聚乳酸-壳聚糖接骨板的体外细胞毒性研究

Cytotoxicity evaluation of biodegradable polylactic acid-chitin plates in osteosynthesis in vitro

目的:利用体外细胞培养技术,对聚乳酸-壳聚糖接骨板的细胞毒性进行研究,为临床应用作前期准备。方法:采用间接接触法(MTT法)评价聚乳酸-壳聚糖接骨板的细胞毒性。MTT法:采用体外培养的小鼠成纤维细胞(L929),在培养液中分别添加不同浓度的聚乳酸-壳聚糖接骨板浸提液(0,50%和100%),逐日用倒置显微镜对各组细胞进行形态学观察,于接种后2、4、7 d通过MTT法显色,在酶标仪450 nm波长处测定吸光度值,计算相对增殖率,对聚乳酸-壳聚糖接骨板的细胞毒性进行评价。结果:随时间推移,不同浓度聚乳酸-壳聚糖接骨板浸提液对L929细胞增殖均有促进作用(P<0.01),在2、4、7 d各浓度之间无明显差异性(P>0.05),提示L929细胞增殖与聚乳酸-壳聚糖接骨板浸提液的浓度无明显相关;随时间推移,实验各组L929细胞数量、形态学变化与空白对照组无明显差异,评分聚乳酸-壳聚糖接骨板的细胞毒性为0级。结论:聚乳酸-壳聚糖接骨板无明显细胞毒性,具有良好细胞相容性,是一种具有发展前景的可吸收骨折内固定材料。

Objective： To evaluate cytotoxicity of polylactic acid - chitin plates in vitro and provide a sound scientific basis for clinical use. Methods ：The cytotoxicity of the polylactic acid -chitin plates was evaluated in extract assay （the MTT assay） using the cell line L929 in vitro. L929 ceils were cultured with different concentrations of the extracts of polylactic acid - chitin plates （0, 50% and 100% , V/V） in vitro. MTT tests were performed to evaluate the cell proliferation in 2, 4 and 7 days. And the morphologic changes of L929 cells were observed by inverted microscope everyday. Results： In the MTT assay, the extracts of Polylactic Acid - Chitin Plates enhanced the cell proliferation as well as the control （P 〈 0. 01 ）. The differences between them were not significant （ P 〉 0. 05 ）. No side effects were found in the growth, proliferation and attachment of L929 cells for the polylaetic acid - chitin plates, compared with the negative control. The cytotoxicity tests showed that the cytotoxicity graduation of the polylactic acid - chitin plates was 0 or 1, and showed nearly no cytotoxicity. Conclusion： The polylactic acid - chitin plates has an ideal cellular compatibility. It is a prospective and applicable internal bone fixation device.