• toothbrush;
  • bacteria;
  • oral microbiology;
  • clinical trial


The aim of this in vivo study was to evaluate the efficacy of three antimicrobial solutions on the disinfection of toothbrushes after storage in closed containers.

Materials and Methods

Sixteen healthy subjects were enroled in this randomized cross-over clinical investigation. The study was conducted in four phases, in which mouthrinses (chlorhexidine gluconate-based or cetilpiridinium-based) and sterile tap water (control group) were used to individually store used toothbrushes in closed containers during 7 days of toothbrushing. Five toothbrushes were used as negative control for bacterial colonisation before contact with oral cavity. Conventional culture and DNA Checkerboard hybridization were used to detect bacterial contamination on the toothbrushes. Subsequently, the number of bacterial species on the bristles was estimated by the DNA Checkerboard method.


One toothbrush presented bacterial contamination in the negative control test. Both culture and DNA Checkerboard showed positive signals of bacterial contamination in the toothbrushes with no differences in the frequency of detection. The control group showed higher total bacterial counts when compared with the mouthrinse groups. Porphyromonas gingivalis had the highest bacterial count followed by Parvimonas micra.


Culture and DNA Checkerboard showed positive signals of bacterial contamination. Mouthrinses that contains 0.12% of chlorhexidine gluconate were more effective in reducing bacterial colonisation on the toothbrushes.