摘要
背景:国产多孔钽植入材料具备三维立体空间结构,而材料的三维立体结构可能会影响细胞的生长及分泌。目的:观察与国产多孔钽复合培养后成骨细胞増殖、细胞周期、成骨性细胞因子分泌功能的变化。方法:提取第3代兔成骨细胞,分3组培养:正常组常规培养,对照组加入多孔钽材料浸提液,实验组加入多孔钽支架材料。培养3,5,7 d,CCK-8法检测各组细胞增殖,扫描电镜观察实验组成骨细胞在多孔钽支架上的生长情况,Elisa试剂盒检测各组成骨细胞内骨钙素与Ⅰ型胶原的分泌;培养7 d时,流式细胞仪检测各组细胞周期。结果与结论:(1)细胞增殖:3组培养不同时间点的细胞增殖无差异;(2)扫描电镜:复合培养第3天,细胞在多孔钽支架表面和孔隙内黏附生长,排列较稀疏,突起较少;复合培养第5天,细胞边缘有突起向四周伸展,相邻细胞间突起相互连接横跨孔隙,连成片状;复合培养第7天,细胞突起融合汇合成片状,分泌的基质覆盖大部分支架表面;(3)Elisa检测:随着培养时间的延长,3组成骨细胞骨钙素分泌逐渐增加,Ⅰ型胶原分泌先升高后降低,实验组培养不同时间点的骨钙素分泌均高于正常组、对照组;(4)细胞周期:3组成骨细胞均是正常的二倍体细胞,无异倍体细胞出现,细胞DNA含量正常,3组细胞周期分布相似;(5)结果表明:多孔钽支架材料具有良好的生物相容性,可能促进成骨细胞的矿化与成骨。
BACKGROUND:The domestic porous tantalum implant material has a three-dimensional spatial structure that may affect the growth and secretion of cells.OBJECTIVE:To investigate the proliferation,cell cycle,osteogenic secretion of osteoblasts co-cultured with domestic porous tantalum scaffolds.METHODS:Passage 3 rabbit osteoblasts were extracted and divided into three groups:normal culture group,control group added with the porous tantalum extract,and experimental group co-cultured with the porous tantalum scaffold.The proliferation of cells in each group was detected by cell counting kit-8 method at 3,5,7 days of culture.The growth of osteoblasts on the porous tantalum scaffold was observed by scanning electron microscopy.ELISA kit assay was applied to detect the levels of osteocalcin and type I collagen in osteoblasts at 3,5,7 days of culture.The cell circle of osteoblasts was detected by flow cytometry at 7 days of culture.RESULTS AND CONCLUSION:(1)Cell proliferation:There was no difference in cell proliferation among the three groups at different time points(P>0.05).(2)Scanning electron microscopy:On the 3rd day of compound culture,the cells adhered to the scaffold surface and pores,and arranged sparsely with few protrusions.On the 5th day,the cells began to extend,and the adjacent cells were interconnected to form a flaky growth.On the 7th day,the cells merged into pieces,and the secreted matrix covered most of the scaffold surface.(3)ELISA detection:With the cultivation time,the osteocalcin secretion from the osteoblasts was gradually increased in all the three groups,while the secretion of type I collagen was increased first and then decreased.The secretion of osteocalcin and type I collagen in the experimental group at different time points were higher than that in normal culture and control groups.(4)Cell cycle:The osteoblasts in the three groups were normal diploid cells,no aneuploid cells appeared,and the cell DNA content was normal.The cell cycle distribution was similar in the three groups.To conclude,the domestic porous tantalum scaffolds have good biocompatibility and may promote osteoblast mineralization and osteogenesis.
作者
王茜
滕雪峰
甘洪全
张辉
崔逸爽
陈婧婧
李琪佳
王志强
Wang Qian;Teng Xue-feng;Gan Hong-quan;Zhang Hui;Cui Yi-shuang;Chen Jing-jing;Li Qi-jia;Wang Zhi-qiang(Medical Experimental Center,North China University of Science and Technology,Tangshan 063210,Hebei Province,China;Department of Orthopedics,Affiliated Hospital of North China University of Science and Technology,Tangshan 063000,Hebei Province,China;First Department of Joint,Second Hospital of Tangshan,Tangshan 063000,Hebei Province,China;Graduate School,North China University of Science and Technology,Tangshan 063009,Hebei Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第30期4757-4762,共6页
Chinese Journal of Tissue Engineering Research
基金
国家科技部科技支撑课题资助项目(2012BAE06B03)
河北省科技支撑资助项目(16277776D)
河北省医学科学研究重点课题计划资助项目(20160225)
华北理工大学博士科研启动基金资助项目(28606299)~~
作者简介
王茜,女,1981年生,河北省衡水市人,汉族,副教授,博士,主要从事神经解剖学、骨组织工程研究。;共同第一作者:滕雪峰,华北理工大学医学实验研究中心,河北省唐山市063210。;通讯作者:王志强,教授,博士生导师,华北理工大学附属医院骨科,河北省唐山市063000。
