PRACTICE SCHOOL REPORT

On
(Transethosomes: A Revolutionary strategy for Enhanced Drug Delivery)

Submitted for Partial Fulfillment of the Requirements

For the Degree of
BACHELOR OF PHARMACY

Submitted by
Manisha Pal
2110920500043

Submitted to
Dr. Ravi Kumar Mittal
(Professor)

Galgotias College of Pharmacy

(Approved by PCI and Affiliated to
Dr. A.P.J. Abdul Kalam Technical
University, Lucknow, U.P.)

Knowledge Park II, Greater Noida, U.P 201310

 GALGOTIAS COLLEGE OF
PHARMACY
(Approved by PCI and Affiliated to Dr. A.P.J. Abdul Kalam Technical
University, Lucknow, U.P.)

STUDENT’S DECLARATION

I Manisha Pal (2110920500043) hereby declare that the presented report on

“Transethosomes: A Revolutionary strategy for Enhanced Drug Delivery”
under the practice school, BP 706 PS is a record of bonafide project work carried out
and submitted by me after completing 150 hr of practice school course work under
the guidance of Dr. Ravi Kumar Mittal.
I also confirm that, the report is only prepared for my academic requirement not for
any other purpose.

Manisha Pal
2110920500043
B.pharm 7th sem.
Date :
 GALGOTIAS COLLEGE OF
PHARMACY
(Approved by PCI and Affiliated to
Dr. A.P.J. Abdul Kalam Technical University, Lucknow, U.P.)

ENDORSEMENT BY THE SUPERVISOR

This is certify that the report entitled the study of “Transethosomes : A Revolutionary
strategy for Enhanced Drug Delivery” is a bonafide work under the subject practice
school, BP 706PS done by Manisha Pal- 2110920500043 in a partial fulfillment of the
requirement of the degree of “Bachelor of Pharmacy” of A.K.T.U. Lucknow, in the
academic year 2023-24.

Date:

Dr. Ravi Kumar Mittal

(Professor)
 GALGOTIAS COLLEGE OF
PHARMACY
(Approved by PCI and Affiliated to
Dr. A.P.J. Abdul Kalam Technical University, Lucknow, U.P.)

ENDORSEMENT BY THE DIRECTOR

This is certify that the report entitled the study of “Transethosomes: A Revolutionary
strategy for Enhanced Drug Delivery” is a bonafide work under the subject practice
school, BP 706PS done by Manisha Pal-2110920500043 in a partial fulfillment of the
requirement of the degree of “Bachelor of Pharmacy” of A.K.T.U. Lucknow, in the
academic year 2023-24.

Date:
Dr. Vikram Sharma
Director
 ACKNOWLEDGEMENT

Presentation inspiration and motivation have always played a key role in the success of
any venture.
I pay my deep sense of gratitude to Dr. Vikram Sharma, Director, Galgotias
College of Pharmacy, Greater Noida for his highest peak of encouragement and for all
the facilities that he provided for this project work.

I feel to acknowledge my indebtedness and deep sense of gratitude to Dr. Ravi

Kumar Mittal Sir whose valuable guidance and kind supervision given to me throughout
the course which shaped the present work as its show.

I am immensely obliged to other staff (Lab technician, Librarian) and my friends for
their elevating inspiration, encouraging guidance and kind supervision in the completion of

my project.

Last, but not the least, my parents are also an important inspiration for me. So with
due regards, I express my gratitude to them.

Manisha pal
2110920500043
B. pharma 7th Sem.
 Table of Contents

Sr.No Particulars Page No.

1. INTRODUCTION 1-7

2. APPROACHES 8-14

3. CHARACTERIZATION 15-18

4. APPLICATIONS 19-22

5. CONCLUSION 23

6. REFERENCE 24-27

INTRODUCTION
The oral route is frequently chosen for drug delivery because of its ease of administration. Still, these
formulations pose significant drawbacks like gastrointestinal discomfort, unpleasant taste, and decreased
bioavailability due to first pass metabolism[1,2]. An alternate strategy, continuous intra venous injection, is
regarded as a high‐dose drug manage ment approach for avoiding hepatic ‘first pass’ metabolism and
maintaining a long‐lasting, consistent drug level. However, this necessitates hospitalization of patients and
careful monitoring under medical help. Hence, the health sector currently reaps significant benefits from
transdermal drug delivery technologies that reduce plasma drug level fluctuations for repeated treatment, avoid
organ toxicity, and early metabolism, limit dose‐based side effects, and prevent gastrointestinal discomfort and
poor bioavailability. It also provides benefits such as controlled drug delivery, lower doses, and improved
patient compliance[3]. The tight junctions in the uppermost layer of the epidermis, stratum corneum (SC),
continue to be a significant barrier to the free penetration of drugs into the body, hence reducing the
bioavailability of trans dermal drugs[4,5,6]. Liposomes for topical administration of therapeutic agents have
ushered in a new era of research in the drug delivery domain. However, their unstable nature of merging with
the skin's lipids, dehydrating, and persisting near the skin surface resulted in poor skin permeability. Hence,
they were only associated with topical drug delivery. To address the limitations of traditional liposomes,
nanocarrier systems such as transfersomes and ethosomes were initiated. Transferosomes were developed by
Cevc et al. [7] in the 1990s containing surfactant (SA) or edge activator (EA)17 to impart elasticity to the
prepared vesicles.
These systems can deform their shape but cannot permeate into deeper layers of the skin. On the other hand,
ethosomal systems were introduced in 1998 containing high levels of ethanol in their formula, which resulted in
improved drug absorption through the skin due to the result of ethanol mixing in lipid bilayer vesicles and
enhancing saturation of SC lipids. To combine the synergistic effect of transfersomes and ethosomes, Song et
al. introduced a new delivery system, that is, transethosomes (TE) (Figure 1) which combines both systems to
conquer transdermal drug delivery systems and can also be administered through ophthalmic, transvaginal and
pulmonary routes[8,9]. Transethosomes also enhance drug penetration across ocular barriers for targeted
delivery to specific ocular tissues and improve bioavailability. They provide sus tained drug release, reducing
the frequency of administra tion and risk of adverse effects in other parts of the body. They are patient‐friendly
as it offers a convenient and noninvasive method for self‐administration. In addition, TEs can be formulated for
targeted delivery to specific regions of the pulmonary system. They enhance drug penetration across lung
tissues, improving therapeutic efficacy. Inhalation of drugs allows for direct delivery to the lungs, where a large
surface area and rich blood supply facilitate rapid absorption into the bloodstream. Furthermore, transvaginal
drug delivery bypassed the gastrointestinal tract and hepatic metabolism, resulting in improved bioavailability
and reduced systemic side effects The vaginal mucosa is highly vascularized, allowing efficient drug absorption
into the bloodstream. This route provides a noninvasive and convenient method of drug administration, with
potential applications in hormone replacement therapy, contraception, and treatment of vaginal infections.1
various approaches to delivering drugs through functio nalization, photodynamic therapy, and multiple routes of
administration. Furthermore, the therapeutic applica tions of transethosome‐based drug delivery systems are
also discussed. These novel ultra‐deformable vesicular systems have drawn much attention in recent years[10].
TEs contain the essential components of ethosomes (phospholipids, etha nol, and water) and the SA/EA in their
formula[11,12]. These nanocarrier systems showed enhanced penetration and biocompatibility, as their main
component in the prepara tion is a phospholipid, the most abundant ingredient of eukaryotic cells[13,14]. The
presence of ethanol together with surfactant provided softness and enhanced penetra tion, promoting their
passage into the biological membrane as measured by deformability and permeation experiments[15]. The SA
in its formulation leads to the transformation of SC lipids, thereby increasing its deform ability into the
biological membrane. Cholesterol can be added to enhance the vesicles' stability and control the drug release
rate. TEs can trap hydrophilic and lipophilic drugs/agents in the aqueous compartment or the lipid bilayer. They
can lower the concentration of drugs in other parts of the body, minimizing the risk of drug toxicity by
delivering pharmaceuticals to the intended site of action. Additionally, TE systems have significant patient
compli ance, stability, and biocompatibility. However, its molecular size must be appropriate for a drug to be
absorbed via the biological membrane. Notably, TEs may cause skin dermatitis like ethosomes since they have
a high alcohol concentration[16]. Moreover, this vesicular formation is easy to scale up and an excellent
industry choice. Following an overview of phospholipids, ethanol effect, edge activator effect on the properties
of trans ethosomal formulation, and effective targeting, this review discusses the production and evaluation
propert ies used to ensure the efficient delivery of drugs. Additionally, this article exclusively highlights the
various approaches to delivering drugs through functio nalization, photodynamic therapy, and multiple routes of
administration. Furthermore, the therapeutic applica tions of transethosome‐based drug delivery systems are
also discussed.

