In situ mechanical behavior of mineralized collagen fibrils in murine cortical bone is altered by aging and disuse.

Mineralized collagen fibrils (MCFs) are the fundamental building blocks of bone, determining its mechanical properties. Aging and disuse are known to impair bone mechanics, but their specific effects on the nanoscale, in situ mechanical behavior of MCFs remain poorly understood. The present study utilized a murine model involving adult and aged mice, with a subset from each age group subjected to disuse through hindlimb unloading (n = 6). To investigate the in situ nanomechanical response of bone, murine tibiae were tensile-loaded within a custom-made axial loading device while being simultaneously scanned with atomic force microscopy (AFM). The bone surface was partially demineralized to expose the collagen fibrils. High-resolution AFM imaging in tapping-mode was then employed to quantitatively assess the morphological changes and nanomechanical properties of MCFs throughout the bone's elastic deformation process. In the adult murine tibia, the initial response to load was characterized by fibril reorientation and an increase in the in situ elastic modulus of MCFs, indicating stretching. It was followed by a sliding phase between adjacent fibrils. In contrast, the aged bone exhibited fibril sliding at the onset of loading, accompanied by a gradual decrease in the elastic modulus of MCFs. The nanomechanical alterations induced by disuse were more pronounced in aged mice compared to adults. The present findings demonstrate that aging and disuse significantly alter the nanoscale deformation mechanisms of bone, shifting the response from fibril stretching to predominant sliding. It provides novel evidence for a unique, age-dependent deformation mechanism at the fibrillar level, enhancing the current understanding of how aging and disuse impair bone quality and mechanics.