小肠粘膜下层与雪旺细胞生物相容性的研究  

Biocompatibility of small intestinal submucosa with Schwann cells in vitro

作　　者：苏琰[1] 张长青[1] 张开刚[1] 谢雪涛[1] 曾炳芳[1] 

机构地区：[1]上海交通大学附属第六人民医院骨科,200233

出　　处：《中华创伤骨科杂志》2007年第2期153-156,共4页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金资助项目（30371444）

摘　　要：目的 研究猪小肠粘膜下层（SIS）与体外培养的鼠雪旺细胞（SCs）的生物相容性。方法 在体外将分离培养的SD乳鼠SCs接种于制备好的猪SIS上进行复合培养，通过相差显微镜和扫描电镜观察不同时间段SCs在SIS上的黏附、生长与增殖情况，用酶联免疫吸附测定和逆转录聚合酶链反应方法检测SCs分泌神经生长因子-β和脑源性神经生长因子的情况。对照组为接种于未平铺SIS的孔中正常培养的SCs。结果培养3～5d后，相差显微镜下见SIS边缘SCs较为密集，黏附良好，多呈长梭形生长；扫描电镜观察见SIS表面SCs增殖黏附良好，胞体突起显著，呈端对端连接或成束排列，细胞表面可见蛋白颗粒分泌。酶联免疫吸附测定和逆转录聚合酶链反应方法检测发现，SCs与SIS复合培养后，神经生长因子-β和脑源性神经生长因子分泌良好，与对照组的SCs差异无统计学意义。结论 猪SIS与鼠SCs有良好的生物相容性，有望用作支架，供SCs黏附生长，构建组织工程神经，用于修复周围神经缺损。Objective To investigate the biocompatibility of porcine small intestinal submucosa （SIS） with rat Schwann cells （SCs） cultured in vitro. Methods The SCs of SD neonate rats were isolated and cultured in vitro, and then seeded on the prepared porcine SIS. At different times, the growth, adhesion and proliferation of SCs on SIS were observed with phase contrast microscope and scanning electron microscope, and their secretion of NGF-β （nerve growth factor-β） and BDNF （brain-derived neurotrophin factor） was examined with ELISA （enzyme linked immunosorbent assay） and RT-PCR （reverse transcription-pelymerase chain reaction）. The SCs were seeded on wells without SIS and cultured as the contrast group. Results When observed under phase contrast microscope 3 to 5 days after culture, SCs were intensive on the edge, well adhered to the SIS and mostly fusiform in shape. The scanning electron microscope showed the SCs grew and adhered actively onto the surface of SIS, the cell body being obviously prominent. The SCs connected end-to-end with each other or aligned in clusters. Secretion of protein granules was seen on the cell surface. ELISA and RT-PCR measurement revealed that, cultured in combination with SIS, SCs secreted NGF-β and BDNF prosperously without obvious difference from those in the contrast group. Condusions Porcine SIS has good biocompatibility with rat SCs. SIS can expectedly be used in repair of peripheral nerve defects as a scaffold where SCs may grow and adhere to construct tissue engineering nerves.

关 键 词：小肠粘膜下层 雪旺细胞 支架 周围神经 

分 类 号：R686[医药卫生—骨科学]