Fig. 1:- Transethosome

COMPONENTS OF TRANSETHOSOME

PROPERTIES To investigate the impact of formulation components on TE preparation, assimilating the role
and influence of these components is crucial for designing effective transethosomal formulations that can
optimize drug delivery across the biological barrier. This section delves into the key formulation components,
such as phospho lipids, ethanol, edge activator, and stabilizer, highlighting their impact on the physicochemical
properties, stability, and performance of Tes[17].

Phospholipids
Phospholipids are the key building blocks and vesicle forming agents derived from different sources. Based on
the sources, phospholipids can be divided into natural and synthetic phospholipids. Natural phospholipids can
be found in various products, including soybeans, egg yolks, and sunflower seeds[18]. Utilizing natural
unsaturated phospholipids cause the SC to fluidize, allowing APIs to permeate deeper layers. However, using
saturated (hydro genated) phospholipids improves or restores the skin's barrier function, facilitating APIs to
remain intact for longer. The phospholipids can be categorized as phospha tidylcholine (PC),
phosphatidylinositol, phosphatidylser ine, phosphatidylethanolamine, phosphatidic acid, and
phosphatidylglycerol. Due to the unsaturated properties of the hydrocarbon chain, compared to synthetic
phospho lipids, natural phospholipids are less stable during liposomal production. Natural lipids can be utilized
to make synthetic phospholipids, and an unlimited number of distinct and specified phospholipids can be
produced by altering the non‐polar and polar portions of phospholipid molecules[19]. Synthetic lipids include
dipalmitoyl phos phatidylcholine (DPPC), dimyristoyl phosphatidylcholine (DMPC), distearoyl
phosphatidylcholine (DSPC), hydroge nated soy phosphatidylcholine, 1,2‐dioleoyl‐sn‐glycero‐3
phosphocholine, 1,2‐distearoyl‐sn‐glycero‐3‐phospho‐1′‐rac glycerol, and 1,2‐dipalmitoyl‐sn‐glycero‐3‐
phosphoglycerol. The type and concentration of phospholipid selection are essential criteria for preparing TEs
because they influence vesicle properties such as entrapment efficiency (EE), size, ζ‐potential, penetration
properties, and stability. In trans ethosomal development, phospholipid concentrations typi cally range from
0.5%–5%. For instance, a study by Garg et al. investigated the effect of various concentrations of PC on the
ketoprofen‐loaded TEs properties.
The study found that increasing PC concentration from 0.5% to 2% resulted in an increase in vesicle size from
174.1 to 264.2 nm and EE from 55.3% to 72.6%, while a further rise in PC concentration to 3% and 4% caused
a decrease in vesicle size to 214.7 and 211.8 nm and EE to 69.3% and 65.9%, respectively. Ahmed et al. study
results demonstrated that when the drug‐to phospholipid's molar dose increases, the particle size increases and
forms multilamellar vesicles, significantly increasing penetration efficiency. Another study by Esposito et al.
evaluated the impact of different types of phospho lipids, including DPPC, DMPC, and DSPC, on the properties
of quercetin‐loaded TEs. The study found that DMPC and DSPC resulted in smaller vesicle sizes and higher
entrap ment efficiency than DPPC. Specifically, DMPC and DSPC led to vesicle sizes of 123.7 nm and
129.8nm, respectively, compared with 228.7nm for DPPC. Furthermore, DMPC and DSPC resulted in
entrapment efficiencies of 54.3% and 58.4%, respectively, compared to 41.7% for DPPC. Moreover, the
unsaturation degree of the phospholipid acyl chain can also impact the properties of TEs. A study by Fang et al.
evaluated the influence of the degree of unsaturation of the phospholipid acyl chain on the properties of TEs
loaded with curcumin. The results showed that TEs prepared with unsaturated phospholipids had higher EE and
skin perme ation than those prepared with saturated phospholipids. This relationship holds good until a specific
concentration; further phospholipid concentration will later not affect EE.

Edge activator
The deformability and flexibility of phospholipid vesicles are imparted by adding an edge activator. The type
and quantity of EA can alter the drug's permeability profile. Surfactants employed in the formation of TEs
include Tween 20, sodium cholate, dipotassium glycyrrhizinate, bile salts, Span 80, oleic acid, and Tween 80.62
As per the study of Hui Song et al. the transethosomal systems containing tween and oleic acid resulted in
increased size, and the sodium deoxycholate system remained virtually unchanged. Still, the zeta potential value
is increased, indicating stability and a threefold increase in EE, which may be due to reduced polarity by
combining deoxycholate anion and drug cation. Salem et al. and Khalid et al. study revealed that increasing
surfactant concentration could cause a substantial decrease in the vesicle size due to interfacial tension being
reduced by high surfactant levels covering the carrier surface. This Tween 80 effect results in its solubilizing
property and inhibition of vesicle cohesion. Due to the poly ethylene oxide groups on the nanovesicle's surface,
tween 20 prevents nanovesicle aggregation[24]. Recently, Fe properties of TEs loaded with a lipophilic drug,
curcumin, was studied by Zhang et al. The study found that the addition of curcumin decreased the particle size
and increased the zeta potential of TEs. Moreover, the TEs showed sustained drug release over 48h and
enhanced skin permeation of curcumin compared to a conventional cream. However, the addition of a drug can
also negatively affect the stability and integrity of TEs, and excessive drug loading can cause aggregation and
drug leakage. The investigations of Zhuang et al. on the effect of different drug‐to‐phospholipid ratios on the
stability and drug release properties of TEs loaded with tacrolimus, an immunosuppressive drug. The findings
showed that high drug‐to‐phospholipid ratios decreased the stability and drug‐release properties of the TEs and
caused drug leakage, which could decrease their therapeutic efficacy.
Ethanol
Ethanol provides softness to the vesicle membrane and acts as a penetration enhancer. Ethanol concentration
influences vesicle size, zeta potential, stability, enhanced skin, mucosal permeability, and entrapment efficacy.
The concentration of ethanol in the TE is 10%–50%. As per the reports of Salem et al. and Abdulbaqi et al., the
size of the vesicle decreases with the increase in ethanol concentration from 10% to 30% w/v and exceeds the
optimum level, leading to a leaky bilayer with a slight increase in the size of the vesicle. Further increase in
ethanol causes the phospholipid to easily dissolve in ethanol, with a significant decrease in entrapment efficacy;
this might be due to the interaction between the lipid layer and ethanol. A study by Tamer et al. found that TEs
containing 10%–20% ethanol showed higher stability and deformability compared to those containing 30%–
50% ethanol. The study also showed that the addition of cholesterol to the TE formulation improved stability
and deformability at higher ethanol concentrations. In another study, Abdallah et showed that TEs containing
30% ethanol significantly increased the skin permeation of 5‐fluorouracil com pared to a control solution. The
study also found that increasing the concentration of PC in the TE formulation enhanced skin permeation.
According to the study of Nayak et al., high ethanol saturation functions as a negative charge on the surface of
the vesicle, preventing the transethosomal system from converging as a result of electrostatic repulsion. Hence,
during the preparation process, the ethanol concentration should be optimized. chlorophyllin TEs were prepared
using 0.1% and 0.3% w/ v concentrations of span 20, tween 20, tween 80, and cremophor A25 as edge
activators with phosphatidylcho line and 20% w/v ethanol. When these surfactants were employed at low
concentrations, the mean vesicle sizes ranged from 456 to 685nm. TEs containing 0.1% w/v cremophor A25
showed a deformability index of 26.2–3.8 mL/s.39 Surfactants at high concentrations may cause skin irritation
and the disruption of the vesicular structure.

