Objective To investigate the effect of Navio robot-assisted unicompartmental knee arthroplasty (UKA) on the biomechanics of knee joint during sitting-up movement, and to determine whether UKA can maintain the biomechanical characteristics of knee joint. Methods The clinical data of 8 patients with medial compartment osteoarthritis treated with medial fixed platform of Navio robot-assisted UKA between January 2018 and January 2019 and had the complete follow-up data were retrospectively analyzed. There were 4 males and 4 females; the age ranged from 58 to 67 years, with an average of 62.3 years. The disease duration was 6-18 months, with an average of 13 months. The varus deformity ranged from 4° to 6°, with an average of 5°; the knee flexion range of motion was 0°-130°, with an average of 110°. All patients had no extension limitation. The imaging data of bilateral knees during sitting-up movement were collected by biplane C-arm X-ray machine at 3 weeks before operation and 7 months after operation. The three-dimensional models of femur and tibia were established by dual-energy CT scanning, and the three-dimensional models of femur and tibia were matched and synchronized with the femur and tibia in X-ray film by automatic matching tracer software. The biomechanical parameters of femur and tibia were measured, including internal rotation/external rotation, varus/valgus, forward/backward displacement of medial and lateral tibia contact center, and lateral compartment joint space. Results Eight patients were followed up 5-7 months, with an average of 6.4 months. In the comparison of the affected side before and after operation, except for the difference of varus/valgus which was significant (t=4.959, P=0.002), the differences in other indicators was not significant (P>0.05). There were significant differences in varus/valgus and internal rotation/external rotation between healthy and affected sides at 3 weeks before operation (P<0.05), and the differences in other indicators was not significant (P>0.05). At 7 months after operation, the difference in the forward and backward displacement of medial tibia contact center was significant (t=3.798, P=0.007), and the differences in other indicators was not significant (P>0.05). Conclusion UKA can effectively correct the varus and valgus of the knee joint, and restore the rotational biomechanical characteristics of the affected knee joint. It does not affect the establishment of the lateral compartment joint space, but the medial and lateral tibia contact center still changes.