不同基台材料对人牙龈上皮细胞半桥粒黏附相关基因及蛋白表达的影响
Effect of different abutment materials on the expression of genes and proteins related to hemidesmosome adhesion in human gingival epithelial cells
摘要目的:探讨聚醚醚酮、氧化锆、纯钛基台材料对人牙龈上皮细胞半桥粒黏附相关基因及蛋白表达的影响,以期筛选出更适合上皮附着的基台材料。方法:制备聚醚醚酮、氧化锆、纯钛试件各48枚,用扫描电镜观察各组表面形貌,用白光干涉仪测量表面粗糙度,光学接触角测量仪测量接触角。采用扫描电镜观察人牙龈上皮细胞在各组试件表面的早期黏附状态,并使用细胞计数试剂盒评估各组人牙龈上皮细胞的增殖能力,实时荧光定量PCR和蛋白质印迹法分别检测各组人牙龈上皮细胞半桥粒黏附相关基因及蛋白的表达水平。结果:3组试件均呈平坦、光滑的表面形貌;聚醚醚酮组、氧化锆组和纯钛组平均粗糙度(Ra值)分别为(95.63±2.06)、(37.93±3.56)、(134.2±4.62)nm( F=368.16, P<0.001),平均最大高度(Rz值)分别为(2.42±0.22)、(0.87±0.10)、(3.77±0.28)nm( F=91.95, P<0.001),组间差异均有统计学意义( P<0.05);接触角总体差异无统计学意义( F=0.45, P=0.658)。人牙龈上皮细胞在3组试件表面均呈扁平伸展的多边形、多角形等不规则形状,表现出典型的铺路石样。培养1和3 d时3组细胞增殖能力( A值)差异均无统计学意义( P>0.05);培养5和7 d时聚醚醚酮组细胞增殖能力( A值)均显著大于氧化锆组和纯钛组( P<0.05)。培养3和7 d时聚醚醚酮组层粘连蛋白α3、整合素β4和胶原蛋白17的mRNA和蛋白表达水平均显著大于同时间点氧化锆组和纯钛组( P<0.05)。 结论:聚醚醚酮相较氧化锆、纯钛基台材料更有利于人牙龈上皮细胞半桥粒的黏附。
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abstractsObjective:To investigate the effects of polyetheretherketone, zirconium dioxide, and titanium abutment materials on the expression of genes and proteins related to hemidesmosome adhesion in human gingival epithelial cells, in order to screen out abutment materials that are easier for epithelial adhesion.Methods:Forty-eight specimens were prepared in each of the three materials, polyetheretherketone, zirconium oxide, and pure titanium specimens. The surface morphology of each group of specimens was observed by scanning electron microscopy, the surface roughness was measured by the white light interferometer, and the contact angle was measured by optical contact angle measuring instrument. The early adhesion status of human gingival epithelial cells on the surface of each group of specimens was observed by scanning electron microscopy, and the proliferation ability of human gingival epithelial cells on the surface of each group of specimens was assessed by using a cell counting kit, and the expression levels of genes and proteins related to the adhesion of human gingival epithelial cells on the surface of each group of specimens were detected by real-time fluorescence quantitative PCR and Western blotting, respectively.Results:The surface morphology of the three groups of specimens was flat and smooth. The mean roughness (Ra value) of the polyetheretherketone, zirconia, and pure titanium groups were (95.63±2.06), (37.93±3.56), and (134.2±4.62) nm ( F=368.16, P<0.001), respectively, and the mean maximum height (Rz value) was (2.42±0.22), (0.87±0.10) and (3.77±0.28) nm ( F=91.95, P<0.001), with statistical significance ( P<0.05). The contact angle results showed no statistical significance between groups ( F=0.45, P=0.658). Human gingival epithelial cells showed irregular shapes such as flattened and extended polygons and polygons on the surface of the three groups of specimens, exhibiting a typical paving stone pattern. The differences in cell proliferation among the three groups at 1 and 3 d of culture were not statistically significant ( P>0.05). Cell proliferation in the polyetheretherketone group was significantly greater those those in the zirconia and pure titanium groups at 5 and 7 d of culture ( P<0.05). The mRNA expression levels and protein expression levels of laminin α3, integrin β4, and collagen ⅩⅦ in the polyetheretheretherketone group at 3 and 7 d of incubation were significantly greater than those in the zirconium oxide and pure titanium groups at the same time points ( P<0.05). Conclusions:Polyetheretherketone is more conducive to the adhesion of hemidesmosome in human gingival epithelial cells than zirconium dioxide and pure titanium abutment materials.
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