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OPN, BSP, and Bone Quality—Structural, Biochemical, and Biomechanical Assessment in OPN−/−, BSP−/−, and DKO Mice

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Abstract

Osteopontin (OPN) and Bone Sialoprotein (BSP), abundantly expressed by osteoblasts and osteoclasts, appear to have important, partly overlapping functions in bone. In gene-knockout (KO, -/-) models of either protein and their double (D)KO in the same CD1/129sv genetic background, we analyzed the morphology, matrix characteristics, and biomechanical properties of femur bone in 2 and 4 month old, male and female mice. OPN−/− mice display inconsistent, perhaps localized hypermineralization, while the BSP−/− are hypomineralized throughout ages and sexes, and the low mineralization of young DKO mice recovers with age. The higher contribution of primary bone remnants in OPN−/− shafts suggests a slow turnover, while their lower percentage in BSP−/− indicates rapid remodeling, despite FTIR-based evidence in this genotype of a high maturity of the mineralized matrix. In 3-point bending assays, OPN−/− bones consistently display higher Maximal Load, Work to Max. Load and in young mice Ultimate Stress, an intrinsic characteristic of the matrix. Young male and old female BSP−/− also display high Work to Max. Load along with low Ultimate Stress. Principal Component Analysis confirms the major role of morphological traits in mechanical competence, and evidences a grouping of the WT phenotype with the OPN−/− and of BSP−/− with DKO, driven by both structural and matrix parameters, suggesting that the presence or absence of BSP has the most profound effects on skeletal properties. Single or double gene KO of OPN and BSP thus have multiple distinct effects on skeletal phenotypes, confirming their importance in bone biology and their interplay in its regulation.

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Acknowledgements

The work was funded by the “Agence Nationale de la Recherche” for the “Mouse_Kosto” project (Grants ANR-13-BSV1-0010-01 to Luc Malaval and ANR-13-BSV1-0010-02 to Hélène Follet), the “Institut National de la Santé et de la Recherche Médicale” (INSERM), the “Université Jean Monnet” of Saint-Etienne, and the “Ministère de l’Enseignement supérieur, de la Recherche et de l’Innovation” (PhD scholarship for Mathieu Maalouf). The authors thank the staff of PLEXAN for skillful technical assistance.

Funding

This article was funded by Agence Nationale de la Recherche, ANR-13-BSV1-0010-01, Luc Malaval, ANR-13-BSV1-0010-02, Hélène Follet, Institut National de la Santé et de la Recherche Médicale (FR), Université Jean Monnet, Saint-Etienne (FR), Ministère de l’Enseignement supérieur, de la Recherche et de l’Innovation (FR).

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  1. LBTO, Pôle Santé Nord – Faculté de Médecine, Inserm, U1059 Sainbiose, Rm 118, Université Jean Monnet, Mines St Etienne, 10 Chemin de La Marandière, St Priest en Jarez, F42270, Saint-Etienne, France

    Luc Malaval, Mathieu Maalouf, Myriam Normand, Arnaud Vanden-Boscche, Norbert Laroche & Laurence Vico

  2. Inserm, Université de Lyon, U1033 Lyos, F69372, Lyon, France

    Hélène Follet, Delphine Farlay, Evelyne Gineyts, Sebastien Rizzo, Charlene Thomas & Brigitte Burt-Pichat

  3. Centre Hospitalier Universitaire de Saint-Etienne, Centre de Cancérologie Universitaire, F42270, Saint Etienne, France

    Wafa Bouleftour

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  1. Luc Malaval
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Contributions

Luc Malaval, Hélène Follet and Delphine Farlay designed the study. Luc Malaval wrote the paper; he is the guarantor. Hélène Follet, Delphine Farlay, Evelyne Gineyts, Sebastien Rizzo, Charlene Thomas, Mathieu Maalouf, Brigitte Burt-Pichat, Wafa Bouleftour-Esquis, Arnaud Vanden-Boscche, and Norbert Laroche contributed to the experimental work. Myriam Normand supervised the statistical analysis of the data. Luc Malaval and Hélène Follet procured funding, along with Laurence Vico who also provided resources and advice for experimental work. All authors revised the paper critically for intellectual content and approved the final version. All authors agree to be accountable for the work and to ensure that any questions relating to the accuracy and integrity of the paper are investigated and properly resolved.

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Correspondence to Luc Malaval.

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Luc Malaval, Hélène Follet, Delphine Farlay, Evelyne Gineyts, Sebastien Rizzo, Charlene Thomas, Mathieu Maalouf, Myriam Normand, Brigitte Burt-Pichat, Wafa Bouleftour-Esquis, Arnaud Vanden-Boscche, Norbert Laroche, Laurence Vico have no conflict of interest.

Human and Animal Rights and Informed Consent

All procedures dealing with animals were in compliance with the European Community Standards on the care and use of laboratory animals (Ministère de l’Agriculture, France, Authorization D18-0801) and approved by the local ethical committee (Comité d’Ethique en Expérimentation Animale de la Loire CEEAL-UJM, agreement n°98) and the Animal Welfare Committee of the PLEXAN facility (Platform for Experiments and Analysis, Faculty of Medicine, University of Saint-Etienne, Saint-Etienne, France).

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Malaval, L., Follet, H., Farlay, D. et al. OPN, BSP, and Bone Quality—Structural, Biochemical, and Biomechanical Assessment in OPN−/−, BSP−/−, and DKO Mice. Calcif Tissue Int (2024). https://doi.org/10.1007/s00223-024-01217-0

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