Enhanced antimicrobial efficacy and energy efficiency of low irradiance 405-nm light for bacterial decontamination.

Due to its increased safety over ultraviolet light, there is interest in the development of antimicrobial violet-blue light technologies for infection control applications. To ensure compatibility with exposed materials and tissue, the light irradiances and dose regimes used must be suitable for the target application. This study investigates the antimicrobial dose responses and germicidal efficiency of 405 nm violet-blue light when applied at a range of irradiance levels, for inactivation of surface-seeded and suspended bacteria. Bacteria were seeded onto agar surfaces (101-108 CFUplate-1) or suspended in PBS (103-109 CFUmL-1) and exposed to increasing doses of 405-nm light (≤ 288 Jcm-2) using various irradiances (0.5-150 mWcm-2), with susceptibility at equivalent light doses compared. Bacterial reductions ≥ 96% were demonstrated in all cases for lower irradiance (≤ 5 mWcm-2) exposures. Comparisons indicated, on a per unit dose basis, that significantly lower doses were required for significant reductions of all species when exposed at lower irradiances: 3-30 Jcm-2/0.5 mWcm-2 compared to 9-75 Jcm-2/50 mWcm-2 for low cell density (102 CFUplate-1) surface exposures and 22.5 Jcm-2/5 mWcm-2 compared to 67.5 Jcm-2/150 mWcm-2 for low density (103 CFUmL-1) liquid exposures (P ≤ 0.05). Similar patterns were observed at higher densities, excluding S. aureus exposed at 109 CFUmL-1, suggesting bacterial density at predictable levels has minimal influence on decontamination efficacy. This study provides fundamental evidence of the greater energy efficacy of 405-nm light for inactivation of clinically-significant pathogens when lower irradiances are employed, further supporting its relevance for practical decontamination applications.

Impact of PVC microplastics in photodynamic inactivation of Staphylococcus aureus and MRSA.

Photodynamic processes have found widespread application in therapies. These processes involve photosensitizers (PSs) that, when excited by specific light wavelengths and in the presence of molecular oxygen, generate reactive oxygen species (ROS), that target cells leading to inactivation. Photodynamic action has gained notable attention in environmental applications, particularly against pathogens and antibiotic-resistant bacteria (ARB) that pose a significant challenge to public health. However, environmental matrices frequently encompass additional contaminants and interferents, including microplastics (MPs), which are pollutants of current concern. Their presence in water and effluents has been extensively documented, highlighting their impact on conventional treatment methods, but this information remains scarce in the context of photodynamic inactivation (PDI) setups. Here, we described the effects of polyvinyl chloride (PVC) microparticles in PDI targeting Staphylococcus aureus and its methicillin-resistant strain (MRSA), using curcumin as a PS under blue light. The presence of PVC microparticles does not hinder ROS formation; however, depending on its concentration, it can impact bacterial inactivation. Our results underscore that PDI remains a potent method for reducing bacterial concentrations in water and wastewater containing ARB, even in highly contaminated scenarios with MPs.

Effectiveness of curcumin-based antimicrobial photodynamic therapy against Staphylococcus aureus.

PURPOSE: This study investigated the effectiveness of curcumin-based antimicrobial photodynamic therapy (aPDT) against Staphylococcus aureus (S. aureus), the causative agent of ventilator-associated pneumonia. METHODS: Curcumin was added to S. aureus culture medium at concentrations of 25, 2.5, and 0.25 µM. After 60 min (20-25°C), each culture was irradiated for 1 and 3 min, and viable bacteria were counted. Curcumin (25 µM) was also added to a bacterial suspension with D-mannitol and sodium azide; microbial counts were determined after irradiation for 3 min. RESULTS: S. aureus was significantly reduced in the 1-min (P = 0.043) and 3-min (P = 0.011) irradiation groups in comparison to the 0-min irradiation group with 25 µM curcumin. No significant differences were observed between the curcumin alone group and the curcumin plus D-mannitol or sodium azide group. CONCLUSION: The findings of this study indicate that prolonged exposure (≥1 min) of S. aureus to LED in 25 µM curcumin solution induces cell wall injury. Curcumin-based aPDT as an adjunct to conventional oral care, employing existing dentistry equipment, offers a promising approach that does not rely on antimicrobial drugs or allows the emergence of resistant bacterial strains. This suggests its potential role in future strategies aimed at preventing ventilator-associated pneumonia.

