Porphyromonas gingivalis bypasses epithelial barrier and modulates fibroblastic inflammatory response in an in vitro 3D spheroid model

Abstract Porphyromonas gingivalis-induced inflammatory effects are mostly investigated in monolayer cultured cells. The aim of this study was to develop a 3D spheroid model of gingiva to take into account epithelio-fibroblastic interactions. Human gingival epithelial cells (ECs) and human oral fibro...

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Autores principales:	Isaac Maximiliano Bugueno, Fareeha Batool, Laetitia Keller, Sabine Kuchler-Bopp, Nadia Benkirane-Jessel, Olivier Huck
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Epithelial Barrier Experimental Periodontitis Human Oral Fibroblasts Epithelium-connective Tissue Interface Gingivalis Infection Medicine R Science Q
Acceso en línea:	https://doaj.org/article/41a1965e25ef41a7941e0b7e8bd6c376
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spelling	oai:doaj.org-article:41a1965e25ef41a7941e0b7e8bd6c3762021-12-02T15:08:36ZPorphyromonas gingivalis bypasses epithelial barrier and modulates fibroblastic inflammatory response in an in vitro 3D spheroid model10.1038/s41598-018-33267-42045-2322https://doaj.org/article/41a1965e25ef41a7941e0b7e8bd6c3762018-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-33267-4https://doaj.org/toc/2045-2322Abstract Porphyromonas gingivalis-induced inflammatory effects are mostly investigated in monolayer cultured cells. The aim of this study was to develop a 3D spheroid model of gingiva to take into account epithelio-fibroblastic interactions. Human gingival epithelial cells (ECs) and human oral fibroblasts (FBs) were cultured by hanging drop method to generate 3D microtissue (MT) whose structure was analyzed on histological sections and the cell-to-cell interactions were observed by scanning and transmission electron microscopy (SEM and TEM). MTs were infected by P. gingivalis and the impact on cell death (Apaf-1, caspase-3), inflammatory markers (TNF-α, IL-6, IL-8) and extracellular matrix components (Col-IV, E-cadherin, integrin β1) was evaluated by immunohistochemistry and RT-qPCR. Results were compared to those observed in situ in experimental periodontitis and in human gingival biopsies. MTs exhibited a well-defined spatial organization where ECs were organized in an external cellular multilayer, while, FBs constituted the core. The infection of MT demonstrated the ability of P. gingivalis to bypass the epithelial barrier in order to reach the fibroblastic core and induce disorganization of the spheroid structure. An increased cell death was observed in fibroblastic core. The development of such 3D model may be useful to define the role of EC–FB interactions on periodontal host-immune response and to assess the efficacy of new therapeutics.Isaac Maximiliano BuguenoFareeha BatoolLaetitia KellerSabine Kuchler-BoppNadia Benkirane-JesselOlivier HuckNature PortfolioarticleEpithelial BarrierExperimental PeriodontitisHuman Oral FibroblastsEpithelium-connective Tissue InterfaceGingivalis InfectionMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-13 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Epithelial Barrier Experimental Periodontitis Human Oral Fibroblasts Epithelium-connective Tissue Interface Gingivalis Infection Medicine R Science Q
spellingShingle	Epithelial Barrier Experimental Periodontitis Human Oral Fibroblasts Epithelium-connective Tissue Interface Gingivalis Infection Medicine R Science Q Isaac Maximiliano Bugueno Fareeha Batool Laetitia Keller Sabine Kuchler-Bopp Nadia Benkirane-Jessel Olivier Huck Porphyromonas gingivalis bypasses epithelial barrier and modulates fibroblastic inflammatory response in an in vitro 3D spheroid model
description	Abstract Porphyromonas gingivalis-induced inflammatory effects are mostly investigated in monolayer cultured cells. The aim of this study was to develop a 3D spheroid model of gingiva to take into account epithelio-fibroblastic interactions. Human gingival epithelial cells (ECs) and human oral fibroblasts (FBs) were cultured by hanging drop method to generate 3D microtissue (MT) whose structure was analyzed on histological sections and the cell-to-cell interactions were observed by scanning and transmission electron microscopy (SEM and TEM). MTs were infected by P. gingivalis and the impact on cell death (Apaf-1, caspase-3), inflammatory markers (TNF-α, IL-6, IL-8) and extracellular matrix components (Col-IV, E-cadherin, integrin β1) was evaluated by immunohistochemistry and RT-qPCR. Results were compared to those observed in situ in experimental periodontitis and in human gingival biopsies. MTs exhibited a well-defined spatial organization where ECs were organized in an external cellular multilayer, while, FBs constituted the core. The infection of MT demonstrated the ability of P. gingivalis to bypass the epithelial barrier in order to reach the fibroblastic core and induce disorganization of the spheroid structure. An increased cell death was observed in fibroblastic core. The development of such 3D model may be useful to define the role of EC–FB interactions on periodontal host-immune response and to assess the efficacy of new therapeutics.
format	article
author	Isaac Maximiliano Bugueno Fareeha Batool Laetitia Keller Sabine Kuchler-Bopp Nadia Benkirane-Jessel Olivier Huck
author_facet	Isaac Maximiliano Bugueno Fareeha Batool Laetitia Keller Sabine Kuchler-Bopp Nadia Benkirane-Jessel Olivier Huck
author_sort	Isaac Maximiliano Bugueno
title	Porphyromonas gingivalis bypasses epithelial barrier and modulates fibroblastic inflammatory response in an in vitro 3D spheroid model
title_short	Porphyromonas gingivalis bypasses epithelial barrier and modulates fibroblastic inflammatory response in an in vitro 3D spheroid model
title_full	Porphyromonas gingivalis bypasses epithelial barrier and modulates fibroblastic inflammatory response in an in vitro 3D spheroid model
title_fullStr	Porphyromonas gingivalis bypasses epithelial barrier and modulates fibroblastic inflammatory response in an in vitro 3D spheroid model
title_full_unstemmed	Porphyromonas gingivalis bypasses epithelial barrier and modulates fibroblastic inflammatory response in an in vitro 3D spheroid model
title_sort	porphyromonas gingivalis bypasses epithelial barrier and modulates fibroblastic inflammatory response in an in vitro 3d spheroid model
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/41a1965e25ef41a7941e0b7e8bd6c376
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_version_	1718388055434330112

Porphyromonas gingivalis bypasses epithelial barrier and modulates fibroblastic inflammatory response in an in vitro 3D spheroid model

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