Antimicrobial photodynamic activity of toluidine blue-carbon nanotube conjugate against Pseudomonas aeruginosa and Staphylococcus aureus - Understanding the mechanism of action.

BACKGROUND: The emergence of drug-resistant bacterial strains has raised the need to develop alternative treatment modalities to combat infectious diseases. Antimicrobial photodynamic therapy (aPDT) is an alternative to conventional treatment modalities. aPDT integrates a photosensitizer, which, after exposure to light of an appropriate wavelength, leads to the generation of cytotoxic reactive oxygen species (ROS). METHODS: The aim of the present study was to synthesize a toluidine blue/multiwalled carbon nanotube conjugate (TBCNT) for enhanced photoinactivation of Pseudomonas aeruginosa and Staphylococcus aureus. Synthesized TBCNT conjugate was characterized and its antibacterial and antibiofilm activity was determined. RESULTS: During TBCNT synthesis, dye loading, and entrapment efficiency of the CNT were 12.04 ±â€¯0.55% and 48.99 ±â€¯2.33%, respectively. The photo-destruction of planktonic cells of the test bacteria was performed by exposure to a 125 mW red laser with a wavelength of 670 nm (radiant exposure of 58.49 J/cm2) for 3 min. Photoinactivation using TBCNT resulted in a 4.91- and 5.47-log10 reduction in P. aeruginosa and S. aureus, respectively. The mechanism of this aPDT was studied by measuring intracellular ROS generation, protein leakage, and lipid peroxidation in the test bacteria after light irradiation. The antibiofilm activity of TBCNT after light exposure was 69.94% and 75.54% for P. aeruginosa and S. aureus, respectively. Photoinactivation of test bacteria treated with TBCNT reduced cell viability and exopolysaccharide production. Confocal laser-scanning microscopy revealed a significant biofilm inhibition efficacy of the TBCNT conjugate. CONCLUSION: Therefore, TBCNT conjugates may be used for the eradication of P. aeruginosa and S. aureus biofilms.

Antimicrobial photodynamic activity of rose bengal conjugated multi walled carbon nanotubes against planktonic cells and biofilm of Escherichia coli.

BACKGROUND: The global threat of antimicrobial resistance especially due to the bacterial biofilms has engaged researchers in the search of new treatment modalities. Antimicrobial photodynamic inactivation (aPDI) is one of the alternative treatment modalities which kills bacteria by generating endogenous reactive oxygen species (ROS). In this work authors evaluated the antibacterial and antibiofilm efficacy of rose Bengal (RB) conjugated to CNT against E. coli. METHODS: The interaction of anionic dye to the CNT was studied using UV-vis spectroscopy, HRTEM, FTIR and spectrofluorometry. Phototoxicity of RBCNT conjugate against E. coli was studied using a green light of 50 mW and radiant exposure of 1674.7 J/ cm2 for 10 min. The antibiofilm activity and mechanism of action of RBCNT conjugate in presence of light was evaluated. RESULTS: The loading and encapsulation was found to be 15.46 ± 1.05 and 61.85 ± 4.23% respectively. The photodynamic inactivation of planktonic cells of E. coli was found to 5.46 and 3.56 log10 CFU/ml reduction on treatment with RBCNT and RB respectively. The conjugate also exhibited efficient and enhanced antibiofilm activity against E. coli. The study of mechanism of action has confirmed cell membrane and DNA damage were the two main targets of aPDI. CONCLUSION: This report has concluded the efficient photodynamic inactivation occurred in Gram negative bacteria E. coli due to the increased production of ROS inside the bacterial cells. Hence, the newly synthesized RBCNT conjugate can be efficiently utilized to control infections caused by E. coli.