珊瑚状钛酸钡纳米压电涂层促进大鼠骨髓间充质干细胞早期成骨分化的体外研究  

Fabrication of a coral-like barium titanate nano-piezoelectric coating and its effect on promoting osteogenic differentiation of rat bone marrow mesenchymal stem cells in vitro

作　　者：李露颖 李颖 李萌 张薇 赵领洲 胡先同 赵彦涛 马楚凡 Li Luying;Li Ying;Li Meng;Zhang Wei;Zhao Lingzhou;Hu Xiantong;Zhao Yantao;Ma Chufan(Department of Stomatology,Air Force Medical Center,CPLA,Beijing 100142,China;Institute of Orthopedics,Fourth Medical Center of the General Hospital of PLA,Beijing 100048,China)

机构地区：[1]中国人民解放军空军特色医学中心口腔科,北京100142 [2]放军总医院第四医学中心骨科医学部,北京100048

出　　处：《中华口腔医学杂志》2024年第6期578-585,共8页Chinese Journal of Stomatology

基　　金：北京市自然科学基金(7232173)。

摘　　要：目的探讨珊瑚状钛酸钡纳米压电涂层的生物相容性,及其于超声激发下产生的压电效应对大鼠骨髓间充质干细胞(BMSC)早期成骨分化的影响,为口腔种植体表面优化提供实验依据。方法通过阳极氧化、水热反应及高温退火在钛表面制备钛酸钡纳米压电涂层(涂层组),以抛光钛试件为对照组。采用扫描电镜、X射线光电子能谱、拉曼光谱和水接触角测量仪分别检测两组钛试件表面形貌、成分、晶相和亲水性;通过压电力显微镜测试涂层组压电性能。培养并鉴定大鼠BMSC并将其接种于两组钛试件表面,以接种于空白培养板的细胞为空白组;施加低强度脉冲超声干预后,检测细胞增殖和死活情况,评价涂层的细胞相容性;使用碱性磷酸酶(ALP)显色试剂盒检测各组碱性磷酸酶活性,并通过实时荧光定量PCR(RT-qPCR)检测成骨相关基因[整合素、骨形态发生蛋白2(BMP-2)、Runt相关转录因子2(RUNX2)]的表达情况,评价涂层促BMSC早期成骨分化的作用。结果涂层组钛表面呈现均匀的珊瑚状形貌,珊瑚触手直径为70~100 nm;主要成分为四方相钛酸钡;涂层组表面亲水性(水接触角10.12°±0.93°)显著优于对照组(水接触角78.32°±0.71°)(F=10165.91,P<0.001);涂层具有稳定的压电性能,压电常数约为5 pC/N。细胞实验显示,培养第3天,无论超声与否,涂层组细胞增殖活性显著小于空白组和对照组(P<0.05);培养第5天,无论超声与否,3组细胞增殖活性差异均无统计学意义(P>0.05);培养第7天,涂层组ALP活性显著大于空白组和对照组(P<0.05);RT-qPCR显示,超声后涂层组整合素和BMP-2 mRNA表达量显著大于其他组,且显著大于未超声涂层组(P<0.05);超声后对照组整合素mRNA表达量显著大于未超声对照组(P<0.05);超声后涂层组RUNX2 mRNA表达量显著大于未超声涂层组(P<0.05)。结论珊瑚状钛酸钡纳米压电涂层具有良好的生物相容性和稳定的压电性能,�ObjectiveTo investigate the biocompatibility of coral-like barium titanate nano-piezoelectric coatings and the influence of ultrasound-excited piezoelectric effect on the early osteogenic differentiation.MethodsThe barium titanate nano-piezoelectric coating(the coating group)was prepared on the surface of titanium metal by anodic oxidation,hydrothermal reaction and high-temperature annealing,and polished titanium specimens were used as control group.The surface morphology,composition,and crystal phase and hydrophilicity of the two groups of titanium specimens were characterized using scanning electron microscopy,X-ray photoelectron spectroscopy,Raman spectroscopy and contact angle meter.The piezoelectric properties of the materials were characterized by piezoresponse force microscopy.Rat bone marrow mesenchymal stem cells(BMSC)were cultured and identified and seeded the surface of titanium specimens in two groups.The cells seeded on blank culture plates were used as blank group.After low intensity pulsed ultrasound intervention,cell proliferation and live/dead staining were detected to evaluate cytocompatibility of the coatings.Alkaline phosphatase(ALP)activity of each group was detected by ALP staining kit,and the expression of osteogenesis-related genes[integrin,bone morphogenetic protein 2(BMP-2),Runt-related transcription factor 2(RUNX2)]was detected by real-time fluorescent quantitative PCR(RT-qPCR)to evaluate the effect of the coating on promoting the early osteogenic differentiation of BMSC.ResultsThe surface of titanium specimens in the coating group showed a uniform coral-like morphology,and the diameter of the coral tentacles was 70-100 nm.The main component was tetragonal barium titanate.The surface hydrophilicity of the coating group(water contact angle 10.12°±0.93°)was significantly better than that of the control group(water contact angle 78.32°±0.71°)(F=10165.91,P<0.001).The coating has a stable piezoelectric property with a piezoelectric constant of about 5 pC/N.Cell experiments showed that,

关 键 词：牙种植体 纳米技术 钛酸钡 涂层 低强度脉冲超声 骨髓间充质干细胞 成骨分化 

分 类 号：R783.6[医药卫生—口腔医学]