Discovery, development and exploitation of steady-state biofilms

Early in vitro biofilm models go back even beyond the invention of the word 'biofilm'. In the dental field, biofilms were simply known as dental plaque and many of the first in vitro models were termed 'artificial mouth microcosm plaques'. The purpose of this review is to highlight important elements of research from over the years regarding in vitro biofilm models, including data from our own laboratories. This helps us to interpret the models and point the way to the future development of biofilm testing. Many hypotheses regarding biofilm phenomena, particularly ecology, metabolism and physiology of volatile sulphur compounds (VSCs) and volatile organic compound (VOC) production could potentially be supported or disproved. In this way, the methods we use for screening biologically active agents including inhibitors, biocides and antimicrobial compounds in general can be improved. Hopefully, any lessons learnt in the past may be of value for the future. In this review, we focus around the need for growth rate controlled long-term biofilms; being continuously monitored using recent technical advances in bioluminescence, selective real-time electrodes, pH electrodes and continuous on-line analysis of the gas phase (both qualitatively and quantitatively). These features allow for accurate determination of growth rate and/or metabolic rate as well as pave the way towards automated assays and fine control of metabolism; impossible to achieve according to conventional biofilm theory. We also attempt to address the questions; can biofilm systems be improved to maintain long term 'real' or 'true' steady states over weeks or months, or are we limited to quasi-steady state systems for a limited period of time.