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bioRxiv · 10.1101/2020.07.10.197673

Visualization of Collagen–Mineral Arrangement using Atom Probe Tomography

Bryan E.J. Lee·Brian Langelier·Kathryn Grandfield
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Abstract

Bone is a complex, hierarchical structure comprised of two distinct phases: the organic, collagen- rich phase and the inorganic mineral-rich phase. This collagen-mineral arrangement has implications for bone function, aging, and disease. However, strategies to extract a single mineralized collagen fibril to investigate the interplay between components with sufficient resolution have been mostly confined to in vitro systems that only approximate the biological environment or transmission electron microscopy studies with lower spatial and chemical resolution. Therefore, there is extensive debate over the location of mineral with respect to collagen in in vivo mineralized tissues as visualization and quantification of the mineral in a living system is difficult or impossible. Herein, we have developed an approach to artificially extract a single mineralized collagen fibril from bone to analyze its composition and structure atom-by-atom with 3D resolution and sub-nanometer accuracy using atom probe tomography. This enables, for the first time, a method to probe fibril-level mineralization and collagen-mineral arrangement from an in vivo system with both the spatial and chemical precision required to comment on collagen- mineral arrangement. Using atom probe tomography, 4D (spatial + chemical) reconstructed volumes of leporine bone were generated with accuracy from correlative scanning electron microscopy. Distinct, winding collagen fibrils were identified with mineralized deposits both encapsulating and incorporated into the collagenous structures. This work demonstrates a novel fibril-level detection method that can be used to probe structural and chemical changes of bone and contribute new insights to the debate on collagen-mineral arrangement in mineralized tissues such as bones, and teeth.

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Bryan E.J. Lee, Brian Langelier, Kathryn Grandfield. 2020-07-10. Visualization of Collagen–Mineral Arrangement using Atom Probe Tomography. https://doi.org/10.1101/2020.07.10.197673

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