In surgical procedures, reusable instruments must undergo cleaning, disinfection, and sterilization. Thus, rigorous attention to sterilization is essential to prevent cross-contamination, and some surgical instruments may also require wear resistance. There is a gap in the literature concerning the development of Zn-incorporated PEO coatings that combine antibacterial activity with tribological properties. The novelty of this study lies in the approach to improve Ti6Al4V wear resistance and antibacterial performance through nanostructured coatings produced by Plasma Electrolytic Oxidation (PEO) with simultaneous incorporation of Zn and Ti oxides. The results indicated that the antibacterial activity of the Zn-free PEO coatings against E. coli was mainly associated with photocatalytic effects related to the anatase phase of TiO₂. In contrast, the inactivation of S. epidermidis was lower, consistent with the higher resistance characteristic of Gram-positive bacteria. For the Zn-incorporated coatings, the reduction in S. epidermidis growth was primarily governed by the leaching of Zn ions, while reactive oxygen species generated by the photocatalytic effects of the coatings also contributed to the observed activity. Coatings with higher proportions of ZnTiO₃ led to a more pronounced inhibition of bacterial growth. Zn-incorporated coatings reduced cell viability by over 70%, altered the area and morphology, and promoted fibroblast adhesion and proliferation. The tribological behavior was influenced by zinc incorporation and microstructure, with TiO₂ contributing to wear resistance and ZnTiO₃ improving lubricity.