Multispecies oral biofilm and identification of components as treatment target

Endodontic infections involve a multispecies biofilm, making it difficult to choose an antimicrobial treatment. Characteristics such as the pathogens involved and number of microorganisms, nutrients, material surface to develop the biofilm, flow and oxygenation conditions are important for biofilm development using in vitro models. To develop a standardized biofilm model, which replicates the main features (chemical, microbiological, and topographical) of an infected root canal tooth to detect components as treatment target. Clinical strains of Enterococcus faecalis, Candida albicans, and Actinomyces israelii were isolated, and a multispecies biofilm was developed using continuous laminar flow reactors under anaerobic conditions in human dental roots. The microbiological composition was determined by counting colony-forming units and scanning electron microscope micrographs. In addition, the chemical composition of the exopolymeric matrix was determined by vibrational Raman spectroscopy and liquid chromatography of biofilm supernatant treated with enzyme. E. faecalis turned out to be the main microorganism in mature biofilm, this was related to the presence of β-galactosidase detected by vibrational Raman spectroscopy. After the enzymatic treatment of the extracellular polymeric substance, the presence of mannose and glucose was established. The present work contributes to better understanding of standard conditions to develop a multispecies biofilm in human dental roots, which could have an impact on the generation of new root canal disinfection techniques in endodontic pathologies. •Standard conditions and anaerobiosis help to develop an oral biofilm as target to inhibition•Knowledge of chemical, microbiological, and topographical composition of biofilm in tooth•Infected root canal tooth to detect components as treatment target