Quantitative flow chamber system for evaluating in vitro biofilms and the kinetics of S. aureus biofilm formation in human plasma media

Abstract Background It has been well established that biofilm formation on orthopaedic implants is a critical event in the pathogenesis of orthopaedic infections, yet the natural history of this process with respect to bacterial adhesion, proliferation, and glycocalyx matrix production remains poorl...

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Autores principales:	Werasak Sutipornpalangkul, Kohei Nishitani, Edward M. Schwarz
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	In vitro biofilm Staphylococcus aureus Scanning Electron microscopy Bioluminescence Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/a968ebfd623240e2b5d22674308cf1c6
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spelling	oai:doaj.org-article:a968ebfd623240e2b5d22674308cf1c62021-11-14T12:08:47ZQuantitative flow chamber system for evaluating in vitro biofilms and the kinetics of S. aureus biofilm formation in human plasma media10.1186/s12866-021-02379-91471-2180https://doaj.org/article/a968ebfd623240e2b5d22674308cf1c62021-11-01T00:00:00Zhttps://doi.org/10.1186/s12866-021-02379-9https://doaj.org/toc/1471-2180Abstract Background It has been well established that biofilm formation on orthopaedic implants is a critical event in the pathogenesis of orthopaedic infections, yet the natural history of this process with respect to bacterial adhesion, proliferation, and glycocalyx matrix production remains poorly understood. Moreover, there are no quantitative methods yet available to assess the differences in biofilm formation between different bacterial strains or implant materials. Consequently, this study aimed to investigate the natural history of S. aureus in in vitro biofilm formation in human plasma media using a flow chamber system. Bioluminescent S. aureus strains were used to better understand the bacterial growth and biofilm formation on orthopaedic materials. Also, the effects of human plasma media were assessed by loading the chamber with Tryptic Soy Broth with 10% human plasma (TSB + HP). Results Scanning electron microscopy (SEM) was utilized to assess the morphological appearance of the biofilms, revealing that S. aureus inoculation was required for biofilm formation, and that the phenotypes of biofilm production after 24 h inoculation with three tested strains (SH1000, UAMS-1, and USA300) were markedly different depending on the culture medium. Time course study of the bioluminescence intensity (BLI) and biofilm production on the implants due to the UAMS-1 and USA300 strains revealed different characteristics, whereby UAMS-1 showed increasing BLI and biofilm growth until peaking at 9 h, while USA300 showed a rapid increase in BLI and biofilm formation at 6 h. The kinetics of biofilm formation for both UAMS-1 and USA300 were also supported and confirmed by qRT-PCR analysis of the 16S rRNA gene. Biofilms grown in our flow chamber in the plasma media were also demonstrated to involve an upregulation of the biofilm-forming-related genes icaA, fnbA, and alt. The BLI and SEM results from K-wire experiments revealed that the in vitro growth and biofilm formation by UAMS-1 and USA300 on stainless-steel and titanium surfaces were virtually identical. Conclusion We demonstrated a novel in vitro model for S. aureus biofilm formation with quantitative BLI and SEM outcome measures, and then used this model to demonstrate the presence of strain-specific phenotypes and its potential use to evaluate anti-microbial surfaces.Werasak SutipornpalangkulKohei NishitaniEdward M. SchwarzBMCarticleIn vitro biofilmStaphylococcus aureusScanning Electron microscopyBioluminescenceMicrobiologyQR1-502ENBMC Microbiology, Vol 21, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	In vitro biofilm Staphylococcus aureus Scanning Electron microscopy Bioluminescence Microbiology QR1-502
spellingShingle	In vitro biofilm Staphylococcus aureus Scanning Electron microscopy Bioluminescence Microbiology QR1-502 Werasak Sutipornpalangkul Kohei Nishitani Edward M. Schwarz Quantitative flow chamber system for evaluating in vitro biofilms and the kinetics of S. aureus biofilm formation in human plasma media
