Mechanism of Action of Oxazoline-Based Antimicrobial Polymers Against Staphylococcus aureus: In Vivo Antimicrobial Activity Evaluation.

Antimicrobial-resistant pathogens have reached alarming levels, becoming one of the most pressing global health issues. Hence, new treatments are necessary for the fight against antimicrobial resistance. Synthetic nanoengineered antimicrobial polymers (SNAPs) have emerged as a promising alternative to antimicrobial peptides, overcoming some of their limitations while keeping their key features. Herein, a library of amphiphilic oxazoline-based SNAPs using cationic ring-opening polymerization (CROP) is designed. Amphipathic compounds with 70% cationic content exhibit the highest activity against clinically relevant Staphylococcus aureus isolates, maintaining good biocompatibility in vitro and in vivo. The mechanism of action of the lead compounds against S. aureus is assessed using various microscopy techniques, indicating cell membrane disruption, while the cell wall remains unaffected. Furthermore, a potential interaction of the compounds with bacterial DNA is shown, with possible implications on bacterial division. Finally, one of the compounds exhibits high efficacy in vivo in an insect infection model.

Synthetic Star Nanoengineered Antimicrobial Polymers as Antibiofilm Agents: Bacterial Membrane Disruption and Cell Aggregation.

Antimicrobial resistance has become a worldwide issue, with multiresistant bacterial strains emerging at an alarming rate. Multivalent antimicrobial polymer architectures such as bottle brush or star polymers have shown great potential, as they could lead to enhanced binding and interaction with the bacterial cell membrane. In this study, a library of amphiphilic star copolymers and their linear copolymer equivalents, based on acrylamide monomers, were synthesized via RAFT polymerization. Their monomer distribution and molecular weight were varied. Subsequently, their antimicrobial activity toward a Gram-negative bacterium (Pseudomonas aeruginosa PA14) and a Gram-positive bacterium (Staphylococcus aureus USA300) and their hemocompatibility were investigated. The statistical star copolymer, S-SP25, showed an improved antimicrobial activity compared to its linear equivalent againstP. aeruginosaPA14. The star architecture enhanced its antimicrobial activity, causing bacterial cell aggregation, as revealed via electron microscopy. However, it also induced increased red blood cell aggregation compared to its linear equivalents. Changing/shifting the position of the cationic block to the core of the structure prevents the cell aggregation effect while maintaining a potent antimicrobial activity for the smallest star copolymer. Finally, this compound showed antibiofilm properties against a robust in vitro biofilm model.

Polymerisation-Induced Self-Assembly of Graft Copolymers.

We report the polymerisation-induced self-assembly of poly(lauryl methacrylate)-graft-poly(benzyl methacrylate) copolymers during reversible addition-fragmentation chain transfer (RAFT) grafting from polymerisation in a backbone-selective solvent. Electron microscopy images suggest the phase separation of grafts to result in a network of spherical particles, due to the ability of the branched architecture to freeze chain entanglements and to bridge core domains. Small-angle X-ray scattering data suggest the architecture promotes the formation of multicore micelles, the core morphology of which transitions from spheres to worms, vesicles, and inverted micelles with increasing volume fraction of the grafts. A time-resolved SAXS study is presented to illustrate the formation of the inverted phase during a polymerisation. The grafted architecture gives access to unusual morphologies and provides exciting new handles for controlling the polymer structure and material properties.

Evaluation of the Antimicrobial Activity in Host-Mimicking Media and In Vivo Toxicity of Antimicrobial Polymers as Functional Mimics of AMPs.

Activity tests for synthetic antimicrobial compounds are often limited to the minimal inhibitory concentration assay using standard media and bacterial strains. In this study, a family of acrylamide copolymers that act as synthetic mimics of antimicrobial peptides were synthesized and shown to have a disruptive effect on bacterial membranes and structural integrity through microscopy techniques and membrane polarization experiments. The polymers were tested for their antimicrobial properties using media that mimic clinically relevant conditions. Additionally, their activity was compared in two different strains of the Gram-positive bacterium Staphylococcus aureus and the Gram-negative bacterium Pseudomonas aeruginosa. We showed that the medium composition can have an important influence on the polymer activity as there was a considerable reduction in minimal inhibitory concentrations against S. aureus grown in synthetic wound fluid (SWF), and against P. aeruginosa grown in synthetic cystic fibrosis sputum media (SCFM), compared to the concentrations in standard testing media. In contrast, we observed a complete loss of activity against P. aeruginosa in the serum-containing SWF. Finally, we made use of an emerging invertebrate in vivo model, using Galleria mellonella larvae, to assess toxicity of the polymeric antimicrobials, showing a good correlation with cell line toxicity measurements and demonstrating its potential in the evaluation of novel antimicrobial materials.