Stabilizer
Stabilizers are commonly used in the formulation of TEs to prevent their aggregation, maintain their size and
structure, and improve their shelf‐life. It imparts stability to the TE system. Cholesterol is the most commonly
used stabilizer. A study by Tamer et al. found that the addition of cholesterol to the TE formulation improved
stability and deformability at higher ethanol concentra tions. The increase in cholesterol concentration declines
the EE due to the low solubility of cholesterol. Liu et at. [21] investigated the effect of different concentrations
of a stabilizer, hydroxypropyl‐β‐cyclodextrin, on the stability and drug release properties of TEs loaded with a
hydrophilic drug, metronidazole. The study found that the addition of hydroxypropyl‐β‐cyclodextrin signifi
cantly improved the stability and drug‐release properties of the TEs. Moreover, the TEs containing a higher
concentration of hydroxypropyl‐β‐cyclodextrin showed enhanced skin permeation of metronidazole. The choice
of a stabilizer depends on the specific drug and formulation, and excessive amounts of a stabilizer can also
negatively affect the stability and integrity of TEs. The effect of different concentrations of sodium deox
ycholate was studied by Cao et at. [22] on the stability and drug release properties of TEs loaded with
ivermectin, an antiparasitic drug. The study found that high concentra tions of sodium deoxycholate (above 1%)
decreased the stability of the TEs and caused drug leakage, which could decrease their therapeutic efficacy.

Drug or active compound

The addition of a drug to the transethosomal system influences the vesicle's mean diameter and zeta potential
without affecting the dispersity index. According to the study of Kabil et plain Nanovesicles showed a negative
charge on the surface. The vesicles displayed a positive charge on the surface upon adding the drug. The effect
of drug loading on the stability and drug release.

Surface functionalization of TE
Functionalization of TEs can be achieved by modifying the surface of the vesicles with different chemical
moieties, such as targeting ligands, peptides, and anti bodies. These modifications can improve the specificity
and selectivity of the TEs, allowing them to target specific cells or tissues. For example, the functionalization of
TEs with folic acid has been shown to enhance their uptake by cancer cells that overexpress folate receptors.
Similarly, the use of transferrin‐conjugated TEs has been proved to enhance the uptake of drugs into cells that
express transferrin receptors[23,24]. Several studies have investigated the impact of functionalization on the
drug delivery efficiency of TEs. Ellis and Hicklin found a peptide that targets the vascular endothelial growth
factor receptor (VEGFR). The VEGFR‐targeted therapy was found to have higher cellular uptake and improved
anticancer activity compared to non‐functionalized therapy in the in vitro studies using breast cancer cells. In
another study by Zhao functionalization with a peptide that targets the αvβ3 integrin, which is over expressed in
several cancer cells. The αvβ3 integrin targeted therapy was found to have enhanced cellular uptake and
improved anticancer activity in in vitro studies using prostate cancer cells. Hui Song developed a sinomenine
hydrochloride‐loaded TE deco rated with an antioxidant surface that achieved targeted drug delivery and
improved hydrophilic drug efficiency, deformability, and permeation efficiency. The antioxi dant surface‐
coated TE is localized at the inflammation site through redox interactions in the presence of highly reactive
oxygen species levels.

APPROACHES

[1]Cold Method
This approach is most accessible and popular method used for the preparation of TEs this technique is usually
performed at lower temperatures[26].It entails preparation of organic and aqueous phase separately. Organic
phase is prepared by dissolving phospholipid s , penetration enhancers and other lipophilic materials in
hydrophobic solvents at room temperature in an enclosed container[27].Then hydrophilic phase is prepared by
taking by-distilled water, standard saline solution or buffer solution in a separate container. Then aqueous phase
is gradually added in hydrophobic phase by using syringe pump. Mixture is continuously agitated for 5-30 min
at 700-2000 rpm speed by using magnetic stirrer.Vivek Gupta et al.[28] Produced TEs by stirring at different
rates and studies revealed that the stirring time affects the size of vesicles. Based of the physicochemical
properties of drug, it will dissolve in either organic or an aqueous phase of TE system[29,30].The final
preparation is kept in refrigerator for furthermore use[31].This approach prevents the drug degradation due to
thermal stress[32].

Oraganic phase is prepared by dissolving lipid in Edge activator is added in mixture & heated upto
ethanol at temperature 20 - 25°C with continous 30°C under vigorous stirring for 5 minutes
stirring

Heated water is added slowly in alcoholic mixture Aqueous phase is prepared by heating water in
separate vessel upto 30°C

Synthesized TEs is stored in refrigerator at

Sonication is performed to reduce the size of temperature 4 - 8°C to maintain stability
synthesized vesicles

Fig 2 :-Method of Preparation of TEs by Cold Method

[2] Hot method
Hot method is an approach which utilized thermal energy for the formulation of TEs. This technique
mainly facilitates lipid fusion, enhance lipids solubility and increase encapsulation efficiency of drug
with within[26].This method includes dispersing of phospholipids in water and heated the mixture in
water bath upto 40°C to get colloidal solution. In separate container, mixture of ethanol and glycol is
taken and maintained at temperature 40°C. then mixture of ethanol and glycol is gradually added in
aqueous phase with continuous stirring for 7-10 min. Based on the physicochemical properties of
drug , it will either dissolve in ethanol (organic phase) or water (aqueous phase). temperature is
maintained at 40°C throughout the procedure. Sonication is used to reduce the size if vesicles of
TEs[33].

Fig 3 :- Hot Method

Organic phase of ethanol

Phospholipid is dispersed in water and heated and glycol mixture is slowly
upto temperature 40°C to form aqueous phase added into aqueous phase
with vigorous mixing for 7 -
10 minutes

Based on the chemical properties, drug will Sonication is performed to

either disslove in aqueous or organic phase reduce the size of
synthesized TEs

Fig 4 :- Method of Preparation of TEs by Hot Method

12
[3] Ethanol injection method
Ethanol injection method is one of the simplest method therefore it has many benefits in comparison
to other methods. Ethanol is mainly used as organic solvent because it is safe to use which makes it a
popular choice[34,35].This technique mainly focuses on the manufacturing of small, unilamillar
vesicles, of dimensions ranging from 30-170 nm. The actual size of vesicles depends upon the
amount of lipids used and the rate of injection administration[36,37].This technique consists of
dissolving phospholipids, SA and the active ingredients in ethanol. At determined flow rate
hydrophobic phase is injected into an hydrophilic phase by using injection system. Then ultrasonic
probe sonicator is used to homogenize the resultant mixture[29,38-40].Salem et al.[41] Prepared an
ultrasound assisted injection process in order to attain the smaller and homogeneous particle
dimensions compared to stirring assisted injection process.Singh et al.[42] Prepared TEs which are
loaded with hydrophilic drugs by utilizing the ethanol injection method followed by hot
procedure(probe sonication). The particle dimensions of vesicles which are sonicated is
comparatively smaller than the non sonicated vesicles.

Phospholipid , SA and Active ingredients is

dissolved in ethanol to form organic phase

This organic phase is injected into the aqueous

phase by using syringe system

Resultant mixture is homogenized by using ultra

sonic probe sonicator

Fig 5 :- Method of Preparation of TEs by Ethanol Injection

13
14
[4]Thin Film Hydration technique (TFH)
TFH is one of the well-known method employed for the preparation of Transethosomes. It consist of
preparation of thin layer of lipid on the internal surface of Rotary evaporator flask. Subsequently, thin film
of lipid is moistened by using water or a buffer solution. It is important to preheat the film of lipid and
water/buffer solution over the transitional temperature of lipids in order to create by-layer. Ultrasonic bath
and vigorous shaking is employed for sonication facilitates the detachment of lipid film from the surface of
flask and forms Transethosomes. This technique always helps to achieve reliable and desirable outcomes
even with small amount of compounds, but it has drawbacks of low encapsulation efficiency[38,43].This
method consist of dissolving EA, Phospholipids and drug in organic phase in RBF. To obtain the
homogeneous dispersion, mixture is placed in a water bath sonicator. For the formation of thin film of lipid
on the internal wall of flask, organic solvent is removed at low pressure and above the lipid transformation
temperature by rotary evaporation[44,45]. Flask is kept in a vaccum oven for overnight to ensure the
removal of organic solvent,aqueous solution of Ethanol or solution of saline phosphate buffer - ethanol is
used to moisten the dried lipid film by rotating the film of lipid. Then, vesicles of TEs are allowed to swell
at room temperature and kept in refrigerator at 4-8°C[46,44]. Sonbaty et al.[42] examined the influence of
methods like ethanol injection method and TFH method on the quality of TEs. The in vitro characterization
demonstrate the dimensions of vesicles manufactured by ethanol injection method and TFH were 245.3 and
62.85 nm respectively. Additionally, value of Zeta potential & encapsulation efficiency for ethanol injection
method were 36.3 mV & 82.35% and for TFH 49.4 mV & 93.88% respectively. Moreover, the percentage of
skin deposition for TE formulation prepared by TFH is 1.5 times greater than compared to TE formulation
prepared by Ethanol injection method and two folds higher than percentage of drug solution deposited in
propylene glycol.

