The important role of microbial biofilms in medical device-related infections is well established. Intervention strategies developed from in vitro biofilm studies often fail to prevent or cure device-related infections, possibly due to limited relevance of the simplified in vitro biofilm models to the much more complex clinical reality. It is important to use in vitro biofilm assays that closely mimic the dynamically changing clinical environment. This review uses ventricular assistant device driveline infections as a model of disease to demonstrate the morphological diversity and dynamics of clinical biofilms that are important for disease pathogenesis. We also provide insights into how to develop in vitro assays to address the complexity of device-related infections, focusing on pathogen-device interactions, infectious microenvironment, and selection of representative microorganisms and biomaterials.

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