Facile synthesis, biofilm disruption properties and biocompatibility study of a poly-cationic peptide functionalized graphene-silver nanocomposite.

Bacterial colonization and biofilm formation is a growing challenge in the biomedical field. Although nanotechnology has emerged as an alternative strategy to combat biofilm formation, the toxicity of nanomaterials is a major concern. In this study, we report a safe-by-design strategy for the synthesis of a poly-cationic peptide functionalized graphene-silver nanocomposite (designated as GAPP) and its enhanced biofilm inhibition and disruption properties to eliminate the biofilm development of Gram-negative bacteria. The graphene-silver (rGOAg) nanocomposite was synthesized by microwave reduction, and subsequently functionalized with an antimicrobial poly-cationic peptide through covalent bonding. The results demonstrated that GAPP effectively killed the planktonic cells and biofilms of Escherichia coli and Pseudomonas aeruginosa depending upon the concentration and duration of the interaction. The complete eradication of preformed biofilm was achieved when treated with 10 μg mL-1 of GAPP for 5 h. The GAPP exerted bactericidal and biofilm inhibition activity through a "contact-kill-release" mode of action, wherein the electrostatic interaction of GAPP with the bacterial cells induced physical disruption accompanied by ROS-mediated biochemical changes. The internalization of GAPP into the cytoplasm through the damaged membrane led to metabolic imbalance in the cells. The peptide functionalization further prevented the dissolution of Ag+ ions, thus minimizing the cytotoxicity of GAPP to adult zebrafish. More importantly, the poly-cationic peptide functionalization enhanced the bioavailability, biofilm inhibition and disruption activities of GAPP, while minimizing its toxicological impact. The results obtained thereby provide an effective strategy in the design of alternative antibacterial agents for fighting biofilms of Gram-negative bacteria.