Analyzing Efficacy and Safety of Anti-Fungal Blue Light Therapy via Kernel-Based Modeling the Reactive Oxygen Species Induced by Light.

OBJECTIVE: The goal of this study is to investigate the efficacy, safety, and mechanism of ABL for inactivating Candida albicans (C. albicans), and to determine the best wavelength for treating candida infected disease, by experimental measurements and dynamic modeling. METHODS: The changes in reactive oxygen species (ROS) in C. albicans and human host cells under the irradiation of 385, 405, and 415 nm wavelengths light with irradiance of 50 mW/cm2 were measured. Moreover, a kernel-based nonlinear dynamic model, i.e., nonlinear autoregressive with exogenous inputs (NARX), was developed and applied to predict the concentration of light-induced ROS, whose kernels were selected by a newly developed algorithm based on particle swarm optimization (PSO). RESULTS: The ROS concentration was increased respectively about 10-12 times in C. albicans and about 3-6 times in human epithelial cells by the ABL treatment with the same fluence of 90 J/cm2. The NARX models were respectively fitted to the data from the experiments on both types of cells. Besides, four different kernel functions, including Gaussian, Laplace, linear and polynomial kernels, were compared in their fitting accuracies. The errors with the Laplace kernel turned out to be only 0.2704 and 0.0593, as respectively fitted to the experimental data of the C. albicans and human host cells. CONCLUSION: The results demonstrated the effectiveness of the NARX modeling approach, and revealed that the 415 nm light was more effective as an anti-fungal treatment with less damage to the host cells than the 405 or 385 nm light. SIGNIFICANCE: The kernel-based NARX model identification algorithm offers opportunities for determining the effective and safe light dosages in treating various fungal infection diseases.

Antimicrobial Blue Light Inactivation of Microbial Isolates in Biofilms.

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

Quinine Improves the Fungicidal Effects of Antimicrobial Blue Light: Implications for the Treatment of Cutaneous Candidiasis.

BACKGROUND AND OBJECTIVE: Candida albicans is an opportunistic fungal pathogen of clinical importance and is the primary cause of fungal-associated wound infections, sepsis, or pneumonia in immunocompromised individuals. With the rise in antimicrobial resistance, it is becoming increasingly difficult to successfully treat fungal infections using traditional antifungals, signifying that alternative non-traditional approaches must be explored for their efficacy. STUDY DESIGN/MATERIALS AND METHODS: We investigated the combination of antimicrobial blue light (aBL) and quinine hydrochloride (Q-HCL) for improved inactivation of C. albicans, in vitro and in vivo, relative to either monotherapy. In addition, we evaluated the safety of this combination therapy in vivo using the TUNEL assay. RESULTS: The combination of aBL (108 J/cm2 ) with Q-HCL (1 mg/mL) resulted in a significant improvement in the inactivation of C. albicans planktonic cells in vitro, where a 7.04 log10 colony forming units (CFU) reduction was achieved, compared with aBL alone that only inactivated 3.06 log10 CFU (P < 0.001) or Q-HCL alone which did not result in a loss of viability. aBL + Q-HCL was also effective at inactivating 48-hour biofilms, with an inactivation 1.73 log10 CFU at the dose of 108 J/cm2 aBL and 1 mg/mL Q-HCL, compared with only a 0.73 or 0.66 log10 CFU by aBL and Q-HCL alone, respectively (P < 0.001). Transmission electron microscopy revealed that aBL + Q-HCL induced morphological and ultrastructural changes consistent with cell wall and cytoplasmic damage. In addition, aBL + Q-HCL was effective at eliminating C. albicans within mouse abrasion wounds, with a 2.47 log10 relative luminescence unit (RLU) reduction at the dose of 324 J/cm2 aBL and 0.4 mg/cm2 Q-HCL, compared with a 1.44 log10 RLU reduction by aBL alone. Q-HCL or nystatin alone did not significantly reduce the RLU. The TUNEL assay revealed some apoptotic cells before and 24 hours following treatment with aBL + Q-HCL. CONCLUSION: The combination of aBL + Q-HCL was effective at eliminating C. albicans both in vitro and in vivo. A comprehensive assessment of toxicity (cytotoxicity and genotoxicity) is required to fully determine the safety of aBL + Q-HCL therapy at different doses. In conclusion, the combination of aBL and Q-HCL may be a viable option for the treatment of cutaneous candidiasis. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.

Characterizing the Antimicrobial Properties of 405 nm Light and the Corning® Light-Diffusing Fiber Delivery System.

