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不同糖浓度对颌骨骨髓间充质干细胞成骨分化的影响
Effects of Different Concentrations of Glucose on the Osteogenic Differentiation of Orofacial Bone Mesenchymal Stem Cells
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摘要: 目的 研究不同糖浓度对颌骨骨髓间充质干细胞（orofacial bonemesenchymal stem cells, OFMSCs）增殖和成骨分化的影响。方法 体外分离、培养OFMSCs，成骨、成脂、成软骨分化诱导及鉴定，并使用不同含糖量培养基（5.5、11、16.5、25、44 mmol/L）培养OFMSCs,以5.5 mmol/L基准糖浓度为对照组，其余组为实验组。采用CCK-8及流式细胞仪检测各组OFMSCs的增殖活性及增殖指数，OFMSCs成骨诱导后4、7 d检测碱性磷酸酶（ALP）活性，21 d进行茜素红染色及矿化定量分析，同时用RT-PCR检测3、7、14及21 d相关成骨基因Runx2、Osterix的表达。结果 培养的OFMSCs成骨诱导21 d后茜素红染色可见钙结节，成脂诱导14 d 后油红O染色可见红色脂滴，成软骨诱导14 d 后阿利新蓝染色可见蓝色胞浆；糖浓度在5.5～25 mmol/L促进OFMSCs的增殖，但随着糖浓度的继续增加（25～44 mmol/L），OFMSCs的增殖活性受抑制；成骨诱导时，随着培养液糖浓度升高，ALP活性呈剂量依赖性降低（P Runx2、Osterix mRNA 表达量高于实验组（P Runx2、Osterix mRNA 表达量均出现先上调再下调的趋势。结论 在一定范围的糖浓度内，糖浓度升高可促进OFMSCs的增殖；而糖浓度升高对成骨分化呈抑制效应。Abstract: Objective To determine the effects of different concentrations of glucose on the proliferation and osteogenic differentiation of orofacial bonemesenchymal stem cells (OFMSCs). Methods OFMSCs were primarily cultured and identified in vitro to undergo osteogenic/adipogenic/chondrogenic differentiation. The cells were exposed to osteogenic medium containing different levels of glucose: 5.5,11,16.5,25,44 mmol/L. The cell activity and proliferation index were measured using a cell counting kit (CCK)-8 and flow cytometry. The alkaline phosphatase (ALP) activity of the cells was measured at the 4th and 7th day. Alizarin red staining was carried out at the 21st day. RT-PCR detecting osteogenesis-related gene Runx2 and Osterix mRNA expression was performed at the 4th, 7th, 14th and 21st day. Results Osteogenesis induced calcium nodes was observed with Alizarin red staining at the 21st day. Adipogenic induced red lipid droplets was observed with Oil Red O staining at the 14th day. Chondrogenic induced blue cytoplasm was observed with Alcian blue staining at the 14th day. With 5.5 to 25 mmol/L glucose,OFMSCs proliferation was promoted.But when the concentration of glucose continued to increase (from 25 to 44 mmol/L),OFMSCs proliferation significantly reduced.The ALP activity decreased with glucose in a concentration-dependent manner (P st day. The calcium nodes and mineralization quantity decreased with glucose in a concentration-dependent manner (P Runx2 and Osterix mRNA than the others (P Runx2 and Osterix mRNA in all groups showed a trend of rising first, followed by an obvious down regulation. Conclusion With certain limits,OFMSC proliferation is promoted by glucose. Osteogenic differentiation is inhibited by glucose in a concentration-dependent manner.
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期刊类型引用(6)
1. 范永晶，王姝，金武龙. 颌骨来源骨髓间充质干细胞成骨分化的特点、优势与应用. 中国组织工程研究. 2024(01): 100-106 . 
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2. 王勤英，孟茂花，陈新，徐星星，陈镜桥，董强. 高糖环境下miRNA对BMSCs成骨分化的调控机制. 中国老年学杂志. 2023(08): 2006-2012 . 百度学术
3. 杨姣姣，胡明，李岩，夏德林. 不同孔径梯度纳米羟基磷灰石三维支架制备及对MC3T3-E1生物学性能的影响. 中国组织工程研究. 2022(21): 3338-3344 . 百度学术
4. 陆玖青，吕佳姝，谢亚佳，甄蕾. 糖尿病大鼠颌骨结构及颌骨骨髓间充质干细胞成骨分化的研究. 口腔颌面外科杂志. 2020(03): 144-149 . 百度学术
5. 唐宇，胡赟，罗丹，丁晓倩，李彩玉，郑雷蕾. 糖尿病环境下成骨细胞KDM6B表达的变化. 四川大学学报(医学版). 2019(05): 660-665 . 百度学术
6. 刘艳，胡赟，张夏，丁俐丹，唐世俊，杨兰，陈杨，郑雷蕾. 双磷酸盐对高糖环境下BMSCs自噬相关因子Beclin1和LC3Ⅱ影响的研究. 四川大学学报(医学版). 2018(05): 694-699 . 百度学术
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