Study Of Tissue Engineeringed Nerve Conduit Constructed By Acellular Muscle And Schwann Cells In Vitro

Objective To investigate the effect of Waller degenerateion on activation of Schwann cells, and the feasibility of mixed culture of autologous acellular muscle which seeded with autologous actived Schwann cells, and to find a new method to repair injuried periphery nerve.Method Left sciatic nerves of SD rats were deligated in operation group to generate Waller degeneration. After 7 days of deligation, ipsilatera sciatic nerves of both operation group and control group were evaluated by S-100 protein immunohistochemical staining, and positive percentage of S-100 protein immunohistochemical staining were statistical compared so as to observe the effect of Waller degeneration on sciatic nerve; Schwann cells, which play an important role in repairation of injuried periphery nerve, became the seed cells of artificial nerve, and this was accepted by researchers. Schwann cells harvested from sciatic nerve of SD rats was cultured, proliferated and purified in vitro, then S-100 protein immunocytochemistry staining was performed and growth curve was drew; To for preparation of acellular muscle, autologous biceps femoris was extracted in 3% Triton X-100 and 4% Deoxycholate, then was observed by scaning electron microscope and HE staining; Finally, Schwann cells and acellular muscle, which pretreated with asepsis polylysine, was mixed cultured for 2 weeks and observed by scaning electron microscope.Result The change of operation group's deligated sciatic nerves were similar to classic Waller degeneration after 1 week, and positive percentage of S-100 protein immunohistochemical staining of experimental group was much higher than the control group, and there was statistical significant difference (P<0.05); Schwann cells harvested from sciatic nerve of SD rats in vitro showed typical positive of S-100 protein immunocytochemistry staining; Regular pores were found in acellular muscle, which is suitable for proliferation of Schwann cells; After mixed culture of Schwann cells and acellular muscle in vitro, Schwann cells can adhere well to the acellular muscle with lining head to tail, then finally form Schwann cells columns which was similar to Büngner's band.Conclusion Activated Schwann cells can be obtained in Waller degeneration of periphery nerve, and can be used in research of tissue engineerined periphery nerve; Schwann cells can adhere well to the acellular muscle, which is suitable for artificial nerve.