Objective To explore the biomechanic effects of multi ple freeze-thaw on human allograft tendons. Methods Thirty tendons (24 flexor digitorum superficial is tendons and 6 flexor poll icis longus tendons) were harvested from 3 fresh cadaver donors and were divided into 6 groups randomly (fresh group; 1 cycle, 2 cycle, 3 cycle, 5 cycle, and 10 cycle freeze-thaw groups). There was 4 flexor digitorum superficial is tendons and 1 flexor poll icis longus tendon in each group. The structural and mechanical properties as well as viscoelastic change were estimated. Results The results of the structural and mechanical properties in 1 cycle, 2 cycle, and 3 cycle freeze-thaw groups were similar to that of the fresh group (P gt;0.05). The tendons in 5 cycle and 10 cycle freeze-thaw groups showed a significantly lower ultimate load and maximum stress when compared with those of fresh group (P lt; 0.05), but there was no significant difference in maximum tensile or maximum strain (P gt; 0.05). Moreover, the tendons in 5 cycle and 10 cycle freeze-thaw groups had a significant increase in viscoelastic properties when compared with fresh group (P lt; 0.05). Conclusion In the cryopreservation of tendon allografts, the cycle of freeze-thaw should not exceed 3 times. Multiple cycle freeze-thaw will weaken the biomechanical properties of tendon allografts, which make grafts easier to fatigue or even rupture.