Viable compositional analysis of an eleven species oral polymicrobial biofilm

Leighann Sherry, Gillian Lappin, Lindsay E. O'Donnell, Emma Millhouse, Owain R. Millington, David J. Bradshaw, Alyson S. Axe, Craig Williams, Christopher J. Nile, Gordon Ramage

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Purpose: Polymicrobial biofilms are abundant in clinical disease, particularly within the oral cavity. Creating complex biofilm models that recapitulate the polymicrobiality of oral disease are important in the development of new chemotherapeutic agents. In order to do this accurately we require the ability to undertake compositional analysis, in addition to determine individual cell viability, which is difficult using conventional microbiology. The aim of this study was to develop a defined multispecies denture biofilm model in vitro, and to assess viable compositional analysis following defined oral hygiene regimens.

Methods: An in vitro multispecies denture biofilm containing various oral commensal and pathogenic bacteria and yeast was created on poly (methyl methacrylate) (PMMA). Denture hygiene regimens tested against the biofilm model included brushing only, denture cleansing only and combinational brushing and denture cleansing. Biofilm composition and viability were assessed by culture (CFU) and molecular (qPCR) methodologies. Scanning electron microscopy and confocal laser scanning microscopy were also employed to visualize changes in denture biofilms following treatment.

Results: Combinational treatment of brushing and denture cleansing had the greatest impact on multispecies denture biofilms, reducing the number of live cells by more than 2 logs, and altering the overall composition in favor of streptococci. This was even more evident during the sequential testing, whereby daily sequential treatment reduced the total and live number of bacteria and yeast more than those treated intermittently. Bacteria and yeast remaining following treatment tended to aggregate in the pores of the PMMA, proving more difficult to fully eradicate the biofilm.

Conclusions: Overall, we are the first to develop a method to enable viable compositional analysis of an 11 species denture biofilm following chemotherapeutic challenge. We were able to demonstrate viable cell reduction and changes in population dynamics following evaluation of various denture cleansing regimens. Specifically, it was demonstrated that daily combinational treatment of brushing and cleansing proved to be the most advantageous denture hygiene regimen, however, residual organisms still remained within the pores of PMMA surface, which could act as a reservoir for further biofilm regrowth. We have identified an industry need for denture cleansing agents with the capacity to penetrate these pores and disaggregate these complex biofilm consortia.
LanguageEnglish
Article number912
Number of pages13
JournalFrontiers in Microbiology
Volume7
DOIs
Publication statusPublished - 10 Jun 2016

Fingerprint

Dentures
Biofilms
Polymethyl Methacrylate
Yeasts
Bacteria
Hygiene
Mouth Diseases
Therapeutics
Oral Hygiene
Population Dynamics
Microbiology
Streptococcus
Confocal Microscopy
Detergents
Electron Scanning Microscopy
Mouth
Cell Survival
Industry
Cell Count

Keywords

  • biofilm
  • polymicrobial
  • viability
  • denture
  • oral diseases
  • oral hygiene regimens
  • denture cleansing regimens
  • viable cell reduction

Cite this

Sherry, L., Lappin, G., O'Donnell, L. E., Millhouse, E., Millington, O. R., Bradshaw, D. J., ... Ramage, G. (2016). Viable compositional analysis of an eleven species oral polymicrobial biofilm. Frontiers in Microbiology, 7, [912]. https://doi.org/10.3389/fmicb.2016.00912
Sherry, Leighann ; Lappin, Gillian ; O'Donnell, Lindsay E. ; Millhouse, Emma ; Millington, Owain R. ; Bradshaw, David J. ; Axe, Alyson S. ; Williams, Craig ; Nile, Christopher J. ; Ramage, Gordon. / Viable compositional analysis of an eleven species oral polymicrobial biofilm. In: Frontiers in Microbiology. 2016 ; Vol. 7.
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Sherry, L, Lappin, G, O'Donnell, LE, Millhouse, E, Millington, OR, Bradshaw, DJ, Axe, AS, Williams, C, Nile, CJ & Ramage, G 2016, 'Viable compositional analysis of an eleven species oral polymicrobial biofilm' Frontiers in Microbiology, vol. 7, 912. https://doi.org/10.3389/fmicb.2016.00912

Viable compositional analysis of an eleven species oral polymicrobial biofilm. / Sherry, Leighann; Lappin, Gillian; O'Donnell, Lindsay E.; Millhouse, Emma; Millington, Owain R.; Bradshaw, David J.; Axe, Alyson S.; Williams, Craig; Nile, Christopher J.; Ramage, Gordon.

In: Frontiers in Microbiology, Vol. 7, 912, 10.06.2016.

Research output: Contribution to journalArticle

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AU - Sherry, Leighann

AU - Lappin, Gillian

AU - O'Donnell, Lindsay E.

AU - Millhouse, Emma

AU - Millington, Owain R.

AU - Bradshaw, David J.

