Real-time monitoring of biofilm formation using a noninvasive impedance-based method

Biofilms are complex three-dimensional microbial communities that adhere to a variety of surfaces and interact with their surroundings. Because of the dynamic nature of biofilm formation, establishing a uniform technique for quantifying and monitoring biofilm volume, shape, and features in real-time is challenging. Herein, we describe a noninvasive electrochemical impedance approach for real-time monitoring of dental plaque-derived multispecies biofilm growth on a range of substrates. A working equation relating electrochemical impedance to live biofilm volume has been developed that is applicable to all three surfaces examined, including glass, dental filling resin, and Ca²⁺-releasing resin composites. Impedance changes of 2.5 %, 35 %, 50 %, and 65 % correlated to biofilm volumes of 0.10 ± 0.01, 16.9 ± 2.2, 29.7 ± 2.3, and 38.6 ± 2.8 µm³/μm², respectively. We discovered that glass, dental filling resin, and Ca²⁺-releasing dental composites required approximately 3.5, 4.5, and 6 days, respectively, to achieve a 50 % change in impedance. The local pH change at the biofilm-substrate interfaces also monitored with potentiometry pH microsensor, and pH change varied according to biofilm volume. This impedance-based technique can be a useful analytical method for monitoring the growth of biofilms on a variety of substrates in real-time. Therefore, this technique may be beneficial for examining antibacterial properties of novel biomaterials.