Mechanism of Action:

Vesicular systems facilitate transdermal drug delivery by enhancing the permeation of the active pharmaceutical ingredient. The vesicles penetrate the skin, allowing the encapsulated drug to traverse the epidermal layers. The inclusion of permeation enhancers such as ethanol, propylene glycol, and isopropyl myristate increases the fluidity of the lipid bilayer, as well as interacts with the lipid components of the stratum corneum, thereby promoting deeper drug penetration.⁹

Figure 3: Mechanism of action of Transethosomes

METHOD OF PREPARATION:

· Cold method

· Hot method

· Reverse phase evaporation method

· Mechanical dispersion method.

1. Cold Method:

This method is commonly employed for the preparation of transethosomes and is particularly suitable for thermolabile drugs that are sensitive to heat. The process is readily scalable for larger production. Initially, the phospholipid is dissolved in ethanol through vigorous agitation to form a uniform solution. This mixture is then heated to 30 °C in a water bath. Separately, water is heated to the same temperature and gradually added to the ethanolic phospholipid solution while maintaining continuous stirring with a magnetic stirrer at 700 rpm to ensure homogeneity. The vesicle size can be further controlled and optimized using probe sonication.¹⁰

Figure 4: Cold method.

2. Hot Method:

The phospholipid is dispersed in water and heated to 40°C, while a combination of ethanol and glycol is similarly heated to the same temperature. The organic phase is then gradually combined with the aqueous phase under continuous stirring to ensure uniform mixing. The choice of solvent system, whether aqueous or ethanolic, is determined based on the solubility of the drug. The temperature is maintained consistently at 40 °C throughout the process. Vesicle size can be further adjusted using probe sonication.¹¹

Figure 5: Hot method.

3.Mechanical Dispersion Technique:

In this technique, a round-bottom flask is employed, and the liquid–surfactant mixture is dissolved in ethanol. The efficiency of the method can be improved by combining the hydrated thin-film approach with ultrasonic homogenization. A thin lipid film is generated using a rotary evaporator, and residual organic solvent is removed by allowing the film to remain under vacuum overnight. During the hydration process, a 10% v/v ethanol solution in phosphate buffer (pH 6.5) is applied at 60rpm. Vesicle size can be further optimized through sonication.¹²

Figure 6: Mechanical dispersion technique

4. Reverse phase evaporation method:

The lipid is initially dissolved in ethanol, while the edge activators are incorporated into the aqueous phase. The aqueous phase is then gradually introduced into the organic phase, and ultra sonication is applied at 0°C to facilitate phase separation. Upon removal of the organic solvent under pressure, gel formation is achieved.¹³

Figure 7: Formulation of Transethosomes by Reverse Phase Evaporation.

Evaluation of Transethosomes:

1. Transmission Electron Microscopy:

Transmission electron microscopy (TEM) was employed to characterize the morphology of the transethosomal vesicles. The selected transethosomal formulation was diluted tenfold with distilled water, and a single drop of the diluted dispersion was deposited onto a 300-mesh carbon-coated copper grid. After standing for approximately one minute to facilitate vesicle adhesion to the carbon surface, excess dispersion was carefully blotted using filter paper. The grid was subsequently rinsed twice with deionized water for 3–5 seconds. Finally, the prepared sample was observed under the transmission electron microscope at an accelerating voltage of 100kV, using magnifications ranging from 10× to 100×.¹⁴

2. Entrapment Efficiency:

The entrapment efficiency of the transethosomal formulation was evaluated using the ultracentrifugation method. Ultracentrifugation was carried out at 1500rpm for 60 minutes at 4°C to separate the vesicular sediment from the supernatant. Following centrifugation, the sediment and supernatant were carefully collected, and the amount of entrapped drug in the sediment was quantified. The percentage entrapment efficiency was subsequently calculated using the appropriate mathematical equation.¹⁵

Amount of entrapped drug

EE= ---------------------------------------------------- X 100

Total Amount

3. Optical Microscopy:

The transethosomal formulation was appropriately diluted, mounted onto glass slides, and examined under an optical microscope at 1200× magnification to assess vesicular morphology. Photomicrographs of the preparation were captured using a digital SLR camera attached to the microscope.¹⁶

4. Vesicle Charge:

The surface charge of the transethosomal vesicles was evaluated by measuring the zeta potential following suitable dilution of the formulation. The morphological characteristics of the vesicles were observed, and photomicrographs were obtained using a digital SLR camera coupled with the microscope.¹⁷

5. Interaction Studies Using FTIR Spectroscopy:

Drug–excipient compatibility was investigated using Fourier transform infrared (FTIR) spectroscopy. Infrared spectra of the pure drug and the powdered transethosomal formulation were obtained using an FTIR spectrophotometer (FTIR 1615, PerkinElmer, USA). Samples were prepared as potassium bromide (KBr) pellets, and the spectra were recorded over the wavenumber range of 3600–400 cm⁻¹.¹⁸

