Abstract:

BACKGROUND: Previous study has investigate the influence of compound enzyme digestion and differential adherence method on culture of Schwann cells, and alkylphenol polyoxyethylene (Triton X-100) preparation for allogenic nerve scaffold.
OBJECTIVE: To prepare an allogenic nerve scaffold using chemical extraction method and culture Schwann cells with the scaffold in vitro.
METHODS: The bilateral sciatic nerves of Wistar rats were treated with 30 g/L trinitrotoluene and 40 g/L sodium deoxycholate for extractions. The middle piece of the extracted samples and the un-extracted samples was harvested for hematoxylin-eosin staining, S-100 and laminin immunohistochemical staining, as well as transmission electron microscope observation. Trypsin and collagenase were used to separate Schwann cells from the double sciatic nerves and brachial plexus of SD fetal rats. Highly purified Schwann cells were achieved with differential adherence and Arab-c to eliminate fibroblast. Finally the Schwann cells were injected into the acellular nerve scaffold and the consequence studies were performed by transmission electron microscope and scanning electron microscope. 
RESULTS AND CONCLUSION: The cells and myelin sheath were removed and basal membrane component was preserved in the sciatic nerve after extraction procedure of trinitrotoluene and sodium deoxycholate. The electron microscopy showed that the extracted nerves are composed of empty basal lamina tubes and collagen fibers between tubes. The residual S-100 protein in nerves was significantly reduced along with the increasing times of extraction, scaffold structure was destroyed after repeated extractions. Acellular allogenic nerve scaffold fits for the growth of Schwann cells, and can transform to aline in vitro. Chemical extraction that uses the detergents of Triton X-100 and deoxycholate is an ideal method to prepare nerve scaffold with biomimetic structure, by the method cells can be removed from the sciatic nerve of Wistar rats while the basal lamina component preserved in the acellular nerve. Allogenous nerve scaffold populated Schwann cells may be an ideal substance to repair the nerve defect after injuries.