Objectives: The main aim of our work was to get one step closer to the in vivo conditions. We started from a multispecies static biofilm model containing five different bacteria, implementing specific enhancements. Our second goal was to improve the analysis of such biofilms regarding collection and identification.

Material and Methods: We started from a multispecies static model with five oral strains growing on hydroxyapatite discs to improve it on multiple points. We modified culture conditions and added two more strains. We also changed bacteria collection, which evolved from manually scrapping the discs surface to the combination of ultrasonic and mechanical harvesting. In a further another step, we developed a dynamic model implementing the above changes with a continuous supply of medium flow and waste disposal. Different methods have been evaluated to monitor the presence of all the species within the biofilms, and to quantify them: gram staining, PCR, MALDI-TOF-MS, qPCR.

Results: The modifications brought to our static model confirmed its reproducibility. Even if improvements need to be made, our dynamic model of oral biofilm is already a good alternative to more sophisticated and expensive models.

Conclusions: This new oral biofilm model represents the premises of another way to study the environmental variations effects on bacterial development, its larger application will result in a better understanding of oral health significant factors.