Development and pyrosequencing analysis of an in-vitro oral biofilm modelReport as inadecuate




Development and pyrosequencing analysis of an in-vitro oral biofilm model - Download this document for free, or read online. Document in PDF available to download.

BMC Microbiology

, 15:24

First Online: 10 February 2015Received: 10 November 2014Accepted: 27 January 2015DOI: 10.1186-s12866-015-0364-1

Cite this article as: Kistler, J.O., Pesaro, M. & Wade, W.G. BMC Microbiol 2015 15: 24. doi:10.1186-s12866-015-0364-1

Abstract

BackgroundDental caries and periodontal disease are the commonest bacterial diseases of man and can result in tooth loss. The principal method of prevention is the mechanical removal of dental plaque augmented by active agents incorporated into toothpastes and mouthrinses. In-vitro assays that include complex oral bacterial biofilms are required to accurately predict the efficacy of novel active agents in vivo. The aim of this study was to develop an oral biofilm model using the Calgary biofilm device CBD seeded with a natural saliva inoculum and analysed by next generation sequencing. The specific objectives were to determine the reproducibility and stability of the model by comparing the composition of the biofilms over time derived from i the same volunteers at different time points, and ii different panels of volunteers.

ResultsPyrosequencing yielded 280,093 sequences with a mean length of 432 bases after filtering. A mean of 320 and 250 OTUs were detected in pooled saliva and biofilm samples, respectively. Principal coordinates analysis PCoA plots based on community membership and structure showed that replicate biofilm samples were highly similar and clustered together. In addition, there were no significant differences between biofilms derived from the same panel at different times using analysis of molecular variance AMOVA. There were significant differences between biofilms from different panels AMOVA, P < 0.002. PCoA revealed that there was a shift in biofilm composition between seven and 14 days AMOVA, P < 0.001. Veillonella parvula, Veillonella atypica-dispar-parvula and Peptostreptococcus stomatis were the predominant OTUs detected in seven-day biofilms, whilst Prevotella oralis, V. parvula and Streptococcus constellatus were predominant in 14-day biofilms.

ConclusionsDiverse oral biofilms were successfully grown and maintained using the CBD. Biofilms derived from the same panel of volunteers were highly reproducible. This model could be used to screen both antimicrobial-containing oral care products and also novel approaches aiming to modify plaque composition, such as pre- or probiotics.

Keywords16S rRNA Bacteria Saliva Plaque Microbiome Microbiota Electronic supplementary materialThe online version of this article doi:10.1186-s12866-015-0364-1 contains supplementary material, which is available to authorized users.

Download fulltext PDF



Author: James O Kistler - Manuel Pesaro - William G Wade

Source: https://link.springer.com/







Related documents