Fig 6 :- Thin Film Hydration Method

15
 Phospholipid , EA , drug is
dissolved in organic phase

Water bath sonicator is used for

homogenous distribution of
organic phase

Organic solvent is removed by

using rotary evaporation to form
thin lipid film

Dried lipid film is diluted by

aqueous solution of ethanol or
solution of saline phosphate
buffer-ethanol by rotating the
lipid film

Synthesized TE vesicles are left

to swell at room temperature 20 -
25°C and stored in refrigerator at
4 - 8°C

Fig 7 :- Method of Preparation of TEs by Thin Film Hydration

16
[5]Mechanical Dispersion Method
This method consists of taking Ethanol & Chloroform mixture in round bottomed flask(RBF). Then Soya
PHOSPHATIDYL CHOLINE is introduced and dissolved in the mixture of Ethanol and Chloroform. Then
organic solvent is removed by employing Rotary vaccum Evaporator. The above step is performed at
temperature over the lipid transformation temperature. It facilitates the deposition of thin film of lipid on the
wall of round bottomed flask. Then RBF is kept overnight to obtain the traces of solvent which got
deposited on the wall of RBF. Then, dried lipid film is hyderated using Hydroethanol by simply rotating
RBF at optimal temperature condition by using different concentration of drug mixture[47].

Fig :- 8 Mechanical Dispersion Method

Chloroform and ethanol Soya phosphtidylcholine is

mixture is taken in round added to chloroform and
bottom flask ethanol mixture

Hydroethanol hydration is
performed by rotating round Organic solvent is removed by
bottom flask at required using rotary vaccum
temperature & using different evaporator at temperature
concentration of drug mixture above the lipid transition
temperature

Traces of solvent is obtained

from deposited thin lipid film
on the surface of round bottom
flask

Fig 9 :- Method of Preparation of TEs by Mechanical Dispersion Method

17
[6] Reverse Phase evaporation method (REV)
REV is a method used to synthesize TEs by preparing oil continuous emulsion, afterwards the non-aqueous
solvent is evaporated to create vesicles. This technique is mainly employed to create unilamillar
vesicle[38].This technique consists of drying of phospho lipids, stabilizers and organic solvent to create thin
lipid film by evaporation under vacuum. The residuary solvent is separated by using nitrogen gas to obtain
dried thin lipid film[38]. Then, in the solvent and solvent mixture, lipid film is suspended and mixed at room
temperature. In the sonication bath, synthesized formulation is sonically disrupted for 10 minutes at 5-6°C in
order to formulate stable emulsion[48,49]. On the removal of solvent at reduced pressure a gel is formed,
leading to formation of colloidal dispersion over mechanical agitation[30].Investigation of Nele et al.
[48]Revealed that the lamillarity of vesicles is affected by the method of formulation. So they performed
different techniques to form vesicles such as REV and film hydration followed by freeze-thaw space
cycles, film hydration followed by agitation on shaker. The outcomes showed that the vesicles produced by
freeze-thaw processes and agitation on shaker method were non-extruded, multi lamillar and varied
unilamillar/bilamillar extruded population. On the other hand, mostly unilamillar extruded vesicle
population produced by REV techniques.

Organic solvent is evaporated

Phospholipid s,stabilizers is under vacuum to create lipid film
dissolved in organic solvent to & Residuary solvent is separated
form oil continuous emulsion by using nitrogen gas

In sonication bath, synthesized

formulation is sonically disrupted Then , in solvent and solvent
for 10 minutes at 5 - 6 °C to form mixture lipid film is suspended
stable emulsion and mixed at room temperature

Fig 10 :- Method of Preparation of TEs by Reverse Phase evaporation method

18
 CHARACTERIZATION
[1] Morphology, vesicular size and poly-diversity index (PDI):
The size and PDI of TEs are most significant features of characterization. It has been observed that
inhalation, parenteral administration and their circulatory half life is influenced by the size of
vesicles[50].The typical shape of vesicular carriers are mostly spherical & imparts flexibility & softness
with an enclosed core.Depending on the formulation,they may be small in size ,unilamellar or
multilamellar[31,32]. As we know, for study of morphology , transmission electrons and scanning electrons
microscopy , nano sized vesicles are mostly employed[51]. Size of vesicles depends upon the equipment and
formulation techniques used in manufacturing and it is important to attain uniform size. For incorporating
the drug in the TEs the typical size of TEs should be between 50-200 nm[52].The mean vesicular size and
size distribution are determined by using techniques such as photon correlation spectroscopy,[53] microtrack
nanotrack [54] wave and dynamic light scattering method[28].
The value of PDI is used to measures the extent of homogeneity in sample, based on the measured size, it
can be mono-dispersed or poly dispersed. The value of PDI can be expandable between 0 - 1 & can be
dimensionless. The value of PDI which is equal to or below 0.3 used in delivery of drug, considered as the
most appropriate and homogeneous vesicle population[55]. On the other hand, high value of PDI denotes the
heterogeneity or multiple diverse vesicular population in the sample[56]. Dynamic light scattering(DLS)
was used to assess PDI[57]. DLS used to study the brownian motion of disperesed particale, particles which
are responsible for scattering of incident light in the solution. The extent of diffusion of lyposome in
suspention is related to the scattering of light. On the basis of quantity of light dispersed, mean particle size
is calculated[58]. Recently, a tool named nano particle tracking analysis was introduced by kim et al.[59] for
the determination of size by measurement of coefficient of particle diffusion from the sample. On the basis
of variations in the scattered light intensity, DLS measures particle diffusion coefficient. On the other hand,
individual movement of particle in consecutive optical video images are established by nano particle
tracking analysis. As DLS a nanoparticle tracking analysis measures the same physical property, so size
calculated by DLS can be validated by employing nanoparticle tracking analysis.

[2]Zeta potential
Zeta potential is commonly described by dispersion of charges on the vesicular carrier’s surface. The
stability of nano particles is majorly established by the presence of surface charge on nanoparticles. The
presence of excipients in formulation establish the nature of charge such as positive of negative on vesicular
carrier. The level of electrostatic repulsion or attraction in the colloidal dispersion system is known as Zeta
potential. It gives information detail relating to each component of formulation and their interaction with
each other.The interaction between the vesicles and biological membrane is also established by Zeta
potential. It provides knowledge related to surface chemistry[60,61].

[3]Entrapment efficiency
The synthesis of TEs with acceptable EE and drug release is only facilitate by the critical evaluation of
properties of vesicles. The TE formulation methodology, its constitution and the hardness of phospholipid
bilayer significantly affects the EE of drug. In the end product of TE a mixture of both encapsulated and
non encapsulated portion of drug have been found. The presence of drug in TE and EE is calculated by first
separating the free drug. There are many approaches used to determine the EE like size exclusion
chromatography which is depends opon size variation, dialysis membrane, centrifugal force and ultra
centrifugation. The EE and drug loading is established an ultracentrifugation method. The non encapsulated
drug from synthesized vesicles is separated by centrifugation of suspension of drug-TE at controlled
19
temperature and speed to prevent the breaking of vesicles[59].The separation of residue and supernatent
liquid occur. The breakdown of vesicles established the concentration of drug molecule in the sediment by
using
appropriate solvent. The spectrophotometric analysis is done to determine the concentration of drug
molecules[62]. The EE can be determined byusingequation:-

%EE = (Total drug added - Free unentraped drug) x 100

Total drug added

[4] Drug content

The vesicles are ruptured to release the content in order to check the amount of active ingredient in the
vesicles.The spectroscopic analysis of solution containing released drug content was performed.Vesicles are
lysed by using solvents like methanol,isopropyl alcohol etc[63].

