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Background: Our previous clinical trial demonstrated that antimicrobial photodynamic therapy (aPDT) with methylene blue (MB) and potassium iodide (KI) effectively killed Candida albicans (C. albicans) in adult AIDS patients with oral candidiasis, regardless of biofilm formation or 25S rDNA genotype. This study evaluated changes in antifungal susceptibility and virulence gene expression in C. albicans before and after aPDT, and explored factors related to clinical aPDT efficacy., Methods: Twenty-one adult AIDS patients with C. albicans oral candidiasis were divided into Group a (400 μM MB, N = 11) and Group b (600 μM MB, N = 10). Both groups received two aPDT treatments, where MB was applied for 5 min, followed by 300 mM KI, and illuminated for 30 min (37.29 J/cm²). C. albicans isolates were collected before and after treatment to assess antifungal susceptibility (fluconazole, itraconazole, flucytosine, amphotericin B) and gene expression (CAT1, HWP1). Peripheral blood tests were analyzed for correlations with aPDT efficacy., Results: aPDT reduced minimum inhibitory concentration (MIC) values for amphotericin B, fluconazole, and flucytosine, with significant reductions primarily after the first treatment. MIC reductions differed between groups, with Group a showing greater decreases in flucytosine and fluconazole MICs, and Group b in amphotericin B MICs. No significant changes in CAT1 or HWP1 expression were observed. Clinical efficacy of aPDT negatively correlated with leukocyte and neutrophil levels., Conclusions: aPDT effectively reduces MICs of antifungal drugs against C. albicans isolated from treated patients, particularly after the first treatment. The concentration of MB required to reduce MICs varies among different antifungal drugs. aPDT does not alter CAT1 or HWP1 expression, and its clinical efficacy in eradicating C. albicans is negatively associated with leukocyte and neutrophil levels., Competing Interests: Declaration of competing interest MRH declares the following potential conflicts of interest. Scientific Advisory Boards: Transdermal Cap Inc, Cleveland, OH; Hologenix Inc. Santa Monica, CA; Vielight, Toronto, Canada; JOOVV Inc, Minneapolis-St. Paul MN; Consulting; USHIO Corp, Japan; Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, Germany. The other authors declare there is no conflict of interest., (Copyright © 2024. Published by Elsevier B.V.)

Background: Chromoblastomycosis (CMB) is a chronic granulomatous fungal infection that affect the skin and subcutaneous tissues. It is clinically problematic due to limited treatment options, low cure rates, and high rates of relapse. This underscores the necessity for innovative treatment approaches. In this study, potassium iodide (KI) combined with Methylene Blue (MB) mediated antimicrobial photodynamic therapy (PDT) were assessed in the treatment of Fonsecaea monophora (F. monophora) both in vitro and in vivo. And the underlying mechanism that contributes to the efficacy of this treatment approach was investigated., Methods: In vitro experiments were conducted using different combinations and concentrations of MB, KI, and 660 nm light (60 mW/cm 2 ) to inhibit F. monophora. The study was carried out using colony-forming unit (CFU) counts and scanning electron microscopy (SEM). The production of singlet oxygen ( 1 O 2 ), free iodine (I 2 ), hydrogen peroxide (H 2 O 2 ), and superoxide anion during the KI combined MB-mediated antimicrobial PDT process was also detected. In vivo experiments were developed using a Balb/c mouse paw infection model with F. monophora and treated with PBS, 10 mM KI, 2 mM MB +100 J/cm² and 10 mM KI+2 mM MB +100 J/cm² respectively. Inflammatory swelling, fungal load and histopathological analyses of the mouse footpads were assessed., Results: KI enhanced the killing effect of MB-mediated antimicrobial PDT on the conidial spores of F. monophora at the cell and infected animal model level. During the process, the main antimicrobial agents in KI combined with MB- mediated antimicrobial PDT could produce stronger toxic active species including free I 2 and H 2 O 2. CONCLUSION: KI combined with MB-mediated antimicrobial PDT could be an effective adjunct therapy for treating CBM., Competing Interests: Declaration of competing interest There are no conflicts of interest to disclose., (Copyright © 2024. Published by Elsevier B.V.)

Background: Tooth discoloration is a common concern in antimicrobial photodynamic therapy (aPDT) using various photosensitizers (PS). Toluidine Blue (TB), Methylene Blue (MB), Phthalocyanine (Pc), and 2-mercaptopyridine-substituted zinc phthalocyanine (TM-ZnPc) are among those studied, but their relative impacts on tooth discoloration remain unclear., Aim: This study aimed to compare the effects of TB, MB, Pc, and TM-ZnPc in aPDT on tooth discoloration, utilizing a controlled experimental setup., Materials and Methods: The study comprised seventy-five single-rooted incisors with root canals. Following meticulous preparation, a standardized area on the crown surface was designated for examination, and precise measurements of the initial tooth colors were recorded. Samples were randomly divided into five groups: Negative control, MB, TM, Pc, and TM-ZnPc. Photoactivation was performed using LED light, and color measurements were taken at multiple time points up to 90 days. Data were converted to Lab* color values of the CIE Lab* color system (International Commission on Illumination, Vienna, Austria), and ΔE values were calculated. Statistical analysis was performed using Two-way ANOVA and Post-Hoc Tukey tests (p < 0.05)., Results: At day 7 and 30, TM-ZnPc and Pc caused less discoloration compared to MB and TB. TM-ZnPc caused more tooth discoloration compared to Pc (p < 0.05). Compared to baseline, MB and TM-ZnPc caused more tooth discoloration at 30 days and TB caused more tooth discoloration at 90 days (p < 0.05). No significant difference was observed in terms of tooth discoloration at all periods evaluated after Pc application (p > 0.05). All photosensitizers tested in the study caused tooth coloration., Conclusion: All PS induced clinically detectable tooth discoloration, with TB and MB causing more significant discoloration compared to Pc and TM-ZnPc at certain time points. TM-ZnPc and Pc demonstrated more stable coloration levels over time, suggesting their potential reliability in aPDT applications. This study highlights the importance of selecting appropriate PS to minimize tooth discoloration in aPDT, with Pc showing promise in this regard., (© 2024. The Author(s).)

Background: One critical issue that impairs the therapeutic effects of antimicrobial photodynamic therapy (aPDT) in root canal disinfection is the insufficient penetration of photosensitizer into dentinal tubules. Therefore, this study aimed to compare the treatments in enhancing photosensitizers' penetrability for aPDT in root canal therapy., Methods: Thirty premolars with a single root canal were randomly divided into three groups (n = 10), using Methylene blue (MB) as a photosensitizer and treated with different approaches: sonic-assisted diffusing group, ultrasonic-assisted diffusing group and the control without treatment. All specimens were observed by stereomicroscope to measure the penetration depth of MB into dentinal tubules., Results: Both sonic and ultrasonic treatments substantially improved the penetrability of MB. The former achieved a deeper penetration depth than the latter did in the same region of root (P<0.05). Regarding the dye penetration depths at different root regions, the control group showed a declining trend from coronal to apical. In contrast, the penetration depths of sonic and ultrasonic-assisted diffusing groups from highest to lowest were middle > coronal > apical (P<0.05)., Conclusions: This study showed that both sonic and ultrasonic treatments remarkably promoted MB's penetration depth into dentinal tubules. Maximum penetration was achieved when treated with a sonic approach., Competing Interests: Declaration of Competing Interest The authors have no conflict of interest to declare., (Copyright © 2022. Published by Elsevier B.V.)

