Experimental Study On The Protective Effect Of Oxygenated Blood Perfusion On Amputated Limbs

Objective:Through the animal experiment,an oxygen-combined blood intermittent perfusion method is adopted to observe the preservation effect on the amputated limbs,and to explore the effectiveness of the preservation method of amputated limbs.To provide an idea for the clinical effective preservation of amputated limbs and prolonged replantation of amputated limbs.Methods: Thirty-two healthy New Zealand white rabbits,male and female were unlimited,2~2.5 kg weight,were randomly divided into four groups: control group,heparin group,oxygenated blood group and hypothermia group(eight in each group).An experimental model of complete amputation of animal limbs was established.32 limbs were amputated and the amputated limbs were recorded as 0h immediately.Control group(Group C): no perfusion treatment and no cryogenic treatment for preservation;Heparin group(Group B): preservation by intermittent perfusion of heparin mixed with normal saline(once an hour after 4 clock,four times in a row,50 ml / h,20 ml);Oxygen-enriched blood group(Group O): preservation by intermittent perfusion of oxygenated blood mixed with heparin(once an hour after 4 clock,four times in a row,50 ml / h,20 ml);Hypothermia group(Group HT)was preserved by low temperature method.The musculi hippicus of each group was harvested for pathological section and biochemical examination at 0h,4h,8h,12 h and 24 h after limb amputation.The histopathological changes of skeletal muscle and microvasculature were observed under light microscope.And the concentration of MDA、SOD、ALT、ALP、MPO and GLU in muscle tissue was measured by Changchun Huili Biotechnology limited company.Finally,these data were analyzed by SPSS19.0 software.Results: As time went on,the pathological changes of the amputated limbs in each group became more and more serious,muscle fiber gap gradually widened,cell edema gradually aggravated,the arrangement of striated muscles gradually disordered,and the horizontal lines of muscle cells gradually dissolved.And there was exudation of red blood cells and infiltration of inflammatory cells in the spatium intermusculare.The pathological changes of the heparin group were similar to those in the control group at 8h,12 h,and 24 h.And the cell edema was aggravated,and the erythrocyte exudation was decreased.In the hypothermia group,the pathological changes were smaller than those in the control group at 4h,8h,12 h,and 24 h.And the spatium intermusculare was wider,the erythrocyte exudation was less,and the infiltration of inflammatory cells around the blood vessel was less than that in the control group.The pathological changes of the oxygen-enriched blood group were better than those in the control group and the heparin group at 8h,12 h,and 24 h.And the spatium intermusculare was wider,the number of erythrocytes in blood vessels was increased.With the extension of time,in every group,the contents of MDA,ALT,ALP,and MPO in the musculi hippicus increased gradually,and the contents of SOD and GLU decreased gradually.There was no statistical difference between 4h and 0h.And compared with 0h,there was significant difference at 8h,12 h and 24 h in the control group and the heparin group(P < 0.05).Compared with 0h,there was significant difference at 12 h and 24 h in the hypothermia group(P < 0.05).And compared with 0h,there was significant difference at 24 h in the oxygen-enriched blood group(P < 0.05).There was no statistical difference between the heparin group and the control group at 8h,12 h,and 24 h.Compared with the control group,the contents of MDA,SOD,ALT,ALP and MPO in the hypothermia group was lower at 4 h,8 h,12 h and 24 h,and the contents of GLU was higher,and the difference is statistically significant(P < 0.05).Compared with the control group,the contents of MDA,SOD,ALT,ALP and MPO in the oxygen-enriched blood group were lower at 8 h,12 h and 24 h(P < 0.05),and the contents of GLU was higher(P < 0.05).Compared with the heparin group,the concentrations of MDA,ALT,ALP and MPO in the oxygen-enriched blood group were lower at 8 h,12 h and 24 h(P < 0.05),and the contents of GLU was higher(P < 0.05).Compared with the hypothermia group,the contents of ALP and MPO in the oxygen-enriched blood group were lower more(P < 0.05).Conclusion:The oxygenated blood perfusion can effectively protect the limbs after 4 clock of amputation though reducing by reducing the damage of free radicals to tissues and providing energy for aerobic metabolism.And compared with the method of pure low temperature protection,it has some advantages.