Experimental Studies On Culture Of Rabbit Schwann Cells In Vitro And Repair Of Facial Nerce Defect Combined With PLGA Conduit

The most common reasons of facial nerve defect are trauma and tumor surgery. The clinical treatment of a severed peripheral nerve involves the use of an autologous nerve graft to bridge the gap when it is difficult to suture it directly. But nerve autografting remains many problems, i.e. sacrificing one or more functional nerves and donor-site sequelaes include loss of sensation, scarring. In addition, the limited availability of donor tissue represents a severe problem. To avoid these problems, more attention has been focused on the development of tissure engineering nerve as a substitute for the nerve autograft.The purpose of this study is to find a new approach to obtain more population and more purification of SCs from neonatal rabbits; Discuss the biological effects of basic fibroblast growth factor (b-FGF) to SCs cultured in vitro ; And the cultured cells are combined with poly lactic acid-co -glycolic acid(PLGA) conduit to construct a tissue engineering artifical nerve to bridge a facial nerve gap.Method: High-density collagenase was used to separate SCs from neonatal rabbits sciatic nerves. Cytosine arabinoside and Geneticin was used to eliminate fibroblast cells; then bFGF was added to promoteproliferation of SCs. The growth and the biological characteristics of SCs were observed by cell morphological survey, cell growth curve, SEM and S-100 stains. The biological effects of b-FGF to SCs were measured by methylthiazol tetrazolium (MTT) method; A tissue engineering bio--artificial nerve constructed by PLGA conduit combining SCs cultured in vitro was to bridge a 1 cm facial nerve gap and the functional recovery was evaluated by means of general observation, histological stain , electrophysiological analyse and TEM examination 8 &16 weeks after surgery.Result: SCs sepreated from sciatic nerves of neonatal rabbits can be cultured , purified and proliferated in vitro ; bFGF can promote the proliferation of SCs; SCs combined PLGA conduit can promote the regeneration of axon and the recovery of function better than empty PLGA conduit does.Conclusion: Schwann Cells harvested from neonatal rabbits can be used as seeded cells in peripheral nerve tissue engineering; b-FGF can promote the proliferation of SCs; PLGA conduit combined autologous SCs is a promising bio-artificial nerve for clinical practice.