Antimicrobial photodynamic therapy (aPDT) has been used as an adjuvant treatment of oral infections as a minimal intervention clinical approach. Its antimicrobial efficacy was demonstrated in several studies; however, there is a lack of evidence on its cytotoxic effect on mouse fibroblasts (NIH/3T3). The aim of this study was to evaluate the cytotoxicity and apoptotic pathways of methylene blue-mediated aPDT on mouse fibroblasts. Cells were treated with 0.1 or 1.0 mg.L -1 methylene blue (MB), and 0.075 or 7.5 J.cm-² LED at 630 nm. Cell viability was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet (CV) assays, while cDNA expression for Bax, Bad, Bcl-2, VDAC-1, cytochrome C and Fas-L was assessed by qRT-PCR (1, 3, 6 and 24 h). The differences between groups were detected by Kruskal-Wallis and post-hoc Dunn's tests for MTT and CV assays, and by ANOVA and post-hoc Tukey test for qPCR (P < 0.05). The combination of 1.0 mg.L -1 MB and 7.5 J.cm-² LED significantly reduced the cellular viability, whereas MB and LED alone were innocuous to fibroblasts. MB-mediated aPDT increased the expression of cytochrome C and Fas-L after 3 h, and Bax/Bcl-2, Bad/Bcl-2, and VDAC-1 after 6 h from treatment. Based on these results, MB-mediated aPDT induced cytotoxicity on mouse fibroblasts, with consequent activation of Bcl-2 apoptosis signaling pathways. Further studies are needed to determine the adequate parameters of aPDT to inactivate microorganisms without damaging fibroblasts., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Antimicrobial Photodynamic therapy (A-PDT) has been used to treat infections. Currently, microbial inactivation data is reported presenting survival fraction averages and standard errors as discrete points instead of a continuous curve of inactivation kinetics. Standardization of this approach would allow clinical protocols to be introduced globally, instead of the piecemeal situation which currently applies., Methods: To this end, we used a power-law function to fit inactivation kinetics and directly report values of lethal doses (LD) and a tolerance factor (T) that informs if inactivation rate varies along the irradiation procedure. A deduced formula was also tested to predict LD for any given survival fraction value. We analyzed the photoantimicrobial effect caused by red light activation of methylene blue (MB-APDT) and by blue light (BL) activation of endogenous microbial pigments against 5 clinically relevant pathogens., Results: Following MB- APDT, Escherichia coli and Staphylococcus aureus cells become increasingly more tolerant to inactivation along the irradiation process (T < 1). Klebsiella pneumoniae presents opposite behavior, i.e., more inactivation is observed towards the end of the process (T > 1). P. aeruginosa and Candida albicans present constant inactivation rate (T˜1). In contrast, all bacterial species presented similar behavior during inactivation caused by BL, i.e., continuously becoming more sensitive to blue light exposure (T > 1)., Conclusion: The power-law function successfully fit all experimental data. Our proposed method precisely predicted LD and T values. We expect that these analytical models may contribute to more standardized methods for comparisons of photodynamic inactivation efficiencies., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Objective: This study evaluated the effect of hydrogen peroxide addition on β-cyclodextrin-conjugated methylene blue in antimicrobial photodynamic therapy(a-PDT) in S. mutans biofilm model using laser or light emitting diode (LED) (λ = 660 nm)., Methods: A preliminary assay was performed to evaluate the cytotoxicity of hydrogen peroxide in oral fibroblasts by the colorimetric method (MTT). Afterwards, groups were divided into (n = 3, in triplicate): C (negative control), CX - chlorhexidine 0.2% (positive control), P (methylene blue/β-cyclodextrin), H (Hydrogen Peroxide at 40 μM), PH, L (Laser), LP, LH (Laser+Hydrogen Peroxide), LPH, LED, LEDP, LEDH, and LEDPH. The biofilm was formed in 24 h with BHI + 1% sucrose (w/v). Light irradiations were conducted with laser, 9 J, 323 J/cm 2 , 113 s or with LED, 8.1 J, 8.1 J/cm 2 for 90 s. Microbial reduction was evaluated by counting the viable microorganisms of the biofilm after the respective treatments, in a selective culture medium, and laser confocal microscopy evaluation., Results: LP, LH, LPH, LEDP, LEDH, and LEDPH groups statistically reduced the counts of S.mutans compared with the C group and the log reductions were of 1.87, 1.94, 2.19, 0.91, 0.92, and 1.33, respectively; the addition of hydrogen peroxide did not potentiate the microbial reductions (LPH and LEDPH) compared with the LP and LEDP groups., Conclusion: The association of hydrogen peroxide with the conjugated β-cyclodextrin nanoparticle as photosensitizer did not result in an enhanced effect of a-PDT; hydrogen peroxide behaved as a photosensitizer, since it reduced the number of S. mutans when associated with laser light., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Photodynamic therapy (PDT) has become an emerging novel therapeutic approach for treating localized microbial infections, particularly those sustained by multidrug-resistant strains. Given the irreplaceable role played by professional phagocytes in limiting infections, such as polymorphonuclear neutrophils, any newly designed antimicrobial therapeutic approach must not interfere with their function. The present investigation presents a detailed analysis of the effect of PDT on the viability and several functional responses of human polymorphonuclear neutrophils loaded with methylene blue (MB), one of the more commonly used photosensitizers in antimicrobial PDT. Taking advantage of the use of a specifically-designed optical LED array for illuminating MB-loaded human polymorphonuclear neutrophils, a number of cell functions have been assayed under miniaturized, strictly controlled and reproducible experimental conditions. The major findings of this study are the following: (1) MB-PDT increases human neutrophils adhesion and does not modify myeloperoxidase release; (2) MB-PDT markedly enhances reactive oxygen species generation that is independent of superoxide-forming phagocytic oxidase and very likely ascribable to LED-dependent excitation of accumulated methylene blue; (3) MB-PDT almost abolishes human neutrophils candidacidal activity by hindering the engulfing machinery. This in vitro study may represent a valuable reference point for future research on PDT applications for treating localized microbial infections., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Antimicrobial photodynamic inactivation (APDI) is a new therapeutic modality which needs more precision during application due to the possibility of exposure of bacteria to sub-lethal doses (sAPDI). In this study, we aimed to evaluate the effect of sAPDI on Pseudomonas aeruginosa quorum sensing (QS) and c-di-GMP signaling which are important virulence factor regulatory systems., Methods: Biofilm formation, pyoverdine, pyocyanin and protease production of P. aeruginosa was evaluated before and after a single sAPDI treatment with 0.8 mM methylene blue (MB) plus 1, 2, and 5-min irradiation with red laser light. Fluorescent lasB, rhlA, pqsA, and cdrA reporters of P. aeruginosa PAO1 and P. aeruginosa ΔmexAB-oprM were treated individually with sAPDI and the regulatory signals were detected. The gene expressions were also assessed after sAPDI using quantitative real-time PCR analysis., Results: Morphological observations and molecular assessments indicated that sAPDI with 0.8 mM MB along with 2- and 5-min irradiation led to an increase in the expression of the Las QS system and c-di-GMP signaling, while 1 min irradiation revealed dissimilar results (increase in lasB expression and decrease in c-di-GMP levels). Expression of rhlA and pqsA did not change in response to sAPDI. Further, a severe lethal effect of sAPDI was observed in P. aeruginosa ΔmexAB-oprM as compared with the wild type strain, whilst there was no difference in QS and c-di-GMP levels as detected by reporters between treated and untreated samples., Conclusion: The results suggest that sAPDI affects QS and c-di-GMP signaling inP. aeruginosa in a time-dependent manner., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Antimicrobial photodynamic therapy (aPDT) shows antimicrobial activity on yeast of the genus Candida. In aPDT, the depth at which the light penetrates the tissue is extremely important for the elaboration of the treatment. The aim of this study was to evaluate the action of aPDT on experimental candidiasis and the laser impact in the tissue using Galleria mellonella as the infection model., Methods: G. mellonella larvae were infected with different Candida albicans strains. After 30 min, they were treated with methylene blue-mediated aPDT and a low intensity laser (660 nm). The larvae were incubated at 37 °C for seven days and monitored daily to determine the survival curve, using the Log-rank test (Mantel Cox). To evaluate the distribution of the laser as well as its depth of action in the larva body, the Interactive 3D surface PLOT of Image J was used. The effects of aPDT on the immune system were also evaluated by the quantification of hemocytes in the hemolymph of G. mellonella after 6 h of Candida infection (ANOVA and Tukey's test)., Results: In both the ATCC 18,804 strain and the C. albicans clinical strain 17, aPDT prolonged the survival of the infected G. mellonella larvae by a lethal fungal dose. There was a statistically significant difference between the aPDT and the control groups in the ATCC strain (P = 0.0056). The depth of laser action in the insect body without the photosensitizer was 2.5 mm and 2.4 mm from the cuticle of the larva with the photosensitizer. In the larvae, a uniform distribution of light occurred along 32% of the body length for the group without the photosensitizer and in 39.5% for the group with the photosensitizer. In the immunological analysis, the infection by C. albicans ATCC 18,804 in G. mellonella led to a reduction in the number of hemocytes in the hemolymph. The aPDT and laser treatment induced a slight increase in the number of hemocytes., Conclusion: Both aPDT and laser treatment positively influenced the treatment of experimental candidiasis. G. mellonella larvae were a useful model for the study of light tissue penetration in antimicrobial photodynamic therapy., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The concentration of methylene blue (MB) photosensitizer could affect the eradication efficacy of antimicrobial photodynamic therapy (aPDT) in the treatment of contaminated implants, which is linked to the osseointegration of the implant. We evaluated osteoblast functions on the contaminated SLA (sandblasting, large-grit and acid-etching) Ti alloy surfaces after the concentration-dependent use of MB-aPDT. Totally 1164 SLA discs were randomly distributed for the analyses of antibacterial efficacy and osteoblast functions. Gram-negative (Aggregatibacter actinomycetemcomitans; A. actinomycetemcomitans) or Gram-positive (Streptococcus mutans; S. mutans) adhered on disc samples was subjected to aPDT with different MB concentrations (200, 250, 300, 350, and 400 μg/mL) using 660 nm diode laser with maximum output 80 mW for 1 min irradiation (4.8 J/cm 2 ). Bactericidal effect was examined by viability, morphology, and lipopolysaccharide (LPS) assays. The disinfected disc surfaces by MB-aPDT to support osteoblast-like MG63 attachment, proliferation, differentiation, and mineralization were assessed for the predetermined culture time intervals. The statistical differences between the means were performed using a one-way analysis of variance (ANOVA) with a post hoc Scheffe test. The results of the morphology observation and bacterial survival examination consistently indicated a remarkably lower quantity of bacterial colonies on biofilm-contaminated surfaces after the aPDT treatment with higher MB concentration. Similarly, the higher MB concentration in aPDT resulted in the lower LPS amounts remaining on the A. actinomycetemcomitans-contaminated surfaces. Intriguingly, the expression of osteoblast cultured on disinfected surfaces using aPDT with higher MB concentration was comparable to the control without contamination. Within the limits of this in vitro model, this formulation of 400 μg/mL MB used in aPDT may be not only the lethal concentration against the 2 bacteria-contaminated implants, but it could also enhance the osteoblast functions on the contaminated implants. Nevertheless, the efficacy in the clinical practice for peri-implantitis therapy remains to be studied., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Candida albicans is an opportunistic commensal microorganism, often associated with severe infections in immunosuppressed individuals. C. albicans has hexose transporters that may favor the intracellular accumulation of photosensitizer (PS). the aims of this study were to investigate the influence of glucose load on photodynamic antimicrobial chemotherapy (PACT); and the role that membrane transport system plays on this therapy in the presence of glucose., Material and Methods: Strains of C. albicans were selected: ATCC 10231, YEM 12, YEM 13, YEM 14 and YEM 15. All strains were grown aerobically on Sabouraud agar and incubated at 30 °C for 24 h. The strains were treated with and without glucose, and divided into Control (no treatment), LED light (660 nm, 166 mW/cm 2 ), Photosensitizer (100 μM methylene blue) and PACT at 1, 3 and 6 min of irradiation groups. The colony forming units were counted and data submitted to statistical analysis (ANOVA) and Tukey's test. The concentration of methylene blue (MB) outside the yeast was measured by fluorescence spectroscopy., Results: PACT inactivate C. albicans and the presence of glucose did not affect the killing effect for most strains. Only YEM12 was partially affected by its presence. Regarding efflux systems, ABC overexpressing strain showed a protective effect on the yeast cells. We observed that yeast with overexpression of major facilitator superfamily (MFS) membrane pore tended to accumulate more MB in its cytoplasm, whereas strains that overexpressed ABC pumps (ATP-binding cassette transporters) tended to decrease MB uptake and survive the photodynamic challenge., Conclusion: Presence of glucose showed a small effect on PACT . The accumulation of MB on yeast induces more photodynamic inactivation; however, the photodynamic efficacy depends on the type and characteristics of the microbial strain., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background and Aim: Streptococcus mutans (S. mutans) is a bacterium that colonizes in the mouth and is a common cause of dental caries and periodontal diseases. This bacterium comprises 70% of the bacteria in the dental plaque. Although tooth decay is a multifactorial complication, S. mutans biofilms are the main cause of cavitated carious lesions. Considering the importance of this microorganism, we aimed at investigating the effect of photodynamic therapy (PDT) using curcumin (CUR) and methylene blue (MB) photosensitizers on S. mutans., Materials and Methods: In this in-vitro experimental study, first, samples of S. mutans were prepared in 110 test tubes and were randomly assigned to 11 groups after colony counting: 1) Positive control group, 2) Negative control group, 3) CUR extract group, 4) 460-nm laser group, 5) 460-nm continuous laser + CUR group, 6) 460-nm discontinues 50% duty cycle (DC) laser + CUR group, 7) 660-nm laser group, 8) 660-nm laser + MB group, 9) MB group, 10) dental light-curing group, and 11) chlorhexidine (CHX) group. After the intervention, cultivation was performed again in blood agar medium, and the bacterial colony-forming units per milliliter (CFU/ml) were counted again. Data were analyzed using analysis of variance (ANOVA) and Tukey's test., Results: CHX and 460-nm low-level continuous laser + CUR had the highest and most significant effect on inhibiting the growth of S. mutans bacterial colonies and showed significant differences with other groups (P < 0.001)., Conclusion: According to the results, MB- and CUR-mediated PDT can significantly eradicate S. mutans colonies., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study reports successful photodynamic inactivation of planktonic and biofilm cells of Enterococcus faecalis using Methylene Blue (MB) in combination with gold nanoparticles synthesized using the cell-free filtrate obtained from 3-day biomass of Trichoderma asperellum strain. Monodispersed colloidal gold nanoparticles were characterized by UV-vis absorption, TEM and DLS to be 13 ± 3 nm spheres. Diode lasers with the peak-power wavelength ʎ = 660 nm (output power of 21, 41 and 68 mW; power density of 55, 108 and 179 mW∙cm -2 , respectively, were used as a light source to study the effects of MB alone, the gold nanoparticles alone (AuNPs) and the MB + AuNPs mixture on the viability of E. faecalis cells. The lethal effect of planktonic cells was achieved for MB after 30 min of laser irradiation with energy fluence of 322 J∙cm -2 . When MB + AuNPs mixture was used as photosensitizer, the lethal effect was achieved with energy fluence of 292 J∙cm -2 . The biofilm culture was more resistant to photo-inactivation and the best bactericidal effect of MB as photosensitizer was found after light dose of 483 J∙cm -2 . The bacterial cell viability was reduced by 99.92%. It was proved that MB + AuNPs mixture synergistically enhances the kill of the studied microorganism as the same light dose resulted in 99.991% kill., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Photodynamic Therapy (PDT) is an alternative to surface decontamination that is based on the interaction between a non-toxic photosensitizer (PS) and a light source to allow for the formation of reactive oxygen species. The objective of this study was to test a new patented device - the "Ultrasonic Photodynamic Inactivation Device" (UPID) under the patent deposit MU-BR 20.2018.00.9356-3 - for the photodynamic inactivation on contaminated acrylic plates and titanium disk. This new low cost device contains light emitting diodes (LEDs) and was built in a stainless-steel container for better light distribution. In addition, 28 waterproof red LEDs plates, with a wavelength of 660 nm were used, containing three irradiators in each plate, for which the irradiation distribution and the spectral irradiance on all 6 internal faces of this device were calculated. The effect of red LED irradiation (660 nm) methylene blue (MB) (100 μmol/L) diluted in water or 70% alcohol on three types of microorganisms: Candida albicans ATCC 10231, Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922. In order to estimate the effects of PDI, acrylic plates and Titanium disks were contaminated by bacterial suspensions (3 × 10 8 CFU/mL), then treated with a solution of MB for 30 min, followed by irradiation for 30 min (0.45 J/cm 2 ). Microbial inhibition was evaluated by counting the number of colony forming units (CFU), compared to the control group. The results showed that the UPID promoted significant reduction (p < 0.001) of the microorganism when compared with the positive control. The new device promoted an effective microbial inhibition on the surfaces tested and, thus, makes possible new studies. The perspective is that this new device may be a low-cost and non-toxic alternative to the disinfection of biomedical devices, non-critical instruments and also for use in the food industry., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Medication-related osteonecrosis of the jaw (MRONJ) is a debilitating complication strongly associated to antiresorptive agents. The present study aimed to describe the use of antimicrobial photodynamic therapy (aPDT) in the prevention of MRONJ., Methods: The sample consisted of 11 non-oncologic osteoporotic subjects in therapy with non-intravenous antiresorptive agents, requiring tooth extractions and/or implant removal. After minimally invasive surgical extractions, each alveolar socket was debrided and bony edges were smoothened. At this point, aPDT was performed using methylene blue-based phenothiazine chloride dye irradiated with a hand-held 100 mW diode laser with a wavelength of 660 ± 10 nm. Flaps were sutured to achieve first intention closure. Soft tissue healing was promoted with weekly applications of low-level laser therapy for 6 weeks. Recall visits were scheduled weekly for the first two months and monthly thereafter up to 6 months. At the 6-month appointment, healing was assessed clinically and radiographically., Results: A total of 62 surgical extractions were performed in both jaws, including 51 natural elements and 11 dental implants. No intraoperative complications were observed. Immediate post-operative period was generally uneventful except for mild pain and ecchymosis that occurred rarely and resolved spontaneously. Healing proceeded uneventfully, with no clinical or radiological prodromal manifestations of MRONJ up to the latest follow-up visit., Conclusions: aPDT might constitute a promising preventive treatment to reduce the risk of MRONJ in non-oncologic osteoporotic patients treated with non-intravenous antiresorptive agents that underwent dentoalveolar surgery., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Candida albicans is an opportunistic fungus, an etiological agent of human infections, presenting high rates of morbidity and mortality. The resistance of C. albicans to conventional therapies has been reported due to the extensive use of conventional antifungals. Photodynamic antimicrobial chemotherapy (PACT) is a technique that combines a visible light with a specific wavelength and a photosensitizer, producing ROS and permanent damages in the treated cells., Methods: In this work, the effects of PACT, using Methylene Blue (MB), as a photosensitizer, on C. albicans development were studied., Results: Significant reduction in both cell growth and biofilm formation after PACT were observed, in a dependent manner on both MB concentration and fluence. In the presence of MB 0.02 mg/mL, it was observed inhibition in biofilm formation of ˜58, 70 and 74%, using fluences of 10, 20 and 30 J/cm 2 , respectively. Also, it was observed inhibition of 54, 66 and 55% in the presence of MB 0.01, 0.02 and 0.05 mg/mL, respectively in the viability of biofilm produced by C. albicans. The number of both yeast and filaments present in the structure of biofilm were reduced after PACT. Furthermore, PACT changed the growth kinetics of C. albicans. Interestingly, we demonstrated increase in the extent of lag phase and an alteration in the profile of the exponential phase after PACT., Conclusions: Taken together, these results indicate the potential PACT effects using MB to decrease the C. albicans development., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: We compared the effectiveness of a single irradiation vs repetitive irradiation of light, for in vitro photodynamic inactivation (PDI) of Candida albicans and Trichophyton mentagrophytes, by using methylene blue (MB) and rose bengal (RB) as photosensitizers (PS)., Methods: MB from 5 to 60 μM and RB from 0.5 to 10 μM, with energy densities from 10 to 60 J/cm 2 , were tested in C. albicans. We further optimize the PDI by reducing the light energy density and PS concentration for the single irradiation experiments by using repetitive doses (two and three times). MB was tested in C. albicans and T. mentagrophytes, and RB was tested in C. albicans., Results: MB-PDI and RB-PDI in C. albicans significantly reduced the number of colony-forming units per milliliter (CFU/mL) when compared to the control groups. Using a single irradiation, over 99% growth inhibition of C. albicans was obtained with MB at 20 μM-60 J/cm 2 , and with RB at 1 μM-30 J/cm 2 and 5 μM-10 J/cm 2 . With repetitive doses, similar results were obtained by reducing several times the light energy density and the PS concentration for C. albicans and T. mentagrophytes., Conclusions: The results showed that RB was more effective than MB for C. albicans inactivation. In addition, it is possible to significantly reduce the amount of PS and light energy density requirements by using repetitive irradiations in both genera tested. It makes the technique less invasive and could reduce the side effects in people extremely sensitive to the PS or the light., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Bacterial elimination using antimicrobial photodynamic therapy (aPDT) has been considered an alternative therapeutic modality in peri-implantitis treatment. The present in vitro study evaluated the dose-dependent and pH-dependent bactericidal effects of methylene blue (MB)-mediated aPDT at eliminating Gram-negative (P. gingivalis and A. actinomycetemcomitans) and Gram-positive (S. mutans) bacteria on sandblasting, large-grit and acid-etching (SLA)-pretreated titanium alloy. The effects of different MB concentrations (50, 100, and 200 μg/mL), the pH of the MB (4, 7, and 10), and irradiation time (0, 30, and 60 s) on the bacterial viability and residual lipopolysaccharide (LPS) levels were examined. The variations in the pH of the MB solution after aPDT for 60 s on the uncontaminated and contaminated specimens were also detected. The experimental results indicated that MB-mediated PDT could effectively kill the majority of bacteria on the titanium alloy surfaces of biofilm-contaminated implants compared with the MB alone. Of note, aPDT exhibited better antibacterial efficacy with increase in the MB concentration and irradiation time. While treated in an acidic solution on the biofilm-contaminated specimens, aPDT caused the pH to increase. By contrast, the initially high alkaline pH decreased to a value of about pH 8.5 after aPDT. Intriguingly, the neutral pH had minor changes, independent of the MB concentration and bacterial species. As expected, aPDT with higher MB concentration at higher pH environment significantly lowered the LPS concentration of A. actinomycetemcomitans and P. gingivalis. On the basis of the data, the aPDT with 200 μg/mL MB at pH 10 for 60 s of irradiation time might be an effectively treatment to eliminate bacteria and LPS adherent to titanium surface, however, the use of the multispecies biofilm model and the evaluation of in vitro osteogenesis needed to be further evaluated., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The objective was to develop, characterize and test a box containing light emission diode (LED), Patent Deposit MU-BR20.2017.002297-3, which was named "Photodynamic Inactivation Device" (PID) and verify if it's suitable in microbial reduction or disinfection action of solid surfaces using PID. The equipment was made in a container of polypropylene with a lid of the same material and, for a better use of irradiation the internal part was covered with a layer of reflective aluminum. In addition, sixty boards of red LED 660 nm wavelength, containing three radiators each, for which the distribution of irradiation and spectral irradiance in all of the six internal faces were calculated in this device. That way, a low cost alternative was tested over three types of microorganisms present on the human microbiota: two strains Gram-positive (Gram +), Staphylococcus aureus and Streptococcus mutans and one strain Gram-negative (Gram -), Escherichia coli, inoculated in orthodontic instruments previously autoclaved. To assess the Photodynamic Inactivation (PDI) over these bacteria, instruments were contaminated by bacterial suspensions (3 × 10⁸ CFU/mL) and ulterior treatment with a solution of 100 μmol/L of MB for 20 min, and irradiated for another 20 min (energy density of 026 J/cm²). Microbial reduction was assessed by number counting of Colony Forming Units (CFU). At the end, microbial reduction of the surface of orthodontic metal instruments was compared with the positive group of each group. Results showed that PID caused a significant reduction (p < 0.05) of the microbial charge stuck in the orthodontic instruments. Thus, the photo prototype of the drawing is appropriate for phototherapy studies, granting it´s advantageous to the low level light therapy as well as for the antimicrobial photodynamic therapy. The perspective is that PID may potentialize the dissemination of phototherapy studies for determining its proper use in health science. And, thus, propose a low cost and atoxic alternative for disinfection of biomedical appliances as non-critical instruments, allowing also for use in the food industry., (Copyright © 2018. Published by Elsevier B.V.)

