Bone Morphogenetic Protein-9 Promotes Osteogenic Differentiation and Mineralization in Human Stem-Cell-Derived Spheroids
<i>Background and Objectives:</i> Alkaline phosphatase activity, mineralized matrix, and osteogenic-related gene expression have been shown to increase in response to bone morphogenetic protein-9 (BMP-9). In this study, spheroids derived from human gingival stem cells were used to determ...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-07-01
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| Series: | Medicina |
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| Online Access: | https://www.mdpi.com/1648-9144/59/7/1315 |
| _version_ | 1797588351173263360 |
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| author | Sung-Bin Lee Hyun-Jin Lee Jun-Beom Park |
| author_facet | Sung-Bin Lee Hyun-Jin Lee Jun-Beom Park |
| author_sort | Sung-Bin Lee |
| collection | DOAJ |
| description | <i>Background and Objectives:</i> Alkaline phosphatase activity, mineralized matrix, and osteogenic-related gene expression have been shown to increase in response to bone morphogenetic protein-9 (BMP-9). In this study, spheroids derived from human gingival stem cells were used to determine the effects of BMP-9 on cell survival, osteogenesis, and mineralization. <i>Materials and Methods</i>: Human gingival stem cells were used to produce spheroids and then grown to concentrations of 0, 0.1, 1, 10, and 100 ng/mL with BMP-9. On days 1, 3, 5, and 7, morphological examination was carried out. A live/dead assay and Cell Counting Kit-8 was used to assess the vitality of cells. On days 7 and 14, alkaline phosphatase activity assays were carried out using a commercially available kit to examine the osteogenic differentiation of cell spheroids. Alizarin Red Staining was performed on the 7th and 14th days to evaluate mineralization, and <i>RUNX2</i> and <i>COL1A1</i> expression levels were evaluated on the 7th and 14th days using real-time polymerase chain reactions. <i>Results:</i> The BMP-9 added at the measured quantities did not appear to alter the shape of the well-formed spheroids produced by stem cells on day 1. In addition, treatment with BMP-9 at doses of 0, 0.1, 1, 10, or 100 ng/mL did not significantly alter cell diameter. Throughout the whole experimental process, viability was maintained. On day 14, the alkaline phosphatase activity in the groups dosed with 0.1, 1, 10, or 100 ng/mL was statistically higher than that in the unloaded control group (<i>p</i> < 0.05). According to qPCR data, the mRNA expression level of <i>RUNX2</i> with 1 ng/mL dosing was higher on day 7 compared to that of the unloaded control group (<i>p</i> < 0.05). <i>Conclusions:</i> These findings suggest that BMP-9 can be employed to stimulate early osteogenic differentiation in stem cell spheroids. |
| first_indexed | 2024-03-11T00:50:44Z |
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| id | doaj.art-4b8cdc3a4b20418398e59b761888c921 |
| institution | Directory Open Access Journal |
| issn | 1010-660X 1648-9144 |
| language | English |
| last_indexed | 2024-03-11T00:50:44Z |
| publishDate | 2023-07-01 |
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| series | Medicina |
| spelling | doaj.art-4b8cdc3a4b20418398e59b761888c9212023-11-18T20:25:00ZengMDPI AGMedicina1010-660X1648-91442023-07-01597131510.3390/medicina59071315Bone Morphogenetic Protein-9 Promotes Osteogenic Differentiation and Mineralization in Human Stem-Cell-Derived SpheroidsSung-Bin Lee0Hyun-Jin Lee1Jun-Beom Park2Dental Implantology, Graduate School of Clinical Dental Science, The Catholic University of Korea, Seoul 06591, Republic of KoreaDepartment of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaDental Implantology, Graduate School of Clinical Dental Science, The Catholic University of Korea, Seoul 06591, Republic of Korea<i>Background and Objectives:</i> Alkaline phosphatase activity, mineralized matrix, and osteogenic-related gene expression have been shown to increase in response to bone morphogenetic protein-9 (BMP-9). In this study, spheroids derived from human gingival stem cells were used to determine the effects of BMP-9 on cell survival, osteogenesis, and mineralization. <i>Materials and Methods</i>: Human gingival stem cells were used to produce spheroids and then grown to concentrations of 0, 0.1, 1, 10, and 100 ng/mL with BMP-9. On days 1, 3, 5, and 7, morphological examination was carried out. A live/dead assay and Cell Counting Kit-8 was used to assess the vitality of cells. On days 7 and 14, alkaline phosphatase activity assays were carried out using a commercially available kit to examine the osteogenic differentiation of cell spheroids. Alizarin Red Staining was performed on the 7th and 14th days to evaluate mineralization, and <i>RUNX2</i> and <i>COL1A1</i> expression levels were evaluated on the 7th and 14th days using real-time polymerase chain reactions. <i>Results:</i> The BMP-9 added at the measured quantities did not appear to alter the shape of the well-formed spheroids produced by stem cells on day 1. In addition, treatment with BMP-9 at doses of 0, 0.1, 1, 10, or 100 ng/mL did not significantly alter cell diameter. Throughout the whole experimental process, viability was maintained. On day 14, the alkaline phosphatase activity in the groups dosed with 0.1, 1, 10, or 100 ng/mL was statistically higher than that in the unloaded control group (<i>p</i> < 0.05). According to qPCR data, the mRNA expression level of <i>RUNX2</i> with 1 ng/mL dosing was higher on day 7 compared to that of the unloaded control group (<i>p</i> < 0.05). <i>Conclusions:</i> These findings suggest that BMP-9 can be employed to stimulate early osteogenic differentiation in stem cell spheroids.https://www.mdpi.com/1648-9144/59/7/1315bone morphogenetic proteinscell differentiationcell survivalgingivagrowth differentiation factor 2stem cells |
| spellingShingle | Sung-Bin Lee Hyun-Jin Lee Jun-Beom Park Bone Morphogenetic Protein-9 Promotes Osteogenic Differentiation and Mineralization in Human Stem-Cell-Derived Spheroids Medicina bone morphogenetic proteins cell differentiation cell survival gingiva growth differentiation factor 2 stem cells |
| title | Bone Morphogenetic Protein-9 Promotes Osteogenic Differentiation and Mineralization in Human Stem-Cell-Derived Spheroids |
| title_full | Bone Morphogenetic Protein-9 Promotes Osteogenic Differentiation and Mineralization in Human Stem-Cell-Derived Spheroids |
| title_fullStr | Bone Morphogenetic Protein-9 Promotes Osteogenic Differentiation and Mineralization in Human Stem-Cell-Derived Spheroids |
| title_full_unstemmed | Bone Morphogenetic Protein-9 Promotes Osteogenic Differentiation and Mineralization in Human Stem-Cell-Derived Spheroids |
| title_short | Bone Morphogenetic Protein-9 Promotes Osteogenic Differentiation and Mineralization in Human Stem-Cell-Derived Spheroids |
| title_sort | bone morphogenetic protein 9 promotes osteogenic differentiation and mineralization in human stem cell derived spheroids |
| topic | bone morphogenetic proteins cell differentiation cell survival gingiva growth differentiation factor 2 stem cells |
| url | https://www.mdpi.com/1648-9144/59/7/1315 |
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