Copper (Cu) could serve as antibacterial coating for Ti6Al4V implants. An additional cell-adhesive layer might compensate Cu cytotoxicity. This study aimed at in vitro and in vivo evaluation of low-temperature plasma treatment of Ti6Al4V plates with Ti/Cu magnetron sputtering (Ti6Al4V-Ti/Cu), plasma-polymerized ethylenediamine (Ti6Al4V-PPEDA), or both (Ti6Al4V-Ti/Cu-PPEDA). Ti6Al4V-Ti/Cu and Ti6Al4V-Ti/Cu-PPEDA had comparable in vitro Cu release and antibacterial effectiveness. Following intramuscular implantation of Ti6Al4V-Ti/Cu, Ti6Al4V-PPEDA, Ti6Al4V-Ti/Cu-PPEDA and Ti6Al4V controls for 7, 14 and 56 days with 8 rats/day, peri-implant tissue was immunohistochemically examined for different inflammatory cells. Ti6Al4V-PPEDA had more mast cells and NK cells than Ti6Al4V, and more tissue macrophages, T lymphocytes, mast cells and NK cells than Ti6Al4V-Ti/Cu-PPEDA. Ti6Al4V-Ti/Cu had more mast cells than Ti6Al4V and Ti6Al4V-Ti/Cu-PPEDA. Results indicate that PPEDA-mediated cell adhesion counteracted Cu cytotoxicity. Ti6Al4V-Ti/Cu-PPEDA differed from Ti6Al4V only for mast cells on day 56. Altogether, implants with both plasma treatments had antibacterial properties and did not increase inflammatory reactions.