[5] Measurement of in vitro skin permeation

Franz diffusion cell is used to determine the in vitro drug release[64,40]. Selection of Dialysis bag
membrane should be done based on drug parameters.The cellophane with particular molecular weight is
commonly employed as dialysis membrane, it is moistened with biological solution mainly Phosphate
buffer saline at pH 7.4.The formulation is loaded in donor site & phosphate buffer saline with pH 7.4 is
loaded in receptor site & mixed by using magnetic stirrer at 300 - 400 rpm at temperature 32+ - 1°C to
imitate the human skin .At periodic intervals ,1ml of portion is collected & at the same time equal amount of
fresh buffer is added to maintain the required volume.Spectrophotometric analysis is performed to analyze
drug samples[31,40].

[6] In vitro study of drug release

The dialysis bag technique is used to analyze the release pattern of drug.In this approach, the formulation of
transethosomes is placed in membrane & then this membrane is kept in a conical flask which contains buffer
solution & incubated.Column centrifugation method is used to centrifuge at predetermined time intervals in
order to extract aliquot.By using appropriate method ,release of drug is evaluated[65]. It was revealed that
the release pattern of drug can be prolonged for the duration of 24 hrs.This may improve patient compliance
by reducing the dosing frequency[66].

[7] Elastic determination

It is crucial element to determine the penetration of synthesized transethosomes through skin.The elastic
properties of bi-layer of vesicles is established by using extrusion method.The vesicles are released by
applying pressure through membrane filter, which is composed of cellulose with optimal pore
size .Dispersion of extruded vesicles is determined by [29]

E = J x {v/p}2

20
 Where, E = Elasticity of membrane
J = Flux rate through cellulose membrane filter

V = Vesicular size after ejection

P = Membrane pore size

The drug permeation of transethosomes found better when compared to ethosomes as it showed better
elasticity index than ethosomes[67].

[8] Phase transformation temperature

The release of drug from vesicles is better understand by determining the temperature at which phase change
takes place . By using constant nitrogen stream, samples are examined at a optimal temperature condition.
The comparison of specimens is performed by employing differential thermal curves[67].

[9] Stability
The stability of synthesized transethosomes can be determined by ensuring that there is any variations in size
of vesicles .In terms of stability , homogeneous preparations attains more stability than heterogeneous
preparations. Preparation stability is also determined by study of its membrane stability, arrangement of
molecules by X ray scattering or differential scanning method.The inner stability of preparation is not
determined by study of particle dimensions because this is just analyzed as Q.C test.This conclusion is made
as this is observed in lyophilized liposomes where cryoprotectant interrupt with the molecular arrangement
of bi-layer membrane & particle size of lyophilized liposome is stable[68].

APPLICATION

[1] Anti fungal drug delivery

Econazole nitratite formulated as transethosomal gel compared to commercialized topical cream of
econazole nitratite to check the potential of antifungal effect of drug.It was observed that transethosomal gel
showed effective skin maintenance and antifungal activity.The release of drug from transethosomal gel in a
regular pattern which cure cutaneous candiadis [69].
[2] Peptide drug delivery
Due to greater particle size ,peptides cannot able to pass through the stratum corneum.As a result ,
transdermal delivery of peptide is tricky.Encapsulated palmitoyl pentapeptide administered in a form of
transethosomal formulation showed increased elasticity , which enhance skin permeation[70].
[3] Transdermal delivery of anti inflamatory bioactives
Paolino et al.[71] Carried out an experiment on human participants by taking ethosomes loaded with
ammonim glycyrrhizinate to examine the efficacy of ethosomal formulation in delivery of antifungal drug.
The ethosomal formulation having ethanol percentage about 45% and lower lecithin concentration gave
better outcomes in both in vivo & in vitro consideration.In case of in vivo consideration it showed enhanced
21
anti inflamatory action in human volunteers & in case of in vitro,it showed better tolerability & increased
percutaneous absorption.Nanoethosomes contain high amount of ethanol which showed better entrapment
efficiency.

[4] Transvaginal drug delivery

The steroidal hormone in female is Progesterone which can be employed to treat Polycystic ovarian
syndrome.But due to oral route of administration ,it undergoes primary metabolism & led to poor solubility
& bioavailability.Salem et al. [72] Prepared transethosomal mucoadhesive gel loaded with Progesterone for
transvaginal delivery of drug.The study found a marked rise in degree of echogenicity thickness of
endometrium, serum progesterone level & fertility rate.Hence transvaginal gel might be promising
preparation for progesterone delivery to enhance the pregnancy rate in polycystic ovarian syndrome.

[5] Anti cancer drug delivery

The trials of dual medications loading into transethosomes to cure cutaneous melanoma completed by
scientists.Two medications such as dacarbazine & tretionin were chosen with synergistic action, that results
in reduced cell toxicity in comparison with other pharmaceutical formulations. Dual drug Transethosomes
showed increased anticancer effectiveness when skin permeation compared to single drug loaded
Transethosomes.Other studies, also revealed that 5 - fluorouracil loaded in transethosomal gel gave good
skin target delivery & high superficial permeation as correlated to ethosomes[73].

[6] Anti arthritis drug delivery

To synthesize antioxidants coated with transethosomes, Transethosomes were loaded with sinomenine
hydrochloride and subsequently enclosed with ascorbic acid.This led to enhanced medication deposition &
dermatological penetration to treat rhematoid oxidative stress[74].

[7] Opthalmic drug delivery

Transethosomes have been displayed to enhance the therapeutic effects & bioavailability of drugs employed
in the treatment of different eye conditions including macular degeneration, dry eye syndrome, glaucoma.
Transethosomes showed the increased drug permeation through corneal epithelium and enhanced drug
retention in ocular tissues.Some studies have also revealed that transethosomes can improve patient
compliance by reducing the frequency of drug administration & potential side effects[75]. Ahmed et al. [76]
Prepared ketoconazole transethosomal vesicles to enhance short biological half life , quick eye clearance and
drug’s ocular permeation which have high permeability into posterior eye. Ciprofloxacin loaded in
transethosomes may demonstrate long lasting antibacterial effect by enhanced ocular bioavailability, which
may give better clinical results in the treatmemt of bacterial endophthalmitis[77].

[8] Transport of drug with action on cardiovascular system

Touitou et al. Investigated effectiveness of minoxidil loaded nanoethosomes to check the efficiency of skin
penetration of cardiovascular drug.The nanoethosomes prepared with 2% phosphatidylcholine & 30% of

22
ethanol showed enhanced permeability than the preparation of hydro-ethanolic & phospholipid minoxidil. In
another experment, Ahad et al. examined the penetration power of valsartan loaded nanoethosomes into the
skin of wistar albino rats. The outcomes showed increased Bio-availability than those of orally administered
drug & enhanced skin penetration when evaluated with liposomal formulation of same drug . Bhosale &
Avachat also documented the enhanced effect of drug which is given transdermally in comparison with oral
administration of same drug[78].

[9] Transport of bioactive agent with antiviral activity

Jain et al. has been prepared lamivudine loaded nanoethosomes to examine the effectiveness of topical
delivery of antiviral drug.The nanoethosomal formulation exhibited 25 times more effective penetration of
drug when compared to Conventional solution.In comparision with free drug solution T- Lymphocytes
exhibited enhanced absorption capacity for nanoethosomes [81].

[10] Aging and androgenic alopecia (AGA)

Wang et al. [49] And Basto et al. [82] manufactured encapsulated vesicles to make effective delivery of
drug niacinamide and cycloastragenol into cutaneous layer. Transethosomes which uses penetration
enhancer like jojoba oil , span 40 , dicetylphosphate and tween 80 have good drug delivery through the skin
barrier. This type of vesicles can be used for synthesis of anti-aging cosmetics to provide protection to
skin.The topical treatment is better for patients in comparison to other therapy choices for androgenic
alopecia as it is very simple to administer & have less side effects because of site-specific
application.Zaafarany et al. [83] Tested coenzyme Q10 (CoQ10) by filling it in various phospholipidic
vesicular system.Results showed that transethosomes were ideal carrier for CoQ10 deposition percentage in
the different layer of skin.Allam et al. [84] Encapsulated drug minoxidil in transethosomes to enhance the
effectiveness & reduce the adverse effect.Transethosomes which contain oleic acid exhibited high skin
deposition and skin permeability.

