Microbial reduction of prebagged human plasma using 405 nm light and its effects on coagulation factors.

Bacterial contamination is the most prevalent infectious complication of blood transfusion in the developed world. To mitigate this, several ultraviolet light-based pathogen reduction technologies (PRTs), some of which require photo-chemicals, have been developed to minimize infection transmission. Relative to UV light, visible 405-nm light is safer and has shown potential to be developed as a PRT for the in situ treatment of ex vivo human plasma and platelet concentrates, without the need for photo-chemicals. This study investigates the effect of 405-nm light on human plasma, with focus on the compatibility of antimicrobial light doses with essential plasma clotting factors. To determine an effective antimicrobial dose that is compatible with plasma, prebagged human plasma (up to 300 mL) was seeded with common microbial contaminants and treated with increasing doses of 405-nm light (16 mW cm-2; ≤ 403 J cm-2). Post-exposure plasma protein integrity was investigated using an AOPP assay, in vitro coagulation tests, and ELISA-based measurement of fibrinogen and Protein S. Microbial contamination in 300 mL prebagged human plasma was significantly reduced (P ≤ 0.05) after exposure to ≤ 288 J cm-2, with microbial loads reduced by > 96.2%. This dose did not significantly affect the plasma protein quality parameters tested (P > 0.05). Increased doses (≥ 345 J cm-2) resulted in a 4.3% increase in clot times with no statistically significant change in protein activity or levels. Overall, this study has demonstrated that the effective microbicidal 405 light dose shows little to no negative effect on plasma quality.

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.

Efficacy of Violet-Blue (405 nm) LED Lamps for Disinfection of High-Environmental-Contact Surfaces in Healthcare Facilities: Leading to the Inactivation of Microorganisms and Reduction of MRSA Contamination.

Effective disinfection procedures in healthcare facilities are essential to prevent transmission. Chemical disinfectants, hydrogen peroxide vapour (HPV) systems and ultraviolet (UV) light are commonly used methods. An emerging method, violet-blue light at 405 nm, has shown promise for surface disinfection. Its antimicrobial properties are based on producing reactive oxygen species (ROS) that lead to the inactivation of pathogens. Studies have shown significant efficacy in reducing bacterial levels on surfaces and in the air, reducing nosocomial infections. The aim of this study was to evaluate the antimicrobial effectiveness of violet-blue (405 nm) LED lamps on high-contact surfaces in a hospital infection-control laboratory. High-contact surfaces were sampled before and after 7 days of exposure to violet-blue light. In addition, the effect of violet-blue light on MRSA-contaminated surfaces was investigated. Exposure to violet-blue light significantly reduced the number of bacteria, yeasts and moulds on the sampled surfaces. The incubator handle showed a low microbial load and no growth after irradiation. The worktable and sink showed an inconsistent reduction due to shaded areas. In the second experiment, violet-blue light significantly reduced the microbial load of MRSA on surfaces, with a greater reduction on steel surfaces than on plastic surfaces. Violet-blue light at 405 nm has proven to be an effective tool for pathogen inactivation in healthcare settings Violet-blue light shows promise as an additional and integrated tool to reduce microbial contamination in hospital environments but must be used in combination with standard cleaning practices and infection control protocols. Further research is needed to optimise the violet-blue, 405 nm disinfection method.

Metabolomics evaluation of the photochemical impact of violet-blue light (405 nm) on ex vivo platelet concentrates.