BACKGROUND AND OBJECTIVES: Hospital-acquired infections (HAIs) and multidrug resistant bacteria pose a significant threat to the U.S. healthcare system. With a dearth of new antibiotic approvals, novel antimicrobial strategies are required to help solve this problem. Violet-blue visible light (400-470 nm) has been shown to elicit strong antimicrobial effects toward many pathogens, including representatives of the ESKAPE bacterial pathogens, which have a high propensity to cause HAIs. However, phototherapeutic solutions to prevention or treating infections are currently limited by efficient and nonobtrusive light-delivery mechanisms. STUDY DESIGN/MATERIALS AND METHODS: Here, we investigate the in vitro antimicrobial properties of flexible Corning® light-diffusing fiber (LDF) toward members of the ESKAPE pathogens in a variety of growth states and in the context of biological materials. Bacteria were grown on agar surfaces, in liquid culture and on abiotic surfaces. We also explored the effects of 405 nm light within the presence of lung surfactant, human serum, and on eukaryotic cells. Pathogens tested include Enterococcus spp, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp., Staphylococcus epidermidis, Streptococcus pyogenes, Candida albicans, and Escherichia coli. RESULTS: Overall, the LDF delivery of 405 nm violet-blue light exerted a significant degree of microbicidal activity against a wide range of pathogens under diverse experimental conditions. CONCLUSIONS: The results exemplify the fiber's promise as a non-traditional approach for the prevention and/or therapeutic intervention of HAIs. Lasers Surg. Med. © 2019 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc.

Daily Phototherapy with Red Light to Regulate Candida albicans Biofilm Growth.

Here, we present a protocol to assess the outcomes of per diem red light treatment on the growth of Candida albicans biofilm. To increase the planktonic growth of C. albicans SN425, the inoculums grew on Yeast Nitrogen Base media. For biofilm formation, RPMI 1640 media, which have high concentrations of amino acids, were applied to help biofilm growth. Biofilms of 48 h were treated twice a day for a period of 1 min with a non-coherent light device (red light; wavelength = 635 nm; energy density = 87.6 J·cm-2). As a positive control (PC), 0.12% chlorhexidine (CHX) was applied, and as a negative control (NC), 0.89% NaCl was applied to the biofilms. Colony forming units (CFU), dry-weight, soluble and insoluble exopolysaccharides were quantified after treatments. Briefly, the protocol presented here is simple, reproducible and provides answers regarding viability, dry-weight and extracellular polysaccharide amounts after red light treatment.

In Vitro Effect of Er:YAG Laser on Different Single and Mixed Microorganisms Being Associated with Endodontic Infections.

Objective: The purpose of this in vitro study was to evaluate the antimicrobial effect of activated irrigation with different modes of erbium-doped yttrium aluminum garnet (Er:YAG) laser application on microorganisms related to secondary endodontic infection. Background: Er:YAG laser has been recommended as an adjuvant tool for root canal disinfection during endodontic treatment. Materials and methods: Laser-activated irrigation (LAI) with 300 or 600 µm tips were tested with or without intermittent irrigation with 0.9% sodium chloride (NaCl) solution against different microorganisms (five single strains and dual species (Streptococcus gordonii combined with Actinomyces oris or Fusobacterium nucleatum) in root canals after 3 days of incubation. In a 21-day infection model, LAI was used together with intermittent rinsing with sodium hypochlorite (NaOCl) against the dual-species mixtures; here the incidence of microbial regrowth after up to 7 days was monitored. Results: In the 3-day root infection model, LAI protocols did not show any significant reduction of the microbial load when compared with manual irrigation with saline solution. In the 21-day infection, S. gordonii combined with A. oris were not detectable anymore after applying the LAI protocol with a 600 µm tip (30 mJ/10 pps) up to 7 days after treatment. Conclusions: Application of LAI with a 600 µm tip by using an Er:YAG laser might be advantageous in treatment of endodontic infections.

Twice-daily red and blue light treatment for Candida albicans biofilm matrix development control.

Phototherapy has been proposed as a direct means of affecting local bacterial infections. However, the use of phototherapy to prevent fungal biofilm development has received comparatively less attention. This study aimed to determine the effects of red light treatment and blue light treatment, without a photosensitizer, on the development of Candida albicans biofilm. During the development of 48-h biofilms of C. albicans SN 425 (n = 10), the biofilms were exposed twice-daily to noncoherent blue and red light (LumaCare; 420 nm and 635 nm). The energy density applied was 72 J cm-2 for blue light and 43.8 J cm2, 87.6 J cm2, and 175.5 J cm2 for red light. Positive control (PC) and negative control (NC) groups were treated twice-daily for 1 min with 0.12% chlorhexidine (CHX) and 0.89% NaCl respectively. Biofilms were analyzed for colony forming units (CFU), dry-weight, and exopolysaccharides (EPS-soluble and EPS-insoluble). Data was analyzed by one-way ANOVA and Tukey post hoc test (α = 0.05). Dry-weight was lower than NC (p < 0.001) and approached PC levels with both red and blue light treatments. CFU were also lower in groups exposed to blue light and higher durations of red light (p < 0.05). EPS-soluble and EPS-insoluble measures were variably reduced by these light exposures. In conclusion, twice-daily exposure to both blue and red lights affect the biofilm development and physiology of polysaccharide production and are potential mechanisms for the control of C. albicans biofilm matrix development.