Introduction: The complex designs of fixed orthodontic appliances hinder proper access for cleaning, favouring the development of gingival inflammation and white spot lesions around brackets. The aim of this study was to evaluate the effectiveness to avoid these undesirable effects by using two prophylactic methods: Photodynamic Therapy (PDT) and conventional ultrasonic scaler (US), in patients during fixed orthodontic treatment., Materials and Methods: Twenty patients under orthodontic treatment for at least 15 months were randomly divided into two groups: PDT mediated by methylene blue (MB) and US. Both treatments were applied in repeated doses (four times in intervals of 2 weeks in the beginning of the study (T0), with booster doses at 3, 6 and 9 months, T1, T2 and T3, respectively). Periodontal clinical parameters (full-mouth plaque score (FMPS), full-mouth bleeding score (FMBS) and probing depth (PD)) were recorded as well as the International Caries Detection and Assessment System (ICDAS) index. Samples of subgingival plaque and saliva (for determination of 4 periodontopathogens and mutans streptococci, respectively), were collected at T0 and at the follow-ups of T1, T2 and T3., Results: The evolution of FMPS, FMBS, PD, ICDAS scores as well as the counts of periodontopathogens and salivary mutans streptococci was similar after US or PDT implementation. Greatest periodontal scores reduction took place at T1 for FMBS and PD and at T2 for FMPS. Main microbial reduction took place at T1. ICDAS index began to increase from T2. No statistically significant intergroup differences were observed., Conclusions: Additional treatment of PDT or US in repeated doses delayed undesired side effects during fixed orthodontic treatment in young patients with low caries risk and signs of gingival inflammation., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objectives: Disinfection of enamel and dentin following cavity preparation is a necessity. Considering the possible effect of antimicrobial agents on tooth structure and dental adhesives, this study aimed to assess the microleakage of composite restorations following disinfection of cavity walls with antimicrobial photodynamic therapy (aPDT)., Materials and Methods: Standard class V cavities were prepared in the buccal surfaces of 68 extracted human premolars. The teeth were then randomly divided into four groups (n = 17) for use of different adhesive systems (Adper Single Bond 2 or One-Step Plus) and disinfection with aPDT. The cavities were filled with Filtek Ultimate composite. After thermocycling and immersion in 0.5% fuchsine, the teeth were sectioned and evaluated under a stereomicroscope to determine the microleakage score. Data were analyzed using the Kruskal Wallis and Wilcoxon tests (P < 0.05)., Results: No significant difference was noted in microleakage in the occlusal and gingival walls among the study groups (P > 0.05). In each study group, microleakage in the gingival wall was significantly higher than that in the occlusal wall (P < 0.05)., Conclusion: Methylene blue-mediated aPDT does not seem to contribute to the microleakage of class V resin-based dental restorations, independently of the composition of bonding agents used., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Antimicrobial photodynamic therapy (aPDT) has been proposed as an adjuvant treatment of dental caries, although there are no well-defined protocols to its clinical application. This study aimed to evaluate the influence of aPDT on the viability of microorganisms, vitality of biofilms, and lactic acid production of dentin caries microcosms. Biofilms were grown on bovine dentin discs in anaerobic conditions at 37 °C for 5 days, inoculating infected carious dentin in modified McBain medium plus 1% sucrose. The biofilms were treated by the combination of deionized water or 100 mg L -1 methylene blue (MB) with 0, 37.5, or 75 J cm -2 LED at 630 nm. The counts of total microorganisms, total streptococci, mutans streptococci, and total lactobacilli were determined by colony-forming units (CFU). The vitality of microbial cells in intact biofilms was measured by confocal laser scanning microscope (CLSM). The lactic acid production was analyzed by enzymatic spectrophotometry at 340 nm. Statistical analysis was conducted by Kruskal-Wallis and post hoc Dunn's tests (P < 0.05). MB and 37.5 J cm -2 LED alone did not interfere in the viability of microorganisms, unlike 75 J cm -2 LED alone that decreased the total microorganism and lactobacillus counts. The combination of MB and 75 J cm -2 LED reduced the viability of all microorganisms and the vitality of intact biofilms. The production of lactic acid was statistically lower in all treatment groups in comparison with that of the control (no treatment), except for MB alone. Therefore, the MB-mediated aPDT was effective in controlling the viability, vitality and the acidogenicity of dentin caries microcosms.

