胶原包埋羟基磷灰石涂层人工机械瓣膜对犬血管内皮细胞的毒性  

Toxicity assessment of hydroxyapatite-coated mechanical heart valve embedded with collagen to vascular endothelial cells

作　　者：程光存[1] 李春生[1] 严宇[1] 王岚[1] 严中亚[1] 罗乐[2] 方晓东 陶汝华 

机构地区：[1]安徽医科大学附属省立医院心脏外科,安徽省心血管病研究所,安徽省合肥市230001 [2]合肥工业大学,安徽省合肥市230009 [3]中国科学院安徽光学精密机械研究所,安徽省合肥市230031

出　　处：《中国组织工程研究》2013年第51期8894-8900,共7页Chinese Journal of Tissue Engineering Research

基　　金：安徽省自然科学基金项目(1308085MH143)~~

摘　　要：背景:前期实验利用脉冲激光沉积方法制作了胶原包埋羟基磷灰石涂层人工机械瓣膜。目的:进一步分析胶原包埋羟基磷灰石涂层人工机械瓣膜的组织相容性和毒性。方法:将传代的犬血管内皮细胞悬液接种在胶原包埋羟基磷灰石涂层人工机械瓣膜材料上,1组置于含体积分数5%CO2、37℃恒温培养箱内静态培养3周,另1组置于含体积分数5%CO2、37℃恒温培养箱动态旋转培养3周,扫描电子显微镜下观察细胞在材料上的附着情况。采用MTT法测定血管内皮细胞与胶原包埋羟基磷灰石涂层人工机械瓣膜材料复合培养的增殖能力。结果与结论:动态旋转培养中见人工机械瓣膜材料的表面有细胞附着,并且数目比静态培养多,细胞分布均匀,在培养第21天可见均匀细胞融合成片,将材料表面均匀覆盖;在静态培养中细胞成堆,分布不均匀。人工机械瓣膜材料对犬血管内皮细胞增殖无明显影响,细胞生长良好。表明胶原包埋羟基磷灰石涂层人工机械瓣膜材料对血管内皮细胞的增殖无明显影响,无明显毒性,具有良好的生物相容性。BACKGROUND： In early experiments, we prepared hydroxyapatite-coated mechanical heart valve embedded with collagen using impulse laser sediment method. OBJECTIVE： To further analyze the histocompatibility and toxicity of hydroxyapatite-coated mechanical heart valve embedded with collagen. METHODS： After passage, canine vascular endothelial cell suspension was inoculated onto the hydroxyapatite-coated mechanical heart valve embedded with collagen. One group was inoculated in 5% CO2, 37 ~C incubator for 3 weeks static culture, and the other group was inoculated in 5% CO2, 37 ~C incubator for 3 weeks spinner culture. Scanning electron microscope was used to observe cell attachment on the material. 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay was used to the proliferative capacity of vascular endothelial cells cultured with the hydroxyapatite-coated mechanical heart valve embedded with collagen. RESULTS AND CONCLUSION： During the spinner culture, adherent cells were found on the surface of mechanical heart valve, and the cells distributed evenly and confluent at 21 days to cover the surface of the material. The number of adherent cells in the spinner culture was higher than that in the static culture. The cellsduring the static culture were aggregated and distributed irregularly. The mechanical heart valve exhibited no effects on the proliferation of canine vascular endothelial cells which grew well. These findings indicate that the hydroxyapatite-coated mechanical heart valve embedded with collagen exert no effect on proliferation of vascular endothelial cells, has no toxicity and has good biocompatibility.

关 键 词：生物材料 细胞外基质材料 组织工程血管材料 脉冲激光沉积 胶原包埋羟基磷灰石涂层人工机械瓣膜 组织工程 犬血管内皮细胞 细胞培养 四唑盐(MTT)比色法 省级基金 

分 类 号：R318[医药卫生—生物医学工程]