INTRODUCTION: Microbicidal violet-blue light in the visible spectrum (405 nm) has been under evaluation for pathogen inactivation in ex vivo human plasma and platelets (PLTs) stored in plasma. Results to date have demonstrated that several blood-borne infectious disease-causing pathogens can be successfully reduced to significantly low levels in the light-treated plasma and PLTs. METHOD: In order to evaluate whether the microbicidal 405 nm light is safe for the treatment of PLT concentrates for pathogen inactivation, LC/MS-based metabolomics analyses were performed to evaluate the overall impact of 405 nm violet-blue light treatment on ex vivo PLT concentrates suspended in plasma and on plasma itself, and to identify metabolome changes in intra-platelet and extra-cellular medium (i.e., plasma). RESULTS: The metabolomics data identified that platelet activating factors (PAFs), agonists and prostaglandins, which can influence PLT basic functions such as integrity, activation, and aggregation potential were unaltered, suggesting that 405 nm light illumination is safe regarding PLT basic functions. Distinct increases in hydroxyl fatty acids and aldehydes, as well as decreases in antioxidant metabolites indicated that reactive oxygen species (ROS) were generated at high levels after only one hour of exposure to 405 nm light. Distinctly changed endogenous photosensitizer metabolites after 1 h of light exposure provided good evidence that 405 nm light was an effective microbicide acting through ROS mechanism and no external additive photosensitizers were required.

Laboratory evaluation of the broad-spectrum antibacterial efficacy of a low-irradiance visible 405-nm light system for surface-simulated decontamination.

Purpose: Lighting systems which use visible light blended with antimicrobial 405-nm violet-blue light have recently been developed for safe continuous decontamination of occupied healthcare environments. This paper characterises the optical output and antibacterial efficacy of a low irradiance 405-nm light system designed for environmental decontamination applications, under controlled laboratory conditions. Methods: In the current study, the irradiance output of a ceiling-mounted 405-nm light source was profiled within a 3×3×2 m (18 m3) test area; with values ranging from 0.001-2.016 mWcm-2. To evaluate antibacterial efficacy of the light source for environmental surface decontamination, irradiance levels within this range (0.021-1 mWcm-2) at various angular (Δ Ï´=0-51.3) and linear (∆s=1.6-2.56 m) displacements from the source were used to generate inactivation kinetics, using the model organism, Staphylococcus aureus. Additionally, twelve bacterial species were surface-seeded and light-exposed at a fixed displacement below the source (1.5 m; 0.5 mWcm-2) to demonstrate broad-spectrum efficacy at heights typical of high touch surfaces within occupied settings. Results: Results demonstrate that significant (P≤0.05) inactivation was successfully achieved at all irradiance values investigated, with spatial positioning from the source affecting inactivation, with greater times required for inactivation as irradiance decreased. Complete/near-complete (≥93.28%) inactivation of all bacteria was achieved following exposure to 0.5 mWcm-2 within exposure times realistic of those utilised practically for whole-room decontamination (2-16 h). Conclusion: This study provides fundamental evidence of the efficacy, and energy efficiency, of low irradiance 405-nm light for bacterial inactivation within a controlled laboratory setting, further justifying its benefits for practical infection control applications.

The microbicidal potential of visible blue light in clinical medicine and public health.

Visible blue light of wavelengths in the 400-470 nm range has been observed to have microbicidal properties. A widely accepted hypothesis for the mechanism of microbial inactivation by visible blue light is that the light causes photoexcitation of either endogenous (present within the microbe) or, exogenous (present in the biological medium surrounding the microbe) photosensitizers such as porphyrins and flavins, which leads to the release of reactive oxygen species that subsequently manifests microbicidal activity. Some of the factors that have been observed to be associated with enhanced microbicidal action include increased duration of exposure, and either pre- or co-treatment with quinine hydrochloride. In case of bacteria, repetitive exposure to the blue light shows no significant evidence of resistance development. Additionally, visible blue light has exhibited the ability to inactivate fungal and viral pathogens and, multidrug-resistant bacteria as well as bacterial biofilms. Visible blue light has demonstrated efficacy in eliminating foodborne pathogens found on food surfaces and exposed surfaces in the food processing environment as well as in the decontamination of surfaces in the clinical environment to minimize the spread of nosocomial infections. We conclude from reviewing existing literature on the application of the blue light in clinical medicine and public health settings that this microbicidal light is emerging as a safer alternative to conventional ultraviolet light-based technologies in multiple settings. However, further comprehensive studies and thorough understanding of the mechanism of microbicidal action of this light in different scenarios is warranted to determine its place in human health and disease.

Effects of Violet-Blue Light-Emitting Diode on Controlling Bacterial Contamination in Boiled Young Sardine.