Aims and Background: The number of pathogenic microorganisms has been increasing over the years, and so as resistance of these microorganisms are developing against various antibiotics. Antimicrobial photodynamic therapy (aPDT), also called photodynamic inactivation, is emerging as a promising alternative to treatments based on conventional antibiotics. Recent patents on structured silver mesoporous silica nanoparticles having antimicrobial activity (WO2010/071831 A2), photosensitiser modified core-shell structure nanocomposites (CN 103536935(A)), and Chitosan-coated magnetic mesoporous silica nanoparticles (MSN) (CN 104785214(A)) helped in selecting method of synthesis of MSN and photosensitizers., Materials and Methods: MSN were synthesised by Sol-Gel method and amino functionalised (APTES). Methylene blue (MB) and ortho-toluidine blue (O-TB) were used as photosensitisers. Different batches were synthesised. The final product was characterised by using FTIR, BET, SEM, time resolved fluorescence. The photosensitiser loaded MSN were illuminated by LED based lamp emitting red light at 620± 20nm for different time lengths viz 15 min and 30 mins. Fluorescence studies and antimicrobial assays were carried out as per 72 well plate method I.P, 2014 using, gram negative E. coli (ATCC no. 8739), S. aureus (ATCC no. 7447) and gram positive P. aeruginosa (ATCC no. 9027) pathogenic microorganisms., Results: MB and O-TB were successfully adsorbed on APTES functionalised MSN. Different exposure time length of the photosensitisers to red light showed different zone of inhibition. MB and O-TB loaded MSN showed significant increase in zone of inhibition after irradiation as compared to MB and O-TB loaded on MSN without exposure to light., Conclusion: MB and O-TB adsorbed on APTES functionalized mesoporous silica nanoparticles were capable of efficiently inactivating E. coli, P. aeruginosa, S. aureus bacteria upon exposure to red light (620± 20nm wavelength) at a much lower concentration. Mesoporous silica nanoparticles played an important role in aPDT due to their high surface area and porous structure. Also, APTES functionalization resulted in the pore expansion of MSN, thereby increasing the loading capacity of the photosensitizer on MSN. From the results obtained it can be concluded that O-TB loaded MSN showed higher activity against gram negative and positive microorganisms microorganism as compared to that of MB., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Antimicrobial photodynamic therapy (aPDT) has shown to exert a bactericidal effect against Streptococcus sanguinis and Streptococcus mutans. However, this efficacy has been reported for either type of bacteria separately. Bacterial suspensions of both strains, separately or together, were treated with concentrations of methylene blue (MB) and rose bengal (RB). Suspensions were irradiated with a light-emitting diode lamp (λ center at 625nm for MB and λ center at 515nm for RB) using a fluence of 18J/cm 2 . RB-aPDT at concentrations of 0.16-0.62 and 0.16-0.31μg/mL, and MB-aPDT at concentrations of 0.62-1.25 and 0.31-1.25μg/mL inhibited the growth of S. mutans and S. sanguinis respectively by 6 log 10 . In suspensions of both strains together, the same 6 log 10 reduction in bacterial growth was achieved using the same concentrations of each photosensiziser. In conclusion, RB-aPDT and MB-aPDT appear to exert the same bactericidal effect against suspensions of S. sanguinis and S. mutans either for single strain treatment or for samples constituted by both bacteria mixed together. RB shows to be slightly more efficient than MB., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Introduction and Objectives: Antimicrobial photodynamic therapy combines the use of a photosensitizing drug with light and oxygen to eradicate pathogens. Trichophyton mentagrophytes is a dermatophytic fungus able to invade the skin and keratinized tissues. We have investigated the use of new methylene blue as the photosensitizing agent for antimicrobial photodynamic therapy to produce the in vitro inactivation of T mentagrophytes., Material and Methods: A full factorial design was employed to optimize the parameters for photoinactivation of the dermatophyte. The parameters studied were new methylene blue concentration, contact time between the photosensitizing agent and the fungus prior to light treatment, and the fluence of red light (wavelength, 620-645nm) applied., Results: The minimum concentration of new methylene blue necessary to induce the death of all T. mentagrophytes cells in the initial suspension (approximate concentration, 10 6 colony forming units per milliliter) was 50μM for a fluence of 81J/cm 2 after a contact time of 10minutes with the photosensitizing-agent. Increasing the concentration to 100μM allowed the fluence to be decreased to 9J/cm 2 ., Conclusions: Comparison of our data with other published data shows that the susceptibility of T. mentagrophytes to antimicrobial photodynamic therapy with new methylene blue is strain-dependent. New methylene blue is a photosensitizing agent that should be considered for the treatment of fungal skin infections caused by this dermatophyte., (Copyright © 2016 AEDV. Publicado por Elsevier España, S.L.U. All rights reserved.)

