Antimicrobial Blue Light Inactivation of Microbial Isolates in Biofilms.

BACKGROUND AND OBJECTIVES: Biofilms cause more than 80% of infections in humans, including more than 90% of all chronic wound infections and are extremely resistant to antimicrobials and the immune system. The situation is exacerbated by the fast spreading of antimicrobial resistance, which has become one of the biggest threats to current public health. There is consequently a critical need for the development of alternative therapeutics. Antimicrobial blue light (aBL) is a light-based approach that exhibits intrinsic antimicrobial effect without the involvement of exogenous photosensitizers. In this study, we investigated the antimicrobial effect of this non-antibiotic approach against biofilms formed by microbial isolates of multidrug-resistant bacteria. STUDY DESIGN/MATERIALS AND METHODS: Microbial isolates of Acinetobacter baumannii, Candida albicans, Escherichia coli, Enterococcus faecalis, MRSA, Neisseria gonorrhoeae, Pseudomonas aeruginosa, and Proteus mirabilis were studied. Biofilms were grown in microtiter plates for 24 or 48 hours or in the CDC biofilm reactor for 48 hours and exposed to aBL at 405 nm (60 mW/cm2 , 60 or 30 minutes). The anti-biofilm activity of aBL was measured by viable counts. RESULTS: The biofilms of A. baumannii, N. gonorrhoeae, and P. aeruginosa were the most susceptible to aBL with between 4 and 8 log10 inactivation after 108 J/cm2 (60 mW/cm2 , 30 minutes) or 216 J/cm2 (60 mW/cm2 , 60 minutes) aBL were delivered in the microplates. On the contrary, the biofilms of C. albicans, E. coli, E. faecalis, and P. mirabilis were the least susceptible to aBL inactivation (-0.30, -0.24, -0.84, and -0.68 log10 inactivation, respectively). The same aBL treatment in biofilms developed in the CDC biofilm reactor, caused -1.68 log10 inactivation in A. baumannii and -1.74 and -1.65 log10 inactivation in two different strains of P. aeruginosa. CONCLUSIONS: aBL exhibits potential against pathogenic microorganisms and could help with the significant need for new antimicrobials in clinical practice to manage multidrug-resistant infections. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.

Antimicrobial Blue Light Inactivation of Neisseria gonorrhoeae: Roles of Wavelength, Endogenous Photosensitizer, Oxygen, and Reactive Oxygen Species.

BACKGROUND AND OBJECTIVES: The aim of this study was to investigate the efficacy, safety, and mechanism of action of antimicrobial blue light (aBL) for the inactivation of Neisseria gonorrhoeae, the etiological agent of gonorrhea. STUDY DESIGN/MATERIALS AND METHODS: The susceptibilities of N. gonorrhoeae (ATCC 700825) in planktonic suspensions to aBL at 405- and 470-nm wavelengths were compared. The roles of oxygen in the anti-gonococcal activity of aBL were studied by examining the effects of hypoxic condition (blowing N2 ) on the anti-gonococcal efficiency of 405-nm aBL. The presence, identification, and quantification of endogenous photosensitizers in N. gonorrhoeae cells and human vaginal epithelial cells (VK2/E6E7 cells) were determined using fluorescence spectroscopy and ultra-performance liquid chromatography (UPLC). Finally, the selectivity of aBL inactivation of N. gonorrhoeae over the host cells were investigated by irradiating the co-cultures of N. gonorrhoeae and human vaginal epithelial cells using 405-nm aBL. RESULTS: About 3.12-log10 reduction of bacterial colony forming units (CFU) was achieved by 27 J/cm 2 exposure at 405 nm, while about 3.70-log10 reduction of bacterial CFU was achieved by 234 J/cm2 exposure at 470 nm. The anti-gonococcal efficacy of 405-nm aBL was significantly suppressed under hypoxic condition. Spectroscopic and UPLC analyses revealed the presence of endogenous porphyrins and flavins in N. gonorrhoeae. The concentrations of endogenous photosensitizers in N. gonorrhoeae (ATCC 700825) cells were more than 10 times higher than those in the VK2/E6E7 cells. In the co-cultures of N. gonorrhoeae and VK2/E6E7 cells, 405-nm aBL at 108 J/cm2 preferentially inactivated N. gonorrhoeae cells while sparing the vaginal epithelial cells. CONCLUSIONS: aBL at 405-nm wavelength is more effective than 470-nm wavelength in inactivating N. gonorrhoeae while sparing the vaginal epithelial cells. Reactive oxygen species generated from the photochemical reactions between aBL and endogenous photosensitizers play a vital role in the anti-gonococcal activity of 405-nm aBL. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.

Photoinactivation of Neisseria gonorrhoeae: A Paradigm-Changing Approach for Combating Antibiotic-Resistant Gonococcal Infection.

Antimicrobial resistance in Neisseria gonorrhoeae is a major issue of public health, and there is a critical need for the development of new antigonococcal strategies. In this study, we investigated the effectiveness of antimicrobial blue light (aBL; wavelength, 405 nm), an innovative nonpharmacological approach, for the inactivation of N. gonorrhoeae. Our findings indicated that aBL preferentially inactivated N. gonorrhoeae, including antibiotic-resistant strains, over human vaginal epithelial cells in vitro. Furthermore, no aBL-induced genotoxicity to the vaginal epithelial cells was observed at the radiant exposure used to inactivate N. gonorrhoeae. aBL also effectively inactivated N. gonorrhoeae that had attached to and invaded into the vaginal epithelial cells in their cocultures. No gonococcal resistance to aBL developed after 15 successive cycles of inactivation induced by subtherapeutic exposure to aBL. Endogenous aBL-activatable photosensitizing porphyrins in N. gonorrhoeae were identified and quantified using ultraperformance liquid chromatography, with coproporphyrin being the most abundant species in all N. gonorrhoeae strains studied. Singlet oxygen was involved in aBL inactivation of N. gonorrhoeae. Together, these findings show that aBL represents a potential potent treatment for antibiotic-resistant gonococcal infection.

Absence of an SOS-like system in Neisseria gonorrhoeae.

The DNA repair capacities of Neisseria gonorrhoeae have not been well characterised, however, it is known that the gonococcus possesses an excision repair system. The fact that genes in this system are part of the SOS regulon in Escherichia coli prompted this investigation into the transcriptional regulation of genes involved in DNA repair in N. gonorrhoeae. Northern (RNA-DNA) dot blot hybridisation was used to investigate potential DNA damage-mediated induction of the gonococcal recA, uvrA and uvrB genes. In contrast to the situation in E. coli, transcription of these genes in N. gonorrhoeae was not induced in response to treatment with methyl methanesulfonate (MMS) and UV light. These data indicated that the gonococcus does not possess an SOS-like system that is induced in response to DNA damage.