Dataset Information

Real-Time Monitoring of Biofilm Formation Using a Noninvasive Impedance-Based Method.

ABSTRACT: 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.

SUBMITTER: Kumar S

PROVIDER: S-EPMC9853957 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature

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Real-Time Monitoring of Biofilm Formation Using a Noninvasive Impedance-Based Method.

Kumar Sriram S Nguyen Anh Tuan AT Goswami Subir S Ferracane Jack J Koley Dipankar D

Sensors and actuators. B, Chemical 20221122 Pt A

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 ...[more]

PMID: 36688105

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Project description:Bacteria found in diverse ecosystems grow in a community of aggregated cells that favors their survival and colonization. Different extracellular polymeric substances are used to entrap this multispecies community forming a biofilm, which can be associated to biotic and abiotic surfaces. This widespread and successful way of bacterial life, however, can lead to negative effects for human activity since many pathogen and spoiling bacteria form biofilms which are not easy to eradicate. Therefore, the search for novel anti-biofilm bio-active molecules is a very active research area for which simple, reliable, and fast screening methods are demanded. In this work we have successfully validated an impedance-based method, initially developed for the study of adherent eukaryotic cells, to monitor the formation of single-species biofilms of three model bacteria in real time. The xCelligence real time cell analyzer (RTCA) equipment uses specific microtiter E-plates coated with gold-microelectrodes that detect the attachment of adherent cells, thus modifying the impedance signal. In the current study, this technology allowed the distinction between biofilm-producers and non-producers of Staphylococcus aureus and Staphylococcus epidermidis, as well as the formation of Streptococcus mutans biofilms only when sucrose was present in the culture medium. Besides, different impedance values permitted discrimination among the biofilm-producing strains tested regardless of the nature of the polymeric biofilm matrix. Finally, we have continuously monitored the inhibition of staphylococcal biofilm formation by the bacteriophage phi-IPLA7 and the bacteriophage-encoded endolysin LysH5, as well as the removal of a preformed biofilm by this last antimicrobial treatment. Results observed with the impedance-based method showed high correlation with those obtained with standard approaches, such as crystal violet staining and bacteria enumeration, as well as with those obtained upon other abiotic surfaces (polystyrene and stainless steel). Therefore, this RTCA technology opens new opportunities in the biofilm research arena and its application could be further explored for other bacterial genera as well as for different bio-active molecules.

Project description:BackgroundPseudomonas aeruginosa is an opportunistic bacterial pathogen responsible for both acute and chronic infections in humans. In particular, its ability to form biofilm, on biotic and abiotic surfaces, makes it particularly resistant to host's immune defenses and current antibiotic therapies as well. Innovative antimicrobial materials, like hydrogel, silver salts or nanoparticles have been used to cover new generation catheters with promising results. Nevertheless, biofilm remains a major health problem. For instance, biofilm produced onto endotracheal tubes (ETT) of ventilated patients plays a relevant role in the onset of ventilation-associated pneumonia. Most of our knowledge on Pseudomonas aeruginosa biofilm derives from in vitro studies carried out on abiotic surfaces, such as polystyrene microplates or plastic materials used for ETT manufacturing. However, these approaches often provide underestimated results since other parameters, in addition to bacterial features (i.e. shape and material composition of ETT) might strongly influence biofilm formation.ResultsWe used an already established biofilm development assay on medically-relevant foreign devices (CVC catheters) by a stably transformed bioluminescent (BLI)-Pseudomonas aeruginosa strain, in order to follow up biofilm formation on ETT by bioluminescence detection. Our results demonstrated that it is possible: i) to monitor BLI-Pseudomonas aeruginosa biofilm development on ETT pieces in real-time, ii) to evaluate the three-dimensional structure of biofilm directly on ETT, iii) to assess metabolic behavior and the production of microbial virulence traits of bacteria embedded on ETT-biofilm.ConclusionsOverall, we were able to standardize a rapid and easy-to-perform in vitro model for real-time monitoring Pseudomonas aeruginosa biofilm formation directly onto ETT pieces, taking into account not only microbial factors, but also ETT shape and material. Our study provides a rapid method for future screening and validation of novel antimicrobial drugs as well as for the evaluation of novel biomaterials employed in the production of new classes of ETT.

Dataset Information

Real-Time Monitoring of Biofilm Formation Using a Noninvasive Impedance-Based Method.

Publications

Real-Time Monitoring of Biofilm Formation Using a Noninvasive Impedance-Based Method.

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