We demonstrate the enhancement of antibacterial and cytocompatibility characteristics of femtosecond laser-treated pure titanium and Ti-6Al-4V titanium alloy samples suitable for orthopedic implant applications. We controlled the wettability of the titanium samples by tailoring the surface geometry using a femtosecond laser. To increase the hydrophobicity, laser-assisted micro-grids patterning was performed on the titanium samples, where we achieved a highest contact angle of 144.6° for a 1 µL de-ionized water droplet. In contrast, the hydrophobic Ti-6Al-4V titanium alloy surfaces were converted to hydrophilic surfaces by fabricating periodic micro-gratings on the samples’ surface, where a lowest contact angle of 19.84° was achieved. Furthermore, we assessed the biocompatibility of the micro-patterned titanium samples by investigating the antibacterial activity against Staphylococcus Aureus bacteria. Moreover, the cytocompatibility of the micro-patterned titanium samples was examined using NCTC Clone 929 (L-929) mouse fibroblasts. The laser-treated titanium samples exhibited enhanced antibacterial performance while maintaining excellent cell compatibility. The experimental results confirmed excellent correlation with the wettability of the laser-patterned samples and their antibacterial characteristics and cytocompatibility. Overall, the findings highlight femtosecond laser surface structuring as a highly effective strategy to simultaneously improve antibacterial behavior and the biocompatibility of implant materials, offering a promising way for the advanced functionalization of orthopedic implants.