Impact of Dexamethasone on Three-Dimensional Stem Cell Spheroids: Morphology, Viability, Osteogenic Differentiation
<i>Background and Objectives</i>: 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...
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MDPI AG
2025-05-01
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| author | Heera Lee Ju-Hwan Kim Hyun-Jin Lee Jun-Beom Park |
| author_facet | Heera Lee Ju-Hwan Kim Hyun-Jin Lee Jun-Beom Park |
| author_sort | Heera Lee |
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| description | <i>Background and Objectives</i>: 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. <i>Materials and Methods</i>: 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. <i>Results</i>: 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. <i>Conclusions</i>: These results indicate that dexamethasone supports cell viability and enhances RUNX2 and COL1A1 expression in stem cell spheroids. |
| format | Article |
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| language | English |
| publishDate | 2025-05-01 |
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| spelling | doaj-art-0fce27e9344047e89abf11455335a13f2025-08-20T02:33:51ZengMDPI AGMedicina1010-660X1648-91442025-05-0161587110.3390/medicina61050871Impact of Dexamethasone on Three-Dimensional Stem Cell Spheroids: Morphology, Viability, Osteogenic DifferentiationHeera Lee0Ju-Hwan Kim1Hyun-Jin Lee2Jun-Beom Park3Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaDepartment of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaDepartment of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaDepartment of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea<i>Background and Objectives</i>: 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. <i>Materials and Methods</i>: 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. <i>Results</i>: 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. <i>Conclusions</i>: These results indicate that dexamethasone supports cell viability and enhances RUNX2 and COL1A1 expression in stem cell spheroids.https://www.mdpi.com/1648-9144/61/5/871cell survivalcell differentiationcellular spheroidsdexamethasoneosteogenesisstem cells |
| spellingShingle | Heera Lee Ju-Hwan Kim Hyun-Jin Lee Jun-Beom Park Impact of Dexamethasone on Three-Dimensional Stem Cell Spheroids: Morphology, Viability, Osteogenic Differentiation Medicina cell survival cell differentiation cellular spheroids dexamethasone osteogenesis stem cells |
| title | Impact of Dexamethasone on Three-Dimensional Stem Cell Spheroids: Morphology, Viability, Osteogenic Differentiation |
| title_full | Impact of Dexamethasone on Three-Dimensional Stem Cell Spheroids: Morphology, Viability, Osteogenic Differentiation |
| title_fullStr | Impact of Dexamethasone on Three-Dimensional Stem Cell Spheroids: Morphology, Viability, Osteogenic Differentiation |
| title_full_unstemmed | Impact of Dexamethasone on Three-Dimensional Stem Cell Spheroids: Morphology, Viability, Osteogenic Differentiation |
| title_short | Impact of Dexamethasone on Three-Dimensional Stem Cell Spheroids: Morphology, Viability, Osteogenic Differentiation |
| title_sort | impact of dexamethasone on three dimensional stem cell spheroids morphology viability osteogenic differentiation |
| topic | cell survival cell differentiation cellular spheroids dexamethasone osteogenesis stem cells |
| url | https://www.mdpi.com/1648-9144/61/5/871 |
| work_keys_str_mv | AT heeralee impactofdexamethasoneonthreedimensionalstemcellspheroidsmorphologyviabilityosteogenicdifferentiation AT juhwankim impactofdexamethasoneonthreedimensionalstemcellspheroidsmorphologyviabilityosteogenicdifferentiation AT hyunjinlee impactofdexamethasoneonthreedimensionalstemcellspheroidsmorphologyviabilityosteogenicdifferentiation AT junbeompark impactofdexamethasoneonthreedimensionalstemcellspheroidsmorphologyviabilityosteogenicdifferentiation |