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Enhanced Bone Remodeling After Fracture Priming

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Abstract

The immune system is an active component of bone repair. Mast cells influence the recruitment of macrophages, osteoclasts and blood vessels into the repair tissue. We hypothesized that if mast cells and other immune cells are sensitized to recognize broken bone, they will mount an increased response to subsequent fractures that may be translated into enhanced healing. To test this, we created a bone defect on the left leg of anesthetized mice and 2 weeks later, a second one on the right leg. Bone repair in the right legs was then compared to control mice that underwent the creation of bilateral window bone defects at the same time. Mice were euthanized at 14 and 56 days. Mineralized tissue quantity and morphometric parameters were assessed using micro-CT and histology. The activity of osteoblasts, osteoclasts, vascular endothelial cells, mast cells, and macrophages was evaluated using histochemistry. Our main findings were (1) no significant differences in the amount of bone produced at 14- or 56 days post-operative between groups; (2) mice exposed to subsequent fractures showed significantly better bone morphometric parameters after 56 days post-operative; and (3) significant increases in the content of blood vessels, osteoclasts, and the number of macrophages in the subsequent fracture group. Our results provide strong evidence that a transient increase in the inflammatory state of a healing injury promotes faster bone remodelling and increased neo-angiogenesis. This phenomenon is also characterized by changes in mast cell and macrophage content that translate into more active recruitment of mesenchymal stromal cells.

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Data Availability

The data that support the findings of this study are openly available in McGill University’s Dataverse Institutional Repository (https://dataverse.scholarsportal.info/dataverse/mcgill).

Code Availability

Not applicable.

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Acknowledgements

The work was performed in the Bone Engineering and the Regenerative Orthopaedics and Innovation Laboratories of the Research Institute-McGill University Health Centre, which are a Fonds de Recherche Québec Santé (FRQS) sponsored Centre de Recherche. The Bone Engineering Labs are supported in part by the Fonds de Recherche Québec Santé-Réseau de recherche en santé buccodentaire et osseuse (FRQS-RSBO). JRGL and KRB are scholars of the Mexican National Council for Science and Technology (CONACYT). JRGL also receives a doctoral fellowship from FRSQ. PAM is FRQS Chercheur boursier clincien Junior 2. The authors would like to acknowledge the contributions of Dr. Christopher Moraes and Miss. Mira Abou-Rjeili for the in vitro studies.

Funding

Canadian Institutes of Health Research (CIHR) and Fonds de recherche du Québec – Santé (FRQS).

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Authors and Affiliations

  1. Bone Engineering Labs, Injury, Repair & Recovery Program, Research Institute, McGill University Health Centre, 1650 Cedar Ave., Montreal, QC, H3G 1A4, Canada

    Jose L. Ramirez-GarciaLuna, Karla Rangel-Berridi, Ore-Oluwa Olasubulumi, Janet E. Henderson & Paul A. Martineau

  2. Biofabrication and Bioengineering Labs, Injury, Repair & Recovery Program, Research Institute, McGill University Health Centre, 1650 Cedar Ave., Montreal, QC, H3G 1A4, Canada

    Karla Rangel-Berridi & Derek H. Rosenzweig

  3. Regenerative Orthopaedics and Innovation Laboratory, Injury, Repair & Recovery Program, Research Institute-McGill University Health Centre, 1650 Cedar Ave., Montreal, QC, H3G 1A4, Canada

    Rahul Gawri & Paul A. Martineau

  4. Experimental Surgery, Faculty of Medicine, McGill University, 3605 Rue de la Montagne, Montreal, QC, H3G 2M1, Canada

    Jose L. Ramirez-GarciaLuna, Karla Rangel-Berridi, Derek H. Rosenzweig, Janet E. Henderson, Rahul Gawri & Paul A. Martineau

  5. Experimental Medicine, Faculty of Medicine, McGill University, 3605 Rue de la Montagne, Montreal, QC, H3G 2M1, Canada

    Janet E. Henderson

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  1. Jose L. Ramirez-GarciaLuna
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Contributions

Conceptualization: JR-GL, JEH, PAM. Data acquisition and curation: JR-G, KR-B, O-OU. Formal analysis: JR-G, DHR, RG, PAM. Funding acquisition: JEH, PAM. Methodology: DHR, JEH, PAM. Resources: DHR, JEH, RG, PAM. Supervision: DHR, JEH, PAM. Manuscript writing, review and editing: all authors.

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Correspondence to Rahul Gawri.

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All authors declare no conflict of interest.

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McGill MGH Facility Animal Care Committee (7016).

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All live animal procedures were conducted in accordance with a protocol approved by McGill MGH Facility Animal Care Committee (7016) and in keeping with the guidelines of the Canada Council on Animal Care.

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Ramirez-GarciaLuna, J.L., Rangel-Berridi, K., Olasubulumi, OO. et al. Enhanced Bone Remodeling After Fracture Priming. Calcif Tissue Int 110, 349–366 (2022). https://doi.org/10.1007/s00223-021-00921-5

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