Lee Heera, Kim Ju-Hwan, Lee Hyun-Jin, Park Jun-Beom
Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea.
Department of Medicine, Graduate School, The Catholic University of Korea, Seoul 06591, Republic of Korea.
Medicina (Kaunas). 2025 May 9;61(5):871. doi: 10.3390/medicina61050871.
: Dexamethasone has been widely researched for its ability to promote osteogenic differentiation in mesenchymal stem cells in basic research. This study focused on examining the effects of dexamethasone on both cell viability and osteogenic differentiation in three-dimensional stem cell spheroids. : These spheroids were created using concave microwells and exposed to dexamethasone at concentrations ranging from 0 μM to 100 μM, including intermediate levels of 0.1 μM, 1 μM, and 10 μM. Microscopic analysis was used to qualitatively assess cellular viability, while a water-soluble tetrazolium salt-based assay provided quantitative viability data. Osteogenic differentiation was evaluated by measuring alkaline phosphatase activity and calcium deposition using Alizarin Red staining. Additionally, the expression levels of genes associated with osteogenesis were measured through quantitative polymerase chain reaction. : The spheroids successfully self-assembled within the first 24 h and maintained their structural integrity over a seven-day period. Analysis of cell viability showed no statistically significant differences across the various dexamethasone concentrations tested. Although there was an observed increase in alkaline phosphatase activity and calcium deposition following dexamethasone treatment, these differences were not statistically significant. RUNX2 gene expression was upregulated in the 1 μM, 10 μM, and 100 μM groups, while COL1A1 expression significantly increased at 0.1 μM and 1 μM. : These results indicate that dexamethasone supports cell viability and enhances RUNX2 and COL1A1 expression in stem cell spheroids.
在基础研究中，地塞米松促进间充质干细胞成骨分化的能力已得到广泛研究。本研究着重考察地塞米松对三维干细胞球中细胞活力和成骨分化的影响。这些细胞球通过凹形微孔板制备，并暴露于浓度范围为0 μM至100 μM的地塞米松中，包括0.1 μM、1 μM和10 μM的中间水平。通过显微镜分析定性评估细胞活力，同时基于水溶性四氮唑盐的检测提供定量活力数据。通过测量碱性磷酸酶活性和使用茜素红染色检测钙沉积来评估成骨分化。此外，通过定量聚合酶链反应测量与成骨相关的基因表达水平。细胞球在前24小时内成功自组装，并在七天内保持其结构完整性。细胞活力分析表明，在所测试的不同地塞米松浓度之间没有统计学上的显著差异。虽然地塞米松处理后观察到碱性磷酸酶活性和钙沉积有所增加，但这些差异无统计学意义。RUNX2基因表达在1 μM、10 μM和100 μM组中上调，而COL1A1表达在0.1 μM和1 μM时显著增加。这些结果表明，地塞米松支持干细胞球中的细胞活力，并增强RUNX2和COL1A1的表达。