Monomer and Oligomer Transition of Zinc Phthalocyanine Is Key for Photobleaching in Photodynamic Therapy.

Photodynamic therapy (PDT) is recognized as a powerful method to inactivate cells. However, the photosensitizer (PS), a key component of PDT, has suffered from undesired photobleaching. Photobleaching reduces reactive oxygen species (ROS) yields, leading to the compromise of and even the loss of the photodynamic effect of the PS. Therefore, much effort has been devoted to minimizing photobleaching in order to ensure that there is no loss of photodynamic efficacy. Here, we report that a type of PS aggregate showed neither photobleaching nor photodynamic action. Upon direct contact with bacteria, the PS aggregate was found to fall apart into PS monomers and thus possessed photodynamic inactivation against bacteria. Interestingly, the disassembly of the bound PS aggregate in the presence of bacteria was intensified by illumination, generating more PS monomers and leading to an enhanced antibacterial photodynamic effect. This demonstrated that on a bacterial surface, the PS aggregate photo-inactivated bacteria via PS monomer during irradiation, where the photodynamic efficiency was retained without photobleaching. Further mechanistic studies showed that PS monomers disrupted bacterial membranes and affected the expression of genes related to cell wall synthesis, bacterial membrane integrity, and oxidative stress. The results obtained here are applicable to other types of PSs in PDT.

Natural photosensitizer-loaded in micellar copolymer to prevent bovine mastitis: A new post-dipping protocol on milking.

Good management practices such as post-dipping applications (post-milking immersion bath) contribute to the dairy cattle health during lactation and minimize the appearance of mastitis (an infection in the mammary gland). The post-dipping procedure is performed conventionally using iodine-based solutions. The search for therapeutic modalities that are not invasive and do not cause resistance to the microorganisms that cause bovine mastitis instigates the interest of the scientific community. In this regard, antimicrobial Photodynamic Therapy (aPDT) is highlighted. The aPDT is based on combining a photosensitizer (PS) compound, light of adequate wavelength, and molecular oxygen (3O2), which triggers a series of photophysical processes and photochemical reactions that generate reactive oxygen species (ROS) responsible for the inactivation of microorganisms. The present investigation explored the photodynamic efficiency of two natural PS: Chlorophyll-rich spinach extract (CHL) and Curcumin (CUR), both incorporated into the Pluronic® F127 micellar copolymer. They were applied in post-dipping procedures in two different experiments. The photoactivity of formulations mediated through aPDT was conducted against Staphylococcus aureus, and obtained a minimum inhibitory concentration (MIC) of 6.8 mg mL-1 for CHL-F127 and 0.25 mg mL-1 for CUR-F127. Only CUR-F127 inhibited Escherichia coli growth with MIC 0.50 mg mL-1. Concerning the count of microorganisms during the days of the application, a significant difference was observed between the treatments and control (Iodine) when the teat surface of cows was evaluated. For CHL-F127 there was a difference for Coliform and Staphylococcus (p < 0.05). For CUR-F127 there was a difference for aerobic mesophilic and Staphylococcus (p < 0.05). Such application decreased bacterial load and maintained the milk quality, being evaluated via total microorganism count, physical-chemical composition, and somatic cell count (SCC).

Effect and Mechanism of Eliminating Staphylococcus aureus by Electron Beam Irradiation and Reducing the Toxicity of Its Metabolites.

The purpose of this study was to evaluate the mechanism of sterilization of Staphylococcus aureus by electron beam irradiation (0.5-, 1-, 2-, 4-, and 6-kGy treatments) and whether it reduces the toxicity of its fermentation supernatant. In this study, we investigated the mechanism of sterilization of S. aureus by electron beam irradiation using colony count, membrane potential, intracellular ATP, and UV absorbance measurements; we used hemolytic, cytotoxic, and suckling mouse wound models to verify that electron beam irradiation reduced the toxicity of the S. aureus fermentation supernatant. The results showed that 2 kGy of electron beam irradiation treatment completely inactivated S. aureus in suspension culture, and 4 kGy inactivated cells in S. aureus biofilms. This study suggests that the bactericidal effect of electron beam irradiation on S. aureus may be attributed to reversible damage to the cytoplasmic membrane, resulting in its leakage and the significant degradation of genomic DNA. The combined results of hemolytic, cytotoxic, and suckling mouse wound models demonstrated that the toxicity of S. aureus metabolites was significantly reduced when the electron beam irradiation dose was 4 kGy. In summary, electron beam irradiation has the potential to control S. aureus and reduce its toxic metabolites in food. IMPORTANCE Electron beam irradiation of >1 kGy damaged the cytoplasmic membrane, and reactive oxygen species (ROS) penetrated the cells. Electron beam irradiation of >4 kGy reduces the combined toxicity of virulent proteins produced by Staphylococcus aureus. Electron beam irradiation of >4 kGy can be used to inactivate Staphylococcus aureus and biofilms on milk.

