釉基质蛋白对人牙囊和牙周膜细胞黏附增殖的影响

Influence of enamel matrix protein on attachment and proliferation ability of dental follicle cells and periodontal ligament cells cultured on titanium

目的 比较釉基质蛋白（EMP）对人牙囊细胞（hDFC）和人牙周膜细胞（hPDLC）在钛片表面黏附和增殖能力的影响。方法 分离培养hDFC和hPDLC,双喷砂加酸蚀处理钛片表面。实验分为4组：hDFC＋EMP诱导组、hDFC组、hPDLC＋EMP诱导组、hPDLC组。采用噻唑蓝（MTT）法和细胞免疫荧光染色法,定量分析细胞在钛片表面的黏附和增殖情况,扫描电子显微镜观察细胞在钛片表面1～7 d的生长情况。结果 1～7 d各组间MTT值的差异无统计学意义（P〉0.05）;加有EMP诱导的2组细胞较2组单纯细胞培养组的形状系数（Sf）值更高（P〈0.05）,hDFC组和hPDLC两组间Sf值差异无统计学意义（P〉0.05）,hDFC＋EMP诱导组较hPDLC＋EMP诱导组Sf值高（P〈0.05）。结论 EMP能促进hPDLC和hDFC在钛片表面的黏附,且对hDFC的促进作用更强;但对接种在钛片表面的hPDLC和hDFC短期增殖没有影响。

Objective The purpose of this study was to compare the adhesion and proliferation ability of human dental follicle cell（hDFC） and human periodontal ligament cell（hPDLC） cultured on titanium, and to investigate their changes of proliferation and growth when induced by enamel matrix protein（EMP）. Methods The hDFCs and hPDLCs were obtained from healthy human impacted third molars and non-impacted premolars extracted for orthodontic reasons, hDFCs and hPDLCs were isolated and cultured by explant technique. Pure titanium disks were prepared by sandblasted and acid-etched technologies. There were four groups in the study： hDFC cultured with EMP group, hDFC group, hPDLC cultured with EMP group, hPDLC group, quantitative analysis of cell proliferation and attachment by methyl thiazolyl tetrazolium（MTT） and fluorescence stain. The growth of cells cultured on titanium disk from day 1 to day 7 was observed by scanning electron microscope. Results MTT assay value showed no significant difference among each group from day 1 to day 7（P〉0.05）. Shape factor（S0 value showed a marked increase in groups induced by EMP than groups without induction（P〈0.05）, no significant difference among hDFC group and hPDLC group（P〉0.05）, and a marked increase in hDFC cultured with EMP group than hPDLC cultured with EMP group（P〈0.05）. Conclusion EMP has played a roal in promoting DSCs and PDLCs attachment, and the effect on hDFCs is more significant than on DFCs. However, EMP has no significant effect on proliferation of PDLCs and DFCs over a short time.