The aim of this study was to evaluate bacterial decontamination of boiled young sardine by treatment with violet-blue light followed by cooling storage of the irradiated boiled sardine. Viable cell count in the samples was evaluated after irradiation with four types of violet-blue light-emitting diodes (LEDs; peak wavelength at 405, 412, 421 or 455 nm) and subsequent cooling storage for two days. LED (405 nm) exhibited bactericidal and growth suppression effects. The irradiation gave a 47% bactericidal rate in comparison with no irradiation samples (control) and the two-day storage suppressed the increase in cell counts to 24%, while the rate of increase was 545% for the control. Integrated viability (IV) based on growth delay analysis was estimated after irradiation of four isolates from boiled sardine with 405 nm light. The irradiation caused growth delay against all isolates, resulting in smaller IV values for three isolates compared to those viabilities estimated from colony forming units. Exposure (405 nm) at 432 J/cm2 fluence resulted in a decrease in water content, resulting in an increase in salinity of the samples. This study demonstrated the advantages of light emitting a narrow violet region as a non-thermal disinfection technology in the processing and storage of boiled sardines.

Photoinhibition of Streptococcus mutans Biofilm-Induced Lesions in Human Dentin by Violet-Blue Light.

This in vitro study determined the effectiveness of violet-blue light on Streptococcus mutans (UA159) biofilm induced dentinal lesions. Biofilm was formed on human dentin specimens in a 96-well microtiter plate and incubated for 13 h in the presence of tryptic soy broth (TSB) or TSB supplemented with 1% sucrose (TSBS). Violet-blue light (405 nm) from quantitative light-induced fluorescence (QLFTM) was used to irradiate the biofilm. Supernatant liquid was removed, and the biofilm was irradiated continuously with QLF for 5 min twice daily with an interval of 6 h for 5 d, except with one treatment on the final day. Colony forming units (CFU) of the treated biofilm, changes in fluorescence (∆F; QLF-Digital BiluminatorTM), lesion depth (L), and integrated mineral loss (∆Z; both transverse microradiography) were quantified at the end of the fifth day. Statistical analysis used analysis of variance (ANOVA), testing at a 5% significance level. In the violet-blue light irradiated groups, there was a significant reduction (p < 0.05) of bacterial viability (CFU) of S. mutans with TSB and TSBS. Violet-blue light irradiation resulted in the reduction of ∆F and L of the dentinal surface with TSBS. These results indicate that violet-blue light has the capacity to reduce S. mutans cell numbers.

Susceptibility of oral bacteria to antibacterial photodynamic therapy.

Effective methods for managing the oral microbiome are necessary to ensure not only the oral but also the systemic health of a human body. The purpose of this study was to determine the sensitivity of four photosensitizers (PSs) to blue light in six representative oral bacterial species that cause intraoral diseases. The following six strains were investigated: Actinomyces israelii, Enterococcus faecium, Fusobacterium nucleatum, Lactobacillus gasseri, Streptococcus mutans, Veillonella parvula. PS stock solutions (1 mg/ml) were prepared by dissolving curcumin and protoporphyrin-IX in dimethyl sulfoxide, and resazurin and riboflavin in distilled water. The inoculation of 20 ml of a bacterial suspension cultured for 24 hours was mixed with 1,980 ml of each test solution, and then a light source was placed in front of the mixture. The irradiation wavelength was 405 nm and its applied energy was 25.3 J. The independent-samples t-test and one-way analysis of variance within groups were performed to compare the antibacterial effects in the four PSs. The antibacterial susceptibility when using different PSs and visible blue-light irradiation differed between the bacterial strains. Antibacterial photodynamic therapy that includes light exposure and PSs can be used to control the oral bacteria strains related to oral disease.

Non-ionizing 405 nm Light as a Potential Bactericidal Technology for Platelet Safety: Evaluation of in vitro Bacterial Inactivation and in vivo Platelet Recovery in Severe Combined Immunodeficient Mice.

