高浓度葡萄糖对人牙周膜成纤维细胞成骨分化能力的影响

摘要

目的：  研究高葡萄糖浓度刺激下牙周膜成纤维细胞（PDLC）的增殖能力和成骨分化能力。方法   体外组织块法培养非糖尿病患者PDLC，分别用0 mg·L-1（C组）、1 100 mg·L-1（L组）、4 500 mg·L-1（H组）浓度葡萄糖刺激PDLC，噻唑蓝（MTT）法检测PDLC增殖能力，矿化诱导后茜素红染色观察矿化结节的形成情况，碱性磷酸酶（ALP）活性检测和实时定量聚合酶链反应（RT-PCR）检测成骨相关基因表达。结果   MTT检测结果显示，H组OD值较L组和C组低（P<0.05）；成骨诱导21 d后，H组矿化结节面积较L组和C组少（P<0.05）；成骨诱导期间， H组ALP活性较L组和C组明显偏低（P<0.05）；H组早期成骨基因Runt相关转录因子2、ALP和Ⅰ型胶原诱导前后相对倍增数较L组和C组低（P<0.05）。结论   高浓度葡萄糖能够抑制PDLC增殖能力和成骨分化能力。%Objective To isolate the periodontal ligament cells(PDLCs) and study the effect of high glucose concentration levels on the cellular activity and osteogenic differentiation of PDLCs from non-diabetic patients in vitro. Methods PDLCs were cultured for 14 days in 0 mg·L-1 glucose medium(C group), 1 100 mg·L-1 glucose medium(L group), and 4 500 mg·L-1 glucose medium(H group). Methyl thiazolyl tetrazolium(MTT) assay for cellular viability was performed. The osteogenic differentiation capacity of PDLCs was evaluated by alizarin red staining and alkaline phosphatase(ALP) activity assaying and real time-transcription polymerase chain reaction. Results H group significantly inhibited the proliferation of PDLCs and reduced the optic density of the MTT and ALP activity assay. The difference was significant(P<0.05). Concerning the mineralized nodule formation, the percentage of the calcified area and the expression of osteogenic genes(ALP, Runx-2, Col-Ⅰ) to the total culture dish of PDLCs in the H group were significantly lower than that in the L and C groups(P<0.05). Conclusion High glucose inhibits the proliferation and differentiation of PDLCs.