静电纺丝聚乳酸-乙醇酸聚合物膜与大鼠嗅鞘细胞的生物相容性研究

目的 探索静电纺丝技术制作的聚乳酸-乙醇酸聚合物（PLGA）膜与大鼠嗅鞘细胞（olfactory ensheathing cells，OECs）的生物相容性。方法 体外培养纯化成年大鼠OECs，将细胞接种于静电纺丝PLGA膜上，使用相差显微镜观察静电纺丝PLGA膜上OECs的细胞形态与分布；用CFDA SE荧光染色检测接种后1～5 d内静电纺丝PLGA膜上OECs的增殖情况；以多聚赖氨酸（PLL）包被细胞做对照。将静电纺丝PLGA膜植入大鼠体内观察其与大鼠的组织相容性。结果 体外培养纯化所得的OECs的细胞纯度大于90%。相差显微镜下示OECs在静电纺丝PLGA膜孔隙中的纳米纤维上黏附良好，细胞状态佳，且特异性地沿纳米纤维方向生长。接种后1～5 d内OECs在静电纺丝PLGA膜上均正常增殖，细胞数目与对照组相比差异无统计学意义。静电纺丝PLGA膜局部移植后大鼠无死亡，PLGA膜与周围组织融合并部分降解。结论 静电纺丝PLGA膜具有良好的生物相容性，OECs在其膜上的黏附和增殖状况良好，是有潜力的神经修复组织工程化神经移植物。

Biocompatibility of electrospun nanofiber-PLGA membrane with rat olfactory ensheathing cells

Objective To explore the biocompatibility of rat olfactory ensheathing cells(OECs) with electrospun nanofiber-PLGA (polyD,L-lactide-co-glycolide]) membrane. Methods The OECs of adult rats were seeded on the electrospun nanofiber-PLGA membrane. The morphology and adhesion to the membrane of OECs were observed by phase contrast microscopy.The proliferation ability of OECs was evaluated by CFDA SE immunofluorescent at 1-5 days after seeding. The control group was seeded into poly-L-lysine-treated plate. Moreover, the electrospun nanofiber-PLGA membrane was also implanted in rats to observe the biocompatibility. Results The purity of OECs obtained from in vitro culture was greater than 90%.The microscopic findings showed that the OECs grew well on the electrospun nanofiber-PLGA membrane, with good adhesion ability and good cell state; the cell also had an orientation growth along the nanofibers.The OECs had a normal proliferation in the electrospun nanofiber-PLGA membrane at 1-5 days after seeding, and the cell number was not significantly different from that of the control group. There was no death in rats after the PLGA film was transplanted, and the PLGA membrane was fused with the surrounding tissues, with some degradation at the fusion sites with the tissues. Conclusion The electrospun nanofiber-PLGA membrane shows a satisfactory biocompatibility, and OECs have good adhesion and proliferation on it, indicating that the membrane might be a potential tissue engineering nerve graft when combined with OECs.