Bacterial contamination of ex vivo stored platelets is a cause of transfusion-transmitted infection. Violet-blue 405 nm light has recently demonstrated efficacy in reducing the bacterial burden in blood plasma, and its operational benefits such as non-ionizing nature, penetrability, and non-requirement for photosensitizing agents, provide a unique opportunity to develop this treatment for in situ treatment of ex vivo stored platelets as a tool for bacterial reduction. Sealed bags of platelet concentrates, seeded with low-level Staphylococcus aureus contamination, were 405 nm light-treated (3-10 mWcm-2) up to 8 h. Antimicrobial efficacy and dose efficiency was evaluated by quantification of the post-treatment surviving bacterial contamination levels. Platelets treated with 10 mWcm-2 for 8 h were further evaluated for survival and recovery in severe combined immunodeficient (SCID) mice. Significant inactivation of bacteria in platelet concentrates was achieved using all irradiance levels, with 99.6-100% inactivation achieved by 8 h (P < 0.05). Analysis of applied dose demonstrated that lower irradiance levels generally resulted in significant decontamination at lower doses: 180 Jcm-2/10 mWcm-2 (P = 0.008) compared to 43.2 Jcm-2/3 mWcm-2 (P = 0.002). Additionally, the recovery of light-treated platelets, compared to non-treated platelets, in the murine model showed no significant differences (P = >0.05). This report paves the way for further comprehensive studies to test 405 nm light treatment as a bactericidal technology for stored platelets.

Effect of phototherapy on the metabolism of Streptococcus mutans biofilm based on a colorimetric tetrazolium assay.

The aim of this in vitro study was to determine the effect of violet-blue light on the metabolic activity of early Streptococcus mutans biofilm, reincubated at 0, 2, and 6 h after 5 min of violet-blue light treatment. S. mutans UA159 biofilm cells were cultured for 12 to 16 h in microtiter plates with Tryptic Soy broth (TSB) or TSB with 1% sucrose (TSBS) and irradiated with violet-blue light for 5 min. After irradiation, the plates were reincubated at 37°C for 0, 2, or 6 h in 5% CO2. Colorimetric tetrazolium salt reduction assay was used to investigate bacterial metabolic activity. Mixed model ANOVA was used to find the difference between the violet-blue light treated and nontreated groups. Bacterial metabolic activity was significantly lower in the violet-blue light group for TSB than in the nontreated group (P < 0.0001) regardless of recovery time. However, the differences between metabolic activity in the treated groups without sucrose decreased over time. For TSBS, metabolic activity was significantly lower with violet-blue light at 0 and 2 h. Violet-blue light inhibited the metabolic activity of S. mutans biofilm cells in the light-treated group. This finding may present a unique treatment method for patients with active caries.

Effect of Violet-Blue Light on Streptococcus mutans-Induced Enamel Demineralization.

BACKGROUND: This in vitro study determined the effectiveness of violet-blue light (405 nm) on inhibiting Streptococcus mutans-induced enamel demineralization. MATERIALS AND METHODS: S. mutans UA159 biofilm was grown on human enamel specimens for 13 h in 5% CO2 at 37 °C with/without 1% sucrose. Wet biofilm was treated twice daily with violet-blue light for five minutes over five days. A six-hour reincubation was included daily between treatments excluding the final day. Biofilms were harvested and colony forming units (CFU) were quantitated. Lesion depth (L) and mineral loss (∆Z) were quantified using transverse microradiography (TMR). Quantitative light-induced fluorescence Biluminator (QLF-D) was used to determine mean fluorescence loss. Data were analyzed using one-way analysis of variance (ANOVA) to compare differences in means. RESULTS: The results demonstrated a significant reduction in CFUs between treated and non-treated groups grown with/without 1% sucrose. ∆Z was significantly reduced for specimens exposed to biofilms grown without sucrose with violet-blue light. There was only a trend on reduction of ∆Z with sucrose and with L on both groups. There were no differences in fluorescence-derived parameters between the groups. CONCLUSIONS: Within the limitations of the study, the results indicate that violet-blue light can serve as an adjunct prophylactic treatment for reducing S. mutans biofilm formation and enamel mineral loss.