Antimicrobial activity of hydroxyl radicals generated by hydrogen peroxide photolysis against Streptococcus mutans biofilm

Int J Antimicrob Agents. 2016 Oct;48(4):373-80. doi: 10.1016/j.ijantimicag.2016.06.007. Epub 2016 Jul 15.

Abstract

Prevention of dental caries with maximum conservation of intact tooth substance remains a challenge in dentistry. The present study aimed to evaluate the antimicrobial effect of H2O2 photolysis on Streptococcus mutans biofilm, which may be a novel antimicrobial chemotherapy for treating caries. S. mutans biofilm was grown on disk-shaped hydroxyapatite specimens. After 1-24 h of incubation, growth was assessed by confocal laser scanning microscopy and viable bacterial counting. Resistance to antibiotics (amoxicillin and erythromycin) was evaluated by comparing bactericidal effects on the biofilm with those on planktonic bacteria. To evaluate the effect of the antimicrobial technique, the biofilm was immersed in 3% H2O2 and was irradiated with an LED at 365 nm for 1 min. Viable bacterial counts in the biofilm were determined by colony counting. The thickness and surface coverage of S. mutans biofilm increased with time, whereas viable bacterial counts plateaued after 6 h. When 12- and 24-h-old biofilms were treated with the minimum concentration of antibiotics that killed viable planktonic bacteria with 3 log reduction, their viable counts were not significantly decreased, suggesting the biofilm acquired antibiotic resistance by increasing its thickness. By contrast, hydroxyl radicals generated by photolysis of 3% H2O2 effectively killed S. mutans in 24-h-old biofilm, with greater than 5 log reduction. The technique based on H2O2 photolysis is a potentially powerful adjunctive antimicrobial chemotherapy for caries treatment.

Keywords: Biofilm; Confocal laser scanning microscopy; Hydrogen peroxide; Hydroxyl radical; Photolysis; Streptococcus mutans.

MeSH terms

  • Anti-Infective Agents, Local / pharmacology*
  • Biofilms / drug effects*
  • Colony Count, Microbial
  • Hydrogen Peroxide / metabolism*
  • Hydroxyl Radical / pharmacology*
  • Microbial Viability / drug effects
  • Microscopy, Confocal
  • Photolysis*
  • Streptococcus mutans / drug effects*
  • Streptococcus mutans / physiology

Substances

  • Anti-Infective Agents, Local
  • Hydroxyl Radical
  • Hydrogen Peroxide