A novel exposure mode based on UVA-LEDs for bacterial inactivation.

As an emerging UV source, ultraviolet light-emitting diodes (UV-LEDs) are increasingly being used for disinfection purposes. UVA-LEDs have a higher output power, lower cost, and stronger penetration and cause less harm than UVC-LEDs. In this study, a novel exposure mode based on UVA was proposed and well demonstrated by various experiments using S. aureus as an indicator. Compared with single-dose exposure, fractionated exposure with a 15 min interval between treatments resulted in increased S. aureus inactivation. A longer interval or lower first irradiation dose was unfavorable for inactivation. Fractionated exposure changed the inactivation rate constant and eliminated the shoulder in the fluence-response curves. This resulted in changing the sensitivity of bacteria to UVA and improving bacterial inactivation. Moreover, the fractioned exposure mode has universality for various bacteria (including gram-positive and gram-negative bacteria). S. aureus was not reactivated by photoreactivation or dark repair after UVA treatment. As expected, the cells were damaged more seriously after fractionated exposure, further suggesting the advantages of this new exposure mode. In addition, the mechanism by which bacteria were inactivated after fractionated exposure was investigated, and it was found that •OH played an important role. A longer interval between treatments showed an adverse effect on inactivation, mainly due to the reduction of •OH and recovery of intracellular GSH. In summary, the current work provides novel ideas for the application of UVA-LEDs, which will give more choices for disinfection treatment.

Methylene Blue/Carbon Dots Composite with Photothermal and Photodynamic Properties: Synthesis, Characterization, and Antibacterial Application.

Photodynamic therapy and photothermal therapy provide new ways to combat antibiotic resistance. In this research, methylene blue (MB) as an effective photosensitizer was conjugated with carbon quantum dots (CQDs), the composite product not only possessed good antibacterial properties against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) due to excellent singlet oxygen (1 O2 ) production rate and light heat transfer performance, but also showed good biocompatibility. Combined with 808 nm and 660 nm laser irradiation, the minimum bactericidal concentration of CQDs-MB towards S. aureus and E. coli was 5 µm. Therefore, this study provides a potential candidate material based on CQDs for clinical applications.

Investigation of diode laser effect on the inactivation of selected Gram-negative bacteria, Gram-positive bacteria and yeast and its disinfection on wastewater and natural milk.

Disinfection can be accomplished by adding external chemical agents to kill harmful microorganisms or by removing them using membranes. However, most chemicals are toxic for humans and animals if it is consumed above a certain concentration. Likewise, membranes have fouling problems. The aim of this study is to investigate the effect of diode laser, which is an environmentally friendly application, on pathogenic microorganisms such as Escherichia coli (ATCC 10536), Staphylococcus aureus (ATCC 6538) and Candida albicans. To reveal the effect of diode laser on aforementioned, various parameters have been studied on how diode laser type, laser irradiation time, laser power density, laser penetration efficiency and biofilm inhibition affect microorganisms. As a result of the study, it was observed that the blue laser was more effective than red and green lasers, and the inhibition rates for 15 min at 0.36 W/cm2 laser power density were 65.9% > 34.52% > 43.63% for S. aureus, E. coli and C. albicans, respectively. After 30 min of blue laser irradiation, the microbial growth inhibitions were found as 85.39%, 41.18% and 54.55% for S. aureus, E. coli and C. albicans, respectively. The highest biofilm inhibition was 94.61% when S. aureus cells were exposed to blue laser irradiation for 60 min. The microbial growth kinetics on three microorganisms were tested by using at 0.54 W/cm2 laser power density for 28 h, and there were not observed any microbial development in microbial cultures. Moreover, blue laser irradiation was successfully disinfected wastewater and natural milk at 0.54 W/cm2 laser power density.

Usefulness of a new DUV-LED device for the control of infection by Escherichia coli, Staphylococcus aureus, mycobacteria and spore-forming bacteria.

