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Modified SHI medium supports growth of a disease‐state subgingival polymicrobial community in vitro

Developing a laboratory model of oral polymicrobial communities is essential for in vitro studies of the transition from healthy to diseased oral plaque. SHI medium is an enriched growth medium capable of supporting in vitro biofilms with similar diversity to healthy supragingival inocula; however,...

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Autores principales: Lamont, Eleanor I., Gadkari, Archita, Kerns, Kristopher A., To, Thao T., Daubert, Diane, Kotsakis, Georgios, Bor, Batbileg, He, Xuesong, McLean, Jeffrey S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7984074/
https://www.ncbi.nlm.nih.gov/pubmed/33174294
http://dx.doi.org/10.1111/omi.12323
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author Lamont, Eleanor I.
Gadkari, Archita
Kerns, Kristopher A.
To, Thao T.
Daubert, Diane
Kotsakis, Georgios
Bor, Batbileg
He, Xuesong
McLean, Jeffrey S.
author_facet Lamont, Eleanor I.
Gadkari, Archita
Kerns, Kristopher A.
To, Thao T.
Daubert, Diane
Kotsakis, Georgios
Bor, Batbileg
He, Xuesong
McLean, Jeffrey S.
author_sort Lamont, Eleanor I.
collection PubMed
description Developing a laboratory model of oral polymicrobial communities is essential for in vitro studies of the transition from healthy to diseased oral plaque. SHI medium is an enriched growth medium capable of supporting in vitro biofilms with similar diversity to healthy supragingival inocula; however, this medium does not maintain the diversity of gram‐negative bacteria more associated with subgingival plaque. Here, we systematically modified SHI medium components to investigate the impacts of varying nutrients and develop a medium capable of supporting a specific disease‐state subgingival community. A diseased subgingival plaque sample was inoculated in SHI medium with increasing concentrations of sucrose (0%, 0.1%, 0.5%), fetal bovine serum (FBS) (0%, 10%, 20%, 30%, 50%), and mucin (0.1, 2.5, 8.0 g/L) and grown for 48 hrs, then the 16S rRNA profiles of the resulting biofilms were examined. In total, these conditions were able to capture 89 of the 119 species and 43 of the 51 genera found in the subgingival inoculum. Interestingly, biofilms grown in high sucrose media, although dominated by acidogenic Firmicutes with a low final pH, contained several uncultured taxa from the genus Treponema, information that may aid culturing these periodontitis‐associated fastidious organisms. Biofilms grown in a modified medium (here named subSHI‐v1 medium) with 0.1% sucrose and 10% FBS had a high diversity closest to the inoculum and maintained greater proportions of many gram‐negative species of interest from the subgingival periodontal pocket (including members of the genera Prevotella and Treponema, and the Candidate Phyla Radiation phylum Saccharibacteria), and therefore best represented the disease community.
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spelling pubmed-79840742021-03-24 Modified SHI medium supports growth of a disease‐state subgingival polymicrobial community in vitro Lamont, Eleanor I. Gadkari, Archita Kerns, Kristopher A. To, Thao T. Daubert, Diane Kotsakis, Georgios Bor, Batbileg He, Xuesong McLean, Jeffrey S. Mol Oral Microbiol Original Articles Developing a laboratory model of oral polymicrobial communities is essential for in vitro studies of the transition from healthy to diseased oral plaque. SHI medium is an enriched growth medium capable of supporting in vitro biofilms with similar diversity to healthy supragingival inocula; however, this medium does not maintain the diversity of gram‐negative bacteria more associated with subgingival plaque. Here, we systematically modified SHI medium components to investigate the impacts of varying nutrients and develop a medium capable of supporting a specific disease‐state subgingival community. A diseased subgingival plaque sample was inoculated in SHI medium with increasing concentrations of sucrose (0%, 0.1%, 0.5%), fetal bovine serum (FBS) (0%, 10%, 20%, 30%, 50%), and mucin (0.1, 2.5, 8.0 g/L) and grown for 48 hrs, then the 16S rRNA profiles of the resulting biofilms were examined. In total, these conditions were able to capture 89 of the 119 species and 43 of the 51 genera found in the subgingival inoculum. Interestingly, biofilms grown in high sucrose media, although dominated by acidogenic Firmicutes with a low final pH, contained several uncultured taxa from the genus Treponema, information that may aid culturing these periodontitis‐associated fastidious organisms. Biofilms grown in a modified medium (here named subSHI‐v1 medium) with 0.1% sucrose and 10% FBS had a high diversity closest to the inoculum and maintained greater proportions of many gram‐negative species of interest from the subgingival periodontal pocket (including members of the genera Prevotella and Treponema, and the Candidate Phyla Radiation phylum Saccharibacteria), and therefore best represented the disease community. John Wiley and Sons Inc. 2020-12-03 2021-02 /pmc/articles/PMC7984074/ /pubmed/33174294 http://dx.doi.org/10.1111/omi.12323 Text en © 2020 The Authors. Molecular Oral Microbiology published by John Wiley & Sons Ltd This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Lamont, Eleanor I.
Gadkari, Archita
Kerns, Kristopher A.
To, Thao T.
Daubert, Diane
Kotsakis, Georgios
Bor, Batbileg
He, Xuesong
McLean, Jeffrey S.
Modified SHI medium supports growth of a disease‐state subgingival polymicrobial community in vitro
title Modified SHI medium supports growth of a disease‐state subgingival polymicrobial community in vitro
title_full Modified SHI medium supports growth of a disease‐state subgingival polymicrobial community in vitro
title_fullStr Modified SHI medium supports growth of a disease‐state subgingival polymicrobial community in vitro
title_full_unstemmed Modified SHI medium supports growth of a disease‐state subgingival polymicrobial community in vitro
title_short Modified SHI medium supports growth of a disease‐state subgingival polymicrobial community in vitro
title_sort modified shi medium supports growth of a disease‐state subgingival polymicrobial community in vitro
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7984074/
https://www.ncbi.nlm.nih.gov/pubmed/33174294
http://dx.doi.org/10.1111/omi.12323
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