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Biofilms are complex three-dimensional communities of microorganisms adhering to a surface and encased in a protective exopolymeric substance. The anatomical complexities in the root canal systems shelter the adhering bacteria in a biofilm from cleaning and shaping procedures. Microbial communities growing in biofilms are remarkably more difficult to eradicate with antimicrobial agents and microorganisms in mature biofilms can be extremely resistant for reasons that have yet to be fully explained. As the influence of root canal dentin and other surfaces on the expression of novel biofilm phenotypes has not yet been touched upon, conclusions and decisions reached from studies of monoculture biofilm in the laboratory must be taken with great caution. While none of the irrigating solutions/disinfecting agents presently used in endodontic treatment are able to do all of the required tasks,many of them can have an impact on biofilms, either by dissolving the film, killing the microbesresiding in the biofilm or by helping to break down or detach the film from the surface.