Sciatic Nerve Preserved By Vitrification Solution Contains Shenfu Injection Repaired Allogenic Nerve Defects In A Rat Model

Objective:To investigate the feasibility of improving sciatic nerve vitrification preservation using Shenfu Injection in rats.Methods:At-80?,SPF male SD rat sciatic nerve segments were preserved in the vitrification solution contains Shenfu Injection(0%,5%,10%,30%)(groups A,B,C and D)for 4 weeks.The sciatic nerves of he vitrification solution contains Shenfu Injection were observed by transmission electron microscope(TEM),the biological activity of the nerve was detected by laser scanning confocal microscope(LSCM)after Calcein-AM and PI double staining,and the expressions of Bcl-2 and Bax of the sciatic nerves were tested by Western blotting.10 mm sciatic nerve defect of the Wistar rats(groups A',B',C'and D')was repaired using the corresponding SD rats sciatic nerves preserved for 4 weeks,After different times to look at the basic situation of rats,at the same time in the cycle(4,8,12,16 weeks)on rat sciatic nerve function testing contrast,in 16 weeks after the transplantation rat nerve conduction velocity and the incubation period for testing,observation of regenerated nerve histological appearance.Result:The results of TEM showed that demyelination of the sciatic nerve in group A was severe,and in groups B,C and D were mild.The results of LSCM showed that there were strong green fluorescent and weak red fluorescent in the groups B,C and D,and weak green fluorescent and strong red fluorescent in the group A.The Bcl-2 protein expression,group A is lower than group B,C,D,B group is lower than C,D group differences were statistically significant(P < 0.05),Bax protein expression,B,C,D group is lower than group A,C,D group is lower than group B,difference between two kinds of protein in group C,D has no statistical significance(P > 0.05)The SFI at different periods after the transplantation,the results of electrophysiological detection,the number of myelinated nerve fibers,and the thickness of myelin at 16 weeks after the surgery,there were significant differences between groups C',D' and groups A',B'(P < 0.05),but there was no significant differences between group A' and group B',and between group C' and group D'(P > 0.05).At 16 weeks after the surgery,The results by TEM observed showed that there were fewer fibers,smaller diameter,sparse distribution,thinner myelin sheath in group A' and group B',but the number of regenerated fibers was more,the distribution was extensive,and the myelin sheath was thicker in group C' and group D'.Conclusion:Shenfu Injection has protective effect on sciatic nerve preserved by vitrification,and can improve nerve regeneration after allograft in rats.