The purpose of this study is to investigate, for the first time, the biocompatibility and antibacterial behavior of coatings obtained by plasma electrolytic oxidation (PEO) on the Ti6Al4V ELI alloy manufactured through the direct metal laser sintering (DMLS) additive manufacturing technique, implanted with two different concentrations of Cu ions by ion plating diversified (IPD). The surface morphology, elemental composition, and phase characteristics of the PEO coatings were investigated by SEM/EDS, GDOES, XRD, and XPS. The cytotoxicity in stem cells and the antibacterial effect were evaluated. The XPS analysis confirmed the successful incorporation of Cu at both concentrations, which appeared in the form of two different chemical components: CuO (92%) and Cu2O (8%). Stem cells were able to adhere and spread on the surface of the Ti6Al4V ELI alloy, with or without PEO coatings and ionic implantation of Cu particles; the materials and coatings were not cytotoxic. Quantitative cell viability assays for Staphylococcus aureus and Pseudomonas aeruginosa bacteria did not show significant differences. However, the staining of the viability wells, mainly for S. aureus, indicates a change in the behavior of these bacteria in coatings that contained the implantation of Cu oxides. This research demonstrated the potential that the union of the properties of the substrate manufactured in additive manufacturing (great versatility), PEO coatings (wear resistance and cell viability), and IPD ion implanted Cu particles can have in different applications.