Background: The aim of this study is to investigate the photoinactivation of C. albicans biofilm on acrylic resin discs (the standard material for dental prosthesis) using the photosensitizers Methylene Blue and a Protoporphyrin IX., Methods: Eighteen thermally activated Methyl Methacrylate Polymers were used for the biofilm growth of Candida albicans ATCC 10231. Two photosensitizers were tested: methylene blue (50μM) and protoporphyrin IX (10μM). Two custom-made LEDs emitting at 660nm and 630nm with approximately 800mW each were used for the irradiation, with duration ranging from 2 to 10min., Results: This study demonstrates that MB decreased the aPDT CFUs by approximately two orders of magnitude, but the protoporphyrin was ineffective., Conclusion: The aPDT with MB significantly reduces (but does not sterilize) the amount of CFU after 10min of irradiation, and it is not dose-dependent. The lack of effect of the protoporphyrin is likely because the negative charges of the proteoglycans present in the extracellular matrix repel the negative charges of the PS, thus preventing its diffusion in the cells., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: Antibacterial photodynamic therapy (aPDT) is a promising adjunctive therapy to the treatment of caries lesions, mainly in the minimally invasive approach to preserve dental tissue and favor its repair. Here we analyzed both the efficacy of aPDT in reducing the bacterial load in cariogenic biofilms and the indirect effect of noxious components produced by aPDT on the viability of dental pulp cells., Methods: The aPDT protocol was established using 0.025g/mL methylene blue (MB) and 5min pre-irradiation time. A continuous-wave diode laser (660nm, 0.04cm(2) spot size, 40mW, 60s, 60J/cm(2) and 2.4J) was used in punctual and distance modes to excite the MB. The protocol was first tested against Streptococcus mutans (U159) biofilms produced in 96-well microplates, and then evaluated on caries-like affected human dentin discs of three thicknesses. The number of colony forming units (CFU) was compared between groups. Discs were then assembled in metallic inserts to produce an artificial pulp chamber and allow investigation of the indirect effects of aPDT on dental pulp cells by the 3-(4,5-dimethylthiazol-2-yl)-diphenyltetrazolium bromide (MTT) assay. Data were analyzed using Student's t test or one-way analysis of variance (ANOVA) followed by the Tukey's test at a significance level of 5%., Results: Bacterial load reduction was observed in biofilms produced both in the microplates (p<0.05) and on the caries-like affected dentin discs (81.01% or mean reduction of log21.010±0.1548; p=0.0029). The cell viability of aPDT and control group was similar (p>0.05)., Conclusions: aPDT may be considered a promise adjunctive therapy for deep carious lesions., (Copyright © 2015 Elsevier B.V. All rights reserved.)

In this work, keratin films doped with different amounts of methylene blue (MB) were developed in order to prepare new biodegradable and biocompatible materials for tissue engineering and wound healing, able to exert antimicrobial photodynamic activity upon irradiation with visible light. Preliminary results indicated that the swelling ratio, as well as the MB release, increases by increasing the pH. Moreover, the generation of reactive oxygen species (ROS) and singlet oxygen can be easily triggered and controlled by a fine-tuning of the irradiation time and MB concentration in the films. As concerns the photodynamic effects on keratin, the ROS attack does not induce any significant photodegradation on the protein, even if a slight photo-oxidation of sulfonated amino acids occurs. Finally, the film with the highest MB concentration (400 μg per gram of keratin) displays a significant photobactericidal activity against Staphylococcus aureus with a bacterial reduction that increases by increasing the irradiation time. In particular, the irradiation of KFMB400 film incubated with S. aureus at a concentration of 10(8) cfu mL(-1) determined the 99.9% killing rate and the killing effect increased proportionally with irradiation time.

