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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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-04-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-87377-7 |
| _version_ | 1818422942417027072 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-14T13:34:16Z |
| format | Article |
| id | doaj.art-29a2b6c563a244f0a96180d08172d1c2 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-14T13:34:16Z |
| publishDate | 2021-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-29a2b6c563a244f0a96180d08172d1c22022-12-21T22:59:38ZengNature PortfolioScientific Reports2045-23222021-04-0111111110.1038/s41598-021-87377-7An optimised GAS-pharyngeal cell biofilm modelHeema K. N. Vyas0Jason D. McArthur1Martina L. Sanderson-Smith2Illawarra Health and Medical Research InstituteSchool of Chemistry and Molecular Bioscience, Molecular Horizons, University of WollongongIllawarra Health and Medical Research InstituteAbstract 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.https://doi.org/10.1038/s41598-021-87377-7 |
| spellingShingle | Heema K. N. Vyas Jason D. McArthur Martina L. Sanderson-Smith An optimised GAS-pharyngeal cell biofilm model Scientific Reports |
| title | 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_short | An optimised GAS-pharyngeal cell biofilm model |
| title_sort | optimised gas pharyngeal cell biofilm model |
| url | https://doi.org/10.1038/s41598-021-87377-7 |
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