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Abstract Background This study aims to explore the relationship between surgically-induced ankle instability and posttraumatic osteoarthritis (PTOA) in a mouse model, and to provide reference for clinical practice. Results Ligamentectomy was performed on 24 eight-week-old male C57BL/6 J mice, which were divided into three groups. Both the anterior talofibular ligament (ATFL) and the calcaneofibular ligament (CFL) were severed in the CFL + ATFL group, while only the CFL was removed in the CFL group. The SHAM group was set as the blank control group. A wheel-running device was used to accelerate the development of ankle osteoarthritis (OA). Balance measurement, footprint analysis, and histological analysis were used to assess the degree of ankle instability and OA. According to the balance test results, the CFL + ATFL group demonstrated the highest number of slips and the longest crossing beam time at 8 weeks postoperatively. The results of gait analysis exhibited that the CFL + ATFL group had the most significant asymmetry in stride length, stance length, and foot base width compared to the CFL and SHAM groups. The OARSI score of the CFL + ATFL group (16.7 ± 2.18) was also much higher than those of the CFL group (5.1 ± 0.96) and the SHAM group (1.6 ± 1.14). Conclusion Based on the mouse model, the findings indicate that severe ankle instability has nearly three times the chance to develop into ankle OA compared to moderate ankle instability.