomes for cancer treatment: from conventional therapy to immunotherapy. Adv Mater. 2020;32(51):2002440.

94. Madni A, Sarfraz M, Rehman M, Ahmad M, Akhtar N, Ahmad S, et al. Liposomal drug delivery: a versatile platform for challenging clinical applications. J Pharm Pharm Sci. 2014;17(3):401–26.

95. García A, De Sanctis JB. An overview of adjuvant formulations and delivery systems. Apmis. 2014;122(4):257–67.

96. Du Y-F, Chen M, Xu J-R, Luo Q, Lu W-L. Preparation and Characterization of DNA Liposomes Vaccine. Liposome-Based Drug Deliv Syst. 2021;259–75.

97. Sharma A, Anghore D, Awasthi R, Kosey S, Jindal S, Gupta N, et al. A review on current carbon nanomaterials and other nanoparticles technology and their applications in biomedicine. World J Pharm Pharm Sci. 2015;4(12):1088–113.

98. Bozzuto G, Molinari A. Liposomes as nanomedical devices. Int J Nanomedicine. 2015;10:975.

99. Gowda GAN, Zhang S, Gu H, Asiago V, Shanaiah N, Raftery D. Metabolomics-based methods for early disease diagnostics. Expert Rev Mol Diagn. 2008;8(5):617–33.

100. Papanicolaou GN, Traut HF. Diagnosis of uterine cancer by the vaginal smear. New York. 1943;46.

101. Mulder WJM, Strijkers GJ, Griffioen AW, van Bloois L, Molema G, Storm G, et al. A liposomal system for contrast-enhanced magnetic resonance imaging of molecular targets. Bioconjug Chem. 2004;15(4):799–806.

102. Mukundan Jr S. Ghaghada KB, Badea CT, Kao CY, Hedlund LW, Provenzale JM, Johnson GA, Chen E, Bellamkonda RV, Annapragada A. A liposomal nanoscale contrast agent for preclinical CT in mice. AJR Am J Roentgenol. 2006;186(2):300–7.

103. Costabile RA, Choyke PL, Frank JA, Girton ME, Diggs R, Billups KL, et al. Dynamic enhanced magnetic resonance imaging of testicular perfusion in the rat. J Urol. 1993;149(5):1195–7.

104. Patravale VB, Mandawgade SD. Novel cosmetic delivery systems: an application update. Int J Cosmet Sci. 2008;30(1):19–33.

105. Budai L, Kaszás N, Gróf P, Lenti K, Maghami K, Antal I, et al. Liposomes for topical use: A physico-chemical comparison of vesicles prepared from egg or soy lecithin. Sci Pharm. 2013;81(4):1151–66.

106. Al-Jamal W, Kostarelos K. Liposomes: from a clinically established drug delivery system to a nanoparticle platform for theranostic nanomedicine. Acc Chem Res. 2011;44(10):1094–104.

107. Svenson S. Theranostics: are we there yet? Mol Pharm. 2013;10(3):848–56.

108. Mukherjee A, Paul M, Mukherjee S. Recent progress in the theranostics application of nanomedicine in lung cancer. Cancers (Basel). 2019;11(5):597.

109. Yin W, Kimbrough CW, Gomez-Gutierrez JG, Burns CT, Chuong P, Grizzle WE, et al. Tumor specific liposomes improve detection of pancreatic adenocarcinoma in vivo using optoacoustic tomography. J Nanobiotechnology. 2015;13(1):1–11.

110. Gurka MK, Pender D, Chuong P, Fouts BL, Sobelov A, McNally MW, et al. Identification of pancreatic tumors in vivo with ligand-targeted, pH responsive mesoporous silica nanoparticles by multispectral optoacoustic tomography. J Control release. 2016;231:60–7.

111. Wallace TL, Larson JL, Bazemore SA, Wilson CW, Cossum PA. The nonclinical safety evaluation of the anticancer drug ATRAGEN®(Liposomal all-trans-retinoic acid). Int J Toxicol. 2000;19(1):33–42.