Oral mucositis (OM) is a debilitating side effect of chemotherapy, which can be relieved by phototherapy. Antimicrobial photodynamic therapy (aPDT) may be used for the treatment of OM, when infection is present. However, there are no studies showing that aPDT affects tissue repair process when used in the treatment of lesions caused by OM. This work aims to evaluate the effect of aPDT in healing OM induced by 5-Fluorouracil (5-FU). Two hundred forty-five hamsters were divided into two groups, control (C) and experimental, which were subdivided into 4 subgroups (Ch, ChP, ChL, aPDT). C group received only the vehicle of chemotherapy and anesthesia, whereas all animals of the experimental groups received anesthesia and chemotherapy agent 5-FU to induce OM. Ch group received no OM treatment; ChP group received an application of methylene blue (MB) 0.01%; ChL received irradiation with low-power-laser (LPL-660 nm/120 J /cm(2)/40 mW/4.4 J per point); and aPDT received MB and LPL irradiation. OM Clinical severity were daily assessed by a blinded examiner. The animals were sacrificed after 5, 7 and 10 days of experiment and their oral mucosa were removed for biochemical (enzymatic activity of SOD and catalase) and histological analyzes (light microscopy). After statistical analysis was performed, results showed that aPDT reduced the severity of OM on the tenth day of the experiment, when compared to the initial OM score (p < 0.05), as well as increased keratinization with organized collagen deposition in the lamina propria. In conclusion, aPDT can be safely used in animals with infected OM because it does not affect lesion-repairing processes., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Objective: The proposed study aims to compare the effectiveness of conventional endodontic treatment (ET) with that of ET associated with antimicrobial photodynamic therapy (aPDT) in patients with apical lesion. Methods: Controlled, double-blind, randomized clinical trial (RCT); superiority study with three parallel arms. Randomization will be conducted in exchange blocks of six, with allocation 1:1:1. The control group will receive conventional ET, while experimental group 1 (EG1) will receive conventional ET + aPDT with laser at 660 nm, fluence of 600 J/cm2; EG2 will receive conventional ET + aPDT with laser at 660 nm, fluence of 1200 J/cm2. The primary outcome will be canal disinfection before treatment, measured by analysis of colony formation (CFU/mL) and the success rate measured after 6 months on the clinical and radiographic evaluations. The mean and standard deviation will be calculated for continuous outcomes, and the CFU/mL mean between groups will be evaluated by ANOVA test. The Chi-squared test will be calculated for binary outcomes. A logistic regression analysis will be performed to assess differences in the success rate between groups, adjusted for the covariates. The Stata 18 software will be used, with a significance threshold of 5%. Conclusions: Few RCTs have evaluated the effectiveness of aPDT in root canal disinfection in patients with permanent dentition presenting apical lesion. New RCTs with larger numbers of participants are needed to support using aPDT as an adjuvant to conventional ET in root canal disinfection for routine use in clinical practice. The trial was registered prospectively in ClinicalTrials.gov (NCT05916859). [ABSTRACT FROM AUTHOR]

Background: Chromoblastomycosis (CMB) is a chronic granulomatous fungal infection that affect the skin and subcutaneous tissues. It is clinically problematic due to limited treatment options, low cure rates, and high rates of relapse. This underscores the necessity for innovative treatment approaches. In this study, potassium iodide (KI) combined with Methylene Blue (MB) mediated antimicrobial photodynamic therapy (PDT) were assessed in the treatment of Fonsecaea monophora (F. monophora) both in vitro and in vivo. And the underlying mechanism that contributes to the efficacy of this treatment approach was investigated. Methods: In vitro experiments were conducted using different combinations and concentrations of MB, KI, and 660 nm light (60 mW/cm2) to inhibit F. monophora. The study was carried out using colony-forming unit (CFU) counts and scanning electron microscopy (SEM). The production of singlet oxygen (1O2), free iodine (I2), hydrogen peroxide (H2O2), and superoxide anion during the KI combined MB-mediated antimicrobial PDT process was also detected. In vivo experiments were developed using a Balb/c mouse paw infection model with F. monophora and treated with PBS, 10 mM KI, 2 mM MB +100 J/cm² and 10 mM KI+2 mM MB +100 J/cm² respectively. Inflammatory swelling, fungal load and histopathological analyses of the mouse footpads were assessed. Results: KI enhanced the killing effect of MB-mediated antimicrobial PDT on the conidial spores of F. monophora at the cell and infected animal model level. During the process, the main antimicrobial agents in KI combined with MB- mediated antimicrobial PDT could produce stronger toxic active species including free I2 and H2O2. Conclusion: KI combined with MB-mediated antimicrobial PDT could be an effective adjunct therapy for treating CBM.

Antifungal resistance in Candida strains has recently increased, especially in C. non-albicans strains, making the development of alternative treatments for fungal infections critical. Antimicrobial photodynamic therapy (aPDT) is a promising alternative treatment that has been successfully used against various infections and microorganisms, including Candida albicans. However, there are still limited studies on C. non-albicans strains, such as Candida tropicalis and Candida glabrata. In this work, the effect of aPDT was evaluated in 18 strains of 3 Candida species: (C. albicans, C. tropicalis and C. glabrata). Synergistic treatments with fluconazole and aPDT using methylene blue (MB) as photosensitizer (PS) were also performed on a C. albicans and a C. tropicalis strain. Resistance to azoles was assessed by Kirby and Bauer disk diffusion method and intracellular MB uptake by cell lysis and spectroscopic analysis. Our results showed that aPDT using MB as PS effectively inhibited the growth of all tested yeast strains in vitro. Even fluconazole-resistant, less-studied strains, such as C. glabrata or C. tropicalis showed significant inhibition, regardless of differences in drug resistance, strain origin, or the MB intracellular concentration. We also observed synergistic effects of aPDT with fluconazole, the combination of sublethal doses of aPDT and fluconazole can effectively eliminate fluconazole-resistant (C. tropicalis) or sensitive (C. albicans) strains. This suggests that aPDT, alone or in combination with antifungal drugs, is a viable alternative treatment for many Candida species. [ABSTRACT FROM AUTHOR]

Antimicrobial photodynamic therapy (aPDT) can be a viable option for management of intranasal infections. However, there are light delivery, fluence, and photosensitizer-related challenges. We report in vitro effectiveness of an easily fabricated, low-cost, portable, LED device and a formulation comprising methylene blue (MB) and potassium iodide (KI) for photoinactivation of pathogens of the nasal cavity, namely, methicillin-resistant Staphylococcus aureus, antibiotic-resistant Klebsiella pneumoniae, multi-antibiotic-resistant Pseudomonas aeruginosa, Candida spp., and SARS-CoV-2. In a 96-well plate, microbial suspensions incubated with 0.005% MB alone or MB and KI formulation were exposed to different red light (~ 660 ± 25 nm) fluence using the LED device fitted to each well. Survival loss in bacteria and fungi was quantified using colony-forming unit assay, and SARS-CoV-2 photodamage was assessed by RT-PCR. The results suggest that KI addition to MB leads to KI concentration-dependent potentiation (up to ~ 5 log10) of photoinactivation in bacteria and fungi. aPDT in the presence of 25 or 50 mM KI shows the following photoinactivation trend; Gm + ve bacteria > Gm − ve bacteria > fungi > virus. aPDT in the presence of 100 mM KI, using 3- or 5-min red light exposure, results in complete eradication of bacteria or fungi, respectively. For SARS-CoV-2, aPDT using MB-KI leads to a ~ 6.5 increase in cycle threshold value. The results demonstrate the photoinactivation effectiveness of the device and MB-KI formulation, which may be helpful in designing of an optimized protocol for future intranasal photoinactivation studies in clinical settings. [ABSTRACT FROM AUTHOR]

Chronic periodontitis is a ubiquitous inflammatory disease in dental healthcare that is challenging to treat due to its impact on bone and tooth loss. Conventional mechanical debridement has been challenging in eliminating complex subgingival biofilms. Hence, adjunctive approaches like low-level laser antimicrobial photodynamic therapy (A-PDT) utilising methylene blue (MB) have been emerging approaches in recent times. This review evaluates the latest research on the use of MB-mediated A-PDT to decrease microbial count and enhance clinical results in chronic periodontitis. Studies have shown the interaction between laser light and MB generates a phototoxic effect thereby, eliminating pathogenic bacteria within periodontal pockets. Moreover, numerous clinical trials have shown that A-PDT using MB can reduce probing depths, improve clinical attachment levels, and decrease bleeding during probing in comparison to traditional treatment approaches. Notably, A-PDT shows superior antibiotic resistance compared to conventional antibiotic treatments. In conclusion, the A-PDT using MB shows promise as an adjunctive treatment for chronic periodontitis. Additional research is required to standardize treatment protocols and assess long-term outcomes of A-PDT with MB in the treatment of periodontitis.

