摘要
目的:观察糖基化终末产物(AGEs)对人牙周膜干细胞(hPDLSCs)及人颌骨骨髓间充质干细胞(hBMMSCs)骨向分化能力的影响。方法:组织块法和有限稀释法克隆化培养hPDLSCs及hBMMSCs,流式细胞仪检测两种细胞表型分子表达,取第3代细胞分别在10μg/mL AGEs刺激下进行矿化诱导,于诱导的21 d,茜素红染色、十六烷基吡啶定量观察钙结节形成情况、诱导7 d,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)和Western blot检测成骨相关基因及蛋白表达情况。结果:流式细胞仪显示两种细胞均阳性表达CD44、CD146、stro-1、CD90。成骨诱导21 d后茜素红染色和定量分析显示,hPDLSCs AGEs组矿化能力低于对照组;而hBMMSCs AGEs组矿化能力高于对照组(P<0.05)。成骨诱导7 d后,ALP染色显示,两种细胞ALP活性变化趋势与茜素红定量分析相同;RT-PCR检测hPDLSCs中ALP、Runx-2、Col-1、Runx-2mRNA表达水平均低于对照组,而hBMMSCs中各基因均高于对照组(P<0.05)。Western blot检测显示,各组Runx-2蛋白表达趋势与RT-PCR结果趋势相同。结论:AGEs抑制hPDLSC的骨向分化,促进hBMMSC骨向分化。
AIM: To investigate the effects of advanced glycation end products (AGEs) on the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) and human bone marrow mesenchymal stem cells (hBMMSCs). METHODS: PDLSCs and BMMSCs were cultured and cloned. Flow cytometery were employed to ex- amine the surface marker of the cells, hPDLSCs and hBMMSCs of passage3 were induced by 10 μg/mL AGEs. The os- teogenic differentiation capacity of the cells were evaluated by alizarin red staining, ALP staining and real-time RT - PCR. RESULTS: CD44, CD146, stro-1 and CI)90 were expressed in AGE-BSA induced hPDLSCs and hBMMSCs. After 21- day induction, the mineralization nodules were fewer in AGEs induced hPDLSCs than in the controls(P 〈 O. 05 ) , but were more in AGEs induced hBMMSCs than in the controls (P 〈 0.05). After 7-day induction ALP, Runx-2 Col-1 and Runx-2 mRNAs were lower in AGEs induced hPDLSCs than in the controls(P 〈 0.05), but were higher in hBMMSCs than in the controls(P 〈0.05). CONCLUSION:AGEs may inhibite the osteogenic differentiation of hPDLSCs and promote the osteogenic differentiation of hBMMSCs.
出处
《牙体牙髓牙周病学杂志》
CAS
北大核心
2013年第6期366-370,375,共6页
Chinese Journal of Conservative Dentistry
基金
国家自然基金(30725042
81020108019)
贵州省省长资金(C_397)
省科技厅自然资金(C_393)
遵义市科技资金资助项目(E_063)
关键词
糖基化终末产物
人牙周膜干细胞
人颌骨骨髓间充质干细胞
骨向分化
advanced glycation end products
human periodontal ligament stem cells
human bone marrow mes-enchymal stem cells
osteogenic differentiation