Journal
JOURNAL OF STRUCTURAL BIOLOGY
Volume 212, Issue 2, Pages -Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jsb.2020.107607
Keywords
Matrix vesicles; Langmuir monolayers; Phosphatidylserine; Biomineralization; Calcium phosphate
Funding
- Fundacao de Amparo a Pesquisa do Estado de Sao Paulo [17/20846-2, 14/24249-0, 2016/21236-6, 2017/08892-9]
- Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior -Brasil (CAPES) [001, 88887.320304/201900]
- Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) [304021/2017-2]
- FAPESP
- CAPES
- National Institutes of Health (USA) [DE12889]
- Brazil-France USP-COFECUB Uc [Sv 184/20]
- Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [14/24249-0, 17/20846-2] Funding Source: FAPESP
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Bone biomineralization is an exquisite process by which a hierarchically organized mineral matrix is formed. Growing evidence has uncovered the involvement of one class of extracellular vesicles, named matrix vesicles (MVs), in the formation and delivery of the first mineral nuclei to direct collagen mineralization. MVs are released by mineralization-competent cells equipped with a specific biochemical machinery to initiate mineral formation. However, little is known about the mechanisms by which MVs can trigger this process. Here, we present a combination of in situ investigations and ex vivo analysis of MVs extracted from growing-femurs of chicken embryos to investigate the role played by phosphatidylserine (PS) in the formation of mineral nuclei. By using self-assembled Langmuir monolayers, we reconstructed the nucleation core - a PS-enriched motif thought to trigger mineral formation in the lumen of MVs. In situ infrared spectroscopy of Langmuir monolayers and ex situ analysis by transmission electron microscopy evidenced that mineralization was achieved in supersaturated solutions only when PS was present. PS nucleated amorphous calcium phosphate that converted into biomimetic apatite. By using monolayers containing lipids extracted from native MVs, mineral formation was also evidenced in a manner that resembles the artificial PS-enriched monolayers. PS-enrichment in lipid monolayers creates nanodomains for local increase of supersaturation, leading to the nucleation of ACP at the interface through a multistep process. We posited that PS-mediated nucleation could be a predominant mechanism to produce the very first mineral nuclei during MV-driven bone/cartilage biomineralization.
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