Abstract Background Bacterial infections in lateral canals pose challenges for root canal treatment. This in vitro study aims to evaluate the antibacterial efficacy of sonic-assisted methylene blue mediated antimicrobial photodynamic therapy (MB-aPDT) against Enterococcus faecalis (E. faecalis) in infected lateral canals. Methods Sixty-five premolars infected with E. faecalis in lateral canals were randomly divided into five groups (n = 13) and treated with : (1) 5.25% NaOCl (positive control); (2) Saline (negative control); (3) Sonic-assisted MB-aPDT; (4) 3% NaOCl + MB-aPDT; (5) 3% NaOCl + sonic-assisted MB-aPDT, respectively. The antibacterial efficacy was evaluated by the colony- counting method (CCM) and scanning electronic microscope (SEM). Results Both 5.25% NaOCl and the 3% NaOCl + sonic-assisted MB-aPDT exhibited the most effective while comparable antibacterial effects without significant statistical difference (P > 0.05). Furthermore, the antibacterial effect of the 3% NaOCl + MB-aPDT group was significantly higher compared to that of the sonic-assisted MB-aPDT group (P

Background: Bacterial infections in lateral canals pose challenges for root canal treatment. This in vitro study aims to evaluate the antibacterial efficacy of sonic-assisted methylene blue mediated antimicrobial photodynamic therapy (MB-aPDT) against Enterococcus faecalis (E. faecalis) in infected lateral canals. Methods: Sixty-five premolars infected with E. faecalis in lateral canals were randomly divided into five groups (n = 13) and treated with : (1) 5.25% NaOCl (positive control); (2) Saline (negative control); (3) Sonic-assisted MB-aPDT; (4) 3% NaOCl + MB-aPDT; (5) 3% NaOCl + sonic-assisted MB-aPDT, respectively. The antibacterial efficacy was evaluated by the colony- counting method (CCM) and scanning electronic microscope (SEM). Results: Both 5.25% NaOCl and the 3% NaOCl + sonic-assisted MB-aPDT exhibited the most effective while comparable antibacterial effects without significant statistical difference (P > 0.05). Furthermore, the antibacterial effect of the 3% NaOCl + MB-aPDT group was significantly higher compared to that of the sonic-assisted MB-aPDT group (P < 0.05). The SEM results demonstrated notable morphological alterations in E. faecalis across all experimental groups, except for the negative control group. Conclusion: The concentration of NaOCl can be reduced to a safe level while preserving its antibacterial efficacy through the synergism with the sonic-assisted MB-aPDT in this study. [ABSTRACT FROM AUTHOR]

Antimicrobial photodynamic therapy (aPDT) used to sterilize carious dentin may irritate pulp tissues because of tissue-penetrating laser and singlet oxygen generation. This study aimed to assess the effects of aPDT on rat pulp tissues. A cavity formed in a rat maxillary first molar was treated with aPDT. The combined photosensitizer and laser irradiation conditions in the aPDT groups were as follows: methylene blue and 100 mW for 60 s, brilliant blue (BB) and 100 mW for 60 s, BB and 50 mW for 120 s, and BB and 200 mW for 30 s. Each cavity was treated with an all-in-one adhesive and filled with flowable resin. aPDT was not applied for the control. In each group, the rats were sacrificed on postoperative days 1 and 14, and thin sections of the treated teeth were prepared. Pulp tissue disorganization (PTD), inflammatory cell infiltration (ICI), and tertiary dentin formation (TDF) were evaluated. At 1-day evaluation, there were significant differences between the aPDT group and controls with respect to PTD and ICI (p < 0.01); 14 days later, almost all specimens showed tertiary dentin formation. The application of aPDT caused reversible damage to the rat pulp, while in the long term, healing occurred with the formation of tertiary dentin. [ABSTRACT FROM AUTHOR]

The optimal combinatorial parameters of antimicrobial photodynamic therapy (aPDT) mediated by methylene blue (MB) with the addition of potassium iodide (KI) against Candida species have never been defined. This study aimed to optimize the combinatorial parameters of aPDT, including the concentrations of MB (X1, 0.1–1.0 mM) and KI (X2, 100–400 mM), light dose (X3, 10–70 J/cm2), and MB's incubation time (X4, 5–35 min) for three Candida species. The best MB + KI-aPDT fungicidal effects (Y) against Candida albicans ATCC 90028 (YCa), Candida parapsilosis ATCC 22019 (YCp), and Candida glabrata ATCC 2950 (YCg) were investigated using a uniform design method. The regression models deduced using this method were YCa = 7.126 + 1.199X1X3 − 1.742X12 + 0.206X22 − 0.361X32; YCp = 10.724 − 0.867X1 − 1.497X2 + 0.560X3 + 1.298X22; and YCg = 0.892 − 0.956X1 + 2.296X3 + 1.299X42 − 3.316X3X4. The optimal combinatorial parameters inferred from the regression equations were MB 0.1 mM, KI 400 mM, a light dose of 20 J/cm2, and a 5-minute incubation time of MB for Candida albicans; MB 0.1 mM, KI 400 mM, a light dose of 70 J/cm2, and a 5-minute incubation time of MB for Candida parapsilosis; MB 0.1 mM, KI 100 mM, a light dose of 10 J/cm2, and a 35-minute incubation time of MB for Candida glabrata. The uniform design method can optimize the combinatorial parameters of aPDT mediated by MB plus KI to obtain the best aPDT fungicidal effects on Candida species, providing a new method to optimize the combinatorial parameters of aPDT for different pathogens in the future. [ABSTRACT FROM AUTHOR]

Antimicrobial photodynamic therapy (aPDT) used to sterilize carious dentin may irritate pulp tissues because of tissue-penetrating laser and singlet oxygen generation. This study aimed to assess the effects of aPDT on rat pulp tissues. A cavity formed in a rat maxillary first molar was treated with aPDT. The combined photosensitizer and laser irradiation conditions in the aPDT groups were as follows: methylene blue and 100 mW for 60 s, brilliant blue (BB) and 100 mW for 60 s, BB and 50 mW for 120 s, and BB and 200 mW for 30 s. Each cavity was treated with an all-in-one adhesive and filled with flowable resin. aPDT was not applied for the control. In each group, the rats were sacrificed on postoperative days 1 and 14, and thin sections of the treated teeth were prepared. Pulp tissue disorganization (PTD), inflammatory cell infiltration (ICI), and tertiary dentin formation (TDF) were evaluated. At 1-day evaluation, there were significant differences between the aPDT group and controls with respect to PTD and ICI (p < 0.01); 14 days later, almost all specimens showed tertiary dentin formation. The application of aPDT caused reversible damage to the rat pulp, while in the long term, healing occurred with the formation of tertiary dentin.

Antibacterial resistance causes around 1.27 million deaths annually around the globe and has been recognized as a top 3 priority health threat. Antimicrobial photodynamic therapy (aPDT) is considered a promising alternative to conventional antibiotic treatments. Algal lipid extracts have shown antibacterial effects when used as photosensitizers (PSs) in aPDT. In this work we assessed the photodynamic efficiency of lipidic extracts of microalgae belonging to different phyla (Bacillariophyta, Chlorophyta, Cyanobacteria, Haptophyta, Ochrophyta and Rhodophyta). All the extracts (at 1 mg mL−1) demonstrated a reduction of Staphylococcus aureus >3 log 10 (CFU mL−1), exhibiting bactericidal activity. Bacillariophyta and Haptophyta extracts were the top-performing phyla against S. aureus , achieving a reduction >6 log 10 (CFU mL−1) with light doses of 60 J cm−2 (Bacillariophyta) and 90 J cm−2 (Haptophyta). The photodynamic properties of the Bacillariophyta Phaeodactylum tricornutum and the Haptophyta Tisochrysis lutea , the best effective microalgae lipid extracts, were also assessed at lower concentrations (75 μg mL−1, 7.5 μg mL−1, and 3.75 μg mL−1), reaching, in general, inactivation rates higher than those obtained with the widely used PSs, such as Methylene Blue and Chlorine e6, at lower concentration and light dose. The presence of chlorophyll c , which can absorb a greater amount of energy than chlorophylls a and b; rich content of polyunsaturated fatty acids (PUFAs) and fucoxanthin, which can also produce ROS, e.g. singlet oxygen (1O 2), when photo-energized; a lack of photoprotective carotenoids such as β-carotene, and low content of tocopherol, were associated with the algal extracts with higher antimicrobial activity against S. aureus. The bactericidal activity exhibited by the extracts seems to result from the photooxidation of microalgae PUFAs by the 1O 2 and/or other ROS produced by irradiated chlorophylls/carotenoids, which eventually led to bacterial lipid peroxidation and cell death, but further studies are needed to confirm this hypothesis. These results revealed the potential of an unexplored source of natural photosensitizers (microalgae lipid extracts) that can be used as PSs in aPDT as an alternative to conventional antibiotic treatments, and even to conventional PSs, to combat antibacterial resistance. • Antibacterial resistance is a top health threat, causing 1.27 M deaths annually worldwide. • This work assessed the photodynamic efficiency of 11 microalgae lipidic extracts. • The lipidic extracts were obtained via Folch extraction and encapsulated in liposomes using a Thin-Film methodology. • Extracts from various microalgae phyla showed significant antibacterial effects. [ABSTRACT FROM AUTHOR]

