高浓度葡萄糖对牙周膜干细胞成骨分化能力的影响

Effects of High Glucose on Proliferation and Osteogenic Differentiation of Periodontal Ligament Stem Cells in Vitro

目的观察高浓度葡萄糖刺激下牙周膜干细胞(PDLSCs)的增殖能力和成骨分化能力。方法组织块法体外培养非糖尿病患者PDLSCs,分别用0mg/L(正常对照组)、1100mg/L(低糖组)、4500mg/L(高糖组)浓度葡萄糖刺激,MTT法检测PDLSCs增殖能力,矿化诱导后茜素红染色观察矿化结节形成,实时定量聚合酶链反应(RT-PCR)检测Runt相关转录因子2(Runx-2)、碱性磷酸酶(ALP)和I型胶原(Col-1)成骨相关基因表达。结果 MTT法检测显示高糖组OD值较低糖组和正常对照组低(P<0.05);21天成骨诱导后,高糖组矿化结节面积少于低糖组和正常对照组(P<0.05,P<0.01);成骨诱导期间,高糖组ALP、Runx2和Col-1诱导前后相对倍增数低于低糖组和正常对照组(P<0.05,P<0.01)。结论高浓度葡萄糖抑制PDLSCs增殖能力和成骨分化能力。

Objective To investigate the effect of high glucose concentration on the cellular activity and os- teogenic differentiation of periodontal ligament stem cells （PDLSCs） in vitro. Methods PDLSCs from non-diabetic patients were obtained and cuhured with tissue block method in a low glucose-concentration medium （1100mg/L of glucose） or in a high glucose-concentration medium（4500mg/L of glucose）, respectively, for 14d. Zero mg/L of glu- cose group served as blank control. MTT assay was used to evaluate cellular viability. The osteogenic differentiation capacity of PDLSCs was determined by alizarin red staining after mineralization induction and real time PCR for detection of Runx-2, alkaline phosphatase （ALP）, and collagen I genes. Results MTF assay results showed that the OD value in high glucose group was lower than that in low glucose group （P〈0.05,P〈0.01）. After 21-day cul- turing, the calcified area was reduced and the expression of Runx-2, ALP, and collagen I genes to the toal culture dish of PDLSCs in high glucose group were lower than those in low glucose group （P〈0.05,P〈0.01）. Conclusion High glucose inhibits the proliferation and differentiation of PDLSCs.