Ultraviolet (UV) rays damage DNA, causing adverse effects such as photoaging and cancer on the skin. For the well-being of individuals, there is a need to develop innovative skin products with high effectiveness using protective and therapeutic agents. In this context, in our study, A nanoemulsion (NE) formulation containing Ylang-ylang essential oil (YO), which has many biological active properties such as antimicrobial, antioxidant, anti-inflammatory, and anticancer, was produced by the ultrasonic emulsification method and characterized. The thermodynamic stability was evaluated, and its in vitro release profile determined the dialysis membrane technique. The cytotoxic effect of YO-NE was examined with the in vitro method on the HacaT cell line using the MTT method and in silico method using the ADMET profile. Dynamic light scattering (DLS) results showed that the average droplet size of the YO-NE formulation was 184.1±2.307 nm, the polydispersity index (PdI) was 0.151±0.006, and the Zeta potential (ζ) -10.8 ± 0.400 mV. As a result of in vitro release studies, it was observed that 99.98± 1.00% of YO release from NE occurred within 5 hours. Based on the thermodynamic stability test results, it was determined that the developed formulation did not show sedimentation or phase separation. Also, it remained stable under three different storage conditions in the three-month stability test. Cytotoxicity results revealed that the YO-NE formulation was safe. All the results indicated that the YO-NE formulation might be considered a non-toxic product candidate with physicochemical properties suitable for topical use.