Hao Yuquan, Li Shujun, Han Xuesong, Hao Yulin, Ai Hongjun
Department of Prosthodontics, School of Stomatology, China Medical University, Shenyang, Liaoning 110002;
Exp Ther Med. 2013 Jul;6(1):241-247. doi: 10.3892/etm.2013.1104. Epub 2013 May 8.
The aim of the present study was to investigate the effects of the surface characteristics of nanoporous titanium oxide films, formed by anodization on Ti-24Nb-4Zr-8Sn (Ti2448) alloy, on the early adhesion of osteoblast-like MG-63 cells. Nanoporous titanium oxide films with two different pore sizes (30 and 90 nm) were formed by anodization in NHF solution on Ti2448 alloy. The surface roughness of the nanoporous titanium oxide films was determined using a Surftest Formtracer and field emission scanning electron microscopy (FESEM). Cell viability was evaluated at different time points using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To investigate the regulatory mechanisms involved in the focal adhesion of osteoblasts to Ti2448 alloy, we quantified the expression levels of integrin β1 and paxillin mRNAs on the nanoporous titanium oxide films during early osteoblast adhesion using real-time RT-PCR. Samples with a 30-nm nanoporous film exhibited a greater number of overlapping microporous structures with microprojections compared with the 90-nm nanoporous film samples. The MTT assay indicated that cell viability on the 30-nm nanoporous surface following 24 and 48 h of cell culture was higher than those observed on the unanodized control and 90-nm nanoporous surfaces. Integrin β1 mRNA expression levels on the 30-nm nanoporous surface following cell culture for 48 h were also significantly higher compared with those on the unanodized control and 90-nm nanoporous surfaces. The results demonstrated that a 30-nm nanoporous titanium oxide film on Ti2448 alloy may provide the optimum bioactive implant surface for the initial adhesion of osteoblasts.
本研究的目的是调查通过阳极氧化在Ti-24Nb-4Zr-8Sn（Ti2448）合金上形成的纳米多孔二氧化钛薄膜的表面特性对成骨样MG-63细胞早期黏附的影响。在Ti2448合金上于NHF溶液中通过阳极氧化形成了具有两种不同孔径（30和90 nm）的纳米多孔二氧化钛薄膜。使用表面粗糙度仪和场发射扫描电子显微镜（FESEM）测定纳米多孔二氧化钛薄膜的表面粗糙度。使用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）法在不同时间点评估细胞活力。为了研究成骨细胞与Ti2448合金黏附斑形成过程中的调控机制，我们使用实时RT-PCR定量了早期成骨细胞黏附过程中纳米多孔二氧化钛薄膜上整合素β1和桩蛋白mRNA的表达水平。与90 nm纳米多孔薄膜样品相比，具有30 nm纳米多孔薄膜的样品表现出更多带有微突起的重叠微孔结构。MTT法表明，细胞培养24和48小时后，30 nm纳米多孔表面上的细胞活力高于未阳极氧化的对照表面和90 nm纳米多孔表面上的细胞活力。细胞培养48小时后，30 nm纳米多孔表面上的整合素β1 mRNA表达水平也显著高于未阳极氧化的对照表面和90 nm纳米多孔表面上的表达水平。结果表明，Ti2448合金上的30 nm纳米多孔二氧化钛薄膜可能为成骨细胞的初始黏附提供最佳的生物活性植入物表面。
