CAJ | 학술논문

目的探讨钛合金表面纳米化对成骨细胞黏附、增殖、分化和成骨矿化能力的影响.方法采用塑性变形和化学处理方法在Ti6Al4V合金上制得一种新型多孔纳米晶体的表面结构.将原代培养的大鼠成骨细胞与纳米表面、光滑表面及粗糙表面钛合金共培养.分别通过细胞计数、扫描电镜、MTT比色法、逆转录聚合酶链反应(reverse transcriptase-polymerase chain reaction,RT-PCR)分析和钙结节荧光染色等方法观察成骨细胞在3组钛合金表面的黏附、增殖、分化及成骨能力,所得数据进行统计学分析.结果黏附能力实验结果显示早期纳米表面黏附的成骨细胞数多于其它两组(P＜0.05).扫描电镜观察显示纳米表面上成骨细胞可以较其它两组更早的伸展黏附.增殖能力实验显示,3组细胞的生长曲线大体形态基本相同,但纳米表面成骨细胞的指数增殖期比光滑表面和粗糙表面长3～10 d,且最高峰更高.RT-PCR研究表明,共培养14 d后,纳米表面黏附的成骨细胞中碱性磷酸酶、Ⅰ型胶原和骨钙素mRNA的转录量均升高(P＜0.05).另外,与光滑和粗糙表面相比,纳米表面形成钙结节面积显著增大(P＜0.05).结论纳米多孔表面技术能促进成骨细胞的体外早期黏附、增殖、分化和成骨能力,为纳米技术进一步应用到人工关节提供新的方向.
목적 탐토태합금표면납미화대성골세포점부、증식、분화화성골광화능력적영향.방법 채용소성변형화화학처리방법재Ti6Al4V합금상제득일충신형다공납미정체적표면결구.장원대배양적대서성골세포여납미표면、광활표면급조조표면태합금공배양.분별통과세포계수、소묘전경、MTT비색법、역전록취합매련반응(reverse transcriptase-polymerase chain reaction,RT-PCR)분석화개결절형광염색등방법관찰성골세포재3조태합금표면적점부、증식、분화급성골능력,소득수거진행통계학분석.결과 점부능력실험결과현시조기납미표면점부적성골세포수다우기타량조(P＜0.05).소묘전경관찰현시납미표면상성골세포가이교기타량조경조적신전점부.증식능력실험현시,3조세포적생장곡선대체형태기본상동,단납미표면성골세포적지수증식기비광활표면화조조표면장3～10 d,차최고봉경고.RT-PCR연구표명,공배양14 d후,납미표면점부적성골세포중감성린산매、Ⅰ형효원화골개소mRNA적전록량균승고(P＜0.05).령외,여광활화조조표면상비,납미표면형성개결절면적현저증대(P＜0.05).결론 납미다공표면기술능촉진성골세포적체외조기점부、증식、분화화성골능력,위납미기술진일보응용도인공관절제공신적방향.
Objective To evaluate the effect of nanoporous surface of titanium alloy on adhesion,proliferation,differentiation and mineralization of osteoblasts.Methods The present study was to prepare a novel nanostructured surface of Ti6Al4V alloy by the severe plastic deformation (SPD).Osteoblasts were cultured on nanoporous, smooth and rough titanium alloy in vitro.The adhesion, proliferation,differentiation and mineralization of osteoblasts were observed by cells counting,scanning electron micrography(SEM),standard MTT assay,reverse transcriptase-polymerase chain reaction(RT-PCR)analysis and fluorescent labeling of calcium deposition.Results The number of cells on the nanoporous titanium alloy was higher than any that of other substrates.The early cellular extension on the nanophase surfaces was observed under SEM.The growth curve of osteoblasts cultured on different surfaces had similar appearance,however,longer exponential phase of growth and higher cell count was observed in osteoblasts cultured on nanoporous surface.Of the examined genes,collagen Ⅰ(COL-Ⅰ),alkaline phosphatase(ALP)and osteocalcin(OC)showed a change in expression with different surfaces.All of them were upregulated on the nanophase surface.In addition, a larger area of calcified nodules on nanophase surface was also observed,compared with the other groups.Conclusion Nanoporous surface technology can improve the early adhesion,proliferation,differential and mineralization of osteoblasts in vitro,which suggests nanometer technology should be further considered for orthopaedic implant applications.