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Background: This study aimed to evaluate post-operative lower limb function following second-generation mechanically aligned medial pivot (MP) TKA implantation. Standard gait analysis was performed to collect kinematic and kinetic data, which were then compared with physiological data from the literature obtained using the same evaluation methodology as the present study. The hypothesis was that this TKA would not fully restore normal knee and adjacent joint motion during walking. Methods: Our cohort comprised 15 patients consecutively enrolled from September 2019 to December 2022 who underwent primary TKA with the second-generation MP Evolution Knee System (MicroPort Orthopaedics Inc., Arlington, TN, USA). Pre-operatively and 6 months post-surgery, gait analysis during level walking was performed on all patients, as well as clinical evaluations using the Knee Society Score (KSS), the Knee Injury and Osteoarthritis Outcome Score (KOOS), and the Visual Analogue Scale (VAS). Results: The clinical scores improved significantly (p < 0.001) after surgery (pre-/post-operative KSS functional, KSS clinical, VAS, and KOOS: 51.7 ± 17.3/84 ± 18.4, 45.3 ± 16.2/74.1 ± 12.6, 6.9 ± 1.8/2.0 ± 1.9, and 33.9 ± 11.8/69.1 ± 16.5, respectively). The statistical parametric mapping (SPM) analysis between the post-operative and reference control data revealed significant differences in the initial and final 20% of the gait cycle for the rotation of the knee in the frontal and transverse planes and for the rotation of the ankle in the sagittal plane. Conclusions: This study shows that new-generation MP TKA with mechanical alignment does not fully restore normal gait biomechanics, particularly in knee rotational movements, indicating a need for improved surgical techniques and prosthetic designs. [ABSTRACT FROM AUTHOR]