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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Main Authors: Heera Lee, Ju-Hwan Kim, Hyun-Jin Lee, Jun-Beom Park
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Medicina
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Online Access:https://www.mdpi.com/1648-9144/61/5/871
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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
collection DOAJ
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.
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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