Journal Article

An oral multispecies biofilm model for high content screening applications

Stumpp, Sascha N., Kommerein, Nadine, Behrens, Peter, Buettner, Falk F. R., Müsken, Mathias, Winkel, Andreas, Ehlert, Nina, Stiesch, Meike, Häussler, Susanne

2017

Overview

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.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Actinomyces / Actinomyces naeslundii / Additives / Antiinfectives and antibacterials / Bacteria / Bacteria - classification / Bacteria - metabolism / Bacterial diseases / Bacterial infections / Bacteriology / Biofilms / Biology and Life Sciences / Biomedical materials / Bone implants / Bone loss / Communities / Complications / Dental implants / Dental plaque / Dental prosthetics / Dental restorative materials / Dentistry / Electron microscopy / Fluorescence / Fluorescence in situ hybridization / Health screening / In Situ Hybridization, Fluorescence / In vitro methods and tests / Infections / Inorganic chemistry / Materials science / Medical schools / Medicine and Health Sciences / Microbial mats / Microscopy / Microscopy, Electron, Scanning / Mouth - microbiology / Physical Sciences / Plaques / Porphyromonas gingivalis / Prostheses / Research and Analysis Methods / Risk factors / Saliva / Scanning electron microscopy / Screening / Sodium chloride / Species / Species Specificity / Streptococcus oralis / Surgical implants / Transplants & implants / Urea / Veillonella dispar