Anin vitromodel ofFusobacterium nucleatumandPorphyromonas gingivalisin single- and dual-species biofilms

Abstract

Purpose: The goal of this study was to develop and validate a standardized in vitro pathogenic biofilm attached onto saliva-coated surfaces. Methods: Fusobacterium nucleatum (F. nucleatum) and Porphyromonas gingivalis (P. gingivalis)strains were grown under anaerobic conditions as single species and in dual-species cultures. Initially, the bacterial biomass was evaluated at 24 and 48 hours to determine the optimal timing for the adhesion phase onto saliva-coated polystyrene surfaces. Thereafter, biofilm development was assessed over time by crystal violet staining and scanning electron microscopy. Results:The data showed no significant difference in the overall biomass after 48 hours for P. gingivalis in single-and dual-species conditions. After adhesion, P. gingivalis in single-and dual-species biofilms accumulated a substantially higher biomass after 7 days of incubation than after 3 days, but no significant difference was found between 5 and 7 days. Although the biomass of the F. nucleatum biofilm was higher at 3 days, no difference was found at 3, 5, or 7 days of incubation. Conclusions: Polystyrene substrates from well plates work as a standard surface and provide reproducible results for in vitro biofilm models. Our biofilm model could serve as a reference point for studies investigating biofilms on different surfaces. Keywords: Bacterial adhesion; Biofilms; Fusobacterium nucleatum; Porphyromonas gingivalis INTRODUCTIONPeriodontal and peri-implant diseases are infections associated with complex biofilm structures that induce an inflammatory response, causing the destruction of connective tissue [1,2], The prevalence of periodontitis in adults is approximately 47% [3], making it the sixth most prevalent oral disease [4], while peri-implantitis was found to be present in 28% of subjects examined in a previous study [5]. Porphyromonas gingivalis is a red complex anaerobic Gram-negative bacterium, strongly associated with the advancement of both types of oral infection [6][7][8]. The mechanisms involved in bacterial colonization of natural and artificial surfaces, as well as the surrounding periodontal tissues, include direct attachment ORCID iDs

Antibiofilm effect of ozonized physiological saline solution on peri‐implant–related biofilm

“…The peri‐implant‐related biofilm model used was based on the Tavares et al study 51 . In this model, authors used two anaerobic bacterial species, P. gingivalis and F. nucleatum .…”

Low‐temperature plasma on peri‐implant–related biofilm and gingival tissue

“…Titanium discs were then incubated in 3 mL of the culture solution and after every 48 hours of incubation, 1.5 mL of the medium was removed, and an equal volume of fresh medium was added. After 5 days of biofilms development, they were treated according to the following groups: LTP1—plasma for 1 minute, LTP3—plasma for 3 minutes, CHX—positive control with 0.2% chlorhexidine for 1 minute, GAS—negative control with argon gas only (no plasma) for 3 minutes, and NEGATIVE—negative control without treatment. The experiment was conducted in duplicates in three independent occasions.…”

The Presence of Periodontal Pathogens in Gastric Cancer

“…Biofilm formation. F. nucleatum is regarded as a central organism for dental biofilm maturation due to its wide ability to aggregate with other microorganisms, such as Porphyromonas gingivalis [Tavares et al 2018]. It is considered as a bridge bacterium between early and late colonizers in dental plaque [He et al 2016].…”