Reliable disinfection and sterilization technologies are needed to deal with the various infectious diseases spreading around the world. Furthermore, bacteria that are difficult to eliminate by ordinary disinfection are also a problem in the medical environment. We examined the germicidal effect of a newly developed deep-ultraviolet light-emitting diode (DUV-LED) prototype device (wavelength of 280 ± 5 nm; power of 0.9 to 1.4 mW/cm2) for floor sterilization against Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), Mycobacterium gordonae (M. gordonae), and Bacillus subtilis (B. subtilis). This prototype device is equipped with highly practical DUV-LEDs with a high output efficiency and a long life, and was designed with consideration of the irradiation distance and the angle of the DUV-LEDs to provide a uniform irradiation rate. We found a statistically significant reduction of ≥90% in the infectious titers of both E. coli and S. aureus after irradiation for 2 s. Although acid-fast bacilli and spore-type bacilli are generally thought to be resistant to UV light irradiation compared to general bacteria, the acid-fast bacillus M. gordonae was inactivated after irradiation for 10 s, and spore-type cells of the bacillus B. subtilis were inactivated by ≥90% after irradiation for 30 s. We also found that the effects were cumulative when irradiation was performed at intervals. In the future, the usefulness of this device as an infection control measure will be evaluated in daily medical practice.

Exposure to b-LED Light While Exerting Antimicrobial Activity on Gram-Negative and -Positive Bacteria Promotes Transient EMT-like Changes and Growth Arrest in Keratinocytes.

While blue LED (b-LED) light is increasingly being studied for its cytotoxic activity towards bacteria in therapy of skin-related infections, its effects on eukaryotic cells plasticity are less well characterized. Moreover, since different protocols are often used, comparing the effect of b-LED towards both microorganisms and epithelial surfaces may be difficult. The aim of this study was to analyze, in the same experimental setting, both the bactericidal activity and the effects on human keratinocytes. Exposure to b-LED induced an intense cytocidal activity against Gram-positive (i.e, Staphylococcus aureus) and Gram-negative (i.e., Pseudomonas aeruginosa) bacteria associated with catheter-related infections. Treatment with b-LED of a human keratinocyte cell line induced a transient cell cycle arrest. At the molecular level, exposure to b-LED induced a transient downregulation of Cyclin D1 and an upregulation of p21, but not signs of apoptosis. Interestingly, a transient induction of phosphor-histone γ-H2Ax, which is associated with genotoxic damages, was observed. At the same time, keratinocytes underwent a transient epithelial to mesenchymal transition (EMT)-like phenotype, characterized by E-cadherin downregulation and SNAIL/SLUG induction. As a functional readout of EMT induction, a scratch assay was performed. Surprisingly, b-LED treatment provoked a delay in the scratch closure. In conclusion, we demonstrated that b-LED microbicidal activity is associated with complex responses in keratinocytes that certainly deserve further analysis.

Inhibition of bacterial growth through LED (light-emitting diode) 465 and 630 nm: in vitro.

Photobiomodulation has been used to inactivate bacterial growth, in different laser or LED protocols. Thus, the aim of this study was to verify the inhibition of Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli, in ATCC strains and bacteria collected from patients with skin burns, after irradiation with LED; 300 µl of saline solution with bacterial suspension was irradiated at a concentration of 0.5-0.63, by the McFarland scale, after five serial dilutions, with evaluation of pre- and post-irradiation pH and temperature control. The cultures were placed in a bacteriological incubator at 37 °C for 24 h for later counting of colony-forming units (CFU). Data were analyzed by Shapiro-Wilk tests and single-factor ANOVA, with Tukey post hoc (p < 0.05). Both wavelengths and energy densities tested showed inhibition of bacterial growth. The comparison of the irradiated groups (ATCC) with the control group showed the following: S. aureus and P. aeruginosa 465 nm (40 J/cm2) and 630 nm (50 J/cm2) and E. coli 465 nm (40 J/cm2) and 630 nm (30 J/cm2). Among the ATCC S. aureus groups, there was a difference for 630 nm (30 J/cm2) and 465 nm (30, 40, 50 J/cm2). The bacteria from the burned patients were S. aureus (30 and 50 J/cm2) and P. aeruginosa (50 J/cm2). We conclude that different bacterial strains were reduced into colony-forming units after LED irradiation.

Expression and Characterization of the Staphylococcus aureus RecA protein: A mapping of canonical functions.