The optimal combinatorial parameters of antimicrobial photodynamic therapy (aPDT) mediated by methylene blue (MB) with the addition of potassium iodide (KI) against Candida species have never been defined. This study aimed to optimize the combinatorial parameters of aPDT, including the concentrations of MB (X1, 0.1–1.0 mM) and KI (X2, 100–400 mM), light dose (X3, 10–70 J/cm2), and MB’s incubation time (X4, 5–35 min) for three Candida species. The best MB + KI-aPDT fungicidal effects (Y) against Candida albicans ATCC 90028 (YCa), Candida parapsilosis ATCC 22019 (YCp), and Candida glabrata ATCC 2950 (YCg) were investigated using a uniform design method. The regression models deduced using this method were YCa = 7.126 + 1.199X1X3 − 1.742X12 + 0.206X22 − 0.361X32; YCp = 10.724 − 0.867X1 − 1.497X2 + 0.560X3 + 1.298X22; and YCg = 0.892 − 0.956X1 + 2.296X3 + 1.299X42 − 3.316X3X4. The optimal combinatorial parameters inferred from the regression equations were MB 0.1 mM, KI 400 mM, a light dose of 20 J/cm2, and a 5-minute incubation time of MB for Candida albicans; MB 0.1 mM, KI 400 mM, a light dose of 70 J/cm2, and a 5-minute incubation time of MB for Candida parapsilosis; MB 0.1 mM, KI 100 mM, a light dose of 10 J/cm2, and a 35-minute incubation time of MB for Candida glabrata. The uniform design method can optimize the combinatorial parameters of aPDT mediated by MB plus KI to obtain the best aPDT fungicidal effects on Candida species, providing a new method to optimize the combinatorial parameters of aPDT for different pathogens in the future.

The laboratorial available methods applied in plasma disinfection can induce damage in other blood components. Antimicrobial photodynamic therapy (aPDT) represents a promising approach and is approved for plasma and platelet disinfection using non-porphyrinic photosensitizers (PSs), such as methylene blue (MB). In this study, the photodynamic action of three cationic porphyrins (Tri-Py(+)-Me, Tetra-Py(+)-Me and Tetra-S-Py(+)-Me) towards viruses was evaluated under white light irradiation at an irradiance of 25 and 150 mW·cm−2, and the results were compared with the efficacy of the approved MB. None of the PSs caused hemolysis at the isotonic conditions, using a T4-like phage as a model of mammalian viruses. All porphyrins were more effective than MB in the photoinactivation of the T4-like phage in plasma. Moreover, the most efficient PS promoted a moderate inactivation rate of the T4-like phage in whole blood. Nevertheless, these porphyrins, such as MB, can be considered promising and safe PSs to photoinactivate viruses in blood plasma. [ABSTRACT FROM AUTHOR]

Objective: Streptococcus mutans is one of the principal causative agents of dental caries (tooth decay) found in the oral cavity. Therefore, this study investigates whether selenium nanoparticles (SeNPs) enhance the efficacy of photodynamic therapy (PDT) against both planktonic communities and the one-day-old biofilm of S. mutans. In this study, the planktonic and 24-h biofilm of S. mutans have been prepared in 96-cell microplates. These forms were treated by methylene blue (MB) and SeNPs and then were exposed to light-emitting diode (LED) lighting. Finally, the results have been reported as CFU/ml. Results: The outcomes demonstrated that MB-induced PDT and SeNPs significantly reduced the number of planktonic bacteria (P-value < 0.001). The comparison between the treated and untreated groups showed that combining therapy with SeNPs and PDT remarkably decreased colony-forming units of one-day-old S. mutans biofilm (P-value < 0.05). The findings revealed that PDT modified by SeNPs had a high potential to destroy S. mutans biofilm. This combination therapy showed promising results to overcome oral infection in dental science. [ABSTRACT FROM AUTHOR]

Antimicrobial photodynamic therapy (APDT) has been studied as a noninvasive therapy for treating cutaneous leishmaniasis to overcome challenges with current treatment, such as toxicity, resistance, and need for in‐patient hospital treatment. Organic light‐emitting diodes (OLEDs) have emerged as an attractive technology that can provide wearable light‐emitting materials that are conformable to human skin. This makes OLEDs ideal candidates for APDT by light‐bandages for ambulatory care. In this work, suitable OLEDs are successfully developed to match the absorbance of three photosensitizers: methylene blue, new methylene blue, and 1,9‐dimethyl‐methylene blue to inactivate two Leishmania species in vitro: Leishmania major and Leishmania amazonensis. Parasites are treated either by LED (20 mW cm−2) or OLED (6.5 mW cm−2) at increasing photosensitizer concentrations at a radiant exposure of 50 J cm−2. 1,9‐Dimethyl‐methylene blue is the most potent photosensitizer, killing both strains at nanomolar concentrations. The effect of different intensities from the OLEDs (0.7, 1.5, and 6.5 mW cm−2) are also explored and it is shown that effective killing of Leishmania occurs even at a very low intensity. These findings demonstrate the great potential of OLEDs as a new approach for ambulatory treatment of cutaneous leishmaniasis by APDT. [ABSTRACT FROM AUTHOR]

Deep tissue abscesses represent a significant source of morbidity and hospital stay, with standard of care drainage failing in many cases. We investigated photodynamic therapy (PDT) as an adjunct to standard of care, to reduce or eliminate bacterial burden. A Phase 1 clinical trial was performed to evaluate safety and feasibility of methylene blue (MB) PDT at the time of abscess drainage. Immediately following drainage, MB-PDT was performed. Diffuse reflectance spectroscopy was performed to evaluate abscess wall optical properties. No study-related adverse events were noted, and no side effects were observed. PDT was well tolerated by all subjects, and infused MB was recovered in all cases. Optical property measurements showed variability in hemoglobin concentration and oxygenation, as well as MB uptake, between subjects. MB-PDT at the time of percutaneous abscess drainage was safe and feasible, and optical property measurements showed wide variance between subjects. [ABSTRACT FROM AUTHOR]

Maryam Pourhajibagher,1–3 Roghayeh Ghorbanzadeh,4 Abbas Bahador1,3,5 1Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran; 2Dental Implant Research Center, Dentistry Research Institute, Tehran University of Medical Sciences, Tehran, Iran; 3Dental Research Center, Dentistry Research Institute, Tehran University of Medical Sciences, Tehran, Iran; 4Private Practice, Tehran, Iran; 5Laser Research Center, Dentistry Research Institute, Tehran University of Medical Sciences, Tehran, Iran Introduction: Porphyromonas gingivalis, an important endodontic pathogen, may be exposed to sublethal doses of photo-activated disinfection (sPAD) during root canal therapy. Such an exposure can affect bacterial survival and virulence features. In this study, we evaluated the effect of sPAD-related oxidative stresses on the expression of oxidative stress response gene (oxyR) in P. gingivalis clinical isolates surviving in vitro photodynamic treatment. Materials and methods: To determine the sPAD, 16 clinical P. gingivalis isolates photosensitized with toluidine blue O (TBO), methylene blue (MB), and indocyanine green (ICG) were irradiated with specific wavelength and energy density of diode laser corresponding to the photosensitizers following bacterial viability measurements. The effects of sPAD on the expression ratio of oxyR of 16 clinical P. gingivalis isolates were then assessed using quantitative real-time PCR (qRT-PCR) assay. Results: Maximum values of sPAD against P. gingivalis were 6.25, 15.6, and 25 µg/mL at fluencies of 171.87, 15.6, and 93.75 J/cm2, respectively, for TBO-, ICG-, and MB-sPAD (P>0.05). ICG-, MB-, and TBO-sPAD could increase the oxyR gene expression of the clinical P. gingivalis isolates 12.3-, 5.6-, and 8.5-fold, respectively. ICG-sPAD increased the expression of oxyR gene in clinical isolates of P. gingivalis ~1.5- and 2-fold higher than TBO- and MB-sPAD, respectively. Conclusion: Our results showed that upregulation of oxyR during sPAD may lead to better survival and increased pathogenicity of P. gingivalis isolates. Therefore, selection of appropriate photo-activated disinfection dosage should be considered for the successful treatment of endodontic infection. Keywords: antimicrobial photodynamic therapy, indocyanine green, methylene blue, oxidative stress, Porphyromonas gingivalis, toluidine blue O, low-level laser therapy