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Modeling ameloblast-matrix interactions using 3D cell culture

The distinct morphology adopted by ameloblasts during amelogenesis is highly stage specific and involved intimately with the development of a hierarchical enamel microstructure. The molecular mechanisms that govern the development of an elongated and polarized secretory ameloblast morphology and the...

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Autores principales: Visakan, Gayathri, Bapat, Rucha Arun, Su, Jingtan, Moradian-Oldak, Janet
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9751369/
https://www.ncbi.nlm.nih.gov/pubmed/36531170
http://dx.doi.org/10.3389/fphys.2022.1069519
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author Visakan, Gayathri
Bapat, Rucha Arun
Su, Jingtan
Moradian-Oldak, Janet
author_facet Visakan, Gayathri
Bapat, Rucha Arun
Su, Jingtan
Moradian-Oldak, Janet
author_sort Visakan, Gayathri
collection PubMed
description The distinct morphology adopted by ameloblasts during amelogenesis is highly stage specific and involved intimately with the development of a hierarchical enamel microstructure. The molecular mechanisms that govern the development of an elongated and polarized secretory ameloblast morphology and the potential roles played by the enamel matrix proteins in this process are not fully understood. Thus far, the in vitro models that have been developed to mimic these early cell-matrix interactions have either been unable to demonstrate direct morphological change or have failed to adapt across ameloblast cell lines. Here, we use a recently established 3D cell culture model to examine the interactions between HAT-7 cells and the major enamel matrix proteins, amelogenin and ameloblastin. We demonstrate that HAT-7 cells selectively respond to functional EMPs in culture by forming clusters of tall cells. Aspect ratio measurements from three-dimensional reconstructions reveal that cell elongation is 5-times greater in the presence of EMPs when compared with controls. Using confocal laser scanning microscopy, we observe that these clusters are polarized with asymmetrical distributions of Par-3 and claudin-1 proteins. The behavior of HAT-7 cells in 3D culture with EMPs is comparable with that of ALC and LS-8 cells. The fact that the 3D model presented here is tunable with respect to gel substrate composition and ameloblast cell type highlights the overall usefulness of this model in studying ameloblast cell morphology in vitro.
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spelling pubmed-97513692022-12-16 Modeling ameloblast-matrix interactions using 3D cell culture Visakan, Gayathri Bapat, Rucha Arun Su, Jingtan Moradian-Oldak, Janet Front Physiol Physiology The distinct morphology adopted by ameloblasts during amelogenesis is highly stage specific and involved intimately with the development of a hierarchical enamel microstructure. The molecular mechanisms that govern the development of an elongated and polarized secretory ameloblast morphology and the potential roles played by the enamel matrix proteins in this process are not fully understood. Thus far, the in vitro models that have been developed to mimic these early cell-matrix interactions have either been unable to demonstrate direct morphological change or have failed to adapt across ameloblast cell lines. Here, we use a recently established 3D cell culture model to examine the interactions between HAT-7 cells and the major enamel matrix proteins, amelogenin and ameloblastin. We demonstrate that HAT-7 cells selectively respond to functional EMPs in culture by forming clusters of tall cells. Aspect ratio measurements from three-dimensional reconstructions reveal that cell elongation is 5-times greater in the presence of EMPs when compared with controls. Using confocal laser scanning microscopy, we observe that these clusters are polarized with asymmetrical distributions of Par-3 and claudin-1 proteins. The behavior of HAT-7 cells in 3D culture with EMPs is comparable with that of ALC and LS-8 cells. The fact that the 3D model presented here is tunable with respect to gel substrate composition and ameloblast cell type highlights the overall usefulness of this model in studying ameloblast cell morphology in vitro. Frontiers Media S.A. 2022-12-01 /pmc/articles/PMC9751369/ /pubmed/36531170 http://dx.doi.org/10.3389/fphys.2022.1069519 Text en Copyright © 2022 Visakan, Bapat, Su and Moradian-Oldak. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Physiology
Visakan, Gayathri
Bapat, Rucha Arun
Su, Jingtan
Moradian-Oldak, Janet
Modeling ameloblast-matrix interactions using 3D cell culture
title Modeling ameloblast-matrix interactions using 3D cell culture
title_full Modeling ameloblast-matrix interactions using 3D cell culture
title_fullStr Modeling ameloblast-matrix interactions using 3D cell culture
title_full_unstemmed Modeling ameloblast-matrix interactions using 3D cell culture
title_short Modeling ameloblast-matrix interactions using 3D cell culture
title_sort modeling ameloblast-matrix interactions using 3d cell culture
topic Physiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9751369/
https://www.ncbi.nlm.nih.gov/pubmed/36531170
http://dx.doi.org/10.3389/fphys.2022.1069519
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