糖基化终末产物对人牙周膜干细胞骨向分化能力影响的研究  

Effects of advanced glycation end products on the osteogenic differentiation of human periodontal ligament stem cells

作　　者：邓超[1] 伍燕[1] 杨琨[1] 崔晓霞[1] 夏佳佳[1] 王岚[1] 刘琪[1] 金岩 

机构地区：[1]遵义医学院附属口腔医院,贵州遵义563003 [2]第四军医大学口腔医院组织工程中心,陕西西安710032

出　　处：《牙体牙髓牙周病学杂志》2012年第5期249-254,共6页Chinese Journal of Conservative Dentistry

基　　金：国家自然基金(30725042;81020108019);国家基础研究项目(973项目)(2010C13944800;2011CB964700);贵州省省长资金(C_397);贵州省科技厅自然资金(C_393);遵义市科技资金(E_063)

摘　　要：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-1、CD90,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 timePCR)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-1、CD90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、200μg/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7 d 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。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 ceils 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 AGEs. 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 induc- tion, Alizarin red staining showed varying degree of mineralization nodules, the expressions of osteogenic genes （ALP,Runx-2, Co1-1 ） 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.

关 键 词：糖基化终末产物 人牙周膜干细胞 骨向分化 

分 类 号：R780.2[医药卫生—口腔医学]