釉基质蛋白对人牙龈上皮细胞增殖的影响

Effects of enamel matrix proteins on the proliferation of human gingival epithelial cells in vitro

目的:研究釉基质蛋白(enamelmatrixproteins,EMPs)对体外培养的人牙龈上皮细胞增殖活性的影响。方法:乙酸提取法获取EMPs,取龈切术中切除的牙龈组织,采用酶消化法培养牙龈上皮细胞。将第1代牙龈上皮细胞按照22500个/孔接种,分为4组:对照组(不加EMPs)及EMPs分别为50、100、200μg/ml的实验组。采用MTT法检测各组细胞数量,对每个时间点的各组实验数据进行方差分析。结果:人牙龈上皮细胞能在EMPs覆盖的平皿上生长。统计学分析表明,不同浓度的EMPs在早期对牙龈上皮细胞的增殖无显著影响;从第3天开始,当培养液中EMPs浓度为200μg/ml时,牙龈上皮细胞增殖显著受抑。结论:EMPs影响牙龈上皮细胞的增殖,并存在剂量和时间依赖性。

PURPOSE： To evaluate the effect of enamel matrix proteins （EMPs） on the proliferation of human gingival epithelial cells in vitro. METHODS： EMPs were extracted from pig tooth germ by acetic acid . The gingival tissues cut off during gingivectomy were separated into two parts through Dispase II digestion and the epithelium part was cultured to acquire the gingival epithelial cells. The first passage epithelial cells were inoculated into 96 -well plate ,22500 cells per well, and exposed to different concentrations of EMPs （50,100,200μg/ml respectively）. The control was epithelial cells cultured in the same medium except without EMPs. The proliferation rates were carried out over a 5-day period and assessed by an MTT assay and the data were analysed by one-way variance. RESULTS： It was shown that gingival epithelial cells well attached and spread on EMPs-coated substrata. There were no significant differences between the control group and various concentrations of EMPs groups at the initial stage,however, EMPs at a concentration of 200μg/ml significantly inhibited gingival epithelial cells proliferation from day 3 over the experiment. CONCLUSIONS： The proliferation of gingival epithelial cells was significantly inhibited by EMPs in a dose- and time- dependent manner, which provides some evidence for the mechanism of EMPs in promoting the periodontal tissue regeneration. Supported by National 863 Project （2002AA205011） and Shanghai Municipal Education Development Foundation （200205）.