6. Stability Study:

Stability studies were conducted to ensure the quality, safety, and efficacy of the transethosomal gel formulation during storage. The formulations were stored for six months in a stability chamber in tightly closed amber-colored glass containers sealed with aluminum foil under three different conditions: room temperature (25±2°C/60% RH±5%), refrigerated conditions (4.0±1.0°C), and accelerated conditions (40± 2°C/75% RH ±5%). Samples were withdrawn at the end of the 1st, 2nd, 3rd, and 6th months and evaluated for in vitro drug release, pH, entrapment efficiency, and drug content.¹⁹

7. Drug Content:

The drug content of the transethosomal gel was quantified using high-performance liquid chromatography (HPLC). An accurately weighed 5g sample of the gel was dissolved in purified water to obtain a final volume of 50mL, followed by sonication in phosphate buffer (pH 7.4) for 15minutes and gentle heating for 5 minutes to ensure complete drug extraction. The analysis was performed in triplicate, and the mean percentage drug content was calculated.²⁰

The % drug content of transethosomal preparation was determined by using the following formula:

Sample absorbance

% Drug content = --------------------------------------

standard absorbance

8. Vesicle Size and Shape:

In the evaluation of transethosomal gel, vesicle size and shape are important parameters that affect drug penetration and stability. Vesicle size is usually measured by Dynamic Light Scattering (DLS), and the ideal size range is 50–300nm with a low polydispersity index (PDI<0.3) indicating uniform distribution. Zeta potential is also determined to check stability. The shape and morphology of vesicles are examined using Transmission Electron Microscopy (TEM) or Scanning Electron Microscopy (SEM), which typically show spherical and smooth vesicles. Proper size and shape ensure better skin permeation and effective drug delivery.

9. Zeta potential:

Zeta potential is an important evaluation parameter of transethosomal gel as it indicates the surface charge and stability of the vesicles. It is measured using a zeta potential analyzer based on electrophoretic mobility. A high absolute zeta potential value (around ±30mV or more) suggests good stability due to electrostatic repulsion between vesicles, which prevents aggregation. Therefore, measuring zeta potential helps to predict the physical stability and shelf life of the transethosomal gel formulation.

10. Stability test:

Stability testing of transethosomal gel is carried out to assess its physical and chemical stability during storage. The formulation is stored at room temperature and accelerated conditions (40°C±2°C/75% RH±5%) as per International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use. During the study, parameters such as appearance, pH, viscosity, drug content, vesicle size, and zeta potential are evaluated at regular intervals. Any significant changes indicate instability of the formulation.

Application:

Compared with conventional liposomes, ethosomes, and nanoethosomes, transethosomes demonstrate superior drug delivery performance. These vesicular systems enhance drug distribution by approximately 65% relative to liposomes, owing to their improved ability to permeate multiple layers of the human skin. The potential effectiveness of transethosomes as transdermal drug delivery carriers is currently being explored using a limited number of bioactive compounds.

Delivery of NSAIDs (Non-steroidal Anti-inflammatory Drugs):

Oral administration of non-steroidal anti-inflammatory drugs (NSAIDs) is frequently associated with gastrointestinal adverse effects. Transethosomal drug delivery systems have been investigated as an alternative approach to overcome these limitations. A transethosomal formulation of ketorolac tromethamine has demonstrated enhanced skin penetration, while a piroxicam-loaded transethosomal gel exhibited superior stability and elasticity when compared with other deformable vesicular systems, as reported by Garg et al.

In a separate study, Paolino et al. evaluated ethosomal formulations containing ammonium glycyrrhizinate in human subjects. The formulation comprising 45% ethanol and a reduced lecithin concentration yielded optimal performance. In vitro findings indicated improved skin tolerability and enhanced percutaneous permeation, while in vivo studies in volunteers showed increased anti-inflammatory activity.²¹

Delivery of Hormones:

Oral administration of hormones is often associated with several limitations, including extensive first-pass metabolism, low oral bioavailability, and the occurrence of dose-dependent adverse effects. Touitou et al. evaluated the transdermal delivery potential of testosterone-loaded ethosomes in comparison with a commercially available testosterone transdermal patch (Testoderm®, Alza Corporation, California) using rabbit pinna skin as the permeation model. The ethosomal formulation exhibited approximately 30-fold higher testosterone skin penetration than the marketed transdermal patch. Furthermore, pharmacokinetic parameters, including the area under the concentration–time curve (AUC) and maximum plasma concentration (Cmax), were significantly greater for the ethosomal system than for Testoderm®, indicating enhanced transdermal absorption.²²

Delivery of Antibiotics:

