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Biomechanical testing is the gold standard to determine bone competence, and has been used extensively. Direct mechanical testing provides detailed information on overall bone mechanical and material properties, but fails in revealing local properties such as local deformations and strains or quantification of fracture progression. Therefore, we incorporated several imaging methods in our mechanical setups in order to get a better insight into bone deformation and failure characteristics. Our aim was to develop an integrative approach for hierarchical investigation of bone, working at different scales of resolution ranging from the whole bone to its ultrastructure. At a macroscopic level, we used high-resolution and high-speed cameras which drastically increased the amount of information obtained from a biomechanical bone test. The new image data proved especially important when dealing with very small bones such as the murine femur. Here the feedback of the camera in the process of aligning and positioning the samples is indispensable for reproducibility. In addition, global failure behavior and fracture initiation can now be visualized with high temporal resolution. At a microscopic level, bone microstructure, i.e. trabecular architecture and cortical porosity, are known to influence bone strength and failure mechanisms significantly. For this reason, we developed an image-guided failure assessment technique, also referred to as functional microimaging, allowing direct time-lapsed 3D visualization and computation of local displacements and strains for better quantification of fracture initiation and progression at the microscopic level. While the resolution of typical desktop micro-computed tomography is around a few micrometers, highly brilliant X-rays from synchrotron radiation permit to explore the nanometer world. This allowed, for the first time, to uncover fully nondestructively the 3D ultrastructure of bone including vascular and cellular structures and to investigate their role in development of bone microcracks in an unprecedented resolution. We conclude that functional microimaging, i.e. the combination of biomechanical testing with non-destructive 3D imaging and visualization are extremely valuable in studying bone failure mechanisms. Functional investigation of microcrack initiation and propagation will lead to a better understanding of the relative contribution of bone mass and bone quality to bone competence.
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