MicroCT-based imaging of microvasculature within the bone tissue

Abstract Angiogenesis is essential for skeletal development, bone healing and regeneration. Various research areas, especially implantology and tissue engineering, would benefit from improved three-dimensional (3D) imaging of the vasculature within bone tissue. X-ray microtomography (microCT) is a well-suited non-destructive 3D imaging technique for bone morphology. For the detection of vessels, a contrast-enhanced microCT-imaging must be used. Limited contrast between perfusion agents and mineralized bone has been the major drawback of this approach, making their distinct segmentation problematic. A decalcification step resolves this issue but inhibits simultaneous assessment of intracortical bone microstructure and vascular morphology. Moreover, the problem of contrasting becomes further compounded in samples with metal implants. This study describes μAngiofil-enhanced microCT-based visualization of vasculature within bone tissue in various small and large animal models, with and without decalcification. We present simultaneous microvascular and bone imaging in murine tibia, murine bone metastatic model, pulp chamber, gingiva and periodontal ligaments. In a large animal model (minipig) we perform visualization and segmentation of different tissue types and vessels in the hemimandible containing metal implants. Our manuscript introduces the first non-destructive approach for 3D imaging of the vasculature within soft and hard tissues in the vicinity of metal implants in a large animal model.