description	Abstract Background It has been well established that biofilm formation on orthopaedic implants is a critical event in the pathogenesis of orthopaedic infections, yet the natural history of this process with respect to bacterial adhesion, proliferation, and glycocalyx matrix production remains poorly understood. Moreover, there are no quantitative methods yet available to assess the differences in biofilm formation between different bacterial strains or implant materials. Consequently, this study aimed to investigate the natural history of S. aureus in in vitro biofilm formation in human plasma media using a flow chamber system. Bioluminescent S. aureus strains were used to better understand the bacterial growth and biofilm formation on orthopaedic materials. Also, the effects of human plasma media were assessed by loading the chamber with Tryptic Soy Broth with 10% human plasma (TSB + HP). Results Scanning electron microscopy (SEM) was utilized to assess the morphological appearance of the biofilms, revealing that S. aureus inoculation was required for biofilm formation, and that the phenotypes of biofilm production after 24 h inoculation with three tested strains (SH1000, UAMS-1, and USA300) were markedly different depending on the culture medium. Time course study of the bioluminescence intensity (BLI) and biofilm production on the implants due to the UAMS-1 and USA300 strains revealed different characteristics, whereby UAMS-1 showed increasing BLI and biofilm growth until peaking at 9 h, while USA300 showed a rapid increase in BLI and biofilm formation at 6 h. The kinetics of biofilm formation for both UAMS-1 and USA300 were also supported and confirmed by qRT-PCR analysis of the 16S rRNA gene. Biofilms grown in our flow chamber in the plasma media were also demonstrated to involve an upregulation of the biofilm-forming-related genes icaA, fnbA, and alt. The BLI and SEM results from K-wire experiments revealed that the in vitro growth and biofilm formation by UAMS-1 and USA300 on stainless-steel and titanium surfaces were virtually identical. Conclusion We demonstrated a novel in vitro model for S. aureus biofilm formation with quantitative BLI and SEM outcome measures, and then used this model to demonstrate the presence of strain-specific phenotypes and its potential use to evaluate anti-microbial surfaces.
format	article
author	Werasak Sutipornpalangkul Kohei Nishitani Edward M. Schwarz
author_facet	Werasak Sutipornpalangkul Kohei Nishitani Edward M. Schwarz
author_sort	Werasak Sutipornpalangkul
title	Quantitative flow chamber system for evaluating in vitro biofilms and the kinetics of S. aureus biofilm formation in human plasma media
title_short	Quantitative flow chamber system for evaluating in vitro biofilms and the kinetics of S. aureus biofilm formation in human plasma media
title_full	Quantitative flow chamber system for evaluating in vitro biofilms and the kinetics of S. aureus biofilm formation in human plasma media
title_fullStr	Quantitative flow chamber system for evaluating in vitro biofilms and the kinetics of S. aureus biofilm formation in human plasma media
title_full_unstemmed	Quantitative flow chamber system for evaluating in vitro biofilms and the kinetics of S. aureus biofilm formation in human plasma media
title_sort	quantitative flow chamber system for evaluating in vitro biofilms and the kinetics of s. aureus biofilm formation in human plasma media
publisher	BMC
publishDate	2021
url	https://doaj.org/article/a968ebfd623240e2b5d22674308cf1c6
work_keys_str_mv	AT werasaksutipornpalangkul quantitativeflowchambersystemforevaluatinginvitrobiofilmsandthekineticsofsaureusbiofilmformationinhumanplasmamedia AT koheinishitani quantitativeflowchambersystemforevaluatinginvitrobiofilmsandthekineticsofsaureusbiofilmformationinhumanplasmamedia AT edwardmschwarz quantitativeflowchambersystemforevaluatinginvitrobiofilmsandthekineticsofsaureusbiofilmformationinhumanplasmamedia
_version_	1718429423038889984

Quantitative flow chamber system for evaluating in vitro biofilms and the kinetics of S. aureus biofilm formation in human plasma media

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