Cecchinato Francesca, Xue Ying, Karlsson Johan, He Wenxiao, Wennerberg Ann, Mustafa Kamal, Andersson Martin, Jimbo Ryo
Department of Prosthodontics, Faculty of Odontology, Malmö University, Malmö, Sweden.
J Biomed Mater Res A. 2014 Nov;102(11):3862-71. doi: 10.1002/jbm.a.35062. Epub 2013 Dec 20.
This work aimed to evaluate the in vitro response of Transfected Human Foetal Osteoblast (hFOB) cultured on a magnesium-loaded mesoporous TiO2 coating. The application of mesoporous films on titanium implant surfaces has shown very promising potential to enhance osseointegration. This type of coating has the ability to act as a framework to sustain bioactive agents and different drugs. Magnesium is the element that, after calcium, is the most frequently used to dope titanium implant surfaces, since it is crucial for protein formation, growth factor expression, and aids for bone mineral deposition on implant surfaces. Mesoporous TiO2 films with an average pore-size of 6 nm were produced by the evaporation-induced self-assembly method (EISA) and deposited onto titanium discs. Magnesium loading was performed by soaking the mesoporous TiO2 discs in a magnesium chloride solution. Surface characterization was conducted by SEM, XPS, optical interferometry, and AFM. Magnesium release profile was assessed at different time points using a Magnesium Detection kit. Cell morphology and spreading were observed with SEM. The cytoskeletal organization was stained with TRITC-conjugated Phalloidin and cell viability was evaluated through a mitochondrial colorimetric (MTT) assay. Furthermore, gene expression of bone markers and cell mineralization were analyzed by real time RT-PCR and alizarin-red staining, respectively. The surface chemical analysis by XPS revealed the successful adsorption of magnesium to the mesoporous coating. The AFM measurements revealed the presence of a nanostructured surface roughness. Osteoblasts viability and adhesion as well as the gene expression were unaffected by the addition of magnesium possibly due to its rapid burst release, however, were enhanced by the 3D nanostructure of the TiO2 layer.
这项工作旨在评估转染的人胎儿成骨细胞（hFOB）在负载镁的介孔TiO₂涂层上的体外反应。介孔膜在钛植入物表面的应用已显示出增强骨整合的非常有前景的潜力。这种类型的涂层有能力作为维持生物活性剂和不同药物的框架。镁是继钙之后最常用于掺杂钛植入物表面的元素，因为它对蛋白质形成、生长因子表达至关重要，并有助于骨矿物质在植入物表面沉积。通过蒸发诱导自组装法（EISA）制备了平均孔径为6nm的介孔TiO₂膜，并将其沉积在钛盘上。通过将介孔TiO₂盘浸泡在氯化镁溶液中来进行镁负载。通过扫描电子显微镜（SEM）、X射线光电子能谱（XPS）、光学干涉测量法和原子力显微镜（AFM）进行表面表征。使用镁检测试剂盒在不同时间点评估镁释放曲线。用SEM观察细胞形态和铺展情况。用TRITC偶联的鬼笔环肽对细胞骨架组织进行染色，并通过线粒体比色法（MTT）测定评估细胞活力。此外，分别通过实时逆转录聚合酶链反应（RT-PCR）和茜素红染色分析骨标志物的基因表达和细胞矿化情况。XPS进行的表面化学分析表明镁成功吸附到介孔涂层上。AFM测量揭示了纳米结构表面粗糙度的存在。成骨细胞的活力和黏附以及基因表达不受镁添加的影响，这可能是由于其快速的突发释放，然而，TiO₂层的三维纳米结构增强了这些指标。
