Microfluidic wound model for studying the behaviors of Pseudomonas aeruginosa in polymicrobial biofilms
Evan Wright1, Suresh Neethirajan2, Xuan Weng1
1BioNano Laboratory, School of Engineering, University of Guelph, Guelph, Canada, N1G 2W1.
Abstract:
Pseudomonas aeruginosa is a particularly problematic opportunistic pathogen due to its capacity to form recalcitrant biofilm structures, while cohabiting with other harmful/pathogenic species and harboring the capability to release toxins that cause tissue necrosis. Although it is now recognized that the majority of biofilm infections are polymicrobial, little is known about the complex interactions that occur within polymicrobial communities and few tools exist for studying these interactions. In this study, we have designed a microfluidic model that mimics the relevant physiological properties of wound microenvironment, while incorporating materials present in the human extracellular matrix/wound environment. Using microfluidics combined with imaging techniques, we have validated the robustness of our model comparing traditional GFP-tagging to new fluorescent staining techniques to visualize/resolve individual species within a polymicrobial habitat. We have also demonstrated that chemotactic stimuli may be incorporated into our model through specialized ports in our chamber. Our system is specifically designed for use with high resolution imaging techniques, allowing for data collection throughout the life of the biofilm and in real-time. Ultimately, this model can be used to investigate the spatio-temporal mechanobiological structures of the wound environment, and the response of the bacteria to the drug transport which will significantly contribute to our understanding of the development and progression of polymicrobial biofilm infections.
Insights
Researchers developed a novel microfluidic model to study polymicrobial biofilm infections in wound environments. This innovative system allows real-time visualization and analysis of bacterial interactions, advancing our understanding of these complex infections.
Area of Science:
- Microbiology
- Biomedical Engineering
- Infectious Diseases
Background:
- Pseudomonas aeruginosa forms difficult-to-treat biofilms and releases toxins.
- Most biofilm infections are polymicrobial, but interactions within these communities are poorly understood.
- Limited tools exist for studying polymicrobial biofilm dynamics.
Purpose of the Study:
- To design and validate a microfluidic model simulating the wound microenvironment for studying polymicrobial biofilms.
- To enable visualization and analysis of individual species within complex bacterial communities.
- To investigate bacterial responses to stimuli and drug transport in real-time.
Main Methods:
- Development of a microfluidic device mimicking wound physiological properties and extracellular matrix components.
- Comparison of traditional GFP-tagging with novel fluorescent staining techniques for species visualization.
- Incorporation of chemotactic stimuli ports for dynamic environmental control.
- High-resolution imaging for real-time data collection throughout biofilm development.
Main Results:
- The microfluidic model successfully mimics key aspects of the wound microenvironment.
- Novel fluorescent staining techniques provide robust visualization of individual species in polymicrobial biofilms.
- The system allows for the introduction of chemotactic stimuli, influencing bacterial behavior.
- Real-time, high-resolution imaging captures biofilm dynamics throughout its lifecycle.
Conclusions:
- The developed microfluidic system is a robust tool for studying polymicrobial biofilm infections.
- This model facilitates investigation into the spatio-temporal mechanobiological structures of wound environments.
- The system aids in understanding bacterial responses to drug transport, crucial for treating polymicrobial biofilm infections.
