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KMID : 1044820180480010012
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Journal of Periodontal & Implant Science
2018 Volume.48 No. 1 p.12 ~ p.21
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An in vitro model of Fusobacterium nucleatum and Porphyromonas gingivalis in single- and dual-species biofilms
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Tavares Livia Jacovassi
Klein Marlise Inez
Panariello Beatriz Helena Dias
de Avila Erica Dorigatti
Pavarina Ana Claudia
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Abstract
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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.
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KEYWORD
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Bacterial adhesion, Biofilms, Fusobacterium nucleatum, Porphyromonas gingivalis
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