Topical administration of antibiotics offers a more effective approach to enhancing their therapeutic efficacy, as conventional oral therapy is often associated with allergic reactions and systemic side effects. Traditional external formulations exhibit limited permeability into deeper skin layers and subdermal tissues. Ethosomal carriers can overcome this limitation by delivering adequate concentrations of antibiotics to the deeper layers of the skin. These vesicles rapidly penetrate the epidermis, ensuring substantial drug deposition in the dermis and targeting infections at their source. In line with this approach, Godin and Touitou developed ethosomal formulations of bacitracin and erythromycin for dermal and intracellular delivery. Their findings demonstrated that ethosomal antibiotics can achieve high therapeutic efficiency and address the limitations of conventional therapies.²³

Delivery of Antifungal Drugs:

Verma et al. investigated the potential of transethosomes as a delivery system for antifungal agents, using econazole nitrate as the model drug. The study compared the performance of an econazole nitrate-loaded transethosomal gel with a commercially available econazole nitrate cream. The results indicated that the transethosomal gel exhibited enhanced ex vivo skin retention and superior in vitro antifungal activity. Additionally, the system provided controlled drug release, contributing to effective management of cutaneous candidiasis.

Delivery of Anti-Parkinsonism Agents:

Trihexyphenidyl hydrochloride (THP), a psychoactive drug commonly used in the management of Parkinson’s disease, was incorporated into an ethosomal formulation by Dayan and Touitou. When compared with conventional liposomal formulations, the ethosomal-THP system demonstrated significantly enhanced skin penetration. These findings suggest that ethosomal delivery of THP may offer improved therapeutic efficacy for the treatment of Parkinson’s disease.

Cosmeceutical Application:

Ethosomes have been effectively incorporated into cosmetic formulations due to their ability to enhance transdermal penetration, improve stability, and minimize skin irritation caused by harsh cosmetic ingredients. Ethosomal creams containing Curcuma longa extract have been developed and investigated for their anti-aging and photoprotective potential. Application of these C. longa extract-loaded ethosomal creams in human volunteers demonstrated promising results in both photoprotection and wrinkle reduction. Additionally, Yeh et al. formulated a transethosome-based hair dye, which was shown to be more effective than a hydroethanolic solution in delivering and enhancing the absorption of black tea extracts onto the hair surface.

Delivery of Anticancer Drugs:

Lei et al. investigated a transethosomal formulation with dual drug loading for the treatment of cutaneous melanoma. Their study demonstrated that the combination of dacarbazine and tretinoin exhibited superior efficacy and lower cytotoxicity compared to other formulations. Dual-drug-loaded transethosomes showed enhanced antitumor activity relative to single-drug systems and improved skin penetration. Similarly, Shaji et al. reported that encapsulating 5-fluorouracil in a transethosomal gel resulted in greater vesicle deformability, enhanced skin penetration, and more effective delivery to deeper skin layers compared to conventional ethosomal formulations.

CONCLUSION:

The advent of ethanol-based ultra-deformable vesicular (UDV) systems has enabled the circumvention of skin barriers that limit the penetration of certain bioactive compounds. This advanced vesicular technology encompasses ethosomes, transferosomes, and transethosomes. Among these, transethosomal systems are particularly noteworthy due to their compatibility with both hydrophilic and hydrophobic therapeutic agents, offering enhanced solubility, flexibility, and skin penetration. Comprising ethanol and edge activators, transethosomes facilitate targeted topical delivery of drugs, including high-molecular-weight molecules such as peptides and proteins, owing to their substantial carrier capacity. Topical application in the form of gels or creams promotes high patient compliance. Furthermore, ethosomal and transethosomal systems have been successfully utilized for the delivery of cosmeceuticals, as well as anticancer, antiviral, and antifungal agents. Overall, transethosomal vesicular systems demonstrate superior efficacy, safety, and patient adherence compared with conventional transdermal drug delivery methods.

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Received on 28.02.2026 Revised on 24.03.2026

Accepted on 14.04.2026 Published on 07.07.2026

Available online from July 10, 2026

Res. J. Pharma. Dosage Forms and Tech.2026; 18(3):227-232.

DOI: 10.52711/0975-4377.2026.00033

This work is licensed under a Creative Commons Attribution-Non Commercial-Share Alike 4.0 International License. Creative Commons License.

Feature	Transethosomal Gel	Liposomal Gel	Niosomal Gel
Skin Penetration	Superior; enhanced by ethanol and surfactants	Moderate; limited by rigid bilayer	Moderate; lacks deformability
Stability	High; resistant to fusion and leakage	Moderate; prone to instability	Moderate; may aggregate over time
Drug Compatibility	Suitable for both hydrophilic and lipophilic drugs	Primarily hydrophilic and hydrophobic drug	Hydrophilic drugs; limited for lipophilic
Patient Compliance	High; semisolid form (gel/cream)	High; semisolid form	High; semisolid form
First-Pass Metabolism	Avoided; direct skin absorption	Avoided; directskin absorption	Avoided; direct skin absorption
Applications	Anti-inflammatory, antifungal, anticancer	Anti-inflammatory, antifungal	Anti-inflammatory, antiviral