An optimised GAS-pharyngeal cell biofilm model

Abstract Group A Streptococcus (GAS) causes 700 million infections and accounts for half a million deaths per year. Biofilm formation has been implicated in both pharyngeal and dermal GAS infections. In vitro, plate-based assays have shown that several GAS M-types form biofilms, and multiple GAS vir...

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Autores principales:	Heema K. N. Vyas, Jason D. McArthur, Martina L. Sanderson-Smith
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/29a2b6c563a244f0a96180d08172d1c2
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spelling	oai:doaj.org-article:29a2b6c563a244f0a96180d08172d1c22021-12-02T15:51:14ZAn optimised GAS-pharyngeal cell biofilm model10.1038/s41598-021-87377-72045-2322https://doaj.org/article/29a2b6c563a244f0a96180d08172d1c22021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-87377-7https://doaj.org/toc/2045-2322Abstract Group A Streptococcus (GAS) causes 700 million infections and accounts for half a million deaths per year. Biofilm formation has been implicated in both pharyngeal and dermal GAS infections. In vitro, plate-based assays have shown that several GAS M-types form biofilms, and multiple GAS virulence factors have been linked to biofilm formation. Although the contributions of these plate-based studies have been valuable, most have failed to mimic the host environment, with many studies utilising abiotic surfaces. GAS is a human specific pathogen, and colonisation and subsequent biofilm formation is likely facilitated by distinct interactions with host tissue surfaces. As such, a host cell-GAS model has been optimised to support and grow GAS biofilms of a variety of GAS M-types. Improvements and adjustments to the crystal violet biofilm biomass assay have also been tailored to reproducibly detect delicate GAS biofilms. We propose 72 h as an optimal growth period for yielding detectable biofilm biomass. GAS biofilms formed are robust and durable, and can be reproducibly assessed via staining/washing intensive assays such as crystal violet with the aid of methanol fixation prior to staining. Lastly, SEM imaging of GAS biofilms formed by this model revealed GAS cocci chains arranged into three-dimensional aggregated structures with EPS matrix material. Taken together, we outline an efficacious GAS biofilm pharyngeal cell model that can support long-term GAS biofilm formation, with biofilms formed closely resembling those seen in vivo.Heema K. N. VyasJason D. McArthurMartina L. Sanderson-SmithNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Heema K. N. Vyas Jason D. McArthur Martina L. Sanderson-Smith An optimised GAS-pharyngeal cell biofilm model
description	Abstract Group A Streptococcus (GAS) causes 700 million infections and accounts for half a million deaths per year. Biofilm formation has been implicated in both pharyngeal and dermal GAS infections. In vitro, plate-based assays have shown that several GAS M-types form biofilms, and multiple GAS virulence factors have been linked to biofilm formation. Although the contributions of these plate-based studies have been valuable, most have failed to mimic the host environment, with many studies utilising abiotic surfaces. GAS is a human specific pathogen, and colonisation and subsequent biofilm formation is likely facilitated by distinct interactions with host tissue surfaces. As such, a host cell-GAS model has been optimised to support and grow GAS biofilms of a variety of GAS M-types. Improvements and adjustments to the crystal violet biofilm biomass assay have also been tailored to reproducibly detect delicate GAS biofilms. We propose 72 h as an optimal growth period for yielding detectable biofilm biomass. GAS biofilms formed are robust and durable, and can be reproducibly assessed via staining/washing intensive assays such as crystal violet with the aid of methanol fixation prior to staining. Lastly, SEM imaging of GAS biofilms formed by this model revealed GAS cocci chains arranged into three-dimensional aggregated structures with EPS matrix material. Taken together, we outline an efficacious GAS biofilm pharyngeal cell model that can support long-term GAS biofilm formation, with biofilms formed closely resembling those seen in vivo.
format	article
author	Heema K. N. Vyas Jason D. McArthur Martina L. Sanderson-Smith
author_facet	Heema K. N. Vyas Jason D. McArthur Martina L. Sanderson-Smith
author_sort	Heema K. N. Vyas
title	An optimised GAS-pharyngeal cell biofilm model
title_short	An optimised GAS-pharyngeal cell biofilm model
title_full	An optimised GAS-pharyngeal cell biofilm model
title_fullStr	An optimised GAS-pharyngeal cell biofilm model
title_full_unstemmed	An optimised GAS-pharyngeal cell biofilm model
title_sort	optimised gas-pharyngeal cell biofilm model
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/29a2b6c563a244f0a96180d08172d1c2
work_keys_str_mv	AT heemaknvyas anoptimisedgaspharyngealcellbiofilmmodel AT jasondmcarthur anoptimisedgaspharyngealcellbiofilmmodel AT martinalsandersonsmith anoptimisedgaspharyngealcellbiofilmmodel AT heemaknvyas optimisedgaspharyngealcellbiofilmmodel AT jasondmcarthur optimisedgaspharyngealcellbiofilmmodel AT martinalsandersonsmith optimisedgaspharyngealcellbiofilmmodel
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An optimised GAS-pharyngeal cell biofilm model

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