Experimental Research On Preservation Of Rat Amputated Limb

Objective: To reduce the ischemic damage of amputated limbs by delaying tissue degenerationand extending the preservative time of amputated limbs with activity, to create conditions forfuture transplant operations in war times, and to promote functional recovery of amputatedlimbs after operations, reduce the rate of disability for increasing the follow-up fighting capacityand enhancing the quality of life.Methods:32Wistar rats (The weight of300-400g) with adult and health were provided by theexperimental animal center of the First Affiliated Hospital of PLA. Experiment One:4Wistarrats were randomly divided into2groups (2rats for each group) for producing the models ofamputated limbs of rats. The first group was treated with UW solution until the pouring outliquid became clear. The second group was the control group without any special treatment.Both of groups were preserved in the refrigerator at4℃. Samples of tissue were taken forpathological examination every6hours since the limbs had been amputated. Experiment Two:16Wistar rats were randomly divided into4groups (4rats for each group) for producing themodels of amputated limbs of rats. The first group was treated with UW solution, the secondgroup was treated with HTK solution, the third group was treated with Ringer solution+dexamethasone+low molecular weight heparin sodium, the fourth group was the control groupwhich was treated with Physiological saline. All of groups were bored continuously in24hoursin the refrigerator at4℃, we collected the irrigation fluids of different times to detect therelated biochemical indexes, and took Statistical analysis by SPSS software. Pathologicalsamples were obtained at the corresponding times to observe the changes of organizationalstructure under light microscope and ultra microstructure under electron microscope.Experiment Three:4Wistar rats were randomly divided into1group for producing the modelsof amputated limbs of rats, this group was bored discontinuously with UW solution in24hoursin the refrigerator at4℃. Sampling method was as same as Experiment Two. We made comparison with the group treated with UW solution continuously by biochemical indexes,changes of organizational structure and ultra microstructure. Experiment Four:4Wistar ratswere divided into1group and bored discontinuously in24hours in the refrigerator at4℃withno-sugar cell culture fluid after producing the models of amputated limbs of rats. Samplingmethod was also as same as Experiment Two. We made comparison with the group treated withUW solution discontinuously by biochemical indexes, changes of organizational structure andultra microstructure.Results: Experiment One: At the same time, changes of organizational structure on muscle werelighter in group1than group2. Experiment Two: After treated with different perfusates,amputated limbs had lighter changes of organizational structure and ultra microstructure thanwhich were treated with Physiological saline. There was significant difference at biochemicalindex. Among three perfusates, UW solution had the lightest changes of organizational structureand ultra microstructure, and there was no significant difference at biochemical index in thesegroups. Experiment Three: Amputated limbs treated with UW solution continuously had thesame changes of organizational structure and ultra microstructure as the group treated with UWsolution discontinuously, there was no significant difference at biochemical index between them.Experiment Four: Amputated limbs treated with UW solution discontinuously had lighterchanges of organizational structure and ultra microstructure than group treated with no-sugarcell culture fluid discontinuously, and there was significant difference at biochemical indexbetween them.Conclusion:1、Amputated limbs treated with perfusates had stronger protective effects thanamputated limbs treated by only cryogenic preservation.2、The UW solution had strongerprotective effects on amputated limbs than any other perfusates.3、It had the same protectiveeffects on amputated limbs treated by boring continuously than boring discontinuously.4、Amputated limbs treated with no-sugar cell culture fluid were much more swelling, it was notcomfortable for no-sugar cell culture fluid to be used on perfusion and preservation ofamputated limbs.