Glucocorticoid (or steroid) induced osteoporosis (GIOP) is the leading form of secondary osteoporosis, affecting up to 50% of patients receiving chronic glucocorticoid therapy. Bone quantity (bone mass) changes in GIOP patients alone are inadequate to explain the increased fracture risk, and bone material changes (bone quality) at multiple levels have been implicated in the reduced mechanics. Quantitative analysis of specific material-level changes is limited. Here, we combined multiscale experimental techniques (scanning small/wide-angle X-ray scattering/diffraction, backscattered electron imaging, and X-ray radiography) to investigate these changes in a mouse model (Crh^−120/+) with chronic endogenous steroid production. Nanoscale degree of orientation, the size distribution of mineral nanocrystals in the bone matrix, the spatial map of mineralization on the femoral cortex, and the microporosity showed significant changes between GIOP and the control, especially in the endosteal cortex. Our work can provide insight into the altered structure-property relationship leading to lowered mechanical properties in GIOP.

Significance statement:​ As a natural nanocomposite with a hierarchical structure, bone undergoes a staggered load transfer mechanism at the nanoscale. Disease and age-related deterioration of bone mechanics are caused by changes in bone structure at multiple length scales. Although clinical tools such as dual-energy X-ray ab- sorptiometry (DXA) can be used to assess the reduction of bone quantity in these cases, little is known about how altered bone quality in diseased bone can increase fracture risk. It is clear that high-resolution diagnostic techniques need to be developed to narrow the gap between the onset and diagnosis of fracture-related changes. Here, by combining several scanning probe methods on a mouse model (Crh^−120/+) of glucocorticoid-induced osteoporosis (GIOP), we developed quantitative and spatially resolved maps of ultrastructural changes in col- lagen fibrils and mineral nanocrystals, mineralization distribution (microscale), and morphology (macroscale) across femoral osteoporotic bone. Our results indicate that the altered bone remodelling in GIOP leads to 1) heterogeneous bone structure and mineralization, 2) reduced degree of orientation of collagen fibrils and mi- neral nanocrystals, and 3) reduced length and increased thickness of mineral nanocrystals, which contribute to mechanical abnormalities. The combined multiscale experimental approach presented here will be used to understand musculoskeletal degeneration in aging and osteoporosis.