Spatiotemporal characterization of microdamage accumulation in rat ulnae in response to uniaxial compressive fatigue loading.

Repetitive fatigue loading can induce microdamage accumulation in bone matrix, which results in impaired mechanical properties and increased fracture susceptibility. However, the spatial distribution and time-variant process of microdamage accumulation in fatigue-loaded skeleton, especially for linear microcracks which are known to initiate bone remodeling, remain not fully understood. In this study, the time-varying process of the morphology and distribution of microcracks in rat ulnae subjected to uniaxial compressive fatigue loading was investigated. Right forelimbs of thirty four-month-old male Sprague-Dawley rats were subjected to one bout of cyclic ramp loading with 0.67 Hz at a normalized peak force of 0.055 N/g body weight for 6000 cycles, and the contralateral left ulnae were not loaded as the control samples. Ten rats were randomly euthanized on Days 3, 5, and 7 post fatigue loading. Our findings via two-dimensional histomorphometric measurements based on basic fuchsin staining and three-dimensional quantifications using contrast-enhanced micro-computed tomography (MicroCT) with precipitated BaSO staining demonstrated that the accumulation of linear microcracks (increase in the amount of linear microcracks) on Day 5 was significantly higher than that on Day 3 and Day 7 post fatigue loading. Our histological and histomorphometric results revealed that linear microcrack density (Cr.Dn) in the tensile cortex at Days 3, 5 and 7 post fatigue loading was significantly higher than that in the compressive side, whereas linear microcrack length (Cr.Le) in the tensile cortex at Day 3 was significantly lower than that in the compressive cortex. Our findings revealed that microcrack accumulation exhibited a non-linear time-varying process at 3, 5 and 7 days post axial compressive fatigue loading (with observable peak Cr.Dn at Day 5). Our findings also revealed distinct distribution of microcrack density and morphology in rat ulnae with tensile and compressive strains, as characterized by more microcracks accumulated in tensile cortices, and longer cracks shown in compressive cortices.