[11] Antihypertensive drug delivery

As we know oral administration of most of antihypertensive drugs results in decreased bioavailability in
body because drug undergoes primary metabolism. For example olmesartan medoxomil was loaded as
active ingrediant in a form of transethosomal gel showed good transdermal permeability [85].

[12] Pulmonery drug delivery

Nasr et al.[86] were studied the vinpocetine & piracetam loaded transethosomes & other vesicular
formulations like transferosomes , microemulsion and other composite system for their healing capability in
memory impairment induced scopolamine.Transethosomes showed improved Entrapment efficiency & more
stability.However,Nano-composite formulations showed better antiinflamatory & antiapoptotic properties
when compared to micro-emulsion and transethosomal formulations.The nanoformulations
(Transethosomes,micro-emulsion & nanocomposite) were highly efficacious due to fine particle size & high
permeability over the nasal mucosa.Lipid nanovesicles loaded with rolapitant created by kabil et al. [47]

23
Which is novel molecular entity that can inhibit the proliferation of lung cancer cells & deliver the drug
through nebulization approach.In vivo study revealed lipid vesicles get placed in the lungs through systemic
circulation.Additionally, transethosomes formulations are non toxic to lung tissues. As the pharmacological
& clinical research of rolapitant has not yet done, the data collected offers an opportunity to study the
potential of rolapitant in treatment of lung cancer.

24
 CONCLUSION

Transethosomes have emerged as a groundbreaking drug delivery system, poised to revolutionize the field
of dermatology and transdermal medicine. By virtue of their unique composition and structure,
transethosomes have demonstrated an unparalleled ability to traverse the skin barrier, releasing drugs in a
controlled, targeted, and efficacious manner.The comprehensive review of the existing literature has
unequivocally highlighted the vast potential of transethosomes in delivering a wide array of drugs,
encompassing small molecules, macromolecules, and even nucleic acids. The versatility of transethosomes
has been further underscored by their successful application in various dermatological and transdermal
contexts, including skin cancer, psoriasis, atopic dermatitis, and pain management.One of the most
significant advantages of transethosomes lies in their ability to enhance skin permeation, thereby facilitating
the delivery of therapeutics to the target site. This is particularly crucial for drugs with poor skin
permeability, which have hitherto been challenging to deliver transdermally.
The use of transethosomes has been shown to significantly improve the bioavailability of such drugs,
leading to enhanced therapeutic outcomes.Furthermore, the flexibility of transethosomes in terms of
formulation and modification has been amply demonstrated. The ability to tailor the composition and
structure of transethosomes to suit specific applications has enabled researchers to optimize their
performance and efficacy. This versatility is expected to play a critical role in the future development of
transethosomes for a wide range of applications.Despite the immense potential of transethosomes, there are
several challenges that need to be addressed to facilitate their translation from bench to bedside. Scalability,
stability, and regulatory considerations are among the key issues that require attention. Moreover,
comprehensive in vivo studies are necessary to fully elucidate the safety, efficacy, and pharmacokinetics of
transethosomes.

25
 REFERENCE

1. Salem HF, Kharshoum RM, Abou‐Taleb HA, AbouTaleb HA, AbouElhassan KM. Progesterone‐loaded
nanosized trans ethosomes for vaginal permeation enhancement: formula tion, statistical optimization, and
clinical evaluation in anovulatory polycystic ovary syndrome. J Liposome Res. 2019;29(2):183‐194.
2. Rahangdale M, Pandey P. Development and characterization of apremilast transethosomal gel for
transdermal delivery. Int J Pharm Sci Nanotechnol. 2021;14(3):5508‐5518.
3. Verma S, Utreja P. Exploring therapeutic potential of invasomes, transfersomes, transethosomes, oleic
acid vesi cles, and cubosomes adopting topical/transdermal route. Micro Nanosyst. 2022;14(1):3‐20.
4. Sguizzato M, Ferrara F, Mariani P, et al. “Plurethosome” as vesicular system for cutaneous administration
of mangiferin: formulative study and 3d skin tissue evaluation. Pharmaceutics. 2021;13(8):1124.
5. Albash R, El‐Nabarawi MA, Refai H, et al. Tailoring of PEGylated bilosomes for promoting the
transdermal delivery of olmesartan medoxomil: in‐vitro characterization, ex‐vivo permeation and in‐vivo
assessment. Int J Nanomedicine. 2019;14:6555‐6574.
6. Panchaxari Gadad A, Patil AS, Singh Y, Mallappa Dandagi P, Bolmal UB, Basu A. Development and
evaluation of flurbiprofen loaded transethosomes to improve transdermal delivery. Indian J Pharm Educ
Res. 2020;54(4):954‐962.
7. Cevc G, Blume G. Lipid vesicles penetrate into intact skin owing to the transdermal osmotic gradients and
hydration force. Biochim Biophys Acta‐ Biomembr. 1992;1104(1): 226‐232.
8. Shaji J, Parab SS. Formulation development of tranexamic acid loaded transethosomal patch for melasma.
Res J Pharm Dosage Forms Technol. 2022;14(1):7‐16.
9. Rodríguez‐Luna A, Talero E, Ávila‐Román J, et al. Prepara tion and in vivo evaluation of rosmarinic
acid‐loaded transethosomes after percutaneous application on a psoriasis animal model. AAPS
PharmSciTech. 2021;22(3):103
10. Esentürk güzel I. Improved skin penetration and deposition of naftifine from transethosomes and
transethosomal gel formulations. Farmacia. 2022;70(3):514‐521.
11. MahmoudDB,ElMeshadAN,FadelM,TawfikA, RamezSA. Photodynamic therapy fortified with topical
oleyl alcohol based transethosomal 8‐methoxypsoralen for ameliorating vitiligo: optimization and clinical
study. Int J Pharm. 2022; 614:121459.
12 . Ansari MD, Ahmed S, Imam SS, et al. CCD based development and characterization of nano‐
transethosome to augment the antidepressant effect of agomelatine on Swiss albino mice. J Drug Delivery
Sci Technol. 2019;54:101234.
13. Al‐Daraji M, Samy N, Fadel M. Topical indocyanine green nano system photodynamic therapy versus
cryotherapy for treatment of common warts: a randomized controlled study. J Nat Remedies. 2021;22(1
(2)):157‐164.
14.. Hesham H, Rady M, Hathout RM, Abdel‐Halim M, Mansour S. The skin delivery of tofacitinib citrate
using transethosomes and hybridized ethosomes/nanostructured lipid carriers for vitiligo therapy:
dermatopharmacokinetics and in vivo assays. Int J Pharm. 2022;629:122387.
15. Ansari SA, Qadir A, Warsi MH, et al. Ethosomes‐based gel formulation of karanjin for treatment of acne
vulgaris: in vitro investigations and preclinical assessment. 3 Biotech. 2021;11(11):456.

