Cargando…

Polymicrobial synergy stimulates Porphyromonas gingivalis survival and gingipain expression in a multi-species subgingival community

BACKGROUND: Dysbiosis in subgingival microbial communities, resulting from increased inflammatory transudate from the gingival tissues, is an important factor in initiation and development of periodontitis. Dysbiotic communities are characterized by increased numbers of bacteria that exploit the ser...

Descripción completa

Detalles Bibliográficos
Autores principales: Davies, Julia R., Kad, Trupti, Neilands, Jessica, Kinnby, Bertil, Prgomet, Zdenka, Bengtsson, Torbjörn, Khalaf, Hazem, Svensäter, Gunnel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8672593/
https://www.ncbi.nlm.nih.gov/pubmed/34911531
http://dx.doi.org/10.1186/s12903-021-01971-9
_version_ 1784615384917213184
author Davies, Julia R.
Kad, Trupti
Neilands, Jessica
Kinnby, Bertil
Prgomet, Zdenka
Bengtsson, Torbjörn
Khalaf, Hazem
Svensäter, Gunnel
author_facet Davies, Julia R.
Kad, Trupti
Neilands, Jessica
Kinnby, Bertil
Prgomet, Zdenka
Bengtsson, Torbjörn
Khalaf, Hazem
Svensäter, Gunnel
author_sort Davies, Julia R.
collection PubMed
description BACKGROUND: Dysbiosis in subgingival microbial communities, resulting from increased inflammatory transudate from the gingival tissues, is an important factor in initiation and development of periodontitis. Dysbiotic communities are characterized by increased numbers of bacteria that exploit the serum-like transudate for nutrients, giving rise to a proteolytic community phenotype. Here we investigate the contribution of interactions between members of a sub-gingival community to survival and development of virulence in a serum environment—modelling that in the subgingival pocket. METHODS: Growth and proteolytic activity of three Porphyromonas gingivalis strains in nutrient broth or a serum environment were assessed using A(600) and a fluorescent protease substrate, respectively. Adherence of P. gingivalis strains to serum-coated surfaces was studied with confocal microscopy and 2D-gel electrophoresis of bacterial supernatants used to investigate extracellular proteins. A model multi-species sub-gingival community containing Fusobacterium nucleatum, Streptococcus constellatus, Parvimonas micra with wild type or isogenic mutants of P. gingivalis was then created and growth and proteolytic activity in serum assessed as above. Community composition over time was monitored using culture techniques and qPCR. RESULTS: The P. gingivalis strains showed different growth rates in nutrient broth related to the level of proteolytic activity (largely gingipains) in the cultures. Despite being able to adhere to serum-coated surfaces, none of the strains was able to grow alone in a serum environment. Together in the subgingival consortium however, all the included species were able to grow in the serum environment and the community adopted a proteolytic phenotype. Inclusion of P. gingivalis strains lacking gingipains in the consortium revealed that community growth was facilitated by Rgp gingipain from P. gingivalis. CONCLUSIONS: In the multi-species consortium, growth was facilitated by the wild-type and Rgp-expressing strains of P. gingivalis, suggesting that Rgp is involved in delivery of nutrients to the whole community through degradation of complex protein substrates in serum. Whereas they are constitutively expressed by P. gingivalis in nutrient broth, gingipain expression in the model periodontal pocket environment (serum) appeared to be orchestrated through signaling to P. gingivalis from other members of the community, a phenomenon which then promoted growth of the whole community. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12903-021-01971-9.
format Online
Article
Text
id pubmed-8672593
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-86725932021-12-17 Polymicrobial synergy stimulates Porphyromonas gingivalis survival and gingipain expression in a multi-species subgingival community Davies, Julia R. Kad, Trupti Neilands, Jessica Kinnby, Bertil Prgomet, Zdenka Bengtsson, Torbjörn Khalaf, Hazem Svensäter, Gunnel BMC Oral Health Research BACKGROUND: Dysbiosis in subgingival microbial communities, resulting from increased inflammatory transudate from the gingival tissues, is an important factor in initiation and development of periodontitis. Dysbiotic communities are characterized by increased numbers of bacteria that exploit the serum-like transudate for nutrients, giving rise to a proteolytic community phenotype. Here we investigate the contribution of interactions between members of a sub-gingival community to survival