The treatment for severe combined anterior cruciate ligament (ACL) and medial collateral ligament (MCL) ruptures is disputed. Using a rabbit model, we examined the effect of insufficiency of medial structures on the reconstructed ACL in combined ACL and MCL injury. 40 rabbits were divided into 2 groups. In both groups, ACL was subjected to in situ freeze-thaw treatment. In group F, only freeze-thaw treatment of ACL was given. In group FM, partial resection of MCL was also done. We killed 5 rabbits on each of 4 occasions: immediately after the operation (time 0), at 6, 12 and 24 weeks postoperatively. At each time, we measured valgus instability and mechanical properties of the ACL. Valgus instability in group FM persisted from time 0 to 24 weeks, and was significantly greater than that in group F. The tensile strength and tangent modulus of the ACL in group FM were lower than those in group F. We found that continuous valgus instability reduces the mechanical properties of the in situ frozen ACL.