112. Brogden RN, Goa KL, Coukell AJ. Amphotericin-B colloidal dispersion. A review of its use against systemic fungal infections and visceral leishmaniasis. Drugs. 1998 Sep;56(3):365–83.

113. Sharma M, Joshi J, Chouhan NK, Talati MN, Vaidya S, Kumar A. Liposome-A Comprehensive Approach for Researchers. In: Molecular Pharmacology. IntechOpen; 2020.

114. Chhikara BS, Parang K. Development of cytarabine prodrugs and delivery systems for leukemia treatment. Expert Opin Drug Deliv. 2010;7(12):1399–414.

115. Porche DJ. Liposomal doxorubicin (Doxil). J Assoc Nurses AIDS Care. 1996;7(2):55–9.

116. Wang R, Billone PS, Mullett WM. Nanomedicine in action: an overview of cancer nanomedicine on the market and in clinical trials. J Nanomater. 2013;2013.

117. Peravali R, Brock R, Bright E, Mills P, Petty D, Alberts J. Enhancing the Enhanced Recovery Program in Colorectal Surgery-use of extended-release epidural morphine (DepoDur®). Ann Coloproctol. 2014;30(4):186.

118. Estradiol-topical--Novavax: Estrasorb. Drugs R D. 2003;4(1):49–51.

119. Lea AP, Balfour JA. Virosomal hepatitis A vaccine (strain RG-SB). BioDrugs. 1997;7(3):232–48.

120. Xiong Y-Q, Kupferwasser LI, Zack PM, Bayer AS. Comparative efficacies of liposomal amikacin (MiKasome) plus oxacillin versus conventional amikacin plus oxacillin in experimental endocarditis induced by Staphylococcus aureus: microbiological and echocardiographic analyses. Antimicrob Agents Chemother. 1999;43(7):1737–42.

121. Dhandapani NV, Thapa A, Sandip G, Shrestha A, Shrestha N, Bhattarai RS. Liposomes as novel drug delivery system: A comprehensive review. 2013;

122. Kalepu S, Nekkanti V. Insoluble drug delivery strategies: review of recent advances and business prospects. Acta Pharm Sin B. 2015;5(5):442–53.

123. Johnson EM, Ojwang JO, Szekely A, Wallace TL, Warnock DW. Comparison of in vitro antifungal activities of free and liposome-encapsulated nystatin with those of four amphotericin B formulations. Antimicrob Agents Chemother. 1998;42(6):1412–6.

124. Passero Jr FC, Grapsa D, Syrigos KN, Saif MW. The safety and efficacy of Onivyde (irinotecan liposome injection) for the treatment of metastatic pancreatic cancer following gemcitabine-based therapy. Expert Rev Anticancer Ther. 2016;16(7):697–703.

Received on 02.04.2022 Modified on 28.05.2022

Asian J. Pharm. Tech. 2022; 12(4):320-328.

DOI: 10.52711/2231-5713.2022.00052

Preparation method	Types of a liposome formed	Entrapment efficiency	Reference
Supercritical fluids method	Monodisperse large unilamellar vesicle	High	⁵⁵
Micro hydrodynamic focusing (MHF)	Monodisperse small unilamellar vesicle and large unilamellar vesicle	Low	⁶⁴
Pulsed jet flow microfluidics method	Monodisperse giant unilamellar vesicle	High	⁷⁴
Thin-film hydration in micro-tubes method	Monodisperse multilamellar vesicle and large unilamellar vesicle	High	⁶⁵
Microfluidic droplets	Monodisperse giant unilamellar vesicle	High	⁶⁷
Modified Electroformation method	Polydisperse giant unilamellar vesicle	High	⁷⁰
Freeze drying of double emulsion method	Monodisperse small unilamellar vesicle	High	⁷²
Hydration of phospholipid deposited on nanostructured material	Monodisperse small unilamellar vesicle and large unilamellar vesicle	Low	⁷⁵