壳聚糖增强型左旋聚乳酸与嗅鞘细胞的生物相容性  

Biocompatibility of olfactory ensheathing cells and poly-L-lactic acid reinforced by chitosan in vitro

作　　者：李卫东[1] 崔志明[1] 徐冠华[1] 范建波[1] 保国锋[1] 孙郁雨[1] 王玲玲[1] 

机构地区：[1]南通大学第二附属医院脊柱外科,江苏省南通市226001

出　　处：《中国组织工程研究》2013年第29期5316-5322,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(81271367);南通市社会事业科技创新与示范计划项目(HS2011009);南通市指令性社会发展科技计划项目(S2010005)~~

摘　　要：背景:既往研究表明壳聚糖和左旋聚乳酸制备的复合支架与一些细胞有着良好的生物相容性。目的:观察壳聚糖增强型左旋聚乳酸支架与大鼠嗅鞘细胞的生物相容性。方法:将出生1-3dSD大鼠的嗅鞘细胞接种于壳聚糖增强型左旋聚乳酸膜上作为实验组,设置嗅鞘细胞与多聚赖氨酸联合培养为对照组,培养1,3,5,7d进行细胞增殖力检测和免疫荧光抗体标记检测。结果与结论:嗅鞘细胞可在壳聚糖增强型左旋聚乳酸膜上存活,细胞毒性评级为Ⅰ级。实验组细胞形态呈圆形和椭圆形,突起少,细胞聚集成团;接种1d后,可见部分细胞团的外围细胞伸出短小突起,并渐向外周散开;接种第3天,见聚集成团的细胞扩散开来,大部分细胞生成突起,呈双极或三极,以双极为主;接种第5天,细胞突起明显伸长,形态仍以双极细胞和三极细胞为主,并可见扁圆细胞和不规则三角形细胞;接种第7天,细胞密度增加,突起伸展。对照组细胞形态与实验组具有类似特征。两组嗅鞘细胞数量、细胞周长和细胞面积差异无显著性意义(P>0.05)。表明壳聚糖增强型左旋聚乳酸支架与嗅鞘细胞具有良好的生物相容性。BACKGROUND：Previous studies have shown that composite scaffold of chitosan and poly-L-lactic acid has good biocompatibility with some cells. OBJECTIVE：To study the biocompatibility of poly-L-lactic acid reinforced by chitosan and olfactory ensheathing cells. METHODS：In experimental group, olfactory ensheathing cells from Sprague-Dawley rats aged 1-3 days were incubated onto chitosan-reinforced poly-L-lactic acid film. And in control group, olfactory ensheathing cells were co-cultured with poly-L-lysine. The proliferative ability of olfactory ensheathing cells was detected and the cells were observed with immunofluorescence histochemical staining at 1, 3, 5, 7 days after culture. RESULTS AND CONCLUSION：Olfactory ensheathing cells could survive on the chitosan-reinforced poly-L-lactic acid film, and the cytotoxic grade wasⅠ. Morphology of the cells in the experimental group was round or oval, with little processes and the cells aggregated into groups. One day after implantation, the periphery cells of the mass extended short projections and gradual y spread outward;3 days after implantation, the cells spread and most of the cells generated projections, most of which were bipolar or tri-polar;5 days after implantation, cel processes significantly extended, most cells were bipolar and tri-polar cells, while some were oval cells and irregular triangular cells;7 days after implantation, the cel density increased, and cel processes extended. Cel morphology of the control group had similar characteristics as the experimental group. There was no obvious difference between the control and the experimental group in number, perimeter or area of the cells （P〉0.05）. It showed that chitosan-reinforced poly-L-lactic acid had good biocompatibility with olfactory ensheathing cells.

关 键 词：生物材料 材料生物相容性 壳聚糖 左旋聚乳酸 多聚赖氨酸 嗅鞘细胞 生物相容性 共培养 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]