Background Osteoarthritis (OA) stands as the most prevalent disability among the elderly population. Assessing functional outcomes after Total Knee Replacement (TKR) typically involves Gait analysis along with other evaluation methods. The objective of this study was to compare Gait results, including temporospatial parameters, joint angles, gait profile score (GPS), and movement analysis profiles (MAP), between conventional and iASSIST TKR techniques. Method The study involved 21 participants (mean age 68.4 ± 4.2 years), with 16 females and 5 males. Among them, 11 patients had traditional surgery (15 TKR), and 10 patients had iASSIST surgery (13 TKR), totaling 28 knees (7 bilateral). The pre-operative Gait analysis was conducted one day before the surgical procedure, whereas the postoperative Gait analysis was performed, on average, 210 ± 20 days after surgery. Gait analysis was conducted using the Qualisys Motion capture system, operating at a rate of 120 Hz. The data were thoroughly analyzed using Visual 3D C-Motion Software. Results An analysis of gait biomechanics metrics, encompassing temporospatial parameters, joint angles, GPS, and MAP, was undertaken. Significant differences were observed in sagittal plane joint angles of the pelvis and hip, transverse plane joint angles of the knee, cadence, and MAP of foot internal/external rotation. However, there were no statistically significant differences between the two TKR techniques in the remaining temporospatial variables, joint angles, GPS, or MAP. Conclusion This study revealed a significant difference between iASSIST-guided TKR and conventional TKR, demonstrating that the iASSIST procedure led to improvements in walking biomechanics. Findings hold potential utility for orthopedic surgeons in their decision-making processes, ultimately contributing to the improvement of functional outcomes following TKR.