羧甲基壳聚糖温敏凝胶对小鼠L929细胞增殖的影响

The Effect of Carboxymethyl Chitosan Thermosensitive Hydrogel on Proliferation of L929 Cells

目的:研究自制羧甲基壳聚糖温敏凝胶对小鼠肺成纤维细胞(L929)增殖的影响及其组织相容性。方法:用羧甲基壳聚糖(CMCS)与甘油磷酸盐(GP)互配,制备羧甲基壳聚糖温敏凝胶,22只小鼠分为正常对照组1只、空白对照组3只,2%凝胶组和5%凝胶组各9只。采用四甲基偶氮唑盐法(MTT)分别测定质量浓度为50、10、2、0.4、0.08倍的羧甲基壳聚糖温敏凝胶浸提液对体外培养的L929细胞的增殖情况;分别将2%和5%的温敏凝胶植入小鼠皮下,于3、7、14d处死,常规组织切片,HE染色观察凝胶周围组织的炎症反应。结果:培养24h^96h,0.4倍及0.08倍浸提液组与阴性和阳性对照组的OD值差异有统计学意义(P<0.05);培养48、72、96h后,除50倍浸提液组外其余各实验组与阴性和阳性对照组的OD值差异有统计学意义(P<0.05),其中0.4倍浸提液促细胞增殖作用最为明显;凝胶植入后前3d,实验组以急性炎症为主,随着时间的延长各实验组炎症细胞逐渐减少至消失。结论:羧甲基壳聚糖温敏凝胶对L929细胞有明显的促进作用,且具有良好的组织相容性,在牙周病治疗中具有潜在的应用价值。

Objective： To investigate the effect of homemade carboxymethyl chitosan thermosensitive hydrogel on the proliferation of mouse lung fibroblasts （L929）, and its tissue compatibility thereof. Methods： Carboxymethyl chitosan （CMCS） interworked with glycerol phosphate （GP） to prepare carboxymethyl chitosan thermosensitive hydrogel. Twenty-two mice were divided into normal control group （n=1）, control group （n=3）, 2% hydrogel group （n=9） and 5% hydrogel group （n=9）. Effects of different concentrations （50, 10, 2, 0.4.and 0.08） of carboxymethyl chitosan thermosensitive hydrogel on proliferation of L929 cells were detected by MTT colorimetric assay. The 2% and 5% of carboxymethyl chitosan thermosensitive hydrogel were implanted subcutaneously in mice. Mice were killed separately on d 3, d 7 and d 14. The regular tissue slices were stained with HE to observe the inflammatory response on the surrounding tissue of hydrogel. Results： After 24 h, there were significant dif- ferences in OD values between 0.4-time and 0.08-time extract groups （P 〈 0.05）. After 48 h, 72 h and 96 h cultivation, there were significant differences in OD values between positive and negative groups except for the 50-time group（P 〈 0.05）. There was the most obvious role in promoting cell proliferation using 0.4-time extracts. On the first three days after histology injecting, the acute inflammation was found in the experimental group. Inflammatory cells decreased to disappear gradually in each experimental group with time. Conclusion： Carboxymethyl chitosan thermosensitive hydrogel could significantly promote the proliferation of L929 ceils and have good biocompatibility, which suggest a potential application for periodontal therapy.