Chromophore-Targeting Precision Antimicrobial Phototherapy.

Phototherapy, encompassing the utilization of both natural and artificial light, has emerged as a dependable and non-invasive strategy for addressing a diverse range of illnesses, diseases, and infections. This therapeutic approach, primarily known for its efficacy in treating skin infections, such as herpes and acne lesions, involves the synergistic use of specific light wavelengths and photosensitizers, like methylene blue. Photodynamic therapy, as it is termed, relies on the generation of antimicrobial reactive oxygen species (ROS) through the interaction between light and externally applied photosensitizers. Recent research, however, has highlighted the intrinsic antimicrobial properties of light itself, marking a paradigm shift in focus from exogenous agents to the inherent photosensitivity of molecules found naturally within pathogens. Chemical analyses have identified specific organic molecular structures and systems, including protoporphyrins and conjugated C=C bonds, as pivotal components in molecular photosensitivity. Given the prevalence of these systems in organic life forms, there is an urgent need to investigate the potential impact of phototherapy on individual molecules expressed within pathogens and discern their contributions to the antimicrobial effects of light. This review delves into the recently unveiled key molecular targets of phototherapy, offering insights into their potential downstream implications and therapeutic applications. By shedding light on these fundamental molecular mechanisms, we aim to advance our understanding of phototherapy's broader therapeutic potential and contribute to the development of innovative treatments for a wide array of microbial infections and diseases.

Blue Light Improves Antimicrobial Efficiency of Silver Sulfadiazine Via Catalase Inactivation.

Background: Blue light exhibits the ability to deactivate catalase present in pathogens, significantly improving the antimicrobial performance of compounds such as hydrogen peroxide (H2O2). However, H2O2 is not used within clinical settings due to its short half-life, limiting its potential applications. In this study, we explore the usage of Food and Drug Administration-approved and clinically used silver sulfadiazine (SSD) as a potential alternative to H2O2, acting as a reactive oxygen species (ROS)-producing agent capable of synergizing with blue light exposure. Materials and methods: For in vitro studies, bacterial strains were exposed to a continuous wave 405 nm light-emitting diode (LED) followed by treatment with SSD for varying incubation times. For in vivo studies, bacteria-infected murine abrasion wounds were treated with daily treatments of 405 nm LED light and 1% SSD cream for up to 4 days. The surviving bacterial population was quantified through agar plating and colony-forming unit quantification. Results: Through a checkerboard assay, blue light and SSD demonstrated synergistic interactions. Against both gram-negative and gram-positive pathogens, blue light significantly improved the antimicrobial response of SSD within both phosphate-buffered saline and nutrient-rich conditions. Examination into the mechanisms reveals that the neutralization of catalase significantly improves the ROS-producing capabilities of SSD at the exterior of the bacterial cell, producing greater amounts of toxic ROS capable of exerting antimicrobial activity against the pathogen. Additional experiments reveal that the incorporation of light improves the antimicrobial performance of SSD within methicillin-resistant Staphylococcus aureus (MRSA)- and Pseudomonas aeruginosa strain 1 (PAO-1)-infected murine abrasion wounds. Conclusions: As an established, clinically used antibiotic, SSD can act as a suitable alternative to H2O2 in synergizing with catalase-deactivating blue light, allowing for better translation of this technology to more clinical settings and further implementation of this treatment to more complex animal models.

Photoinactivation of catalase sensitizes a wide range of bacteria to ROS-producing agents and immune cells.

Bacteria have evolved to cope with the detrimental effects of ROS using their essential molecular components. Catalase, a heme-containing tetramer protein expressed universally in most aerobic bacteria, plays an indispensable role in scavenging excess hydrogen peroxide (H2O2). Here, through use of wild-type and catalase-deficient mutants, we identified catalase as an endogenous therapeutic target of 400-420 nm blue light. Catalase residing inside bacteria could be effectively inactivated by blue light, subsequently rendering the pathogens extremely vulnerable to H2O2 and H2O2-producing agents. As a result, photoinactivation of catalase and H2O2 synergistically eliminated a wide range of catalase-positive planktonic bacteria and P. aeruginosa inside biofilms. In addition, photoinactivation of catalase was shown to facilitate macrophage defense against intracellular pathogens. The antimicrobial efficacy of catalase photoinactivation was validated using a Pseudomonas aeruginosa-induced mouse abrasion model. Taken together, our findings offer a catalase-targeting phototherapy approach against multidrug-resistant bacterial infections.

Photo-Disassembly of Membrane Microdomains Revives Conventional Antibiotics against MRSA.

Confronted with the rapid evolution and dissemination of antibiotic resistance, there is an urgent need to develop alternative treatment strategies for drug-resistant pathogens. Here, an unconventional approach is presented to restore the susceptibility of methicillin-resistant S. aureus (MRSA) to a broad spectrum of conventional antibiotics via photo-disassembly of functional membrane microdomains. The photo-disassembly of microdomains is based on effective photolysis of staphyloxanthin, the golden carotenoid pigment that gives its name. Upon pulsed laser treatment, cell membranes are found severely disorganized and malfunctioned to defense antibiotics, as unveiled by membrane permeabilization, membrane fluidification, and detachment of membrane protein, PBP2a. Consequently, the photolysis approach increases susceptibility and inhibits development of resistance to a broad spectrum of antibiotics including penicillins, quinolones, tetracyclines, aminoglycosides, lipopeptides, and oxazolidinones. The synergistic therapy, without phototoxicity to the host, is effective in combating MRSA both in vitro and in vivo in a mice skin infection model. Collectively, this endogenous chromophore-targeted phototherapy concept paves a novel platform to revive conventional antibiotics to combat drug-resistant S. aureus infections as well as to screen new lead compounds.