二氧化钛纳米管阵列加载重组人骨形成蛋白2的体外生物活性研究  

TiO2 nanotubes functionalized with recombinant human bone morphogenetic protein-2 enhance biological activity in vitro

作　　者：孙子环 夏荣[1] 孙磊[1] 胡小晔 闵曦 徐基亮[1] 

机构地区：[1]安徽医科大学第二附属医院口腔科,合肥230601 [2]中国科学院固体物理研究所安徽省纳米材料与纳米结构重点实验室

出　　处：《中华口腔医学杂志》2015年第8期478-482,共5页Chinese Journal of Stomatology

基　　金：基金项目：安徽省自然科学基金（1508085MH156）;安徽省学术和技术带头^、科研活动经费资助项目（2014H030）

摘　　要：目的 探讨二氧化钛纳米管阵列加载重组人骨形成蛋白2 （recombinant human bone morphogenetic protein-2,rhBMP-2）对小鼠骨髓间充质干细胞（bone mesenchymal stem cell,BMSC）早期活性的影响,为钛种植体表面生物化学改性提供实验依据.方法 利用阳极氧化技术在纯钛片表面制备双层二氧化钛纳米管阵列,化学接枝rhBMP-2（实验组）,以机械抛光的纯钛为空白对照组,阴性对照A组为二氧化钛纳米管组、阴性对照B组为二氧化钛纳米管＋羰基二咪唑组.用场发射扫描电镜观察各组形貌并用X射线光电子能谱仪检测各组元素.各组试件与BMSC共培养,检测第1天各组细胞黏附铺展情况（每组样本量为3）,第1、3、5天各组细胞增殖A值及第5、7、11天各组碱性磷酸酶活性（每组每个时间点样本量为3）.结果 场发射扫描电镜示实验组表面可见粟粒状颗粒物,X射线光电子能谱仪示实验组氮峰明显增高.场发射扫描电镜示第1天实验组细胞黏附铺展良好,细胞间连接广泛而紧密,均优于其他3组.第1天各组细胞增殖效果不明显;第3、5天实验组A值（3.295±0.153、3.823±0.059）均显著高于空白对照组（2.479±0.064、3.131±0.096）、阴性对照A组（2.715±0.075、3.371±0.047）及阴性对照B组（2.756±0.132、3.637±0.047） （P＜0.05）;第5、7、11天实验组碱性磷酸酶活性（0.0477±0.0287、0.0615±0.0016、0.0667±0.0018）均显著高于其他3组（P＜0.05）.结论 二氧化钛纳米管阵列可通过生物化学法加载rhBMP-2并具备良好的生物相容性.Objective To investigate the effect of TiO2 nanotube arrays covalently modified by recombinant human bone morphogenetic protein-2（rhBMP-2） on the early bioactivity of mesenchymal stem cells（MSC） in vitro and to provide experimental evidence for the biochemical modification of titanium implants.Methods In the experiment group,double titanium nanotube arrays were prepared by anodization,and were chemically grafted with rhBMP-2.Mechanically polished pure titanium was used as blank control group,and titanium dioxide nanotubes was used as negative control A group,and titanium dioxide nanotubes ＋ carbonyldiimidazole as negative control B group.Field emission scanning electron microscope（FE-SEM） and X-ray photoelectron spectroscopy（XPS） were used to detect the morphology and physicochemical properties of the experiment group,blank control group and the negative control group.Cell adhesion on the specimen surface of the experiment group,blank control group and negative control group on the 1st day was tested.Cell proliferation on the 1st,3rd and 5th day and alkaline phosphatase activity on the 5th,7th and 11th day was also tested.Results FE-SEM showed that the surface of titanium nanotubes loaded with rhBMP-2 possessed visible miliary particulate matter.XPS showed that nitrogen peak in the group of titanium nanotubes loaded with rhBMP-2 was significantly greater that those in the other groups.FE-SEM showed that the cells on the surface of the experimental group on the 1st day spread well,better than those in the control group and negative control group.Cell proliferation activity on the 1st day in different groups was not obvious（P〉0.05）,the A value of the experimental group on the 3rd and 5th day （3.295±0.153,3.823±0.059） were significantly higher than those in the control group（2.479±0.064,3.131±0.096） and negative control A group（2.715±0.075,3.371±0.047） and negative control B group（2.756±0.132,3.637±0.047）（P〈0.05）.Alkaline phosphatase activity on the 5th,7th and 1

关 键 词：钛 纳米管 间质干细胞 骨形成蛋白 

分 类 号：R783.1[医药卫生—口腔医学]