Preparation Of Scaffold Of Acellular Nerve And Study Of Planting Isogenous Schwann Cells Into Scaffold In Vitro

Objective:To prepare an allogenous nerve scaffold with chemical extraction and to look for an ideal substance to repair large gap of nerve defect after injuries by cultured population of Schwann cells(Scs) and acellular allogenous nerve grafts.Method:The double sciatic nerves of rats were treated with a solution of Triton X-100 and a solution of sodium deoxycholate at room temperature.HE staining was performed to visualize cells and immunohistochemical staining was performed to visualize the presence of S-100 and laminin.The ultrastructure of acellular and fresh nerves were observed and photographed in a transmission electron microscope.Trypsin and collagenase were used to separate Schwann cells from the double sciatic nerves and brachial plexus of SD rat .It was used to achieve high purified Scs with Arab-c to eliminate fibroblast.Finally the Scs were injected into the acellular nerve grafts and the consequence studied.Results:The cells and myelin sheath were removed and basal membrane component was preserved in the sciatic nerve after extraction procedure. The transmission electron microscopy showed that the myelin sheaths were absent in the extracted nerve segments and empty basal lamina tubes remained in the endoneurium.Allogenous nerve grafts and high purified Scs were acquired, which could be integrated from each other well. Scs could survive and transform to aline in vitro.Conclusion:Chemical extraction is an ideal method to prepare nerve scaffold that the detergents of Triton X-100 and deoxycholate, by which cells and myelin sheaths could be removed from the sciatic of Wistar rats while the basal lamina component preserved in the acellular nerve.Allogenous nerve scaffold populated...