Ostrich oil, a natural bird fat oil, has significant biological activities in preventing various pains and inflammation. The aim of the current study was to develop ostrich oil loaded nanoemulsions to check the anti-inflammatory efficacy. The developed nanoemulsions(F1-F3) were evaluated in vitro for various physicochemical parameters including thermodynamic stability, nanoemulsifying potential, mean droplet size, polydispersity index (PDI), zeta potential (ZP), viscosity (η), refractive index (RI) and transmittance (%T).Optimizednanoemulsions(F2) had mean droplet size (99.8 nm), PDI (0.084), ZP (-84.2 mV), viscosity (43.5cp), RI (1.344) and transmittance (94.1%).TEM photomicrograph of F2 nanoemulsion were spherical in shape with smooth surface, supporting DLS measured droplet size. Furthermore, anti-inflammatory activity of optimized nanoemulsion (F2) in rats was much better as compared to pure ostrich oil.