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Roles of Plasminogen Activator Inhibitor-1 in Heterotopic Ossification Induced by Achilles Tenotomy in Thermal Injured Mice

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Abstract

Heterotopic ossification (HO) is the process by which ectopic bone forms at an extraskeletal site. Inflammatory conditions induce plasminogen activator inhibitor 1 (PAI-1), an inhibitor of fibrinolysis, which regulates osteogenesis. In the present study, we investigated the roles of PAI-1 in the pathophysiology of HO induced by trauma/burn treatment using PAI-1-deficient mice. PAI-1 deficiency significantly promoted HO and increased the number of alkaline phosphatase (ALP)-positive cells in Achilles tendons after trauma/burn treatment. The mRNA levels of inflammation markers were elevated in Achilles tendons of both wild-type and PAI-1-deficient mice after trauma/burn treatment and PAI-1 mRNA levels were elevated in Achilles tendons of wild-type mice. PAI-1 deficiency significantly up-regulated the expression of Runx2, Osterix, and type 1 collagen in Achilles tendons 9 weeks after trauma/burn treatment in mice. In in vitro experiments, PAI-1 deficiency significantly increased ALP activity and mineralization in mouse osteoblasts. Moreover, PAI-1 deficiency significantly increased ALP activity and up-regulated osteocalcin expression during osteoblastic differentiation from mouse adipose-tissue-derived stem cells, but suppressed the chondrogenic differentiation of these cells. In conclusion, the present study showed that PAI-1 deficiency promoted HO in Achilles tendons after trauma/burn treatment partly by enhancing osteoblast differentiation and ALP activity in mice. Endogenous PAI-1 may play protective roles against HO after injury and inflammation.

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Acknowledgements

We thank Dr Shigeaki Yamanaka for his technical expertise.

Funding

The present study was supported by the following grants: A JSPS KAKENHI Grant-in-Aid for Early Career Scientists (No. 22K16755) to Y. M and Grants-in-Aid for Scientific Research (No. C:20K09514; No. C:KK230021) to H.K.; The Salt Science Research Foundation (No. 22C1) to H.K.

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Authors and Affiliations

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

    Yuya Mizukami, Naoyuki Kawao, Takashi Ohira, Daiki Hashimoto, Kiyotaka Okada, Osamu Matsuo & Hiroshi Kaji

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  1. Yuya Mizukami
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  2. Naoyuki Kawao
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Correspondence to Hiroshi Kaji.

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All mouse experiments were performed according to the Guide for the Care and Use of Laboratory Animals from the National Institutes of Health and the institutional guidelines for the use and care of laboratory animals at Kindai University. The protocol was approved by the Experimental Animal Welfare Committee of Kinki University (permit numbers: KAME-2022–073).

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Mizukami, Y., Kawao, N., Ohira, T. et al. Roles of Plasminogen Activator Inhibitor-1 in Heterotopic Ossification Induced by Achilles Tenotomy in Thermal Injured Mice. Calcif Tissue Int 114, 535–549 (2024). https://doi.org/10.1007/s00223-024-01193-5

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