Micro-damage formation within the skeleton is an important stimulant for bone remodeling, however abnormal build-up of micro-damage can lead to skeletal fragility. In this study, µCT imaging based micro finite element (μFE) models were used to evaluate tissue level damage criteria in whole healthy and metastatically-involved vertebrae. T13-L2 spinal segments were excised from osteolytic (n=3) and healthy (n=3) female athymic rnu/rnu rats. Osteolytic metastasis was generated by intercardiac injection of HeLa cancer cells. Micro-mechanical axial loading was applied to the spinal motion segments under μCT imaging. Vertebral samples underwent BaSO4 staining and sequential calcein/fuchsin staining to identify load induced micro-damage. μCT imaging was used generate specimen specific μFE models of the healthy and osteolytic whole rat vertebrae. Model boundary conditions were generated through deformable image registration of loaded and unloaded scans. Elevated stresses and strains were detected in regions of micro-damage identified through histological and BaSO4 staining within healthy and osteolytic vertebral models, as compared to undamaged regions. Additionally, damaged regions of metastatic vertebrae experienced significantly higher local stresses and strains than those in the damaged regions of healthy specimens. Areas identified by BaSO4 staining, however, yielded lower levels of stress and strain in damaged and undamaged regions of healthy and metastatic vertebrae as compared to fuschin staining. The multimodal (experimental, image-based and computational) techniques used in this study demonstrated the ability of local stresses and strains computed through µFE analysis to identify trabecular micro-damage, that can be applied to biomechanical analyses of healthy and diseased whole bones.

Three-dimensional image-based strain measurement in whole bones allows representation of physiological, albeit quasi-static, loading conditions. However, such work to date has been limited to specimens postmortem. The main purpose of this study is to verify the efficacy of deformable image registration of post-euthanasia strain to characterize the in vivo mechanical behavior of rat vertebrae. A micro-computed tomography-compatible custom loading device was used to apply 75 N load to a three-level caudal motion segment of a healthy rat. Loaded and unloaded micro-computed tomography scans were acquired in vivo and post-sacrifice. A micro-computed tomography-based deformable image registration algorithm was used to calculate vertebral strains live and post-euthanasia. No significant difference was found in the in vivo strains (-0.011 ± 0.001) and ex vivo strains (-0.012 ± 0.001) obtained from the comparisons of loaded and unloaded images (p = 0.3). Comparisons between unloaded-unloaded and loaded-loaded scans yielded significantly lower axial strains, representing the error of the method. Qualitatively, high strains were observed adjacent to growth plate regions in evaluating the loaded-unloaded images. Strain patterns in the loaded-loaded and unloaded-unloaded scans were inconsistent as would be expected in representing noise. Overall, live and dead loaded to unloaded comparisons yielded similar strain patterns and magnitudes. Point-wise differences in axial strain fields also supported this observation. This study demonstrated a proof of concept, suggesting that post-euthanasia micro-computed tomography-based strain analysis is able to represent the in vivo quasi-static behavior of rat tail vertebrae.