title: An oral multispecies biofilm model for high content screening applications.
abstract: Peri-implantitis caused by multispecies biofilms is a major complication in dental implant treatment. The bacterial infection surrounding dental implants can lead to bone loss and, in turn, to implant failure. A promising strategy to prevent these common complications is the development of implant surfaces that inhibit biofilm development. A reproducible and easy-to-use biofilm model as a test system for large scale screening of new implant surfaces with putative antibacterial potency is therefore of major importance. In the present study, we developed a highly reproducible in vitro four-species biofilm model consisting of the highly relevant oral bacterial species Streptococcus oralis, Actinomyces naeslundii, Veillonella dispar and Porphyromonas gingivalis. The application of live/dead staining, quantitative real time PCR (qRT-PCR), scanning electron microscopy (SEM) and urea-NaCl fluorescence in situ hybridization (urea-NaCl-FISH) revealed that the four-species biofilm community is robust in terms of biovolume, live/dead distribution and individual species distribution over time. The biofilm community is dominated by S. oralis, followed by V. dispar, A. naeslundii and P. gingivalis. The percentage distribution in this model closely reflects the situation in early native plaques and is therefore well suited as an in vitro model test system. Furthermore, despite its nearly native composition, the multispecies model does not depend on nutrient additives, such as native human saliva or serum, and is an inexpensive, easy to handle and highly reproducible alternative to the available model systems. The 96-well plate format enables high content screening for optimized implant surfaces impeding biofilm formation or the testing of multiple antimicrobial treatment strategies to fight multispecies biofilm infections, both exemplary proven in the manuscript.
authors.affiliation: Department of Prosthetic Dentistry and Biomedical Materials Science, Hannover Medical School, Hannover, Germany.
authors.forename: Nadine
authors.initials: N
authors.lastname: Kommerein
authors.affiliation: Department of Prosthetic Dentistry and Biomedical Materials Science, Hannover Medical School, Hannover, Germany.
authors.forename: Sascha N
authors.initials: SN
authors.lastname: Stumpp
authors.affiliation: Institute of Molecular Bacteriology, TWINCORE, Centre of Experimental and Clinical Infection Research, Hannover, Germany.|Department of Molecular Bacteriology, Helmholtz Centre for Infection Research, Braunschweig, Germany.
authors.forename: Mathias
authors.initials: M
authors.lastname: Müsken
authors.affiliation: Institute for Inorganic Chemistry, Leibniz University of Hannover, Hannover, Germany.
authors.forename: Nina
authors.initials: N
authors.lastname: Ehlert
authors.affiliation: Department of Prosthetic Dentistry and Biomedical Materials Science, Hannover Medical School, Hannover, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Winkel
authors.affiliation: Institute of Molecular Bacteriology, TWINCORE, Centre of Experimental and Clinical Infection Research, Hannover, Germany.|Department of Molecular Bacteriology, Helmholtz Centre for Infection Research, Braunschweig, Germany.
authors.forename: Susanne
authors.initials: S
authors.lastname: Häussler
authors.affiliation: Institute for Inorganic Chemistry, Leibniz University of Hannover, Hannover, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Behrens
authors.affiliation: Institute of Clinical Biochemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Falk F R
authors.initials: FF
authors.lastname: Buettner
authors.affiliation: Department of Prosthetic Dentistry and Biomedical Materials Science, Hannover Medical School, Hannover, Germany.
authors.forename: Meike
authors.initials: M
authors.lastname: Stiesch
mesh_terms: D001419:Bacteria / Q000145:classification / Q000378:metabolism*; D018441:Biofilms; D017404:In Situ Hybridization, Fluorescence; D008855:Microscopy, Electron, Scanning; D009055:Mouth / Q000382:microbiology*; D013045:Species Specificity