Experimental research on preservation of amputated limb of rat with University of Wisconsin (UW)solution

Wang Nan; Han Yan; Chen Miao

doi:10.3969/j.issn.1672-8521.2014.02.006

Abstract：Objective:To investigate the effect of University of Wisconsin (UW)solution in prolonging the vital-ity of amputated limbs. Methods:Both hindlegs were amputated in 20 adult male and healthy Wistar rats weighing of 300-400 g. In part one of the experiment,10 Wistar rats were randomly divided into 2 groups (5 rats in each group).The femoral artery of each of the amputated limbs was perfused with UW solution (4 ℃)until the liquid escaping from the femoral vein became clear. The amputated limbs of the control group were devoid of any special treatment. Limbs in both groups were preserved in the refrigerator at 4 ℃. Samples of tissue were taken for path-ological examination every 6 hours after the amputation. In part two of the experiment,10 Wistar rats were ran-domly divided into 2 groups (5 rats in each group).In the continuous perfusion group the amputated limb was per-fused with UW solution continuously for 24 hours at 4 ℃ with the speed of 12 ml/h.In the intermittent perfusion group the amputated limb was perfused with UW solution periodically with 12 ml/h for 20 minutes every 4 hours for 24 hours after amputation. Limbs in both groups were preserved in the refrigerator at 4 ℃. The irrigation fluid from each perfusion was collected to determine the contents of alkaline phosphatase (ALP),alanine aminotrans-ferase (ALT)and glucose,and muscles were sectioned to observe the changes in muscles harvested every 1 2 hours under light microscope and electron microscope after the limbs had been amputated. Results:In the experiment one,the degree of edema,fragmentation,dissolution of striped muscles were less marked in perfusion group than that of the control group at the same time point. In the experiment two,amputated limbs perfused with UW solu-tion continuously showed the same changes in structure and microstructure as the group treated with UW solution discontinuously,including fiber edema,fragmentation and horizontal dissolution in muscle.There were also swell-ing and fracture of mitochondria. There was no significant difference in contents of ALP,ALT and GLU between two groups (P>0.05 ). Conclusions:Perfusion with UW solution and cryopreservation has better protective effects on dismembered limbs compared with that undergoing cryogenic preservation only.The protective effects were same between continuous and periodic perfusion with UW solution on dismembered limbs for 24 hours.