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目的:通过体外实验,研究喷砂酸蚀钛表面对成骨细胞分化行为的影响,并对喷砂酸蚀钛表面影响成骨细胞分化的信号通路进行研究,探讨粗糙钛表面调控成骨细胞分化的作用机制。方法:采用扫描电子显微镜(SEM)对光滑钛表面、普通喷砂酸蚀钛表面进行表征。将成骨细胞接种到各样本表面,采用实时荧光定量PCR检测成骨细胞在不同材料表面骨功能基因表达的差异。采用Western免疫印迹法研究普通喷砂酸蚀钛表面对成骨细胞内ERK1/2蛋白磷酸化水平的影响。采用SAS9.0软件包对数据进行统计学分析。结果:SEM结果显示,光滑钛表面的粗糙度只有0.2μm,普通喷砂酸蚀钛表面有不规则孔隙,其平均粗糙度约为3.2μm。与光滑钛表面相比,普通喷砂酸蚀钛表面能通过促进骨功能基因的表达显著促进成骨细胞分化(P<0.05)。普通喷砂酸蚀钛表面加入ERK1/2通路特异性抑制剂(PD98095),成骨细胞骨功能基因表达显著增强。Western印迹结果显示,与光滑钛表面相比,普通喷砂酸蚀钛表面组成骨细胞内ERK1/2蛋白的磷酸化水平降低。加入PD98095的光滑钛表面组和未加入该抑制剂的喷砂酸蚀钛表面组ERK1/2蛋白磷酸化水平均降低,但后者不如前者效应显著。结论:表面粗糙度是影响成骨细胞生物学行为的重要因素,粗糙钛表面能显著促进成骨细胞分化,并且其促进成骨细胞分化与抑制ERK1/2蛋白磷酸化有关。
OBJECTIVE: To study the effect of sandblast-etched titanium surface on the differentiation of osteoblasts in vitro and to study the signal pathways that affect the differentiation of osteoblasts on the surface of sand-blasted acid-etched titanium, and to investigate the effect of rough titanium surface on the differentiation of osteoblasts The mechanism of action. Methods: Scanning electron microscope (SEM) was used to characterize the surfaces of smooth titanium and ordinary sandblasted titanium. Osteoblasts were seeded on the surface of each sample, and real-time fluorescent quantitative PCR was used to detect the difference of osteoblast gene expression between different materials. Western blotting was used to study the effect of common sandblasting titanate on the phosphorylation of ERK1 / 2 in osteoblasts. SAS9.0 software package for statistical analysis of the data. Results: The SEM results showed that the roughness of the smooth titanium surface was only 0.2μm, the surface of the ordinary sandblasting titanium had irregular pores and the average roughness was about 3.2μm. Compared with smooth titanium surface, ordinary sandblasting titania surface can significantly promote osteoblast differentiation by promoting the expression of osteogenic genes (P <0.05). Ordinary sandblasting titanium surface by adding ERK1 / 2 pathway specific inhibitor (PD98095), osteoblast function gene expression was significantly enhanced. Western blot results showed that compared with the smooth titanium surface, the level of phosphorylation of ERK1 / 2 protein in osteoblasts decreased with the conventional sandblasting surface. The phosphorylation level of ERK1 / 2 in the surface of smooth titanium group added with PD98095 and the surface of sandblast coated titanium without the inhibitor decreased, but the latter was not as effective as the former. Conclusion: The surface roughness is an important factor affecting the biological behavior of osteoblasts. The rough titanium surface can significantly promote the differentiation of osteoblasts, and promote the differentiation of osteoblasts and inhibit the phosphorylation of ERK1 / 2 protein.