26
16. Mosallam S, Albash R, Abdelbari MA. Advanced vesicular systems for antifungal drug delivery. AAPS
PharmSciTech. 2022;23(6):206.
17. El Kayal M. Optimization of the colloidal properties of different vesicular systems aiming to encapsulate
(‐) epigallocatechin‐3‐gallate. Farmacia. 2020;68(1):97‐110.
18. Monteiro N, Martins A, Reis RL, Neves NM. Liposomes in tissue engineering and regenerative
medicine. J R Soc Interface. 2014;11(101):20140459.
19. Van Hoogevest P, Wendel A. The use of natural and synthetic phospholipids as pharmaceutical
excipients. Eur J Lipid Sci Technol (EJLST). 2014;116(9):1088‐1107.
20. Torchilin VP. Recent advances with liposomes as pharma ceutical carriers. Nat Rev Drug Discov.
2005;4(2):145‐160
21. Liu R. Formulation optimization of transethosomes for enhanced skin delivery of metronidazole. J
Liposome Res. 2019;29(4):335‐344.
22. Cao Y. Transethosomes containing sodium deoxycholate for dermal delivery of ivermectin:
optimization, characteriza tion, and in vitro/in vivo evaluation. Drug Des Devel Ther. 2019;13:2465‐2478.
23. Limongi T, Susa F, Marini M, et al. Lipid‐based nanovesi cular drug delivery systems. Nanomaterials.
2021;11(12): 3391.
24. Ellis LM, Hicklin DJ. VEGF‐targeted therapy: mechanisms of anti‐tumour activity. Nat Rev Cancer.
2008;8(8):579‐591.
25. Natsheh H, Touitou E. Phospholipid vesicles for dermal/ transdermal and nasal administration of active
molecules: the effect of surfactants and alcohols on the fluidity of their lipid bilayers and penetration
enhancement properties. Molecules. 2020;25(13):2959.
26.Shah S, Dhawan V, Holm R, Nagarsenker MS, Perrie Y. Liposomes: Advancements and innovation in
the manufacturing process. Advanced drug delivery reviews. 2020 Jan 1;154:102-22.
27.Sundar VD, Divya P, Dhanaraju MD. Design development and characterisation of tramadol
hydrochloride loaded transethosomal gel formulation for effective pain management. Indian J. Pharm. Educ.
Res. 2020 Apr 1;54(2):88-97
28.Gupta V, Joshi NK. Formulation, development and evaluation of ketoprofen loaded transethosomes gel.
Journal of Drug Delivery and Therapeutics. 2022 Jan 15;12(1):86-90.

29. Song CK, Balakrishnan P, Shim CK, Chung SJ, Chong S, Kim DD. A novel vesicular carrier,
transethosome, for enhanced skin delivery of voriconazole: characterization and in vitro/in vivo evaluation.
Colloids and surfaces B: biointerfaces. 2012 Apr 1;92:299-304.
30.Mishra KK, Kaur CD, Gupta A. Development of itraconazole loaded ultra-deformable transethosomes
containing oleic-acid for effective treatment of dermatophytosis: Box-Behnken design, ex-vivo and in-vivo
studies. Journal of Drug Delivery Science and Technology. 2022 Jan 1;67:102998.
31.Hallan SS, Sguizzato M, Mariani P, Cortesi R, Huang N, Simelière F, Marchetti N, Drechsler M, Ruzgas
T, Esposito E. Design and characterization of ethosomes for transdermal delivery of caffeic acid.
Pharmaceutics. 2020 Aug 6;12(8):740.
32.Costanzo M, Esposito E, Sguizzato M, Lacavalla MA, Drechsler M, Valacchi G, Zancanaro C, Malatesta
M. Formulative study and intracellular fate evaluation of ethosomes and transethosomes for vitamin D3
delivery. International journal of molecular sciences. 2021 May 19;22(10):5341..

27
33. Kumar L,Verma S,Singh K, Prasad DN, Jain AK.Ethanol Based Vesicular Carriers in Transdermal Drug
Delivery: Nanoethosomes and Transethosomes in Focus. Nano World J. 2016;2(3):41-51

34. Chen ZX, Li B, Liu T, Wang X, Zhu Y, Wang L, Wang XH, Niu X, Xiao Y, Sun Q. Evaluation of
paeonol-loaded transethosomes as transdermal delivery carriers. European journal of pharmaceutical
sciences. 2017 Mar 1;99:240-5.
35.Garg V, Singh H, Bhatia A, Raza K, Singh SK, Singh B, Beg S. Systematic development of
transethosomal gel system of piroxicam: formulation optimization, in vitro evaluation, and ex vivo
assessment. AAPS pharmscitech. 2017 Jan;18:58-71.
36.Abdulbaqi IM, Darwis Y, Khan NA, Assi RA, Khan AA. Ethosomal nanocarriers: the impact of
constituents and formulation techniques on ethosomal properties, in vivo studies, and clinical trials.
International journal of nanomedicine. 2016 May 25:2279-304.
37.Šturm L, Poklar Ulrih N. Basic methods for preparation of liposomes and studying their interactions with
different compounds, with the emphasis on polyphenols. International journal of molecular sciences. 2021
Jun 18;22(12):6547.

38.Pérez JV, Cortés DM, y Gómez YG. Potential use of transethosomes as a transdermal delivery system for
metabolites from Chenopodium murale. Materials Today Communications. 2022 Mar 1;30:103165.
39.Kaul S, Jain N, Nagaich U. Ultra deformable vesicles for boosting transdermal delivery of 2-
arylpropionic acid class drug for management of musculoskeletal pain. Journal of Pharmaceutical
Investigation. 2022 Mar;52(2):217-31.
40. El-Sonbaty MM, Akl MA, Khalid M, Kassem AA. Does the technical methodology influence the quality
attributes and the potential of skin permeation of Luliconazole loaded transethosomes?. Journal of Drug
Delivery Science and Technology. 2022 Feb 1;68:103096.
41. Salem HF, Nafady MM, Kharshoum RM, Abd el-Ghafar OA, Farouk HO. Mitigation of rheumatic
arthritis in a rat model via transdermal delivery of dapoxetine HCl amalgamated as a nanoplatform: in vitro
and in vivo assessment. International journal of nanomedicine. 2020 Mar 6:1517-35.
42. Singh M, Lee KE, Vinayagam R, Kang SG. Synthesis and assessment of lipid nanovesicles for efficient
transdermal delivery of hydrophilic molecules. Nano. 2022 Apr 7;17(04):2250031.
43. Fu Y, Saraswat A, Vartak R, Patki M, Patel K. Liposomal formulation: Opportunities, challenges, and
industrial applicability. Multifunctional Nanocarriers. 2022 Jan 1:79-102.
44. Kabil MF, Nasr M, Ibrahim IT, Hassan YA, El-Sherbiny IM. New repurposed rolapitant in
nanovesicular systems for lung cancer treatment: Development, in-vitro assessment and in-vivo
biodistribution study. European journal of pharmaceutical sciences. 2022 Apr 1;171:106119.
45. Khan AU, Jamshaid H, ud Din F, Zeb A, Khan GM. Designing, optimization and characterization of
trifluralin transfersomal gel to passively target cutaneous leishmaniasis. Journal of Pharmaceutical Sciences.
2022 Jun 1;111(6):1798-811.
46. Wang FC, Hudson PL, Burk K, Marangoni AG. Encapsulation of cycloastragenol in phospholipid
vesicles enhances transport and delivery across the skin barrier. Journal of Colloid and Interface Science.
2022 Feb 15;608:1222-8.

47. Dubey V, Mishra D, Jain NK. Melatonin loaded ethanolic liposomes: physicochemical characterization

28
and enhanced transdermal delivery. European Journal of Pharmaceutics and Biopharmaceutics. 2007 Sep
1;67(2):398-405.
48. Nele V, Holme MN, Kauscher U, Thomas MR, Doutch JJ, Stevens MM. Effect of formulation method,
lipid composition, and PEGylation on vesicle lamellarity: a small-angle neutron scattering study. Langmuir.
2019 Apr 12;35(18):6064-74.
49.Zare Kazemabadi F, Heydarinasab A, Akbarzadeh A, Ardjmand M. Preparation, characterization and in
vitro evaluation of PEGylated nanoliposomal containing etoposide on lung cancer. Artificial cells,
nanomedicine, and biotechnology. 2019 Dec 4;47(1):3222-30.
50. Elsana H, Olusanya TO, Carr-Wilkinson J, Darby S, Faheem A, Elkordy AA. Evaluation of novel
cationic gene based liposomes with cyclodextrin prepared by thin film hydration and microfluidic systems.
Scientific reports. 2019 Oct 22;9(1):15120.
51. Wang Y, Wang R, Qi X, Li W, Guan Q, Wang R, Li X, Li Y, Yang Z, Feng Y. Novel transethosomes
for the delivery of brucine and strychnine: Formulation optimization, characterization and in vitro evaluation
in hepatoma cells. Journal of drug delivery science and technology. 2021 Aug 1;64:102425.

52. William B, Noemie P, Brigitte E, Geraldine P. Supercritical fluid methods: An alternative to

conventional methods to prepare liposomes. Chemical Engineering Journal. 2020 Mar 1;383:123106.
53.Torchilin VP. Recent advances with liposomes as pharmaceutical carriers. Nature reviews Drug
discovery. 2005 Feb 1;4(2):145-60.

