掺镁微弧氧化钛表面促进大鼠BMSCs成骨分化的研究  

Magnesium-doped TiO_(2) coating on titanium by micro-arc oxidation promoting osteogenic differentiation of rat BMSCs

作　　者：王桂芳[1] 蒋欣泉[1] WANG Guifang;JIANG Xinquan(Department of Prosthodontics,Shanghai Ninth People s Hospital,Shanghai Jiao Tong University School of Medicine,National Clinical Research Center for Oral Diseases,Shanghai Engineering Research Center of Advanced Dental Technology and Materials,Oral Bioengineering Lab,Shanghai Research Institute of Stomatology,Shanghai Key Laboratory of Stomatology,Shanghai 200011,China)

机构地区：[1]上海交通大学医学院附属第九人民医院·口腔医学院口腔修复科,国家口腔疾病临床医学研究中心,上海口腔医学先进技术与材料工程技术研究中心,上海市口腔医学重点实验室,上海市口腔医学研究所,上海200011

出　　处：《口腔生物医学》2021年第2期77-81,86,共6页Oral Biomedicine

基　　金：国家自然科学基金青年项目(82001011)。

摘　　要：目的:探讨掺镁微弧氧化钛表面对大鼠骨髓间充质干细胞(BMSCs)成骨分化活性的影响。方法:以含钙、磷电解液微弧氧化处理钛表面(MAO)为对照组,通过在电解液中加入不同浓度镁构建低镁(Mg-1)和高镁(Mg-2)含量微弧氧化处理钛表面;采用场发射扫描电子显微镜(SEM)观察表面相貌结构,X射线光电子能谱仪(EDS)分析材料表面化学元素,MTT法检测钛表面大鼠BMSCs细胞代谢活性,细胞骨架染色观察钛表面细胞生长情况,碱性磷酸酶(ALP)染色/半定量分析细胞ALP蛋白的表达,实时定量RT-PCR检测Alp、Runt相关转录因子2(Runx2)、骨钙素(Ocn)、骨形态发生蛋白2(Bmp2)、骨涎蛋白(Bsp)、骨桥蛋白(Opn)等成骨分化基因的表达,免疫荧光检测OCN蛋白的表达。结果:BMSCs在MAO、Mg-1、Mg-2三组钛表面生长状态均良好,Mg-2组BMSCs增殖代谢活性最佳(P<0.05);与MAO组相比,Mg-1组和Mg-2组提高了材料表面细胞成骨分化相关基因和ALP、OCN蛋白的表达,其中Mg-1组促进Runx2、Alp基因和ALP蛋白表达的作用更佳,Mg-2组上调Ocn、Bmp2、Bsp、Opn基因和OCN蛋白表达更明显(P<0.05)。结论:镁离子掺入微弧氧化钛表面具有良好生物学相容性,可促进大鼠BMSCs增殖活性,显著提高钛表面的成骨分化性能,具有潜在的促骨结合作用。Objective:To investigate the effect of magnesium-doped TiO_(2)coating on titanium by micro-arc oxidation on promoting osteogenic differentiation of rat bone marrow mesenchymal stem cells(BMSCs).Methods:The micro-arc oxidation(MAO)treatment of titanium surface in calcium and phosphorus electrolyte was set as the control group.By adding different concentrations of magnesium into the electrolyte,low(Mg-1)and high magnesium(Mg-2)content MAO treated titanium surfaces were constructed.The surface morphology was observed by field emission scanning electron microscope(SEM),the chemical elements on the surface were analyzed by X-ray photoelectron spectroscopy(EDS).MTT method was applied to detect the metabolic activity of BMSCs,cytoskeleton staining was used to observe the growth of BMSCs,alkaline phosphatase(ALP)staining/semi quantitative analysis was used to analyze the expression of ALP protein.The osteogenic differentiation related genes,including Alp,Runx2,Ocn,Bmp2,Bsp,Opn,were analyzed by real-time PCR.The expression of OCN protein was detected by immunofluorescence.Results:BMSCs grew well on the titanium surface of MAO,Mg-1 and Mg-2 groups.The metabolism activity of BMSCs in Mg-2 group was the best(P<0.05).Compared with MAO,the expressions of osteogenic differentiation related genes were increased in both Mg-1 and Mg-2 groups,and the expressions of ALP and OCN protein were enhanced as well.The expression of Runx2,Alp gene expression and ALP protein expression was significantly higher in Mg-1 group when compared with Mg-2 group,while the osteogenic related genes such as Ocn,Bmp2,Bsp,Opn and protein expression of OCN were significantly up-regulated in Mg-2 when compared to Mg-1 group(P<0.05).Conclusions:The incorporation of magnesium ion into the micro-arc oxidation treated titanium surface possessed good biocompatibility,promoted the proliferation of rat BMSCs,significantly improved the osteogenic differentiation of titanium surface,and had a potential in promoting bone implant contact.

关 键 词：镁离子 微弧氧化 钛 骨髓间充质干细胞 成骨分化 

分 类 号：R780.2[医药卫生—口腔医学]