成骨生长肽对人牙周膜细胞增殖和碱性磷酸酶活性的影响

Osteoblastic growth peptide regulates proliferation of human periodontal ligament cells and alkaline phosphatase activities

背景:成骨生长肽是新近发现的一种生长因子,具有促进有丝分裂、促进成骨等作用,在组织损伤后的修复过程中发挥了重要的生理作用。目的:观察成骨生长肽对体外培养的人牙周膜细胞增殖及其碱性磷酸酶活性的影响。设计、时间及地点:细胞水平的对比观察实验,于2006-02/06在教育部口腔生物医学工程重点实验室完成。材料:临床上因正畸需要拔除的牙周健康、无龋的新鲜第1前磨牙牙周膜组织。成骨生长肽购于美国Sigma公司。方法:在体外培养的人牙周膜细胞中加入10-11,10-10,10-9,10-8及10-7mol/L不同浓度的成骨生长肽,用四甲基偶氮唑盐比色法检测细胞的增殖率;以流式细胞仪法检测细胞的增殖周期;应用酶联免疫法检测细胞内碱性磷酸酶活性。主要观察指标:细胞培养第1,2,3天观察细胞增殖情况;第6天观察细胞内碱性磷酸酶活性。结果:成骨生长肽浓度在10-11～10-7mol/L时,对人牙周膜细胞的增殖率有明显的促进作用(P<0.05),最佳作用浓度为10-9mol/L。当成骨生长肽浓度在10-9mol/L时,能明显提高细胞S期所占的比例,从而加速细胞的增殖活动。成骨生长肽在10-11～10-7mol/L时,能显著提高细胞内碱性磷酸酶活性(P<0.05),最佳作用浓度为10-9mol/L。结论:成骨生长肽可促进人牙周膜细胞的增殖活性,同时可以提高细胞碱性磷酸酶的活性。

BACKGROUND： Osteogenic growth peptide （OGP） is a newly discovered growth factor to promote mitosis and bone formation, it is of great significance on the repair process of tissue injury. OBJECTIVE： To elucidate the effect of OGP on proliferation of human periodontal ligament cells cultured in vitro and alkaline phosphatase activities. DESIGN, TIME AND SETTING： A randomized control observational experiment based on ceils was completed at the Key Laboratory for Oral Biomedical Engineering by the State Ministry of Education from February 2006 to June 2006. MATERIALS： Periodontal ligament tissues were harvested from periodontal health caries-free fresh first molar, requiring extraction because of clinical orthodontic needs. OGP was purchased from Sigma Company. METHODS： Human periodontal ligament cells cultured in vitro were added into medium containing various concentrations of OGP （10^-11, 10^-10, 10^-9, 10^-8 and 10^-7 mol/L）; Cell proliferation was determined using M-IT method; Cell cycle was assayed with flow cytometry; Alkaline phosphatase activity was examined with enzyme-linked immunosorbent assay. MAIN OUTCOME MEASURES： Cellular proliferation at the 1^st, 2^nd, 3^rd days of cell culture; Alkaline phosphatase activity at the 6th day of cell culture. RESULTS： All OGP concentrations used （10 11 10 7 mol/L） were effective to promote the proliferation of human periodontal ligament cells （P 〈 0.05）, and the maximum response was obtained with 10 9 mol/L, at which of OGP could notably increase the percentage of cells at S phase and thus accelerating cellular proliferation. OGP at various concentrations （10 11 10 7 mol/L） could significantly enhance alkaline phosphatase activity （P 〈 0.05） and the optimal concentration was 104 mol/L. CONCLUSION： OGP stimulates proliferation of human periodontal ligament cells and alkaline phosphatase activity.