This work aims to evaluate the characterization, antibacterial activity, and mesenchymal stem cell (MSC) viability in films obtained by cold plasma polymerization, from the essential oil of Eucalyptus globulus on Ti6Al4V alloy. The film presented low roughness values and a contact angle of 85°. The process incorporated a coating with oxygenated functional groups (OH and C═O). The effect of ultraviolet radiation, ethylene oxide, autoclave, and air plasma sterilization on polymer degradation was evaluated. Biofilm formation by Staphylococcus aureus was reduced on the film surface with no toxicity to MSCs. These effects can be attributed to functional groups, combined with the surface roughness and contact angle. These results suggest a potential for the application of this coating in biomaterials.