This research focused on tribological and antibacterial studies of 3D-printed Ti6Al4V alloys based on the selective laser melting method vs conventional technology after gas nitriding for biomedical applications. Gas nitriding reduced friction and wear in the following order: 3D-printed AM Ti6Al4V > wrought CM Ti6Al4V > CP-Ti. The best tribological performance was demonstrated on nitrided Ti6Al4V alloy using 3D printing. The antibacterial activity of the samples was estimated for the Gram-positive Staphylococcus aureus and the Gram-negative bacteria Escherichia coli. AM Ti6Al4V showed a positive antibacterial effect of nitriding against S. aureus bacteria by up to 30 %. Gas nitriding of 3D-printed Ti6Al4V alloys suggests an effective strategy to improve the antibacterial activity and mechanical resistance of biomedical implants.