title: A dual-species microbial model for studying the dynamics between oral streptococci and periodontal pathogens during biofilm development on titanium surfaces by flow cytometry.
abstract: The association of the pioneer organisms Streptococcus mutans ATCC 25175 or Streptococcus oralis ATCC 9811 with secondary colonizers Fusobacterium nucleatum ATCC 25586 or Porphyromonas gingivalis ATCC 33277 during biofilm development on titanium surfaces was evaluated by flow cytometry (FCM) using specific polyclonal antibodies. ELISA and FCM were employed, revealing high antibody sensitivity and specificity. Biofilm formation of four dual-species combinations was analyzed by crystal violet staining, while the association between streptococci and periodontal pathogens was assessed using FCM. Dual-species association between S. oralis and P. gingivalis or F. nucleatum showed a proportional decrease in S. oralis during biofilm development, with a concomitant increase in P. gingivalis or F. nucleatum. This trend was not observed in either of the dual-species associations of S. mutans with the periodontal pathogens. Our dual-species microbial model, which employed FCM, proved to be useful in the study of partnerships between bacteria in oral associations, showing that the presence of primary colonizers is required for the establishment of secondary colonizers in biofilms. The proposed experimental approach is technically simple to prepare and analyze, and also proved to be reproducible; hence, it is well-suited for investigating the development and dynamics of oral communities.
authors.affiliation: Department of Biomolecular Science, Via Saffi 2, University of Urbino "Carlo Bo", 61029 Urbino, Italy.
authors.forename: Anita
authors.initials: A
authors.lastname: Manti
authors.affiliation: Department of Biomolecular Science, Via Saffi 2, University of Urbino "Carlo Bo", 61029 Urbino, Italy.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Ciandrini
authors.affiliation: Department of Biomolecular Science, Via Saffi 2, University of Urbino "Carlo Bo", 61029 Urbino, Italy.
authors.forename: Raffaella
authors.initials: R
authors.lastname: Campana
authors.affiliation: Department of Biomolecular Science, Via Saffi 2, University of Urbino "Carlo Bo", 61029 Urbino, Italy.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Dominici
authors.affiliation: Department of Biomolecular Science, Via Saffi 2, University of Urbino "Carlo Bo", 61029 Urbino, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Ciacci
authors.affiliation: CRB - Centro di Ricerche Biotecnologiche, Università del Sacro Cuore, via Milano 24, 26100 Cremona, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Federici
authors.affiliation: Department of Biomolecular Science, Via Saffi 2, University of Urbino "Carlo Bo", 61029 Urbino, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Sisti
authors.affiliation: Department of Biomolecular Science, Via Saffi 2, University of Urbino "Carlo Bo", 61029 Urbino, Italy.
authors.forename: Marco B L
authors.initials: MB
authors.lastname: Rocchi
authors.affiliation: Department of Biomolecular Science, Via Saffi 2, University of Urbino "Carlo Bo", 61029 Urbino, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Papa
authors.affiliation: Department of Biomolecular Science, Via Saffi 2, University of Urbino "Carlo Bo", 61029 Urbino, Italy. Electronic address: wally.baffone@uniurb.it.
authors.forename: Wally
authors.initials: W
authors.lastname: Baffone
mesh_terms: D018441:Biofilms / Q000254:growth & development*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D016967:Fusobacterium nucleatum / Q000254:growth & development / Q000502:physiology*; D005840:Gentian Violet / Q000032:analysis; D056265:Microbial Interactions; D009055:Mouth / Q000382:microbiology*; D016966:Porphyromonas gingivalis / Q000254:growth & development / Q000502:physiology*; D013194:Staining and Labeling; D013291:Streptococcus / Q000254:growth & development / Q000502:physiology*; D014025:Titanium
chemical_list: D014025:Titanium; D005840:Gentian Violet
keywords: Biofilm; Flow cytometry; Oral streptococci; Periodontal pathogens; Titanium