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Consistent and reproducible long-term in vitro growth of health and disease-associated oral subgingival biofilms

BACKGROUND: Several in vitro oral biofilm growth systems can reliably construct oral microbiome communities in culture, yet their stability and reproducibility through time has not been well characterized. Long-term in vitro growth of natural biofilms would enable use of these biofilms in both in vi...

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Autores principales: Velsko, Irina M., Shaddox, Luciana M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6042318/
https://www.ncbi.nlm.nih.gov/pubmed/29996764
http://dx.doi.org/10.1186/s12866-018-1212-x
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author Velsko, Irina M.
Shaddox, Luciana M.
author_facet Velsko, Irina M.
Shaddox, Luciana M.
author_sort Velsko, Irina M.
collection PubMed
description BACKGROUND: Several in vitro oral biofilm growth systems can reliably construct oral microbiome communities in culture, yet their stability and reproducibility through time has not been well characterized. Long-term in vitro growth of natural biofilms would enable use of these biofilms in both in vitro and in vivo studies that require complex microbial communities with minimal variation over a period of time. Understanding biofilm community dynamics in continuous culture, and whether they maintain distinct signatures of health and disease, is necessary to determine the reliability and applicability of such models to broader studies. To this end, we performed next-generation sequencing on biofilms grown from healthy and disease-site subgingival plaque for 80 days to assess stability and reliability of continuous oral biofilm growth. RESULTS: Biofilms were grown from subgingival plaque collected from periodontitis-affected sites and healthy individuals for ten eight-day long generations, using hydroxyapatite disks. The bacterial community in each generation was determined using Human Oral Microbe Identification by Next-Generation Sequencing (HOMINGS) technology, and analyzed in QIIME. Profiles were steady through the ten generations, as determined by species abundance and prevalence, Spearman’s correlation coefficient, and Faith’s phylogenetic distance, with slight variation predominantly in low abundance species. Community profiles were distinct between healthy and disease site-derived biofilms as demonstrated by weighted UniFrac distance throughout the ten generations. Differentially abundant species between healthy and disease site-derived biofilms were consistent throughout the generations. CONCLUSIONS: Healthy and disease site-derived biofilms can reliably maintain consistent communities through ten generations of in vitro growth. These communities maintain signatures of health and disease and of individual donors despite culture in identical environments. This subgingival oral biofilm growth and perpetuation model may prove useful to studies involving oral infection or cell stimulation, or those measuring microbial interactions, which require the same biofilms over a period of time. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12866-018-1212-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-60423182018-07-13 Consistent and reproducible long-term in vitro growth of health and disease-associated oral subgingival biofilms Velsko, Irina M. Shaddox, Luciana M. BMC Microbiol Research Article BACKGROUND: Several in vitro oral biofilm growth systems can reliably construct oral microbiome communities in culture, yet their stability and reproducibility through time has not been well characterized. Long-term in vitro growth of natural biofilms would enable use of these biofilms in both in vitro and in vivo studies that require complex microbial communities with minimal variation over a period of time. Understanding biofilm community dynamics in continuous culture, and whether they maintain distinct signatures of health and disease, is necessary to determine the reliability and applicability of such models to broader studies. To this end, we performed next-generation sequencing on biofilms grown from healthy and disease-site subgingival plaque for 80 days to assess stability and reliability of continuous oral biofilm growth. RESULTS: Biofilms were grown from subgingival plaque collected from periodontitis-affected sites and healthy individuals for ten eight-day long generations, using hydroxyapatite disks. The bacterial community in each generation was determined using Human Oral Microbe Identification by Next-Generation Sequencing (HOMINGS) technology, and analyzed in QIIME. Profiles were steady through the ten generations, as determined by species abundance and prevalence, Spearman’s correlation coefficient, and Faith’s phylogenetic distance, with slight variation predominantly in low abundance species. Community profiles were distinct between healthy and disease site-derived biofilms as demonstrated by weighted UniFrac distance throughout the ten generations. Differentially abundant species between healthy and disease site-derived biofilms were consistent throughout the generations. CONCLUSIONS: Healthy and disease site-derived biofilms can reliably maintain consistent communities through ten generations of in vitro growth. These communities maintain signatures of health and disease and of individual donors despite culture in identical environments. This subgingival oral biofilm growth and perpetuation model may prove useful to studies involving oral infection or cell stimulation, or those measuring microbial interactions, which require the same biofilms over a period of time. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12866-018-1212-x) contains supplementary material, which is available to authorized users. BioMed Central 2018-07-11 /pmc/articles/PMC6042318/ /pubmed/29996764 http://dx.doi.org/10.1186/s12866-018-1212-x Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Velsko, Irina M.
Shaddox, Luciana M.
Consistent and reproducible long-term in vitro growth of health and disease-associated oral subgingival biofilms
title Consistent and reproducible long-term in vitro growth of health and disease-associated oral subgingival biofilms
title_full Consistent and reproducible long-term in vitro growth of health and disease-associated oral subgingival biofilms
title_fullStr Consistent and reproducible long-term in vitro growth of health and disease-associated oral subgingival biofilms
title_full_unstemmed Consistent and reproducible long-term in vitro growth of health and disease-associated oral subgingival biofilms
title_short Consistent and reproducible long-term in vitro growth of health and disease-associated oral subgingival biofilms
title_sort consistent and reproducible long-term in vitro growth of health and disease-associated oral subgingival biofilms
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6042318/
https://www.ncbi.nlm.nih.gov/pubmed/29996764
http://dx.doi.org/10.1186/s12866-018-1212-x
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