Bone hierarchical organization through the lens of materials science: Present opportunities and future challenges

Multiscale characterization is essential to better understand the hierarchical architecture of bone and an array of analytical methods contributes to exploring the various structural and compositional aspects. Incorporating X-ray tomography, X-ray scattering, vibrational spectroscopy, and atom probe tomography alongside electron microscopy provides a comprehensive approach, offering insights into the diverse levels of organization within bone. X-ray scattering techniques reveal information about collagen-mineral spatial relationships, while X-ray tomography captures 3D structural details, especially at the microscale. Electron microscopy, such as scanning and transmission electron microscopy, extends resolution to the nanoscale, showcasing intricate features such as collagen fibril organization. Additionally, atom probe tomography achieves sub-nanoscale resolution and high chemical sensitivity, enabling detailed examination of bone composition. Despite various technical challenges, a correlative approach allows for a comprehensive understanding of bone material properties. Real-time investigations through in situ and in operando approaches shed light on the dynamic processes in bone. Recently developed techniques such as liquid, in situ transmission electron microscopy provide insights into calcium phosphate formation and collagen mineralization. Mechanical models developed in the effort to link structure, composition, and function currently remain oversimplified but can be improved. In conclusion, correlative analytical platforms provide a holistic perspective of bone extracellular matrix and are essential for unraveling the intricate interplay between structure and composition within bone.