Germ‐Free C57BL/6 Mice Have Increased Bone Mass and Altered Matrix Properties but Not Decreased Bone Fracture Resistance

Author:

Vahidi Ghazal1ORCID,Moody Maya2,Welhaven Hope D.2,Davidson Leah3,Rezaee Taraneh4,Behzad Ramina4,Karim Lamya4,Roggenbeck Barbara A.5,Walk Seth T.5,Martin Stephen A.6,June Ronald K.1,Heveran Chelsea M.1ORCID

Affiliation:

1. Department of Mechanical & Industrial Engineering Montana State University Bozeman Montana USA

2. Department of Chemistry & Biochemistry Montana State University Bozeman Montana USA

3. Department of Chemical and Biological Engineering University of Idaho Moscow Idaho USA

4. Department of Bioengineering University of Massachusetts Dartmouth Massachusetts USA

5. Department of Microbiology & Cell Biology Montana State University Bozeman Montana USA

6. Translational Biomarkers Core Laboratory, Center for American Indian and Rural Health Equity Montana State University Bozeman Montana USA

Abstract

ABSTRACTThe gut microbiome impacts bone mass, which implies a disruption to bone homeostasis. However, it is not yet clear how the gut microbiome affects the regulation of bone mass and bone quality. We hypothesized that germ‐free (GF) mice have increased bone mass and decreased bone toughness compared with conventionally housed mice. We tested this hypothesis using adult (20‐ to 21‐week‐old) C57BL/6J GF and conventionally raised female and male mice (n = 6–10/group). Trabecular microarchitecture and cortical geometry were measured from micro–CT of the femur distal metaphysis and cortical midshaft. Whole‐femur strength and estimated material properties were measured using three‐point bending and notched fracture toughness. Bone matrix properties were measured for the cortical femur by quantitative back‐scattered electron imaging and nanoindentation, and, for the humerus, by Raman spectroscopy and fluorescent advanced glycation end product (fAGE) assay. Shifts in cortical tissue metabolism were measured from the contralateral humerus. GF mice had reduced bone resorption, increased trabecular bone microarchitecture, increased tissue strength and decreased whole‐bone strength that was not explained by differences in bone size, increased tissue mineralization and fAGEs, and altered collagen structure that did not decrease fracture toughness. We observed several sex differences in GF mice, most notably for bone tissue metabolism. Male GF mice had a greater signature of amino acid metabolism, and female GF mice had a greater signature of lipid metabolism, exceeding the metabolic sex differences of the conventional mice. Together, these data demonstrate that the GF state in C57BL/6J mice alters bone mass and matrix properties but does not decrease bone fracture resistance. © 2023 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

Funder

National Institutes of Health

National Science Foundation

Publisher

Oxford University Press (OUP)

Subject

Orthopedics and Sports Medicine,Endocrinology, Diabetes and Metabolism

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