Sola-Ruiz M Fernanda, Perez-Martinez Carolina, Labaig-Rueda Carlos, Carda Carmen, Martín De Llano J Javier
Department of Stomatology, Faculty of Medicine and Dentistry, University of Valencia, 46010 Valencia, Spain.
Department of Pathology and Health Research Institute of the Hospital Clínico (INCLIVA), Faculty of Medicine and Dentistry, University of Valencia, 46010 Valencia, Spain.
Int J Mol Sci. 2017 Apr 13;18(4):823. doi: 10.3390/ijms18040823.
The aim of this work was to observe the behavior of osteoblast cells cultured in vitro on titanium discs in relation to disc surface roughness and the addition of melatonin to the culture medium. MG63 osteoblast cells were cultivated on 120 Grade 5 Ti divided into three groups: Group E, treated with dual acid etch; Group EP, treated with dual acid etch and calcium phosphate; and Group M, machined. Surface roughness was examined under a laser scanning confocal microscope (CLSM) and scanning electron microscopy (SEM). The proliferation and morphology of cells were determined under fluorescence microscopy and SEM. Messenger ribonucleic acid (mRNA) of different genes related to osteoblastic differentiation was quantified by means of real-time quantitative polymerase chain reaction (RT-PCR) assay. The greatest surface roughness was found in Group EP (Ra 0.354 µm), followed by Group E (Ra 0.266 µm), and Group M (Ra 0.131 µm), with statistically significant differences between the groups ( < 0.001). In the presence of melatonin a trend to a higher cell proliferation was observed in all groups although significant differences were only found in Group M ( = 0.0079). Among the genes studied, a significant increase in phosphate-regulating neutral endopeptidase, X-linked (PHEX) expression was observed in cells cultured on EP discs. The addition of melatonin increased osteoblast cell proliferation and differentiation, and may favor the osseointegration of dental implants.
本研究的目的是观察体外培养于钛盘上的成骨细胞行为与盘表面粗糙度以及向培养基中添加褪黑素之间的关系。将MG63成骨细胞培养在分为三组的120级5钛上:E组,用双酸蚀刻处理;EP组,用双酸蚀刻和磷酸钙处理;M组,机械加工。在激光扫描共聚焦显微镜(CLSM)和扫描电子显微镜(SEM)下检查表面粗糙度。在荧光显微镜和SEM下测定细胞的增殖和形态。通过实时定量聚合酶链反应(RT-PCR)测定法定量与成骨细胞分化相关的不同基因的信使核糖核酸(mRNA)。在EP组中发现最大表面粗糙度(Ra 0.354 µm),其次是E组(Ra 0.266 µm)和M组(Ra 0.131 µm),各组之间存在统计学显著差异(<0.001)。在褪黑素存在的情况下,所有组均观察到细胞增殖有升高趋势,尽管仅在M组中发现显著差异(=0.0079)。在所研究的基因中,在EP盘上培养的细胞中观察到磷酸调节中性内肽酶X连锁(PHEX)表达显著增加。褪黑素的添加增加了成骨细胞的增殖和分化,并可能有利于牙种植体的骨整合。