AIM: To investigate the effect of advanced glycation end products (AGEs) on the osteogenic differentiation of human periodontal ligament stem cells (HPDLSCs). METHODS: PDLSCs were isolated by limited dilution of culture cells for single cell clone. Flow cytometry was employed to study the surface marker of the cloned cells. Passage 3 PDLSCs were induced with different concentrations of ACEs. The osteogenic differentiation capacity of hPDLSCs was evaluated by Alizarin red staining, ALP and real-time quantitative reverse transcription polymerase chain reaction (realtion RT-PCR). RESULTS: Expression of cell surface molecules CD44, CD146, stor-1 and CD90 of AGEs - induced group was significantly higher than that of the control group by flow cytometry. After 21 -day induction, Alizarin red staining showed varying degree of mineralization nodules, the expressions of osteogenic genes (ALP,rn Runx-2, Col-1) of unstimulated group were significantly higher than those of AGEs-PDLSCs group by real time PCR after 7-day induction (P < 0.05). CONCLUSION: AGEs reduce the osteogenic differentiation capacity of PDLSCs in a concentration-dependent manner.%目的:探讨糖基化终末产物(AGEs-HSA)对人牙周膜干细胞(HPDLSC)骨向分化能力的影响.方法:体外组织块法和有限稀释法克隆化培养牙周膜干细胞,流式细胞仪检测牙周膜细胞表型分子CD44、CD146、stor-l、CD9O,对其进行干细胞鉴定,将培养出的牙周膜干细胞与不同浓度的AGE-HSA共同培养,并矿化诱导后茜素红染色观察钙结节形成情况、碱性磷酸酶染色观察ALP活性、实时定量聚合酶链反应(real time PCR)检测成骨基因表达情况.结果:流式细胞仪细胞表型分析CD44、CD146、stor-l、CDg0表达呈阳性.成骨诱导21 d后茜素红染色,对照组和实验组均出现不同程度的矿化结节,定量分析显示1、10、100、200 μg/mLAGEs组矿化能力均比对照组低,差异均有统计学意义(P＜0.05)；不同浓度的实验组之间矿化能力均有统计学差异(P＜0.05),且随着AGEs浓度的升高,矿化能力逐渐降低.成骨诱导7d ALP染色比较发现各实验组均比对照组减弱.成骨诱导1周后RT-PCR检测各实验组的成骨基因ALP、Runx-2、Col-1、Runx-2 mRNA表达水平均较对照组低,差异有统计学意义(P＜0.05)；不同浓度的实验组之间各成骨基因的表达也有统计学差异(P＜0.05).结论:AGEs能抑制HPDLSC的骨向分化,并在一定范围内呈浓度依赖性.
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