Antimicrobial Photodynamic Therapy Mediated by Curcumin-Loaded Polymeric Nanoparticles in a Murine Model of Oral Candidiasis.

Antimicrobial photodynamic therapy (aPDT) has been proposed as an alternative method for oral candidiasis (OC), while nanocarriers have been used to improve the water solubility of curcumin (CUR). The aim of this study is to encapsulate CUR in polymeric nanoparticles (NPs) and to evaluate its photodynamic effects on a murine model of OC. Anionic and cationic CUR-NP is synthesized using poly-lactic acid and dextran sulfate and then characterized. Female mice are immunosuppressed and inoculated with Candida albicans (Ca) to induce OC. aPDT is performed by applying CUR-NP or free CUR on the dorsum of the tongue, followed by blue light irradiation for five consecutive days. Nystatin is used as positive control. Afterward, Ca are recovered and cultivated. Animals are euthanized for histological, immunohistochemical, and DNA damage evaluation. Encapsulation in NP improves the water solubility of CUR. Nystatin shows the highest reduction of Ca, followed by aPDT mediated by free CUR, which results in immunolabelling of cytokeratins closer to those observed for healthy animals. Anionic CUR-NP does not show antifungal effect, and cationic CUR-NP reduces Ca even in the absence of light. DNA damage is associated with Ca infection. Consecutive aPDT application is a safe treatment for OC.

An in Vitro Study on the Effect of Combined Treatment with Photodynamic and Chemical Therapies on Candida albicans.

Candida albicans is the most commonly encountered human fungal pathogen, and it is traditionally treated with antimicrobial chemical agents. The antimicrobial effect of these agents is largely weakened by drug resistance and biofilm-associated virulence. Enhancement of the antimicrobial activity of existing agents is needed for effective candidiasis treatment. Our aim was to develop a therapy that combined biofilm disruption with existing antimicrobial agents. Photodynamic therapy (PDT) utilizing curcumin and blue light was tested as an independent therapy and in combination with fluconazole treatment. Viability assays and morphology analysis were used to assess the effectiveness of C. albicans treatment. Results showed that fluconazole treatment decreased the viability of planktonic C. albicans, but the decrease was not as pronounced in adherent C. albicans because its biofilm form was markedly more resistant to the antimicrobiotic. PDT effectively eradicated C. albicans biofilms, and when combined with fluconazole, PDT significantly inhibited C. albicans to a greater extent. This study suggests that the addition of PDT to fluconazole to treat C. albicans infection enhances its effectiveness and can potentially be used clinically.

Influence of pre-irradiation time employed in antimicrobial photodynamic therapy with diode laser.

The aim of the present study was to evaluate, in vitro, the effect of different pre-irradiation times of the photosensitizer in photodynamic therapy in biofilms formed by Streptococcus mutans and Candida albicans, through the evaluation of the microbial load. The factors under study were as follows: times of pre-irradiation of the photosensitizer in three levels (1, 2, or 5 min). For the control of the cariogenic dental biofilm with antimicrobial photodynamic therapy (aPDT), methylene blue (0.01%) was used in association with the diode laser (InGaAlP) with a wavelength of 660 nm. Chlorhexidine digluconate (0.12% CHX) and saline were used as positive and negative controls, respectively. The study design was carried out in complete and randomized blocks. The sample consisted of 15 S. mutans biofilms cultures, randomly divided into five groups and 15 C. albicans cultures, also divided into five groups. The experiment was performed in triplicate (n = 3) and the response variables were obtained through quantitative analysis of bacterial viability, expressed in colony-forming units (CFU) per square millimeter of the specimen area. The data were analyzed with the aid of the ANOVA one-way test and Tukey's post-test. All analyses were performed using the Graph Pad Prism 4.0 program, with a significance level of 5%. For the S. mutans group, only the saline solution presented a statistically significant difference when compared to the other treatments (p < 0.05), that is, the treatment with aPDT, irrespective of the irradiation time applied, was similar to the treatment with CHX and both were more effective in reducing cariogenic biofilm compared to saline. For the group of C. albicans, there was no statistical difference between the groups (p > 0.05). Therefore, it can be concluded that the treatment with aPDT reduced the number of CFUs of S. mutans in a similar way to CHX, independently of the pre-irradiation time applied. No effect of this therapy or of the different pre-irradiation times on the C. albicans biofilm could be observed. In this way, the pre-irradiation time of 1 min can be used to reduce the microbial load of S. mutans.

