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目的 :探讨烟草浸提液(smokeless tobacco,ST)对人牙龈成纤维细胞(human gingival fibroblasts,HGFs)在纯钛片上附着及增殖的影响及机制。方法:原代HGFs与不同浓度ST共同培养2 h和2、4、6、8 d;采用CCK-8法测定细胞黏附及增殖;采用双抗夹心酶联免疫吸附法检测不同时间培养上清液中纤维连接蛋白(fibronectin,FN)的含量。采用SPSS19.0软件包对数据进行统计学分析。结果:HGFs在钛片上的黏附率随ST浓度升高呈下降趋势,5.0 g/L和10 g/L的ST组黏附率分别为(34.316±7.725)%和(25.478±10.651)%,显著低于对照组(100%,P<0.01)。细胞在钛片上增殖至第2天和第4天,对照组显著大于2.5、5.0、10 g/L ST组(P<0.05);第6天和第8天时,对照组均显著大于0.625~10 g/L各ST组(P<0.05)。ST与HGFs培养到第8天,0.625、1.25、2.5 g/L的ST组FN浓度分别为(69.352±31.640)、(23.595±8.625)和(7.292±2.865)ng/m L,显著低于对照组(142.188±28.126)ng/m L(P<0.05)。结论 :ST能显著抑制HGFs在钛片上的黏附和增殖,其机制可能与细胞产生FN减少有关。
Objective: To investigate the effect and mechanism of tobaccoless tobacco (ST) on attachment and proliferation of human gingival fibroblasts (HGFs) on pure titanium plate. Methods: Primary HGFs were incubated with different concentrations of ST for 2 h, 2, 4, 6 and 8 d. Cell adhesion and proliferation were measured by CCK-8 assay. The supernatants of culture supernatants were detected by double-antibody sandwich enzyme-linked immunosorbent assay The content of fibronectin (FN). SPSS19.0 software package for statistical analysis of the data. Results: The adhesion rate of HGFs on titanium plate decreased with the increase of ST concentration. The adherence rates of HGFs in ST group at 5.0 g / L and 10 g / L were (34.316 ± 7.725)% and (25.478 ± 10.651)%, respectively, In the control group (100%, P <0.01). Cells were proliferated on day 2 and day 4 in the control group, and were significantly larger than those in the control group at 2.5, 5.0 and 10 g / L ST (P <0.05). On day 6 and day 8, the control group was significantly larger than 0.625 ~ 10 g / L in each ST group (P <0.05). ST and HGFs were cultured on the 8th day. The concentrations of FN in ST group were (69.352 ± 31.640), (23.595 ± 8.625) and (7.292 ± 2.865) ng / m L at 0.625, 1.25 and 2.5 g / L, Group (142.188 ± 28.126) ng / m L (P <0.05). CONCLUSION: ST can significantly inhibit the adhesion and proliferation of HGFs on titanium plate, which may be related to the decrease of FN in cells.