牙骨质附着蛋白对牙周膜细胞黏附、伸展及增殖的影响

Effects of cementum attachment proteins on attachment, spreading and proliferation of human periodontal ligament cells

目的观察牙骨质附着蛋白（CAP）对体外培养的人牙周膜细胞（hPDLCs）黏附、伸展和增殖活性的影响。方法组织块培养法获得人hPDLCs。培养液中分别加入不同浓度的CAP（0、0.125、0.25、0.5、1.0、2.0μg/mL）。细胞计数法检测不同浓度CAP对hPDLCs黏附的影响;同时通过计数检测视野中伸展的细胞数,计算hPDLCs在培养1、4、7h后的伸展率;MTT比色法测定、比较不同浓度CAP处理组hPDLCs细胞的增殖活性。结果0.5、1.0μg/mLCAP处理组hPDLCs的黏附力明显强于空白对照组（CAP浓度为0）（P〈0.01）;不同浓度CAP处理组与空白对照组对细胞伸展影响的比较无明显差异。CAP对hPDLCs的促增殖作用呈浓度和时间依赖性,0.5-2.0μg/mL的CAP均能显著促进hPDLCs的增殖（P〈0.05）。结论CAP对体外培养的hPDLCs具有促黏附及促增殖作用,但对其细胞伸展无显著影响。

Objective To investigate the effects of cementum attachment protein （CAP） on the attachment, spreading and proliferation of cultured human periodontal ligament cells （hPDLCs） in vitro. Methods hPDLCs were obtained from periodontal ligament explants and cultured. Various concentrations of CAP （0, 0. 125, 0.25, 0.5, 1.0 and 2.0 μg/mL） was added to the culture fluid. The effects of CAP on the attachment ability of hPDLCs were detected by cytometry. Cell spreading rates were calculated after culturing for 1, 4 and 7 h by analysis of micrographs taken at predetermined sites of each wells. Cell proliferation rates were assessed by MTT assay and comparisons were made among groups with different concentrations of CAP. Results The attachment of hPDLCs was significantly stimulated by CAP at concentrations of 0.5 and 1.0 μg/mL than the blank control group （no CAP, P 〈 0.01 ）. However, there was no significant change in cell spreading rate between the control group and experimental groups. The proliferation of hPDLCs was significantly stimulated by CAP in a dose-and tlme-dependent manner, and CAP at a concentration of 0.5 to 2.0 μg/mL significantly enhanced the proliferation of hPDLCs （P 〈 0.05）. Conclusion CAP can promote the attachment and proliferation ability of human hPDLCs cultured in vitro, but have no marked effects on their spreading.