Vitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells

<i>Background and Objectives:</i> Vitamin D is a bone modulator widely used in regenerative medicine. This study aimed to analyze the effects of vitamin D on the osteogenic differentiation and mineralization of human mesenchymal stem cells. <i>Materials and Methods:</i> Spher...

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Autores principales:	Hyun-Jin Lee, Young-Min Song, Seunghoon Baek, Yoon-Hee Park, Jun-Beom Park
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	cell differentiation osteogenesis stem cells vitamin D Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/704b258579a74fe1b61b03b1ab7dcb7f
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spelling	oai:doaj.org-article:704b258579a74fe1b61b03b1ab7dcb7f2021-11-25T18:19:08ZVitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells10.3390/medicina571112711648-91441010-660Xhttps://doaj.org/article/704b258579a74fe1b61b03b1ab7dcb7f2021-11-01T00:00:00Zhttps://www.mdpi.com/1648-9144/57/11/1271https://doaj.org/toc/1010-660Xhttps://doaj.org/toc/1648-9144<i>Background and Objectives:</i> Vitamin D is a bone modulator widely used in regenerative medicine. This study aimed to analyze the effects of vitamin D on the osteogenic differentiation and mineralization of human mesenchymal stem cells. <i>Materials and Methods:</i> Spheroids were fabricated using human bone marrow-derived stem cells, and were cultured in the presence of vitamin D at concentrations of 0, 0.1, 1, 10, and 100 nM. Stem cell spheroids were fabricated and the morphological evaluation was conducted on days 1, 3, 7 and 14. Determination of qualitative cellular viability was performed with Live/Dead Kit assay on days 1 and 7. Quantitative cellular viability was evaluated with Cell Counting Kit-8 on days 1, 3, 7, and 14. To analyze the osteogenic differentiation of cell spheroids, alkaline phosphatase activity assays were performed with commercially available kit on days 7 and 14. Real-time polymerase chain reaction was used to determine the expression levels of RUNX2, BSP, OCN, and COL1A1 on days 7 and 14. <i>Results:</i> The stem cells produced well-formed spheroids, and addition of vitamin D did not result in any noticeable changes in the shape. The addition of vitamin D did not significantly change the diameter of the spheroids at 0, 0.1, 1, 10, or 100 nM concentrations. Quantitative cell viability results from days 1, 3, 7 and 14 showed no significant difference between groups (<i>p</i> > 0.05). There was significantly higher alkaline phosphatase activity in the 0.1 nM group when compared with the control group on day 14 (<i>p</i> < 0.05). Real-time polymerase chain reaction results demonstrated that the mRNA expression levels of RUNX2, OCN, and COL1A1 were significantly increased when vitamin D was added to the culture. <i>Conclusions:</i> Based on these findings, we concluded that vitamin D could be applied to the increased osteogenicity of stem cell spheroids.Hyun-Jin LeeYoung-Min SongSeunghoon BaekYoon-Hee ParkJun-Beom ParkMDPI AGarticlecell differentiationosteogenesisstem cellsvitamin DMedicine (General)R5-920ENMedicina, Vol 57, Iss 1271, p 1271 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	cell differentiation osteogenesis stem cells vitamin D Medicine (General) R5-920
spellingShingle	cell differentiation osteogenesis stem cells vitamin D Medicine (General) R5-920 Hyun-Jin Lee Young-Min Song Seunghoon Baek Yoon-Hee Park Jun-Beom Park Vitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells
description	<i>Background and Objectives:</i> Vitamin D is a bone modulator widely used in regenerative medicine. This study aimed to analyze the effects of vitamin D on the osteogenic differentiation and mineralization of human mesenchymal stem cells. <i>Materials and Methods:</i> Spheroids were fabricated using human bone marrow-derived stem cells, and were cultured in the presence of vitamin D at concentrations of 0, 0.1, 1, 10, and 100 nM. Stem cell spheroids were fabricated and the morphological evaluation was conducted on days 1, 3, 7 and 14. Determination of qualitative cellular viability was performed with Live/Dead Kit assay on days 1 and 7. Quantitative cellular viability was evaluated with Cell Counting Kit-8 on days 1, 3, 7, and 14. To analyze the osteogenic differentiation of cell spheroids, alkaline phosphatase activity assays were performed with commercially available kit on days 7 and 14. Real-time polymerase chain reaction was used to determine the expression levels of RUNX2, BSP, OCN, and COL1A1 on days 7 and 14. <i>Results:</i> The stem cells produced well-formed spheroids, and addition of vitamin D did not result in any noticeable changes in the shape. The addition of vitamin D did not significantly change the diameter of the spheroids at 0, 0.1, 1, 10, or 100 nM concentrations. Quantitative cell viability results from days 1, 3, 7 and 14 showed no significant difference between groups (<i>p</i> > 0.05). There was significantly higher alkaline phosphatase activity in the 0.1 nM group when compared with the control group on day 14 (<i>p</i> < 0.05). Real-time polymerase chain reaction results demonstrated that the mRNA expression levels of RUNX2, OCN, and COL1A1 were significantly increased when vitamin D was added to the culture. <i>Conclusions:</i> Based on these findings, we concluded that vitamin D could be applied to the increased osteogenicity of stem cell spheroids.
format	article
author	Hyun-Jin Lee Young-Min Song Seunghoon Baek Yoon-Hee Park Jun-Beom Park
author_facet	Hyun-Jin Lee Young-Min Song Seunghoon Baek Yoon-Hee Park Jun-Beom Park
author_sort	Hyun-Jin Lee
title	Vitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells
title_short	Vitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells
title_full	Vitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells
title_fullStr	Vitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells
title_full_unstemmed	Vitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells
title_sort	vitamin d enhanced the osteogenic differentiation of cell spheroids composed of bone marrow stem cells
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/704b258579a74fe1b61b03b1ab7dcb7f
work_keys_str_mv	AT hyunjinlee vitamindenhancedtheosteogenicdifferentiationofcellspheroidscomposedofbonemarrowstemcells AT youngminsong vitamindenhancedtheosteogenicdifferentiationofcellspheroidscomposedofbonemarrowstemcells AT seunghoonbaek vitamindenhancedtheosteogenicdifferentiationofcellspheroidscomposedofbonemarrowstemcells AT yoonheepark vitamindenhancedtheosteogenicdifferentiationofcellspheroidscomposedofbonemarrowstemcells AT junbeompark vitamindenhancedtheosteogenicdifferentiationofcellspheroidscomposedofbonemarrowstemcells
_version_	1718411325130932224

Vitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells

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