Laser-based culturomics and single cell force spectroscopy for studying interspecies interactions in implant-associated biofilms

Abstract text (incl. figure legends and references)

Introduction

Polymicrobial oral biofilms cause severe implant-associated infections. A profound understanding of formation and structure of these biofilms would facilitate the development of improved methods to diagnose, prevent and treat implant-associated infections. However, microbial isolates for fastidious species are limited and the impact of interspecies interactions on the development of biofilms and implant-associated pathologies is barely understood.

Objectives

The aim of the study was to develop methods for the high-throughput isolation of oral microorganisms and in-depth characterization of biofilm ecology.

Material and Methods

Laser bioprinting was adapted for high-throughput cultivation, characterization, storage and transfer of oral microbiota. Fluidic Force Microscopy (FluidFM) was applied to characterize physical interspecies interactions within biofilms at single cell level.

Results

Laser-based culturomics generated a first extensive culture collection for microorganisms from human dental implants and detected the microbial consortia co-localized in biofilms. Physical interactions of selected co-isolates were characterized at single cell level. According to parameters, which describe physical contact in FluidFM (attachment points, maximum adhesion force and detachment distance), different biophysical profiles were observed for different pairs of strains. Effects of taxonomy, specific adhesion-receptor recognition and co-aggregation inhibitors on physical interactions were described.

Conclusion

Culturomics based on laser bioprinting can effectively design, generate, reproduce and preserve complex biofilm micropatterns. Biophysics of physical interactions between different species within biofilms can be studied at single cell level with FluidFM system.