Plasma Electrolytic Oxidation (PEO) layers were produced on Ti-6Al-4V in different conditions, so as to assess the influence of layer structure, current mode, duty cycle and surface finishing on microstructural features and tribological behaviour. In DC regime, the double-layer structure (silicate bath followed by phosphate bath) beneficially affected wear resistance. In unipolar pulsed DC (phosphate bath), the wear resistance of single layers improved with increasing duty cycle, due to improved microstructure and adhesion: high duty cycle single layers can be considered an alternative to double-layer deposition. Surface finishing by abrasive blasting with spheroidal glass beads leads to surface roughness decrease and hence to decreased friction and improved wear resistance. The best-performing PEO layers showed promising results in the comparison with reference materials such as CoCrMo (both uncoated and (Ti,Nb)N PVD-coated) and PVD-coated Ti-6Al-4V up to 30 N normal load.