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bioRxiv · 10.1101/2023.06.19.545614

Liquid Transmission Electron Microscopy for Probing Collagen Biomineralization

DiCecco, L.-A.·Gao, R.·Gray, J. L.·Kelly, D. F.·Sone, E. D.·Grandfield, K.
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Abstract

Collagen biomineralization is foundational to hard tissue assembly. While studied extensively, collagen mineralization processes are not fully understood as the majority of theories are derived from electron microscopy (EM) in static, dehydrated, or frozen conditions, unlike the liquid phase environment where mineralization occurs dynamically. Herein, novel liquid transmission EM (TEM) strategies are presented, where collagen mineralization was explored in liquid conditions for the first time. Custom thin-film enclosures were employed to visualize the mineralization of reconstituted collagen fibrils in a calcium-phosphate and polyaspartic acid solution to promote intrafibrillar mineralization. TEM highlighted that at early time points, precursor mineral particles attached to collagen and progressed to crystalline mineral platelets aligned with fibrils at later time points. This aligns with observations from other techniques and validates this liquid TEM approach. This work provides a new liquid imaging approach for exploring collagen biomineralization, advancing toward understanding disease pathogenesis and remineralization strategies for hard tissues.

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DiCecco, L.-A., Gao, R., Gray, J. L., Kelly, D. F., Sone, E. D., Grandfield, K.. 2023-06-22. Liquid Transmission Electron Microscopy for Probing Collagen Biomineralization. https://doi.org/10.1101/2023.06.19.545614

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