Recombinases are responsible for homologous recombination (HR), proper genome maintenance, and accurate deoxyribonucleic acid (DNA) duplication. Moreover, HR plays a determining role in DNA transaction processes such as DNA replication, repair, recombination, and transcription. Staphylococcus aureus, an opportunistic pathogen, usually causes respiratory infections such as sinusitis, skin infections, and food poisoning. To date, the role of the RecA gene product in S. aureus remains obscure. In this study, we attempted to map the functional properties of the RecA protein. S. aureus expresses the recA gene product in vivo upon exposure to the DNA-damaging agents, ultraviolet radiation, and methyl methanesulfonate. The recombinant purified S. aureus RecA protein displayed strong single-stranded DNA affinity compared to feeble binding to double-stranded DNA. Interestingly, the RecA protein is capable of invasion and formed displacement loops and readily performed strand-exchange activities with an oligonucleotide-based substrate. Notably, the S. aureus RecA protein hydrolyzed the DNA-dependent adenosine triphosphate and cleaved LexA, showing the conserved function of coprotease. This study provides the functional characterization of the S. aureus RecA protein and sheds light on the canonical processes of homologous recombination, which are conserved in the gram-positive foodborne pathogen S. aureus.

Shifts in the Skin Microbiota after UVB Treatment in Adult Atopic Dermatitis.

BACKGROUND: The pathophysiology in atopic dermatitis (AD) is not fully understood, but immune dysfunction, skin barrier defects, and alterations of the skin microbiota are thought to play important roles. AD skin is frequently colonized with Staphylococcus aureus (S. aureus) and microbial diversity on lesional skin (LS) is reduced compared to on healthy skin. Treatment with narrow-band ultraviolet B (nb-UVB) leads to clinical improvement of the eczema and reduced abundance of S. aureus. However, in-depth knowledge of the temporal dynamics of the skin microbiota in AD in response to nb-UVB treatment is lacking and could provide important clues to decipher whether the microbial changes are primary drivers of the disease, or secondary to the inflammatory process. OBJECTIVES: To map the temporal shifts in the microbiota of the skin, nose, and throat in adult AD patients after nb-UVB treatment. METHODS: Skin swabs were taken from lesional AD skin (n = 16) before and after 3 treatments of nb-UVB, and after 6-8 weeks of full-body treatment. We also obtained samples from non-lesional skin (NLS) and from the nose and throat. All samples were characterized by 16S rRNA gene sequencing. RESULTS: We observed shifts towards higher diversity in the microbiota of lesional AD skin after 6-8 weeks of treatment, while the microbiota of NLS and of the nose/throat remained unchanged. After only 3 treatments with nb-UVB, there were no significant changes in the microbiota. CONCLUSION: Nb-UVB induces changes in the skin microbiota towards higher diversity, but the microbiota of the nose and throat are not altered.

Alternating magnetic fields and antibiotics eradicate biofilm on metal in a synergistic fashion.

Hundreds of thousands of human implant procedures require surgical revision each year due to infection. Infections are difficult to treat with conventional antibiotics due to the formation of biofilm on the implant surface. We have developed a noninvasive method to eliminate biofilm on metal implants using heat generated by intermittent alternating magnetic fields (iAMF). Here, we demonstrate that heat and antibiotics are synergistic in biofilm elimination. For Pseudomonas aeruginosa biofilm, bacterial burden was reduced >3 log with iAMF and ciprofloxacin after 24 h compared with either treatment alone (p < 0.0001). This effect was not limited by pathogen or antibiotic as similar biofilm reductions were seen with iAMF and either linezolid or ceftriaxone in Staphylococcus aureus. iAMF and antibiotic efficacy was seen across various iAMF settings, including different iAMF target temperatures, dose durations, and dosing intervals. Initial mechanistic studies revealed membrane disruption as one factor important for AMF enhanced antibacterial activity in the biofilm setting. This study demonstrates the potential of utilizing a noninvasive approach to reduce biofilm off of metallic implants.

Assessing the photodynamic efficacy of different photosensitizer-light treatments against foodborne bacteria based on the number of absorbed photons.

