Viable compositional analysis of an eleven species oral polymicrobial biofilm

Sherry, L. , Lappin, G., O'Donnell, L. E., Millhouse, E., Millington, O. R., Bradshaw, D. J., Axe, A. S., Williams, C., Nile, C. J. and Ramage, G. (2016) Viable compositional analysis of an eleven species oral polymicrobial biofilm. Frontiers in Microbiology, 7, 912. (doi: 10.3389/fmicb.2016.00912) (PMID:27375612) (PMCID:PMC4902011)

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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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Ramage, Professor Gordon and Williams, Dr Craig and Nile, Dr Christopher and Millhouse, Dr Emma and Sherry, Dr Leighann and Lappin, Mrs Gillian
Authors: Sherry, L., Lappin, G., O'Donnell, L. E., Millhouse, E., Millington, O. R., Bradshaw, D. J., Axe, A. S., Williams, C., Nile, C. J., and Ramage, G.
College/School:College of Medical Veterinary and Life Sciences > School of Infection & Immunity
College of Medical Veterinary and Life Sciences > School of Medicine, Dentistry & Nursing > Dental School
College of Medical Veterinary and Life Sciences > School of Molecular Biosciences
Journal Name:Frontiers in Microbiology
Publisher:Frontiers Media
ISSN:1664-302X
ISSN (Online):1664-302X
Published Online:10 June 2016
Copyright Holders:Copyright © 2016 Sherry, Lappin, O’Donnell, Millhouse, Millington, Bradshaw, Axe, Williams, Nile and Ramage
First Published:First published in Frontiers in Microbiology 7:912
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
614821The Natural History of Dentures: A Microbiological PerspectiveGordon RamageBiotechnology and Biological Sciences Research Council (BBSRC)BB/K501013/1SM - DENTAL SCHOOL