Titanium and its alloys face limitations in biomedical applications due to inadequate wear resistance, corrosion resistance, and biocompatibility. This study investigates the effects of pulsed laser frequency and duration on the properties of a remelted Ti6Al4V alloy layer. Results demonstrate that pulsed laser surface remelting (PLSR) generates 2–5 μm microgrooves and a gradient-modified “texture-melting layer.” The remelted layer undergoes martensitic transformation, increasing microhardness from 370 Hv to 575 Hv. The friction coefficient decreases from 0.63 to 0.31, and the wear rate reduces by 36.2 % (from 11.2 × 10 −6 mm 3·N −1·m −1 to 7.15 × 10 −6 mm 3·N −1·m −1), attributed to the martensitic structure and surface ripples. The electrochemical test results show that the passivation film formed on the surface improves the corrosion resistance. The open-circuit potential rising from −0.2331 V to 0.0228 V and corrosion current density decreasing by 59.4 % (from 5.463 × 10 −6 A·cm −2 to 2.217 × 10 −6 A·cm −2). Additionally, PLSR enhances surface hydrophilicity, reducing the water contact angle from 68.79° to 40.91°, which significantly promotes MC3T3-E1 cell proliferation and biocompatibility. These findings establish PLSR as a simple and effective method to enhance the wear resistance, corrosion resistance, and biocompatibility of Ti6Al4V alloys for biomedical applications.