Fabrication of Nontoxic Reduced Graphene Oxide Protein Nanoframework as Sustained Antimicrobial Coating for Biomedical Application

Bacterial colonization on medical devices is a major concern in the healthcare industry. In the present study, we report synthesis of environmental sustainable reduced graphene oxide (rGO) on the large scale through biosynthetic route and its potential application for antibacterial coating on medical devices. HRTEM image depicts formation of graphene nanosheet, while DLS and ζ potential studies reveal that in aqueous medium the average hydrodynamic size and surface charge of rGO are 4410 ± 116 nm and −25.2 ± 3.2 mV, respectively. The Raman, FTIR, and XPS data suggest in situ conjugation of protein with rGO. The as-synthesized rGO protein nanoframework exhibits dose-dependent antibacterial activity and potential of killing of 94% of Escherichia coli when treated with 80 μg/mL of rGO for 4 h. The hemolytic and cytotoxicity studies demonstrate that rGO protein nanoframework is highly biocompatible at the same concentration showing significant antimicrobial properties. The rGO coated on the glass surface obtained through covalent bonding exhibits potent antibacterial activity. Antibacterial mechanism further demonstrates that rGO-protein nanoframework in dispersed state (rGO solution) exerts bactericidal effect through physical disruption accompanied by ROS-mediated biochemical responses. The rGO subsequently entering into the cytoplasm through the damaged membrane causes metabolic imbalance in the cells. In sharp contrast, physical damage of the cell membrane is the dominant antibacterial mechanism of rGO in the immobilized state (rGO coated glass). The obtained results help indepth understanding of the antibacterial mechanism of the biosynthesized rGO and a novel way to develop nontoxic antibacterial coating on medical devices to prevent bacterial infection.