Original Full Length ArticleMulti-level femoral morphology and mechanical properties of rats of different ages

•Macro-micro-nano-multi-level study is conducted to study bone properties of rat.•Age-related structural and mechanical properties of bone are explored.•Femur morphology and mechanical property are correlated with the animal age.•Mineral content and material properties explain age-related bone fragility.

A macro-micro-nano-multi-level study was conducted to explore age-related structural and mechanical properties of bone, as well as the effects of aging on bone properties. A total of 70 male Wistar rats were used, ranging in the ages of 1, 3, 5, 7, 9, 11, 14, 15, 16, and 17 months (n = 7/age group). After micro-computed tomography (CT) scanning, longitudinal cortical bone specimens with a length of 5 mm were cut along the femoral shaft axis from left femur shafts for mechanical testing, and the cross-sectional areas were measured. The macro-mechanical properties obtained in mechanical testing and microarchitecture parameters measured by micro-CT were significantly correlated with the animal age (r2 = 0.96, p < 0.001). Scanning electron microscopy was used for detecting the microarchitecture features of the fractured surfaces, which exhibited age-related plate-fibrous-mixed fibrous-plate texture, resulting in changes in macro-mechanical properties (r2 > 0.90, p < 0.001). The mineral phase of the left femoral shaft and head was analyzed by atomic force microscopy. Longitudinal and transverse trabecular bone tissues, as well as longitudinal cortical bone tissue, were used for nanoindentation test, and the chemical composition was evaluated by quantitative chemical analyses. The correlations between mineral content and bone material properties (i.e., elastic properties of the bone tissue and size and roughness of bone mineral grains) were highly significant (r > 0.95, p < 0.001). Multi-level femur morphology, mechanical property, and mineral content were significantly correlated with the animal age. The correlations between bone mineral content and bone material morphological and mechanical properties may partly explain the increase in bone fragility with aging, which will provide a theoretical basis for the investigation of age-related bone properties in clinics.