芒柄花素促进牙囊干细胞成骨向分化

Formononetin Motivates Osteogenic Differentiation of Dental Follicle Stem Cells

目的:研究芒柄花素对牙囊干细胞(human dental follicle stem cells,hDFSCs)成骨分化的影响。方法:首先对hDFSCs进行分离、培养及鉴定,并使用含不同浓度芒柄花素(0、0.1、1、10μmol/L)培养基培养细胞。通过CCK-8实验、结晶紫染色和活/死细胞荧光染色评估细胞的活性状况;通过细胞骨架荧光染色观察细胞骨架状况;采用RT-qPCR检测细胞成骨相关基因的表达变化;利用碱性磷酸酶(alkaline phosphatase,ALP)染色及活性检测法评估细胞成骨分化过程中ALP活性的影响;通过茜素红染色法评估细胞成骨分化过程中钙化结节数量的影响。结果:hDFSCs具有干细胞特性;不同浓度的芒柄花素对hDFSCs活性无显著性影响;1μmol/L的芒柄花素可显著促进Runx2、OCN、Col-Iα1的mRNA表达;并能够提高的细胞内ALP活性和钙化结节的数量。结论:1μmol/L芒柄花素可促进hDFSCs的成骨分化。

Objective:To elucidate the effect of formononetin on osteogenic differentiation of human dental follicle stem cells(hDFSCs).Methods:hDFSCs were isolated,cultured,and identified.The cells were treated with different concentrations of formononetin(0,0.1,1,and 10μmol/L)in the culture medium.Cell viability was assessed by CCK-8 assay,crystal violet staining,and live/dead cell fluorescence staining.The cell cytoskeleton was examined using fluorescent staining.The expression of osteogenic-related genes was analyzed through RT-qPCR.Alkaline phosphatase(ALP)activity was assessed to evaluate the osteogenic differentiation of cells.The influence of formononetin on calcium nodule formation during osteogenic differentiation was evaluated using alizarin red staining.Results:hDFSCs exhibited stem cell characteristics.Different concentrations of formononetin had no significant effect on hDFSCs viability.Whereas,1μmol/L formononetin facillitated the expression of Runx2,OCN,and Col-Iα1 mRNA.Additionally,1μmol/L formononetin enhanced the intracellular ALP activity and the number of calcium nodules.Conclusion:Formononetin at 1μmol/L promotes the osteogenic differentiation of hDFSCs.