掺镁纳米多孔钛涂层的生物特性及促成骨分化作用  

Biological characteristics and osteogenic differentiation of magnesium-doped nanoporous titanium coating

作　　者：赵姗 张凌[3] 李胜男 康楠 孟箭[1] 李晓东 ZHAO Shan;ZHANG Lin;LI Sheng-nan;KANG Nan;MENG Jian;LI Xiao-dong(Department of Stomatology,Central Hospital of Xuzhou.Xuzhou 221000,Jiangsu Province;Department of Stomatology,Boluo County People's Hospital,Huizhou 516199,Guangdong Province;Department of Oromaxillofacial Head and Neck Oncology,Shanghai Ninth People′s Hospital,College of Stomatology,Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China)

机构地区：[1]徐州市中心医院口腔科,江苏徐州221000 [2]博罗县人民医院口腔科,广东惠州516199 [3]上海交通大学医学院附属第九人民医院口腔颌面-头颈肿瘤科,上海200011

出　　处：《上海口腔医学》2024年第1期6-12,共7页Shanghai Journal of Stomatology

基　　金：国家口腔疾病临床医学研究中心开放课题(NCRCO-202101);徐州市科技计划项目(KC21187);徐州医科大学附属医院发展基金资助项目(XYFY2020035)。

摘　　要：目的:通过微弧氧化(micro-arc oxidation,MAO)将生物活性镁离子掺入纳米多孔钛基础涂层,探讨其物理特性及对成骨的影响。方法:通过改变MAO电解液成分,控制多孔钛涂层中掺镁量,制备非含镁和含镁钛多孔钛涂层(MAO、MAO-mg)。采用扫描电镜(SEM)、粗糙度及接触角和能量色散X射线光谱仪(EDS)对样品进行表征分析,电感耦合等离子体/光学发射光谱仪(ICP-OES)测定掺镁纳米多孔钛涂层的Mg^(2+)释放能力。通过活/死双染、CCK-8检测材料增殖-毒性,并使用FITC-鬼笔环肽通过染色β肌动蛋白,确定细胞骨架结构。通过茜素红(ARS)、碱性磷酸酶(ALP)染色及实时聚合酶链式反应(qRT-PCR)确定涂层对体外成骨分化的影响。采用SPSS 25.0软件包对数据进行统计学分析。结果:加入镁离子的MAO电解液不会改变多孔钛涂层物表面特征,MAO制备出的各组涂层具有相似的微孔结构(P>0.05),MAO处理组(MAO、MAO-mg)表面粗糙度和接触角无显著差异(P>0.05),但显著高于Ti组(P<0.05)。EDS及ICP分析显示,镁离子成功掺入并从材料中释放。活/死双染及细胞增殖测定显示,与Ti组相比,MAO处理组无毒(P>0.05),且随着细胞培养时间延长,MAO-mg显著促进细胞增殖(P<0.05)。MAO-mg组ALP、ARS染色效果显著高于其他组;qRT-PCR显示,MAO-mg组的Runx2 mRNA(P<0.05)、ALP(P<0.05)和骨钙素OCN(P<0.05)表达显著高于Ti、MAO组。结论:MAO成功制备含镁纳米多孔钛涂层,且表现出明显的促成骨分化作用。PURPOSE:Bioactive magnesium ions were successfully incorporated into the nanoporous titanium base coating by micro-arc oxidation(MAO),and its physical properties and osteogenic effects were explored.METHODS:Nonmagnesium-containing and magnesium-containing titanium porous titanium coatings(MAO,MAO-mg)were prepared by changing the composition of MAO electrolyte and controlling the doping of magnesium in porous titanium coatings.The samples were characterized by scanning electron microscope(SEM),roughness,contact angle and energy dispersive X-ray spectrometer(EDS).Mg^(2+) release ability of magnesium-doped nanoporous titanium coatings was determined by inductively coupled plasma/optical emission spectrometer(ICP-OES).The structure of the cytoskeleton was determined by live/dead double staining,CCK-8 detection of material proliferation-toxicity,and staining ofβ-actin using FITC-phalloidin.The effects of the coating on osteogenic differentiation in vitro were determined by alizarin red(ARS),alkaline phosphatase(ALP)staining and real-time polymerase chain reaction(qRT-PCR).SPSS 25.0 software package was used for statistical analysis.RESULTS:The MAO electrolyte with magnesium ions did not change the surface characteristics of the porous titanium coating.Each group prepared by MAO had similar microporous structure(P>0.05).There was no significant difference in surface roughness and contact angle between MAO treatment group(MAO,MAO-mg)(P>0.05),but significantly higher than that of Ti group(P<0.05).With the passage of cell culture time,MAO-mg group promoted cell proliferation(P<0.05).MAO-mg group was significantly higher than other groups in ALP and ARS staining.The expression of Runx2 mRNA(P<0.05),ALP(P<0.05)and osteocalcin OCN(P<0.05)in MAO-mg group was significantly higher than that in Ti and MAO groups.CONCLUSIONS:MAO successfully prepared magnesium-containing nanoporous titanium coating,and showed a significant role in promoting osteogenic differentiation.

关 键 词：微弧氧化 掺镁纳米多孔涂层 成骨整合 镁离子 骨诱导性 

分 类 号：R783.1[医药卫生—口腔医学]