脱细胞基质面神经修复材料的制备

目的 制备具有天然神经组织结构的支架,构建组织工程化面神经用于修复面神经损伤。方法 取家兔面神经,改良化学萃取法制备脱细胞神经基质,HE染色形态学观察去细胞及脱髓鞘情况,荧光分光光度计测定支架内细胞经Quant-iT PicoGreen工作液染色后的DNA含量。MTT法检测细胞在支架上的相对生长率从而检测支架的细胞毒性。结果 支架移植体呈圆柱形,弹性与正常神经基本一致,组织观察显示细胞结构未见残余完整细胞及细胞碎片残留,未见神经髓鞘及轴突结构,细胞外基质形成纵向排列结构,结构之间可见空隙。兔脱细胞面神经基质支架内残留的DNA含量较正常兔面神经明显下降(P<0.01)。神经基质供体无细胞毒性。结论 改良化学萃取法可有效去除面神经细胞,天然结构保存完好,细胞毒性低,可作为组织工程化面神经的支架。

Preparation of acellular matrix facial nerve repair material

Objective To prepare the acellular matrix facial nerve repair material and construct a tissue-engineered facial nerve with a natural neural tissue structure which can be used for the repair of peripheral nerve injury.Methods The rabbit facial nerve was taken, and acellular nerve matrix was prepared by chemical extraction, the degree of decellation and demyelination was observed by HE staining.The DNA content of the cells in the scaffold after staining with Quant-iT PicoGreen working solution were measured by fluorescence spectrophotometer. The relative growth rate of cells on the scaffold was determined by MTT to detect the cytotoxicity.Results The scaffold graft was cylindrical, and its elasticity was basically consistent with normal nerves. Tissue observation showed that the structure of cell, the nerve myelin sheath and axon had been completely removed. The extracellular matrix formed a longitudinal arrangement structure and was filled with voids. The residual DNA content in the rabbit acellular facial nerve matrix scaffold was significantly lower than that in the normal rabbit facial nerve. Nerve matrix donors were non-cytotoxic.Conclusion The modified chemical extraction method can effectively remove facial nerve cells, preserve the natural structure intact and have low cytotoxicity. It can be used as a scaffold for tissue engineering facial nerves.