54. Abdulbaqi IM, Darwis Y, Assi RA, Khan NA. Transethosomal gels as carriers for the transdermal
delivery of colchicine: statistical optimization, characterization, and ex vivo evaluation. Drug design,
development and therapy. 2018 Apr 9:795-813.

55.Danaei MR, Dehghankhold M, Ataei S, Hasanzadeh Davarani F, Javanmard R, Dokhani A, Khorasani S,

Mozafari MR. Impact of particle size and polydispersity index on the clinical applications of lipidic
nanocarrier systems. Pharmaceutics. 2018 May 18;10(2):57.
56. Gaumet M, Vargas A, Gurny R, Delie F. Nanoparticles for drug delivery: the need for precision in
reporting particle size parameters. European journal of pharmaceutics and biopharmaceutics. 2008 May
1;69(1):1-9.
57.Bektay HŞ, Kahraman E, Güngör S. Design of skin-simulating nanoformulations for ceramide
replacement in the skin: a preliminary study. Maced. Pharm. Bull.. 2020;66:101-2.
58.Nkanga CI, Bapolisi AM, Okafor NI, Krause RW. General perception of liposomes: formation,
manufacturing and applications. Liposomes-advances and perspectives. 2019 Mar 26.
59.Kim A, Ng WB, Bernt W, Cho NJ. Validation of size estimation of nanoparticle tracking analysis on
polydisperse macromolecule assembly. Scientific reports. 2019 Feb 25;9(1):2639.
60. Radomska-Soukharev A. Stability of lipid excipients in solid lipid nanoparticles. Advanced drug
delivery reviews. 2007 Jul 10;59(6):411-
61. Severino P, Szymanski M, Favaro M, Azzoni AR, Chaud MV, Santana MH, Silva AM, Souto EB.
Development and characterization of a cationic lipid nanocarrier as non-viral vector for gene therapy.
European Journal of Pharmaceutical Sciences. 2015 Jan 23;66:78-82.

29
62.Bi Y, Xia H, Li L, Lee RJ, Xie J, Liu Z, Qiu Z, Teng L. Liposomal Vitamin D3 as an Anti-aging Agent
for the Skin. Pharmaceutics. 2019 Jul 3;11(7):311.
63. Mbah C, Builders P, Nzekwe I, Kunle O, Adikwu M, Attama A. Formulation and in vitro evaluation of
pH-responsive ethosomes for vaginal delivery of metronidazole. Journal of Drug Delivery Science and
Technology. 2014 Jan 1;24(6):565-71

64. Sudhakar K, Mishra V, Jain S, Rompicherla NC, Malviya N, Tambuwala MM. Development and
evaluation of the effect of ethanol and surfactant in vesicular carriers on Lamivudine permeation through the
skin. International journal of pharmaceutics. 2021 Dec 15;610:121226.
65. United State Pharmacopeial/National Formul. Validation of Compendial Methods Section. USP38/NF33
2015:2256. The United States Pharmacopeial Convention 12601, Twinbrook Parkway, Rockville, MD
20852.
66.Kumar L, Utreja P. Formulation and characterization of transethosomes for enhanced transdermal
delivery of propranolol hydrochloride. Micro and nanosystems. 2020 Apr 1;12(1):38-47.
67. Shaji J, Bajaj R. Transethosomes: A new prospect for enhanced transdermal delivery. International
journal of pharmaceutical sciences and research. 2018 Jul 1;9(7):2681-5

68. Stark B, Pabst G, Prassl R. Long-term stability of sterically stabilized liposomes by freezing and freeze-
drying: Effects of cryoprotectants on structure. European journal of pharmaceutical sciences. 2010 Nov
20;41(3-4):546-55.
69.Verma S, Utreja P. Transethosomes of econazole nitrate for transdermal delivery: Development, in-vitro
characterization, and ex-vivo assessment. Pharmaceutical Nanotechnology. 2018 Sep 1;6(3):171-9.
70.Kim JE, Oh GH, Jang GH, Kim YM, Park YJ. Transformer-ethosomes with palmitoyl pentapeptide for
improved transdermal delivery. Journal of drug delivery science and technology. 2019 Aug 1;52:460-7.
71.Paolino D, Lucania G, Mardente D, Alhaique F, Fresta M. Ethosomes for skin delivery of ammonium
glycyrrhizinate: in vitro percutaneous permeation through human skin and in vivo anti-inflammatory activity
on human volunteers. Journal of controlled release. 2005 Aug 18;106(1-2):99-110.
72.Salem HF, Kharshoum RM, Abou-Taleb HA, AbouTaleb HA, AbouElhassan KM. Progesterone-loaded
nanosized transethosomes for vaginal permeation enhancement: formulation, statistical optimization, and
clinical evaluation in anovulatory polycystic ovary syndrome. Journal of liposome research. 2019 Apr
3;29(2):183-94.
73.Shaji J, Bajaj R. Formulation development of 5-fluorouracil transethosomes for skin cancer therapy. Int.
J. Pharm. Pharm. Res. 2017;11:454-64.
74. Song H, Wen J, Li H, Meng Y, Zhang Y, Zhang N, Zheng W. Enhanced transdermal permeability and
drug deposition of rheumatoid arthritis via sinomenine hydrochloride-loaded antioxidant surface
transethosome. International journal of nanomedicine. 2019 May 1:3177-88.
75.Zazo H, Colino CI, Lanao JM. Current applications of nanoparticles in infectious diseases. Journal of
Controlled Release. 2016 Feb 28;224:86-102.
76.Ahmed TA, Alzahrani MM, Sirwi A, Alhakamy NA. Study the antifungal and ocular permeation of
ketoconazole from ophthalmic formulations containing trans-ethosomes nanoparticles. Pharmaceutics. 2021
Jan 24;13(2):151.

30
77.Youssef A, Dudhipala N, Majumdar S. Ciprofloxacin loaded nanostructured lipid carriers incorporated
into in-situ gels to improve management of bacterial endophthalmitis. Pharmaceutics. 2020 Jun;12(6):572.
78.Ahad A, Aqil M, Kohli K, Sultana Y, Mujeeb M. Enhanced transdermal delivery of an anti-hypertensive
agent via nanoethosomes: statistical optimization, characterization and pharmacokinetic assessment.
International journal of pharmaceutics. 2013 Feb 25;443(1-2):26-38.
79.Nayak D, Tawale RM, Aranjani JM, Tippavajhala VK. Formulation, optimization and evaluation of
novel ultra-deformable vesicular drug delivery system for an anti-fungal drug. AAPS PharmSciTech. 2020
Jul;21:1-0.
80.Gadad AP, Patil AS, Singh Y, Dandagi PM, Bolmal UB, Basu A. Development and evaluation of
flurbiprofen loaded transethosomes to improve transdermal delivery. Indian J of Pharmaceutical Education
and Research. 2020 Oct 1;54(4):954-62.
81.Jain S, Tiwary AK, Sapra B, Jain NK. Formulation and evaluation of ethosomes for transdermal delivery
of lamivudine. Aaps Pharmscitech. 2007 Oct;8:249-57.
82. Basto R, Andrade R, Nunes C, Lima SA, Reis S. Topical delivery of niacinamide to skin using hybrid
nanogels enhances photoprotection effect. Pharmaceutics. 2021 Nov 20;13(11):1968.
83. El-Zaafarany GM, Abdel-Aziz RT, Montaser MH, Nasr M. Coenzyme Q10 phospholipidic vesicular
formulations for treatment of androgenic alopecia: ex vivo permeation and clinical appraisal. Expert opinion
on drug delivery. 2021 Oct 3;18(10):1513-22.
84.Allam AA, Fathalla D, Safwat MA, Soliman GM. Transferosomes versus transethosomes for the dermal
delivery for minoxidil: Preparation and in vitro/ex vivo appraisal. Journal of Drug Delivery Science and
Technology. 2022 Oct 1;76:103790.
85. Albash R, Abdelbary AA, Refai H, El-Nabarawi MA. Use of transethosomes for enhancing the
transdermal delivery of olmesartan medoxomil: in vitro, ex vivo, and in vivo evaluation. International
journal of nanomedicine. 2019 Mar 15:1953-68.
86. Nasr M, Wahdan SA. Neuroprotective effects of novel nanosystems simultaneously loaded with
vinpocetine and piracetam after intranasal administration. Life sciences. 2019 Jun 1;226:117-29.

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