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TGF-β in dentin matrix extract induces osteoclastogenesis in vitro

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Abstract

Previously, we have demonstrated that the extracellular matrix from dentin affects osteoclastic activity in co-culture between osteoclast and osteoblast-rich fraction from mouse marrow cells. In the present study, we aimed to investigate the mechanisms of dentin matrix extract-induced osteoclastogenesis in mouse bone marrow macrophages (BMMs). Dentin proteins were extracted from bovine incisor root dentin using 0.6 M HCl. BMMs were cultured in α-MEM containing macrophage colony-stimulating factor/receptor activator of nuclear factor kappa-B ligand in the presence or absence of dentin matrix extract. Tartrate-resistant acid phosphatase (TRAP)-positive cell number, total TRAP activity, and the mRNA levels of osteoclast-related genes, assayed by real-time RT-PCR, were determined as markers of osteoclastogenesis. A neutralizing antibody against transforming growth factor-β1 (TGF-β1), SB431542, a TGF-β receptor inhibitor, and ELISA were used to determine the role of TGF-β1. We observed increases in TRAP-positive cell number, TRAP activity, and the mRNA levels of osteoclast-related genes of BMMs cultured with dentin extract. The use of a neutralizing antibody against TGF-β1 or SB431542 inhibited the inductive effect of dentin extract, suggesting TGF-β1 involvement. The addition of exogenous TGF-β1, but not bone morphogenic protein-2, also increased osteoclastogenesis, corresponding to the ELISA determination of TGF-β1 in the dentin extract. In conclusion, our results indicate that proteins from dentin matrix have an inductive effect in osteoclastogenesis, which is mediated, in part, by TGF-β1.

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Acknowledgments

This study was supported by a Grant-in-Aid for Scientific Research (20592392) from the Ministry of Education, Culture, Sports, Science and Technology, Japan and the Chulalongkorn University Centenary Academic Development Project (to WS), Thailand.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Pediatric Dentistry, Faculty of Dentistry, Chulalongkorn University, 34, Henri-Dunant Road, Pathumwan, Bangkok, 10330, Thailand

    Wannakorn Sriarj

  2. Pharmacology, Department of Bio-matrix, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan

    Kazuhiro Aoki, Keiichi Ohya & Mariko Takahashi

  3. Pediatric Dentistry, Department of Oral Health Sciences, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan

    Yuzo Takagi & Hitoyata Shimokawa

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  1. Wannakorn Sriarj
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  2. Kazuhiro Aoki
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Correspondence to Wannakorn Sriarj.

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Sriarj, W., Aoki, K., Ohya, K. et al. TGF-β in dentin matrix extract induces osteoclastogenesis in vitro. Odontology 103, 9–18 (2015). https://doi.org/10.1007/s10266-013-0140-3

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