AU - Axe, Alyson S.

AU - Williams, Craig

AU - Nile, Christopher J.

AU - Ramage, Gordon

N1 - This Document is Protected by copyright and was first published by Frontiers. All rights reserved. it is reproduced with permission

PY - 2016/6/10

Y1 - 2016/6/10

N2 - Purpose: Polymicrobial biofilms are abundant in clinical disease, particularly within the oral cavity. Creating complex biofilm models that recapitulate the polymicrobiality of oral disease are important in the development of new chemotherapeutic agents. In order to do this accurately we require the ability to undertake compositional analysis, in addition to determine individual cell viability, which is difficult using conventional microbiology. The aim of this study was to develop a defined multispecies denture biofilm model in vitro, and to assess viable compositional analysis following defined oral hygiene regimens.Methods: An in vitro multispecies denture biofilm containing various oral commensal and pathogenic bacteria and yeast was created on poly (methyl methacrylate) (PMMA). Denture hygiene regimens tested against the biofilm model included brushing only, denture cleansing only and combinational brushing and denture cleansing. Biofilm composition and viability were assessed by culture (CFU) and molecular (qPCR) methodologies. Scanning electron microscopy and confocal laser scanning microscopy were also employed to visualize changes in denture biofilms following treatment.Results: Combinational treatment of brushing and denture cleansing had the greatest impact on multispecies denture biofilms, reducing the number of live cells by more than 2 logs, and altering the overall composition in favor of streptococci. This was even more evident during the sequential testing, whereby daily sequential treatment reduced the total and live number of bacteria and yeast more than those treated intermittently. Bacteria and yeast remaining following treatment tended to aggregate in the pores of the PMMA, proving more difficult to fully eradicate the biofilm.Conclusions: Overall, we are the first to develop a method to enable viable compositional analysis of an 11 species denture biofilm following chemotherapeutic challenge. We were able to demonstrate viable cell reduction and changes in population dynamics following evaluation of various denture cleansing regimens. Specifically, it was demonstrated that daily combinational treatment of brushing and cleansing proved to be the most advantageous denture hygiene regimen, however, residual organisms still remained within the pores of PMMA surface, which could act as a reservoir for further biofilm regrowth. We have identified an industry need for denture cleansing agents with the capacity to penetrate these pores and disaggregate these complex biofilm consortia.

AB - Purpose: Polymicrobial biofilms are abundant in clinical disease, particularly within the oral cavity. Creating complex biofilm models that recapitulate the polymicrobiality of oral disease are important in the development of new chemotherapeutic agents. In order to do this accurately we require the ability to undertake compositional analysis, in addition to determine individual cell viability, which is difficult using conventional microbiology. The aim of this study was to develop a defined multispecies denture biofilm model in vitro, and to assess viable compositional analysis following defined oral hygiene regimens.Methods: An in vitro multispecies denture biofilm containing various oral commensal and pathogenic bacteria and yeast was created on poly (methyl methacrylate) (PMMA). Denture hygiene regimens tested against the biofilm model included brushing only, denture cleansing only and combinational brushing and denture cleansing. Biofilm composition and viability were assessed by culture (CFU) and molecular (qPCR) methodologies. Scanning electron microscopy and confocal laser scanning microscopy were also employed to visualize changes in denture biofilms following treatment.Results: Combinational treatment of brushing and denture cleansing had the greatest impact on multispecies denture biofilms, reducing the number of live cells by more than 2 logs, and altering the overall composition in favor of streptococci. This was even more evident during the sequential testing, whereby daily sequential treatment reduced the total and live number of bacteria and yeast more than those treated intermittently. Bacteria and yeast remaining following treatment tended to aggregate in the pores of the PMMA, proving more difficult to fully eradicate the biofilm.Conclusions: Overall, we are the first to develop a method to enable viable compositional analysis of an 11 species denture biofilm following chemotherapeutic challenge. We were able to demonstrate viable cell reduction and changes in population dynamics following evaluation of various denture cleansing regimens. Specifically, it was demonstrated that daily combinational treatment of brushing and cleansing proved to be the most advantageous denture hygiene regimen, however, residual organisms still remained within the pores of PMMA surface, which could act as a reservoir for further biofilm regrowth. We have identified an industry need for denture cleansing agents with the capacity to penetrate these pores and disaggregate these complex biofilm consortia.

KW - biofilm

KW - polymicrobial

KW - viability

KW - denture

KW - oral diseases

KW - oral hygiene regimens

KW - denture cleansing regimens

KW - viable cell reduction

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SN - 1664-302X

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Sherry L, Lappin G, O'Donnell LE, Millhouse E, Millington OR, Bradshaw DJ et al. Viable compositional analysis of an eleven species oral polymicrobial biofilm. Frontiers in Microbiology. 2016 Jun 10;7. 912. https://doi.org/10.3389/fmicb.2016.00912