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Role of osteoclasts in heterotopic ossification enhanced by fibrodysplasia ossificans progressiva-related activin-like kinase 2 mutation in mice

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Abstract

Fibrodysplasia ossificans progressiva (FOP) is a disorder of skeletal malformations and progressive heterotopic ossification. The constitutively activating mutation (R206H) of the bone morphogenetic protein type 1 receptor, activin-like kinase 2 (ALK2), is responsible for the pathogenesis of FOP. Although transfection of the causal mutation of FOP into myoblasts enhances osteoclast formation by transforming growth factor-β (TGF-β), the role of osteoclasts in heterotopic ossification is unknown. We therefore examined the effects of alendronate, SB431542 and SB203580 on heterotopic ossification induced by the causal mutation of FOP. Total bone mineral content as well as numbers of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated and alkaline phosphatase (ALP)-positive cells in heterotopic bone were significantly higher in muscle tissues implanted with ALK2 (R206H)-transfected mouse myoblastic C2C12 cells than in the tissues implanted with empty vector-transfected cells in nude mice. Alendronate, an aminobisphosphonate, did not affect total mineral content or numbers of TRAP-positive multinucleated and ALP-positive cells in heterotopic bone, which were enhanced by the implantation of ALK2 (R206H)-transfected C2C12 cells, although it significantly decreased serum levels of cross-linked C-telopeptide of type I collagen, a bone resorption index. Moreover, neither SB431542, an inhibitor of TGF-β receptor type I kinase, nor SB203580, an inhibitor of p38 mitogen-activated protein kinase, affected the increase in heterotopic ossification due to the implantation of ALK2 (R206H)-transfected C2C12 cells. In conclusion, the present study indicates that osteoclast inhibition does not affect heterotopic ossification enhanced by FOP-related mutation.

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Acknowledgments

We thank Prof. Takenobu Katagiri (Saitama Medical University) for ALK2 (R206H) plasmids. This study was partly supported by a grant from the Osaka Medical Research Foundation for Intractable Diseases, a Kinki University Research and Grant-in-Aid for Scientific Research (C: 25460305) to NK and (C: 24590289) to HK from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.

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All authors have no conflicts of interest.

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Authors and Affiliations

  1. Department of Physiology and Regenerative Medicine, Kinki University Faculty of Medicine, 377-2 Ohnohigashi, Osakasayama, 589-8511, Osaka, Japan

    Naoyuki Kawao, Masato Yano, Yukinori Tamura, Kiyotaka Okada & Hiroshi Kaji

  2. Life Science Research Institute, Kinki University, Osaka, Japan

    Katsumi Okumoto

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  1. Naoyuki Kawao
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  2. Masato Yano
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  3. Yukinori Tamura
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  4. Katsumi Okumoto
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  6. Hiroshi Kaji
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Correspondence to Hiroshi Kaji.

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Kawao, N., Yano, M., Tamura, Y. et al. Role of osteoclasts in heterotopic ossification enhanced by fibrodysplasia ossificans progressiva-related activin-like kinase 2 mutation in mice . J Bone Miner Metab 34, 517–525 (2016). https://doi.org/10.1007/s00774-015-0701-3

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