Increasing interests in photodynamic treatment (PDT) for food preservation require a holistic method to evaluate and compare different photosensitizer (PS)-light treatments. In this report, the absorbed photons were used as the basis to assess the antimicrobial photodynamic efficacy of two PSs, chlorophyllin sodium magnesium salt (Chl-Mg) and chlorophyllin sodium copper salt (Chl-Cu), under blue and white light against two typical foodborne pathogens, Gram-negative Escherichia coli, and Gram-positive Staphylococcus aureus. The results showed that the phototoxicity of a PS was predominantly decided by the absorbed photons rather than the characteristics of light sources. Photosensitized Chl-Mg exhibited superior antimicrobial activity as compared to that of ChlCu. The applied treatments were found to be more effective against S. aureus than E. coli. Bacterial inactivation kinetics as a function of the number of absorbed photons could be described by Weibull model with R2 from 0.947-0.962, and kinetics constants D in the range of 0.202 × 1017 photons/cm2-2.409 × 1018 photons/cm2. The kinetics models may find promising applications in the design, assessment, and optimization of PDT processes.

Aerosol-based antimicrobial photoinactivation in the lungs: an action spectrum study.

Chronic lung infections are among the most diffused human infections, being often associated with multidrug-resistant bacteria. In this framework, the European project "Light4Lungs" aims at synthesizing and testing an inhalable light source to control lung infections by antimicrobial photoinactivation (aPDI), addressing endogenous photosensitizers only (porphyrins) in the representative case of S. aureus and P. aeruginosa. In the search for the best emission characteristics for the aerosolized light source, this work defines and calculates the photo-killing action spectrum for lung aPDI in the exemplary case of cystic fibrosis. This was obtained by applying a semi-theoretical modelling with Monte Carlo simulations, according to previously published methodology related to stomach infections and applied to the infected trachea, bronchi, bronchioles and alveoli. In each of these regions, the two low and high oxygen concentration cases were considered to account for the variability of in vivo conditions, together with the presence of endogenous porphyrins and other relevant absorbers/diffusers inside the illuminated biofilm/mucous layer. Furthermore, an a priori method to obtain the "best illumination wavelengths" was defined, starting from maximizing porphyrin and light absorption at any depth. The obtained action spectrum is peaked at 394 nm and mostly follows porphyrin extinction coefficient behavior. This is confirmed by the results from the best illumination wavelengths, which reinforces the robustness of our approach. These results can offer important indications for the synthesis of the aerosolized light source and definition of its most effective emission spectrum, suggesting a flexible platform to be considered in further applications.

Bacterial Photoinactivation Using PLGA Electrospun Scaffolds.

The use of ultraviolet (UV) and blue irradiation to sterilize surfaces is well established, but commercial applications would be enhanced if the light source is replaced with ambient light. In this paper, it is shown that nanofibers can be explored as an alternative methodology to UV and blue irradiation for bacterial inactivation. It is demonstrated that this is indeed possible using spun nanofibers of poly[lactic-co-(glycolic acid)] (PLGA). This work shows that PLGA spun scaffolds can promote photoinactivation of Staphylococcus aureus and Escherichia coli bacteria with ambient light or with laser irradiation at 630 nm. With the optimized scaffold composition of PLGA85:15 nanofibers, the minimum intensity required to kill the bacteria is much lower than in antimicrobial blue light applications. The enhanced effect introduced by PLGA scaffolds is due to their nanofiber structures since PLGA spun nanofibers were able to inactivate both S. aureus and E. coli bacteria, but cast films had no effect. These findings pave the way for an entirely different method to sterilize surfaces, which is less costly and environmentally friendly than current procedures. In addition, the scaffolds could also be used in cancer treatment with fewer side effects since photosensitizers are not required.

The effect of femtosecond laser irradiation on the growth kinetics of Staphylococcus aureus: An in vitro study.

We investigated the effect of femtosecond laser irradiation on the growth kinetics of Staphylococcus aureus. In order to improve laser-based antimicrobial therapy and develop a clinically viable modality, various laser parameters such as laser light wavelength, laser power, exposure time, and energy density were studied. The INSPIRE HF100 laser system (Spectra Physics) provided the femtosecond laser light, which was pumped by a mode-locked femtosecond Ti: sapphire laser MAI TAI HP (Spectra Physics). The survival of the bacterial cells was monitored after irradiation by determination of growth rate using optical density, which is a rapid, simple, and reliable method. The growth rate of laser-exposed cultures was compared to control cultures. Fifteen minutes of exposure to femtosecond laser radiation with a wavelength of 390 nm and 400 nm at an average power of 50 mW was enough to significantly reduce bacterial viability, with a lag in the growth phase of 5 h longer than the control culture (P < 0.0001 by ANOVA and Tukey test).