Validation of an in vitro biofilm model of supragingival plaque.
The study of biofilm structure and function mandates the use of model systems for which a host of environmental variables can be rigorously controlled. We describe a model of supragingival plaque containing Actinomyces naeslundii, Veillonella dispar, Fusobacterium nucleatum, Streptococcus sobrinus, and Streptococcus oralis wherein cells are cultivated anaerobically in a saliva-based medium on hydroxyapatite discs coated with a salivary pellicle, with material and pieces of apparatus common to all microbiology laboratories. After 0.5 hr, 16.5 hrs, 40.5 hrs, and 64.5 hrs, the composition of adherent biofilms was analyzed by culture techniques, live/dead fluorescence staining, and confocal laser scanning microscopy. Repeated independent trials demonstrated the repeatability of biofilm formation after 40.5 hrs and 64.5 hrs. Brief exposures of biofilms to chlorhexidine or Triclosan produced losses in viability similar to those observed in vivo. This biofilm model should prove very useful for pre-clinical testing of prospective anti-plaque agents at clinically relevant concentrations.
Institute for Oral Microbiology and General Immunology, Center for Dentistry, Oral Medicine, and Maxillofacial Surgery, University of Zürich, Switzerland. guggenhe@zzmk.unizh.ch
B
B
Guggenheim
E
E
Giertsen
P
P
Schüpbach
S
S
Shapiro
D000190:Actinomyces / Q000187:drug effects / Q000254:growth & development; D000704:Analysis of Variance; D000891:Anti-Infective Agents, Local / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects / Q000254:growth & development*; D002710:Chlorhexidine / Q000494:pharmacology; D015169:Colony Count, Microbial; D044622:Dental Pellicle; D003773:Dental Plaque / Q000382:microbiology*; D017886:Durapatite; D016967:Fusobacterium nucleatum / Q000187:drug effects / Q000254:growth & development; D006801:Humans; D008826:Microbial Sensitivity Tests; D018613:Microscopy, Confocal; D008954:Models, Biological; D015203:Reproducibility of Results; D012463:Saliva / Q000382:microbiology; D019533:Streptococcus oralis / Q000187:drug effects / Q000254:growth & development; D016986:Streptococcus sobrinus / Q000187:drug effects / Q000254:growth & development; D014260:Triclosan / Q000494:pharmacology; D014678:Veillonella / Q000187:drug effects / Q000254:growth & development
D000891:Anti-Infective Agents, Local; D014260:Triclosan; D017886:Durapatite; D002710:Chlorhexidine