Perpetuation of subgingival biofilms in an in vitro model

This study evaluated the reproducibility of in-vitro-grown biofilms, initiated with subgingival plaque from patients with periodontal disease, and continued through several cycles by re-inoculating new biofilms from previously grown biofilms. Subgingival plaque samples from bleeding pockets along with saliva samples were collected from three patients with chronic periodontitis and perpetuated through seven cycles. Calcium hydroxyapatite disks were coated with sterilized saliva inoculated with dispersed subgingival plaque. The biofilms were grown anaerobically at 37°C for 10 days, and at specific intervals total viable bacteria were enumerated and the species present were analysed by DNA-DNA checkerboard hybridization. All cycles of biofilm growth occurred at similar rates and reached steady-state at day 7. No statistically or microbially significant differences were found for viable counts or species present, at the same period of maturation, among the different cycles. This study demonstrated that growth of certain target subgingival periodontal species in this biofilm model was reproducible and could be perpetuated in vitro through several cycles. The model could be useful in future studies to characterize different periodontopathogenic properties and biofilm interactions, especially in recolonization studies.