Objective: Capsaicin is an active compound found in chili pepper and has been shown to have analgesic, antioxidant, anticancer, and anti-obesity properties. To improve its penetration into the skin, capsaicin was prepared in a transethosome vesicle. Importantly, transethosomes are vesicles that consist of phosphatidylcholine, surfactant, and ethanol. In this study, capsaicin was prepared in a transethosome vesicle using two different methods: Direct transethosome formation and thin layer hydration. The aims of this study were to determine the effects of various methods of transethosome formation on capsaicin characteristics and to evaluate the penetration capabilities of transethosome capsaicin gel. Methods: Ultimately, transethosome formation via the thin layer method yielded more favorable characteristics; these formations had particle sizes of 174.9±2.02 nm and an entrapment efficiency of 84.85±1.15%. The transethosome suspension was then developed into a gel formulation using 1% carbomer. An in vitro penetration test was performed using a Franz diffusion cell of mice abdomen skin, and the performance of the transethosome capsaicin gel was compared to that of the standard capsaicin gel. Results: Penetration rate of capsaicin from either the transethosome gel preparation and the standard gel substance was 1549.68±49.6 and 846.05±10.1 µg/cm2, respectively. Conclusions: According to these results, it can be concluded that gel preparations containing transethosome increase capsaicin penetration into the skin. © 2017 The Authors. Published by Innovare Academic Sciences Pvt Ltd.