【摘要】 背景:羧甲基壳聚糖钙做为一种新型生物学材料,其生物学特征方面的研究较少。目的:评价羧甲基壳聚糖钙体外成骨细胞毒性。设计、时间及地点:对照观察实验,于2009-02-18/03-30在滨州医学院中心实验室完成。材料:羧甲基壳聚糖溶液与氯化钙溶液按一定比例混合、搅拌生成白色絮状沉淀,反复离心、冲洗沉淀后干燥,得羧甲基壳聚糖钙样品,呈白色粉末状固体;红外光谱分析特征性波峰发生了改变,提示羧甲基壳聚糖与钙离子发生了络合。方法:通过MTT法检测羧甲基壳聚糖钙浸提液对原代培养SD大鼠成骨细胞的毒性作用,以第1,2,3,5,7天为检测时间点,计算细胞相对增殖率,对材料毒性进行分级。正常培养基作为对照组。主要观察指标:①不同时间点各组细胞形态变化。②不同时间点各组细胞相对增殖率及材料毒性分级。结果:成骨细胞培养过程中,实验组细胞生长良好,细胞呈梭形、多边形等形态,细胞形态饱满,与对照组细胞形态无明显差别。在第1,2,3,5,7天时羧甲基壳聚糖钙浸提液对成骨细胞的相对增殖率均在97%以上,毒性分级为0或1级。结论:羧甲基壳聚糖钙具有良好的细胞相容性,细胞毒性在合格范围内,是一种有应用潜力的新型生物材料。更多还原

【Abstract】 BACKGROUND:The synthesis of carboxymethyl-chitosan calcium (CMCS-Ca2+) as a new biomaterial compound was widely studied,but there is few report regarding the biological characteristics. OBJECTIVE:To evaluate the cytotoxicity of CMCS-Ca2+ on osteoblasts in vitro. DESIGN,TIME AND SETTING:A controlled observation was performed at the Central Laboratory of Binzhou Medical University from February 18th to March 30th,2009. MATERIALS:Carboxymethyl-chitosan was mixed with calcium chloride to a certain degree to obtain white flocculent precipitate which was centrifuged,washed,dried again and again to obtain CMCS-Ca2+,a white power solid. Infrared spectrum indicated that wave crest of CMCS-Ca2+ had been changed,suggesting that a complexation occurred between carboxymethyl-chitosan and calcium ion. METHODS:The cytotoxicity of CMCS-Ca2+ leaching liquor to primary-cultured osteoblasts was detected using MTT assay. The relative growth rate was calculated and the cytotoxicity was graded at day 1,2,3,5,and 7. Normal culture media was considered as control group. MAIN OUTCOME MEASURES:Cell morphology,relative growth rate,and cytotoxicity grade at different time points. RESULTS:Cells grew well in the experimental group,shaping fusiform and polygon. In addition,the cells were well-stacked. There was no significant difference in cell morphology between the two groups. The relative growth rate was over 97%,and cytotoxicity grade was 0 or 1 at day 1,2,3,5 and 7. CONCLUSION:CMCS-Ca2+ has a good biocompatibility. The cytotoxicity of the new calcium compound satisfies the safety standard.更多还原