Real-time assessment of Streptococcus mutans biofilm metabolism on resin composite

Fernando Luis Esteban Florez, Rochelle Denise Hiers, Kristin Smart, Jens Kreth, Fengxia Qi, Justin Merritt, Sharukh Soli Khajotia

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Objective The release of unpolymerized monomers and by-products of resin composites influences biofilm growth and confounds the measurement of metabolic activity. Current assays to measure biofilm viability have critical limitations and are typically not performed on relevant substrates. The objective of the present study was to determine the utility of firefly luciferase assay for quantification of the viability of intact biofilms on a resin composite substrate, and correlate the results with a standard method (viable colony counts). Methods Disk-shaped specimens of a dental resin composite were fabricated, wet-polished, UV-sterilized, and stored in water. Biofilms of Streptococcus mutans (strain UA159 modified by insertion of constitutively expressed firefly luc gene) were grown (1:500 dilution; anaerobic conditions, 24 h, 37 °C) in two media concentrations (0.35x and 0.65x THY medium supplemented with 0.1% sucrose; n = 15/group). An additional group of specimens with biofilms grown in 0.65x + sucrose media was treated with chlorhexidine gluconate solution to serve as the control group. Bioluminescence measurements of non-disrupted biofilms were obtained after addition of D-Luciferin substrate. The adherent biofilms were removed by sonication, and bioluminescence of sonicated bacteria was then measured. Viable colony counts were performed after plating sonicated bacteria on THY agar plates supplemented with spectinomycin. Bioluminescence values and cell counts were correlated using Spearman correlation tests (α = 0.05). Results Strong positive correlations between viable colony counts and bioluminescence values, both before- and after-sonication, validated the utility of this assay. Significance A novel non-disruptive, real-time bioluminescence assay is presented for quantification of intact S. mutans biofilms grown on a resin composite, and potentially on antibacterial materials and other types of dental biomaterials.

Original languageEnglish (US)
Pages (from-to)1263-1269
Number of pages7
JournalDental Materials
Volume32
Issue number10
DOIs
Publication statusPublished - Oct 1 2016

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Keywords

  • Biofilms
  • Bioluminescent assays
  • Colony count
  • Composite resins
  • Dental materials
  • Microbial
  • Streptococcus mutans

ASJC Scopus subject areas

  • Materials Science(all)
  • Dentistry(all)
  • Mechanics of Materials

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