Research On The Reconstruction Of Rats Sciatic Nerve Defect Via Plasma Modified Nerve Conduits Seeded With Rats Bmscs

The crucial part of peripheral nerve reconstruction in tissue engineering was the fabrication of nerve substitution and the raw material should be easily acquired. Two aspects were involved, one was the preparation of biomaterials, and the other was the cells for the biostructure. Previous research had revealed that after plasma treatment, biomaterials would have good hydrophilicity, and bone mesenchymal stem cells (BMSCs) were the most easily acquired one of all the stem cells that we knew up to date. Our research focused on the plasma treated Poly lactic-co-glycolic Acid (PLGA) nerve conduit seeded with BMSCs, and we tried to reveal that whether BMSCs had good cell affinity with plasma treated PLGA conduits, could BMSCs be induced to schwann cells (SCs) in vivo, and the feasibility of the cell seeded nerve conduit to repair the sciatic nerve defect efficiently.Our research meant to develop a method for the proliferation and purification of the BMSCs, tried to fabricate the plasma treated PLGA nerve conduit, aimed to explore the cell consistent of the plasma treated PLGA nerve conduit with the BMSCs, planed to illustrate the role of the plasma treated PLGA nerve conduit seeded with BMSCs played in the reconstruction of rat sciatic nerve defect.Many research reports the methods for the acquirement of the BMSCs, but all these methods had limitations. Because in order to acquire the BMSCs at relatively higher purification, the cells should be passaged for so many times that the cells were no longer fit for the research. We develop a method to isolate,purify and proliferate the BMSCs by sort the Passage 0 (P0) cells via flow cytosorting (FCS) and passage the cells by collagenase NB4.The cells were identified by the specific marks on the membrane and their abilities to differenciate into adipocytes,condrocytes and osteocytes. Although many researches use PLGA scaffold as nerve conduit, the physical and chemical character of these scaffold was not satisfying. We tried to modify the PLGA by pretreated the PLGA fiber and braided the nerve conduit triaxially. The mechanical property,hydrophilicity and decomposability of the nerve conduit was assessed.In our research, we fabricated the plasma treated PLGA conduit seeded with BMSCs and tried to explore whether the cell adherence and proliferation rate was decreased via MTT assay. Scanning electron microscope and confocal laser microscope were used to observe the cell numbers,morphology and behavior on the PLGA conduit surface.In vivo evaluation of the plasma treated PLGA nerve conduits seeded with BMSCs complex was conducted by detection of their role in the reconstruction of the rat sciatic nerve defect. Electrophysiology,immunohistology and axons calculation of the artificial nerve were conducted to illustrate the function of this complex Based on our studies, several conclutions have been drawn: BMSCs could be acquired and purified by P0 cells sortation via FCS and passaged by collagenase NB4; plasma pretreated and braided triaxially PLGA nerve conduit had nice mechanical property,hydrophilicity and decomposability; BMSCs could proliferate and adhere better on plasma treated PLGA scaffold; the plasma treated PLGA nerve conduits seeded with BMSCs complex could repair the rat sciatic nerve defect and had similar results compared with autologous nerve.