In this study, TiO₂ nanotube layers were prepared by anodic oxidation on the Ti6Al4V laser shock peening surface. Under simulated body fluid conditions, the surface properties, microstructure, and friction and wear properties were studied. The results showed that the surface of Ti6Al4V had plastic deformation after laser shock peening, and grain refinement improved the surface’s microhardness and wear resistance. The length of TiO₂ nanotubes prepared by laser shock peening increased, the wall of TiO₂ nanotubes became thicker, the microhardness of the surface decreased, and the surface roughness increased. The TiO₂ nanotube layer’s adhesion to the laser shock peening surface increased by 17.7% compared to the substrate. The higher hydrophilicity of the TiO₂ nanotube layer improved lubrication, which contributed to a decrease in wear mass and friction coefficient. The wear mechanism was a combination of abrasive wear, fatigue wear, and adhesive wear.