Building an Antifouling Zwitterionic Coating on Urinary Catheters Using an Enzymatically Triggered Bottom-Up Approach

Catheter associated urinary tract infections are common during hospitalization due to the formation of bacterial biofilms on the indwelling device. In this study, we report an innovative biotechnology-based approach for the covalent functionalization of silicone catheters with antifouling zwitterionic moieties to prevent biofilm formation. Our approach combines the potential bioactivity of a natural phenolics layer biocatalytically conjugated to sulfobetaine-acrylic residues in an enzymatically initiated surface radical polymerization with laccase. To ensure sufficient coating stability in urine, the silicone catheter is plasma-activated. In contrast to industrial chemical methods, the methacrylate-containing zwitterionic monomers are polymerized at pH 5 and 50 °C using as an initiator the phenoxy radicals solely generated by laccase on the phenolics-coated catheter surface. The coated catheters are characterized by X-ray photoelectron spectroscopy (XPS), Fourier transformed infrared (FTIR) analysis, atomic force microscopy (AFM), and colorimetrically. Contact angle and protein adsorption measurements, coupled with in vitro tests with the Gram-negative Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus in static and dynamic conditions, mimicking the operational conditions to be faced by the catheters, demonstrate reduced biofilm formation by about 80% when compared to that of unmodified urinary catheters. The zwitterionic coating did not affect the viability of the human fibroblasts (BJ-5ta) over seven days, corresponding to the extended useful life of urinary catheters

Quorum-Quenching and Matrix-Degrading Enzymes in Multilayer Coatings Synergistically Prevent Bacterial Biofilm Formation on Urinary Catheters

Bacteria often colonize in-dwelling medical devices and grow as complex biofilm communities of cells embedded in a self-produced extracellular polymeric matrix, which increases their resistance to antibiotics and the host immune system. During biofilm growth, bacterial cells cooperate through specific quorum-sensing (QS) signals. Taking advantage of this mechanism of biofilm formation, we hypothesized that interrupting the communication among bacteria and simultaneously degrading the extracellular matrix would inhibit biofilm growth. To this end, coatings composed of the enzymes acylase and α-amylase, able to degrade bacterial QS molecules and polysaccharides, respectively, were built on silicone urinary catheters using a layer-by-layer deposition technique. Multilayer coatings of either acylase or amylase alone suppressed the biofilm formation of corresponding Gram-negative Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus. Further assembly of both enzymes in hybrid nanocoatings resulted in stronger biofilm inhibition as a function of acylase or amylase position in the layers. Hybrid coatings, with the QS-signal-degrading acylase as outermost layer, demonstrated 30% higher antibiofilm efficiency against medically relevant Gram-negative bacteria compared to that of the other assemblies. These nanocoatings significantly reduced the occurrence of single-species (P. aeruginosa) and mixed-species (P. aeruginosa and Escherichia coli) biofilms on silicone catheters under both static and dynamic conditions. Moreover, in an in vivo animal model, the quorum quenching and matrix degrading enzyme assemblies delayed the biofilm growth up to 7 days