In Vitro Models for Candida Biofilm Development.

Development of Candida spp. biofilms on medical devices such as catheters and voice prosthesis has been recognized as an increasing clinical problem. Different in vitro models are presented with increasing complexity. Each model system can be utilized for analysis of new active compounds to prevent or treat Candida biofilms as well as to study molecular processes involved in biofilm formation. Susceptibility studies of clinical isolates are generally performed in a simple 96-well model system similar to the CLSI standard. In the present chapter, optimized conditions that promote biofilm formation within individual wells of microtiter plates are described. In addition, the method has proven useful in preparing C. albicans biofilms for investigation by a variety of microscopic and molecular techniques. A more realistic and more complex biofilm system is presented by the Amsterdam Active Attachment (AAA) model. In this 24-well model all crucial steps of biofilm formation: adhesion, proliferation, and maturation, can be simulated on various surfaces, while still allowing a medium throughput approach. This model has been applied to study susceptibility, complex molecular mechanisms as well as interspecies (Candida-bacterium) interactions. Finally, a realistic microfluidics channel system is presented to follow dynamic processes in biofilm formation. In this Bioflux-based system, molecular mechanisms as well as dynamic processes can be studied at a high time-resolution.