To develop bone implant material with excellent antibacterial and biocompatible properties, nanotubular titanium surface was coated with hydroxyapatite (HA) and graphene oxide (GO). Layer-by-layer deposition was achieved by coating HA on an anodic-grown titanium dioxide nanotube array (ATi) with electrolytic deposition, followed by coating with GO using anodic-electrophoretic deposition. The antibacterial activity against both Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria was determined based on the percentage of surviving bacteria and the amount of ribonucleic acid (RNA) leakage and correlated with membrane disruption. The oxidative stress induced in both strains of bacteria by GO was determined by cyclic voltammetry and is discussed. Importantly, the antibacterial GO coatings on HA-ATi were not cytotoxic to preosteoblasts and promoted osteoblast proliferation after 5 days and calcium deposition after 21 days in standard cell culture conditions.