复合壳聚糖微球-丝素基载药α-磷酸三钙骨水泥的细胞相容性及毒性  

Cytocompatibility and cytotoxicity of a novel composite bone cement material composed of chitosan microsphere, alpha-tricalcium phosphate and silk fibroin

作　　者：王金宁[1] 皮斌[1] 王鹏[1] 朱雪松[1] 杨惠林[1] 

机构地区：[1]苏州大学附属第一医院骨科,江苏省苏州市215006

出　　处：《中国组织工程研究》2014年第16期2519-2525,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(81071451;81171689;81301559);江苏省自然科学基金资助项目(BK2011264;SBK201341421);江苏省教育厅研究生创新计划(CXZZ11_0123)~~

摘　　要：背景:磷酸钙骨水泥具有良好的生物相容性和骨传导性,已被应用于临床,但其不良的力学性能和缺乏骨诱导性限制了其进一步发展和应用。目的:制备壳聚糖微球-丝素基载药α-磷酸三钙骨水泥,验证其细胞相容性及细胞毒性。方法:分别以含体积分数10%胎牛血清和1%双抗的α-MEM培养基、苯酚、100%及50%复合壳聚糖微球-丝素基载药α-磷酸三钙骨水泥材料的浸提液培养MC3T3-E1细胞,采用MTT法评估细胞生长增殖情况,采用乳酸脱氢酶活性检测法判断复合壳聚糖微球-丝素基载药α-磷酸三钙骨水泥材料的毒性。将MC3T3-E1细胞系与复合壳聚糖微球-丝素基载药α-磷酸三钙骨水泥材料共培养,扫描电镜观察细胞在材料表面的附着及生长。结果与结论:复合壳聚糖微球-丝素基载药α-磷酸三钙骨水泥材料浸提液对MC3T3-E1细胞的生长增殖无明显影响,无明显细胞毒性。MC3T3-E1细胞在复合壳聚糖微球-丝素基载药α-磷酸三钙骨水泥材料表面生长良好,伸展充分,在材料表面伸出伪足,与材料贴附紧密,表明壳聚糖微球-丝素基载药α-磷酸三钙骨水泥细胞相容性良好。BACKGROUND：With excelent biocompatibility and osteoconduction, calcium phosphate bone cement has been used in clinic, but the poor mechanical properties and lack of osteoinduction restrict its further use. OBJECTIVE：To investigate the cytocompatibility and cytotoxicity of a novel drug-carrying composite of bone cement composed of chitosan microsphere, α-tricalcium phosphate and silk fibroin. METHODS：MC3T3-E1 cels were cultured in vitroin minimum essential medium alpha medium （α-MEM）, which was supplemented with 10% fetal bovine serum, and 1% streptomycin sulfate, extract of the cement material at concentrations of 100% and 50%, and 6.4 mL/L phenol. 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay was used to measure cellproliferation and the cytotoxicity was assessed by the activity of lactate dehydrogenase. The MC3T3-E1 cels culturedin vitro were colected and seeded on the composite cement material, and cellmorphology was observed by scanning electron microscope. RESULTS AND CONCLUSION：The extract of composite cement material had no influences on the MC3T3-E1 cellproliferation, showing no obvious cytotoxicity. The scanning electron microscope image showed MC3T3-E1 cels adhered and proliferated wel on the composite cement material composed of chitosan microsphere, α-tricalcium phosphate and silk fibroin, and pseudopodia out of the cels were closely attached to the material surface. In conclusion, the cement composite was proved to have satisfactory cytocompatibility and no obvious cytotoxicity.

关 键 词：生物材料 骨生物材料 丝素基磷酸钙骨水泥 壳聚糖微球 浸提液 MC3T3-E1细胞 细胞增殖 MTT法 乳酸脱氢酶活性 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]