壳聚糖与大鼠嗅鞘细胞的生物相容性

Biocompatibility of chitosan films and olfactory ensheathing cells

背景:众多研究证实,嗅鞘细胞移植和生物导管在修复中枢神经系统损伤中发挥了显著的作用,课题组前期研究已证实了壳聚糖在大鼠体内神经系统具有良好的生物相容性。目的:观察壳聚糖与大鼠嗅鞘细胞的生物相容性。设计、时间及地点:体外细胞学对比观察,于2005-12/2006-05在武汉协和医院泌尿外科实验室完成。材料:将壳聚糖粉溶于10g/L的乙酸溶液中,配制成10g/L的壳聚糖溶液;显微剥离成年大鼠嗅球表面层,按常规方法行原代细胞培养。方法:实验分为3组,每组10孔。壳聚糖组将制备好的壳聚糖溶液0.1mL加入到培养板中,50℃烘箱至成膜干燥,加入40g/L氢氧化钠溶液中和乙酸,双蒸水漂洗,晾干即可。多聚赖氨酸组按常规方法赖氨酸包被培养板。未包被组不用任何材料包被。将原代培养的嗅鞘细胞分别接种至壳聚糖包被、多聚赖氨酸包被和未包被的培养板上进行培养。主要观察指标:于培养第3,5天显微镜下观察细胞形态学变化,MTT比色实验检测细胞存活和增殖状况。结果:嗅鞘细胞可在壳聚糖膜上贴壁生长,细胞形态及数量与未包被组相比差异无显著性意义,但与多聚赖氨酸组相比细胞数量偏低。结论:嗅鞘细胞与壳聚糖有良好的生物相容性。

BACKGROUND： Olfactory ensheathing cell transplantation and biological canal play an important role in repairing central nervous system injury; in addition, previous studies have shown that chitosan has a great biocompatibility in rat nervous system. OBJECTIVE： To research the biocompatibility of chitosan film and olfactory ensheathing cells of rats. DESIGN, TIME AND SETTIN： An in vitro control trial based on cytology level was performed in the Urology Laboratory of Wuhan Union Hospital from December 2005 to May 2006. MATERIALS： Chitosan powder （10 g/L） was dissolved in acid solution, and olfactory ensheathing cells derived from surface layer were normally primary-cultured. METHODS： The olfactory ensheathing cells were assigned into three groups, with 10 wells for each group. Chitosan solution （0.1 mL） was incubated onto culture plate and dried at 50℃. Then, the cells were incubated with sodium hydroxide solution （40 g/L） to neutralize acid solution, rinsed with double-distilled water, and open-air dried. Cells were harvested onto culture plate coating with poly-L-lysine, and any materials were not used in the control group. The primary-cultured olfactory ensheathing cells were incubated onto culture plate coating with chitosan, poly-L-lysine, and nothing. MAIN OUTCOME MEASURES： The morphological changes were observed using microscopy at day 3 and 5 post-culture, and MTT method was used to determinate survival and proliferation of olfactory ensheathing cells. RESULTS： Olfactory ensheathing cells could attach and grow on chitosan film. The shape and quantity of cells had no significant difference compared to control group. But the amount was lower than poly-L-lysine group. CONCLUSION： Olfactory ensheathing cells and chitosan have a good biocompatibility.