烟草烟雾提取物对人牙龈成纤维细胞在3Y-TZP表面生长状态的影响

目的：观察比较不同浓度烟草烟雾提取物（Cigarette Smoke Extract,CSE）对人牙龈成纤维细胞（HumanGingival Fibroblasts,HGFs）在3mol%氧化钇作为稳定剂的四方氧化锆多晶体陶瓷（3Y-TZP）种植材料表面粘附、增殖、形态伸展及细胞状态的影响。方法：原代培养HGFs并进行细胞来源鉴定;SRB法检测不同浓度CSE作用下HGFs在材料表面粘附、增殖情况;免疫荧光染色,病理图像分析系统计数评价HGFs细胞铺展面积及形状系数改变;激光共聚焦显微镜下观察拍照,观察凋亡形态;扫描电镜观察细胞粘附的形态结构改变。结果：细胞波形丝蛋白染色阳性,角蛋白染色阴性;随CSE浓度升高,HGFs在材料表面粘附、增殖、铺展面积、形状系数及形态伸展均呈下降趋势,各实验组与对照组间均存在显著差异（P〈0.05）,抑制作用随浓度升高更加明显,呈浓度依赖性。结论：CSE对HGFs在3Y-TZP种植材料表面的生长可产生明显抑制作用。提示吸烟对陶瓷种植体植入后的软组织界面产生不良影响,其损害程度可能与烟草有害物质的接触量有相关性。

Effects of Cigarette Smoke Extract on the Growth State of HGFs on the Surfaces of 3Y-TZP

Objective： To observe the effects of the cigarette smoke extract（CSE）on the attachment,proliferation,spreading shape and growth state of human gingival fibroblasts（HGFs） to 3Y-TZP in vitro.Methods： HGFs were obtained from human normal gingival tissues and the origin of cells were identified by immunochemistry of vimentin and cytokeratin.HGFs to 3Y-TZP were cultured in the presence of CSE at various concentrations.The attachment and growth of cells attached to 3Y-TZP were measured by SRB method.Immunofluorescence was used to detect and analyze the shapes of HGFs attached to 3Y-TZP.Apoptosis morphological observation were observed under LSCM,and structure changes was showed by SEM.Results： Immunochemical study showed that vimentin was expressed in HGFs while cytokeratin was negative.With the increasing of CSE concentration,the attachment,proliferation,and spreading shape of HGFs in all groups decreased in a concentration-dependent manner.The difference between CSE groups and control group was statistically significant（P0.05）.Conclusion： CSE can inhibit the attachment,proliferation,and spreading shape of HGFs on the surface of 3Y-TZP,suggesting that smoking may have influence on the result of oral implant operation and the damage may be associated with the amount of tobacco hazardous substances.