纯钛表面微米坑复合纳米管形貌对骨髓间充质干细胞增殖及成骨分化的影响  

Acid etching and anodic oxidation of titanium surface of nanotubes promotes the proliferation and osteogenesis of bone marrow mesenchymal stem cells

作　　者：马燕 张华 商建平[2] 姜琦[3] Ma Yan;Zhang Hua;Shang Jian-ping;Jiang Qi(Department of Stomatology,Health Care Hospital of Linyi,Linyi 276000,Shandong Province,China;Department of Stomatology,Central People’s Hospital of Siping,Siping 136000,Jilin Province,China;China-Japan Union Hospital,Jilin University,Changchun 130033,Jilin Province,China)

机构地区：[1]临沂市妇女儿童医院口腔科,山东省临沂市276000 [2]四平市中心人民医院口腔科,吉林省四平市136000 [3]吉林大学中日联谊医院,吉林省长春市130033

出　　处：《中国组织工程研究》2018年第29期4614-4619,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:近年来对钛种植体材料表面微米纳米的仿生设计成为了生物材料领域的研究热点。目的:观察纯钛试件表面微米坑复合纳米管形貌对骨髓间充质干细胞增殖和成骨分化的影响。方法:采用酸蚀阳极氧化方法,在纯钛表面构建微米坑复合纳米管形貌,作为实验组,以抛光处理的光滑钛试样为对照组,采用场发射扫描电子显微镜观察两组试样表面形貌。将骨髓间充质干细胞接种于两组钛试样表面,常规培养7 d内,MTT法检测细胞增殖能力;成骨诱导培养3,7,14 d后,检测碱性磷酸酶活性;成骨诱导培养7,14,21 d,RT-qPCR和Western blot检测成骨标志的mRNA及蛋白表达。结果与结论:(1)对照组试样表面无微米及纳米级结构,实验组试样表面可见为凹凸不平的微米坑形貌与均匀分布的纳米管阵列;(2)随着培养时间的增长,两组试样表面的骨髓间充质干细胞数量逐渐增加,实验组培养3,5,7 d的细胞数量明显多于对照组(P<0.05);(3)实验组成骨诱导培养7,14 d的碱性磷酸酶活性高于对照组(P<0.05);(4)实验组成骨诱导培养7,14,21 d的骨桥蛋白、骨特异性转录因子、骨钙素的mRNA及蛋白表达均高于对照组;(5)结果表明,钛表面微米坑复合纳米管形貌有利于骨髓间充干细胞的增殖与成骨分化。BACKGROUND:In recent years,the biomimetic design of titanium implant surface at micron-nano level has become a research hotspot in the biomaterial research.OBJECTIVE:To observe the effect of acid etching and anodic oxidation on the surface morphology of pure titanium and the proliferation and osteogenesis of bone marrow mesenchymal stem cells(BMSCs).METHODS:The nanotubes with micron roughness were prepared on the titanium surface by acid etching and anodizing method.The surface morphology of nanotubes with micron roughness was used as experimental group and smooth titanium sample treated by polishing as control group.BMSCs were seeded on the surface of titanium samples and observed using field emission scanning electron microscope.Proliferation of the cells was detected by MTT assay within 7 days of conventional culture.Alkaline phosphatase activity was measured At 3,7,14 days after osteogenic induction.Osteogenic marker genes and proteins were detected using RT-qPCR and western blot at 7,14,21 days after osteogenic induction.RESULTS AND CONCLUSION:The control group had no micron and nano structure surface of the samples,while uneven surface,micron-sized holes,and uniformly distributed nanotube arrays were visible in the experimental group.With the culture time,the number of BMSCs on the sample surface in the two groups was gradually increased,and the cell number in the experimental group was significantly higher than that in the control group at 3,5,7 days after culture(P<0.05).The alkaline phosphatase activity in the experimental group was also higher than that in the control group at 7 and 14 days after osteogenic induction(P<0.05).Compared with the control group,the expression levels of osteopontin,bone-specific transcription factors,osteocalcin mRNAs and proteins were increased in the experimental group at 7,14,21 days after culture.These results suggest that the acid etching and anodic oxidation of the titanium surface of nanotubes can promote BMSCs proliferation and osteogenic differentiation.

关 键 词：钛种植体 酸蚀阳极氧化处理 微米坑 纳米管 骨髓间充质干细胞 细胞增殖 成骨分化 干细胞 牙种植体 钛 骨髓 间质干细胞 细胞增殖 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]