Abstract
Medical device-related biofilms are a major cause of hospital-acquired infections, especially chronic infections. Numerous diverse models to study surface-associated biofilms have been developed; however, their usability varies. Often, a simple method is desired without sacrificing throughput and biological relevance. Here, we present an in-house developed 3D-printed device (FlexiPeg) for biofilm growth, conceptually similar to the Calgary Biofilm device but aimed at increasing ease of use and versatility. Our device is modular with the lid and pegs as separate units, enabling flexible assembly with up- or down-scaling depending on the aims of the study. It also allows easy handling of individual pegs, especially when disruption of biofilm populations is needed for downstream analysis. The pegs can be printed in, or coated with, different materials to create surfaces relevant to the study of interest. We experimentally validated the use of the device by exploring the biofilms formed by clinical strains of Escherichia coli and Klebsiella pneumoniae, commonly associated with device-related infections. The biofilms were characterized by viable cell counts, biomass staining, and scanning electron microscopy (SEM) imaging. We evaluated the effects of different additive manufacturing technologies, 3D printing resins, and coatings with, for example, silicone, to mimic a medical device surface. The biofilms formed on our custom-made pegs could be clearly distinguished based on species or strain across all performed assays, and they corresponded well with observations made in other models and clinical settings, for example, on urinary catheters. Overall, our biofilm device is a robust, easy-to-use, and relevant assay, suitable for a wide range of applications in surface-associated biofilm studies, including materials testing, screening for biofilm formation capacity, and antibiotic susceptibility testing.
Highlights
With advances in medical procedures, the use of diverse medical devices has become a new norm in temporary situations, for example, intravenous or urinary catheterization, as well as providing longterm solutions, such as implants
We demonstrate the usability of the system by viable cell counts, biomass staining, and scanning electron microscopy (SEM) imaging, and we demonstrate that this biofilm device is a robust, easy-to-use, and relevant assay adjustable to the user’s needs
The concept of the FlexiPeg biofilm device was inspired by the CBD/MBECTM device with the aim of keeping its high-throughput characteristics while simplifying the handling of pegs, reducing the cost of material, and making the system more versatile in terms of applicability to different experimental setups
Summary
With advances in medical procedures, the use of diverse medical devices has become a new norm in temporary situations, for example, intravenous or urinary catheterization, as well as providing longterm solutions, such as implants. Both temporary medical devices and implants can serve as a platform for bacterial biofilm growth (Costerton et al, 2005). 3D-Printed Peg Biofilm Device defined as microbial communities where the cells are attached to a surface or to each other and enclosed in an extracellular matrix (ECM) (Costerton et al, 1995), and they represent the predominant state of growth in the environment and during chronic infections (Hall-Stoodley et al, 2004). Established biofilms are difficult, or even impossible, to eradicate without the removal of the device due to their high tolerance to both antimicrobial treatment and immune factors (Høiby et al, 2015)
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