Antibacterial and Antibiofouling Activities of Antimicrobial Peptide-Functionalized Graphene–Silver Nanocomposites for the Inhibition and Disruption of Staphylococcus aureus Biofilms

Owing to the emergence of antibiotic-resistant strains, bacterial infection and biofilm formation are growing concerns in healthcare management. Herein, we report an eco-benign strategy for the synthesis and functionalization of graphene-silver (rGOAg) nanocomposites with an antimicrobial peptide (AMP) for the treatment of Staphylococcus aureus infection. The synthesis of rGOAg nanocomposites was carried out by simple microwave reduction, and the as-synthesized rGOAg was covalently functionalized with an AMP. As a natural AMP, poly-l-lysine (PLL) functionalization of rGOAg enhanced the antibacterial efficacy and target specificity against the S. aureus biofilm. The robust bactericidal efficiency and biofilm disruption by AMP-functionalized rGOAg (designated as GAAP) occurred through the "contact-kill-release" mode of action, where the electrostatic interaction with bacterial cells together with intracellular ROS generation induced physical disruption to the cell membrane. The internalization of GAAP into the cytoplasm through the damaged cell membrane caused an outburst of intracellular proteins and DNA. Crystal violet staining along with fluorescence and confocal microscopic images showed an effective inhibition and disruption of the S. aureus biofilm upon treatment with GAAP. PLL functionalization also prevented the dissolution of Ag+ ions and thereby minimized the in vitro toxicity of GAAP to the 3 T6 fibroblast and human red blood cells. The ex vivo rat skin disinfection model further demonstrated the potency of GAAP in eliminating the biofilm formation and disruption of the S. aureus biofilm. The obtained results demonstrated a general approach for designing a functional nanocomposite material to disrupt the mature biofilm and provided a promising strategy for treating bacterial infection.