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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Bibliographic Details
Main Authors: Sung-Bin Lee, Hyun-Jin Lee, Jun-Beom Park
Format: Article
Language:English
Published: MDPI AG 2023-07-01
Series:Medicina
Subjects:
Online Access:https://www.mdpi.com/1648-9144/59/7/1315
Description
Summary:<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.
ISSN:1010-660X
1648-9144