Abstract: Background and Objective: Erectile dysfunction (ED) is a common disease in diabetic patients. Alprostadil (Prostaglandin E1, PGE1) has shown higher success rates compared with phosphodiesterase type 5 inhibitors (PDE-5) in the treatment of ED. The combined formula of PGE1 with D-tocopheryl polyethylene glycol succinate (TPGS) was aimed to find a novel treatment for diabetic patients suffering from ED via protecting epithelial cells that lined penile vessels from the oxidative stress. Materials and Methods: PGE1-TPGS was prepared either with labrasol, polyethylene glycol 200 (PEG 200) or a mixture of labrasol and PEG 200, F1, F2 and F3 respectively. The prepared formulations were characterized for morphology and zeta potential and then were added to 1.5% of hydroxypropyl methyl cellulose solution to form topical hydrogel. Drug release study for all investigated formulations was conducted in comparison with raw-PGE1 and tested in endothelial cells (EA. hy926) under stress condition. Results: The obtained data revealed that PGE1-TPGS prepared with labrasol achieved the highest particle size while, PGE1-TPGS formula of PEG 200 and labrasol mixture showed the smallest particle size. Transmission electron microscope (TEM) images of the prepared formulations showed spherical shape micelles. Oxidative stress investigation showed a dramatic improvement in epithelial cell survival that attenuated the permeation enhancer’s cytotoxicity. F1, F2 and F3 treatments showed improved cell viability when compared with blank formulations. At concentration 10 μg mL–1, results showed that the investigated formulations improved cell viability from 9.13% (F1 blank), 7.55% (F2 blank) and 8.82% (F3 blank) to 90.52, 91.27 and 93.94% for F1, F2 and F3 formulations, respectively. Conclusion: The study revealed that combination of PGE1 with TPGS micelles formula loaded topical gel could be a successful strategy for the treatment of ED in diabetic patients.