Abstract
Human synovial mesenchymal stem cells (hSMSCs) are a promising cell source for cartilage regeneration because of their superior chondrogenic potential in vitro. This study aimed to further optimize the conditions for inducing chondrogenesis of hSMSCs, focusing on the dose of dexamethasone in combination with transforming growth factor-β3 (TGFβ3) and/or bone morphogenetic protein-2 (BMP2). When hSMSCs-derived aggregates were cultured with TGFβ3, dexamethasone up to 10 nM promoted chondrogenesis, but attenuated it with heterogeneous tissue formation when used at concentrations over than 100 nM. On the other hands, BMP2-induced chondrogenesis was remarkably disturbed in the presence of more than 10 nM dexamethasone along with unexpected adipogenic differentiation. In the presence of both TGFβ3 and BMP2, dexamethasone dose dependently promoted cartilaginous tissue formation as judged by tissue volume, proteoglycan content, and type 2 collagen expression, whereas few adipocytes were detected in the formed tissue when cultures were supplemented with over 100 nM dexamethasone. Even in chondrogenic conditions, dexamethasone thus affected hSMSCs differentiation not only toward chondrocytes, but also towards adipocytes dependent on the dose and combined growth factor. These findings have important implications regarding the use of glucocorticoids in in vitro tissue engineering for cartilage regeneration using hSMSCs.
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The authors thank Fumiko Hirayama and Yukiko Eguchi for their technical support.
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Supplementary Fig.
1. Results obtained using hSMSCs isolated from other two donors; related to Fig. 2. Scale bars = 500 μm. (TIFF 11793 kb)
Supplementary Fig.
2. Results obtained using hSMSCs isolated from other two donors; related to Fig. 4. Scale bars = 500 μm (left panels) and 50 μm (right panels). (TIFF 15848 kb)
Supplementary Fig.
3. Effect of dexamethasone compared to that of other types of glucocorticoids (prednisolone, betamethasone, and fluocinolone acetonide) on two-dimension micromass cultured with BMP2 (50 ng/mL). Alcian blue and Oil red O staining at day 7 culture. Scale bars = 1 mm (left panels) and 100 μm (right panels). (TIFF 26588 kb)
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Chijimatsu, R., Kobayashi, M., Ebina, K. et al. Impact of dexamethasone concentration on cartilage tissue formation from human synovial derived stem cells in vitro. Cytotechnology 70, 819–829 (2018). https://doi.org/10.1007/s10616-018-0191-y
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DOI: https://doi.org/10.1007/s10616-018-0191-y