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.

Development of an in vitro system to study oral biofilms in real time through impedance technology: validation and potential applications

“…Different in vitro model systems have been developed to study complex biofilms. Oral biofilm models can roughly be divided into those that use natural samples such as saliva or dental plaque [9,10], or approaches in which defined bacterial populations are employed [11]. The biofilm models that most closely resemble in vivo microbial communities are certainly those that apply real samples with whole ecosystem diversity, but these microcosms are also more complex to interpret and standardize due to biological heterogeneity.…”

Evaluation of oral microbial corrosion on the surface degradation of dental implant materials

“…Despite improvements in material composition and implant design, implant failures do occur because of individual or synergistic factors including bacterial contamination, premature loading, impaired healing, surgical‐related factors resulting in trauma such as compression necrosis, residual cement, and excessive occlusal stresses . However, bacterial biofilm formation remains an etiological agent in both early‐ and late‐stage failures . Early‐stage failures can occur when bacterial biofilm develops shortly after implant placement, triggering an enhanced inflammatory response while preventing host tissue cells from integrating with the implant surface .…”

“…10,11 However, bacterial biofilm formation remains an etiological agent in both early-and late-stage failures. 12 Early-stage failures can occur when bacterial biofilm develops shortly after implant placement, triggering an enhanced inflammatory response while preventing host tissue cells from integrating with the implant surface. 13,14 In contrast, late-stage failure can occur after previous establishment of osseointegration by periimplantitis where pathogenic bacteria penetrate the soft-tissue seal with the implant surface, triggering eventual tissue necrosis and bone loss around the implant.…”

Evaluation of Biological Activity of Mastic Extracts Based on Chemotherapeutic Indices

“…This microorganism has been reported to dominate the biofilm community (27), colonize the microbial flora around dental implants (28), be present into the saliva and pooled subgingival plaque samples of aggressive periodontitis (29) and Fanconi's anemia patients (30). Toothpaste that contains of mastic is available in Japan.…”