and development of virulence in a serum environment—modelling that in the subgingival pocket. METHODS: Growth and proteolytic activity of three Porphyromonas gingivalis strains in nutrient broth or a serum environment were assessed using A(600) and a fluorescent protease substrate, respectively. Adherence of P. gingivalis strains to serum-coated surfaces was studied with confocal microscopy and 2D-gel electrophoresis of bacterial supernatants used to investigate extracellular proteins. A model multi-species sub-gingival community containing Fusobacterium nucleatum, Streptococcus constellatus, Parvimonas micra with wild type or isogenic mutants of P. gingivalis was then created and growth and proteolytic activity in serum assessed as above. Community composition over time was monitored using culture techniques and qPCR. RESULTS: The P. gingivalis strains showed different growth rates in nutrient broth related to the level of proteolytic activity (largely gingipains) in the cultures. Despite being able to adhere to serum-coated surfaces, none of the strains was able to grow alone in a serum environment. Together in the subgingival consortium however, all the included species were able to grow in the serum environment and the community adopted a proteolytic phenotype. Inclusion of P. gingivalis strains lacking gingipains in the consortium revealed that community growth was facilitated by Rgp gingipain from P. gingivalis. CONCLUSIONS: In the multi-species consortium, growth was facilitated by the wild-type and Rgp-expressing strains of P. gingivalis, suggesting that Rgp is involved in delivery of nutrients to the whole community through degradation of complex protein substrates in serum. Whereas they are constitutively expressed by P. gingivalis in nutrient broth, gingipain expression in the model periodontal pocket environment (serum) appeared to be orchestrated through signaling to P. gingivalis from other members of the community, a phenomenon which then promoted growth of the whole community. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12903-021-01971-9. BioMed Central 2021-12-15 /pmc/articles/PMC8672593/ /pubmed/34911531 http://dx.doi.org/10.1186/s12903-021-01971-9 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Davies, Julia R.
Kad, Trupti
Neilands, Jessica
Kinnby, Bertil
Prgomet, Zdenka
Bengtsson, Torbjörn
Khalaf, Hazem
Svensäter, Gunnel
Polymicrobial synergy stimulates Porphyromonas gingivalis survival and gingipain expression in a multi-species subgingival community
title Polymicrobial synergy stimulates Porphyromonas gingivalis survival and gingipain expression in a multi-species subgingival community
title_full Polymicrobial synergy stimulates Porphyromonas gingivalis survival and gingipain expression in a multi-species subgingival community
title_fullStr Polymicrobial synergy stimulates Porphyromonas gingivalis survival and gingipain expression in a multi-species subgingival community
title_full_unstemmed Polymicrobial synergy stimulates Porphyromonas gingivalis survival and gingipain expression in a multi-species subgingival community
title_short Polymicrobial synergy stimulates Porphyromonas gingivalis survival and gingipain expression in a multi-species subgingival community
title_sort polymicrobial synergy stimulates porphyromonas gingivalis survival and gingipain expression in a multi-species subgingival community
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8672593/
https://www.ncbi.nlm.nih.gov/pubmed/34911531
http://dx.doi.org/10.1186/s12903-021-01971-9
work_keys_str_mv AT daviesjuliar polymicrobialsynergystimulatesporphyromonasgingivalissurvivalandgingipainexpressioninamultispeciessubgingivalcommunity
AT kadtrupti polymicrobialsynergystimulatesporphyromonasgingivalissurvivalandgingipainexpressioninamultispeciessubgingivalcommunity
AT neilandsjessica polymicrobialsynergystimulatesporphyromonasgingivalissurvivalandgingipainexpressioninamultispeciessubgingivalcommunity
AT kinnbybertil polymicrobialsynergystimulatesporphyromonasgingivalissurvivalandgingipainexpressioninamultispeciessubgingivalcommunity
AT prgometzdenka polymicrobialsynergystimulatesporphyromonasgingivalissurvivalandgingipainexpressioninamultispeciessubgingivalcommunity
AT bengtssontorbjorn polymicrobialsynergystimulatesporphyromonasgingivalissurvivalandgingipainexpressioninamultispeciessubgingivalcommunity
AT khalafhazem polymicrobialsynergystimulatesporphyromonasgingivalissurvivalandgingipainexpressioninamultispeciessubgingivalcommunity
AT svensatergunnel polymicrobialsynergystimulatesporphyromonasgingivalissurvivalandgingipainexpressioninamultispeciessubgingivalcommunity