中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2015年
8期
478-482
,共5页
孙子环%夏荣%孙磊%胡小晔%闵曦%徐基亮
孫子環%夏榮%孫磊%鬍小曄%閔晞%徐基亮
손자배%하영%손뢰%호소엽%민희%서기량
钛%纳米管%间质干细胞%骨形成蛋白
鈦%納米管%間質榦細胞%骨形成蛋白
태%납미관%간질간세포%골형성단백
Titanium%Nanotubes%Mesenchymal stem cells%Bone morphogenetic protein
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.
目的 探討二氧化鈦納米管陣列加載重組人骨形成蛋白2 (recombinant human bone morphogenetic protein-2,rhBMP-2)對小鼠骨髓間充質榦細胞(bone mesenchymal stem cell,BMSC)早期活性的影響,為鈦種植體錶麵生物化學改性提供實驗依據.方法 利用暘極氧化技術在純鈦片錶麵製備雙層二氧化鈦納米管陣列,化學接枝rhBMP-2(實驗組),以機械拋光的純鈦為空白對照組,陰性對照A組為二氧化鈦納米管組、陰性對照B組為二氧化鈦納米管+羰基二咪唑組.用場髮射掃描電鏡觀察各組形貌併用X射線光電子能譜儀檢測各組元素.各組試件與BMSC共培養,檢測第1天各組細胞黏附鋪展情況(每組樣本量為3),第1、3、5天各組細胞增殖A值及第5、7、11天各組堿性燐痠酶活性(每組每箇時間點樣本量為3).結果 場髮射掃描電鏡示實驗組錶麵可見粟粒狀顆粒物,X射線光電子能譜儀示實驗組氮峰明顯增高.場髮射掃描電鏡示第1天實驗組細胞黏附鋪展良好,細胞間連接廣汎而緊密,均優于其他3組.第1天各組細胞增殖效果不明顯;第3、5天實驗組A值(3.295±0.153、3.823±0.059)均顯著高于空白對照組(2.479±0.064、3.131±0.096)、陰性對照A組(2.715±0.075、3.371±0.047)及陰性對照B組(2.756±0.132、3.637±0.047) (P＜0.05);第5、7、11天實驗組堿性燐痠酶活性(0.0477±0.0287、0.0615±0.0016、0.0667±0.0018)均顯著高于其他3組(P＜0.05).結論 二氧化鈦納米管陣列可通過生物化學法加載rhBMP-2併具備良好的生物相容性.
목적 탐토이양화태납미관진렬가재중조인골형성단백2 (recombinant human bone morphogenetic protein-2,rhBMP-2)대소서골수간충질간세포(bone mesenchymal stem cell,BMSC)조기활성적영향,위태충식체표면생물화학개성제공실험의거.방법 이용양겁양화기술재순태편표면제비쌍층이양화태납미관진렬,화학접지rhBMP-2(실험조),이궤계포광적순태위공백대조조,음성대조A조위이양화태납미관조、음성대조B조위이양화태납미관+탄기이미서조.용장발사소묘전경관찰각조형모병용X사선광전자능보의검측각조원소.각조시건여BMSC공배양,검측제1천각조세포점부포전정황(매조양본량위3),제1、3、5천각조세포증식A치급제5、7、11천각조감성린산매활성(매조매개시간점양본량위3).결과 장발사소묘전경시실험조표면가견속립상과립물,X사선광전자능보의시실험조담봉명현증고.장발사소묘전경시제1천실험조세포점부포전량호,세포간련접엄범이긴밀,균우우기타3조.제1천각조세포증식효과불명현;제3、5천실험조A치(3.295±0.153、3.823±0.059)균현저고우공백대조조(2.479±0.064、3.131±0.096)、음성대조A조(2.715±0.075、3.371±0.047)급음성대조B조(2.756±0.132、3.637±0.047) (P＜0.05);제5、7、11천실험조감성린산매활성(0.0477±0.0287、0.0615±0.0016、0.0667±0.0018)균현저고우기타3조(P＜0.05).결론 이양화태납미관진렬가통과생물화학법가재rhBMP-2병구비량호적생물상용성.
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 1th day in the experimental group (0.0477 ± 0.0287,0.0615 ± 0.0016,0.0667 ± 0.0018) were better than those in the control group,negative control A group and negative control B group(P＜0.05).Conclusions Titanium nanotube arrays can be loaded with rhBMP-2 by biochemical methods and have good biocompatibility.
