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Inhibition of NF-κB Signaling-Mediated Crosstalk Between Macrophages and Preosteoblasts by Metformin Alleviates Trauma-Induced Heterotopic Ossification

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Abstract

Heterotopic ossification (HO) is a pathological condition that occurs in soft tissues following severe trauma. The exact pathogenesis of HO remains unclear. Studies have shown that inflammation predisposes patients to the development of HO and triggers ectopic bone formation. Macrophages are crucial mediators of inflammation and are involved in HO development. The present study investigated the inhibitory effect and underlying mechanism of metformin on macrophage infiltration and traumatic HO in mice. Our results found that abundant levels of macrophages were recruited to the injury site during early HO progression and that early administration of metformin prevented traumatic HO in mice. Furthermore, we found that metformin attenuated macrophage infiltration and the NF-κB signaling pathway in injured tissue. The monocyte-to-macrophage transition in vitro was suppressed by metformin and this event was mediated by AMPK. Finally, we showed that inflammatory mediator’s regulation by macrophages targeted preosteoblasts, leading to elevated BMP signaling, and osteogenic differentiation and driving HO formation, and this effect was blocked after the activation of AMPK in macrophages. Collectively, our study suggests that metformin prevents traumatic HO by inhibiting of NF-κB signaling in macrophages and subsequently attenuating BMP signaling and osteogenic differentiation in preosteoblasts. Therefore, metformin may serve as a therapeutic drug for traumatic HO by targeting NF-κB signaling in macrophages.

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DATA AVAILABILITY

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Abbreviations

HO:

Heterotopic ossification

NSAID:

Non-steroidal anti-inflammatory drugs

MCP1:

Monocyte chemokine

MIP1-α:

Macrophage inflammatory protein 1α

AMPK:

AMP-activated protein kinase

MAPK:

Mitogen-activated protein kinase

JAK/STAT:

JAKs signaling and activator of transcription

BMP:

Bone morphogenetic protein

LPS:

Lipopolysaccharides

ALP:

Alkaline phosphatase

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Funding

This work was supported by National Nature Science Foundation of China (31900852 to HL), Nature Science Foundation of Jiangxi Province of China (20192ACB21026 and 20224ACB206024 to HL).

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Author notes

  1. Jia Hou and Jie Chen contributed equally to this article.

Authors and Affiliations

  1. Department of Pathophysiology, School of Basic Medical Sciences, Nanchang University, Nanchang, 330006, Jiangxi, China

    Jia Hou, Jie Chen, Jingjing Fan, Zhimin Tang, Wenwen Zhou & Hui Lin

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  1. Jia Hou
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Contributions

Study conception and design: HL, JH, JC, ZM T, WW Z, and JJ F. JH and JC performed experiments, provided data analysis, and interpretation. ZM T, WW Z, and JJ F performed experiments on animal study and co-culture study. Drafting of the article: JH, JC, and HL. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Hui Lin.

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Ethics Approval

All the animal protocol was reviewed and approved by the Animal Care Committee of Nanchang University (NCDXSYDWFL-2015097). The animal procedures were performed strictly in accordance with National Institutes of Health Guide for Animal Care.

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Hou, J., Chen, J., Fan, J. et al. Inhibition of NF-κB Signaling-Mediated Crosstalk Between Macrophages and Preosteoblasts by Metformin Alleviates Trauma-Induced Heterotopic Ossification. Inflammation 46, 1414–1429 (2023). https://doi.org/10.1007/s10753-023-01817-2

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