Properties of halogenated and sulfonated porphyrins relevant for the selection of photosensitizers in anticancer and antimicrobial therapies.

The impact of substituents on the photochemical and biological properties of tetraphenylporphyrin-based photosensitizers for photodynamic therapy of cancer (PDT) as well as photodynamic inactivation of microorganisms (PDI) was examined. Spectroscopic and physicochemical properties were related with therapeutic efficacy in PDT of cancer and PDI of microbial cells in vitro. Less polar halogenated, sulfonamide porphyrins were most readily taken up by cells compared to hydrophilic and anionic porphyrins. The uptake and PDT of a hydrophilic porphyrin was significantly enhanced with incorporation in polymeric micelles (Pluronic L121). Photodynamic inactivation studies were performed against Gram-positive (S. aureus, E. faecalis), Gram-negative bacteria (E. coli, P. aeruginosa, S. marcescens) and fungal yeast (C. albicans). We observed a 6 logs reduction of S. aureus after irradiation (10 J/cm2) in the presence of 20 µM of hydrophilic porphyrin, but this was not improved with incorporation in Pluronic L121. A 2-3 logs reduction was obtained for E. coli using similar doses, and a decrease of 3-4 logs was achieved for C. albicans. Rational substitution of tetraphenylporphyrins improves their photodynamic properties and informs on strategies to obtain photosensitizers for efficient PDT and PDI. However, the design of the photosensitizers must be accompanied by the development of tailored drug formulations.

Genotoxic effect of photodynamic therapy mediated by curcumin on Candida albicans.

Photodynamic therapy (PDT) is a promising method for localized and specific inactivation of fungi and bacteria. A nontoxic light-sensitive compound is taken up by cells, which are then exposed selectively to light, which activates toxicity of the compound. We investigated the potential of sublethal PDT using light-sensitive curcumin (CUR) in combination with blue (455 nm) light to promote reactive oxygen species (ROS) formation in the form of singlet oxygen and DNA damage of Candida albicans. Surprisingly, CUR-mediated PDT but also light alone caused significantly longer comet tails, an indication of DNA damage of C. albicans when compared with the negative control. The intracellular ROS production was also significantly higher for the group treated only with light. However, PDT compared to blue light alone significantly slowed DNA repair. Comet tails decreased during 30 min visualized as a 90% reduction in length in the absence of light for cells treated with light alone, while comet tails of cells treated with PDT only diminished in size about 45%. These results indicate that complex mechanisms may result in PDT in a way that should be considered when choosing the photosensitive compound and other aspects of the treatment design.

A new model of in vitro fungal biofilms formed on human nail fragments allows reliable testing of laser and light therapies against onychomycosis.

Onychomycoses represent approximately 50 % of all nail diseases worldwide. In warmer and more humid countries like Brazil, the incidence of onychomycoses caused by non-dermatophyte molds (NDM, including Fusarium spp.) or yeasts (including Candida albicans) has been increasing. Traditional antifungal treatments used for the dermatophyte-borne disease are less effective against onychomycoses caused by NDM. Although some laser and light treatments have demonstrated clinical efficacy against onychomycosis, their US Food and Drug Administration (FDA) approval as "first-line" therapy is pending, partly due to the lack of well-demonstrated fungicidal activity in a reliable in vitro model. Here, we describe a reliable new in vitro model to determine the fungicidal activity of laser and light therapies against onychomycosis caused by Fusarium oxysporum and C. albicans. Biofilms formed in vitro on sterile human nail fragments were treated with 1064 nm neodymium-doped yttrium aluminum garnet laser (Nd:YAG), 420 nm intense pulsed light (IPL) IPL 420, followed by Nd:YAG, or near-infrared light ((NIR) 700-1400 nm). Light and laser antibiofilm effects were evaluated using cell viability assay and scanning electron microscopy (SEM). All treatments were highly effective against C. albicans and F. oxysporum biofilms, resulting in decreases in cell viability of 45-60 % for C. albicans and 92-100 % for F. oxysporum. The model described here yielded fungicidal activities that matched more closely to those observed in the clinic, when compared to published in vitro models for laser and light therapies. Thus, our model might represent an important tool for the initial testing, validation, and "fine-tuning" of laser and light therapies against onychomycosis.

In vitro combination therapy using low dose clotrimazole and photodynamic therapy leads to enhanced killing of the dermatophyte Trichophyton rubrum.

BACKGROUND: Superficial infections of the skin and mucous membranes caused by dermatophyte fungi are amongst the most common and challenging infections to treat. Previously we demonstrated the phototoxic effects of photodynamic therapy (PDT) towards Trichophyton rubrum, using a green laser to photoactivate Rose Bengal (RB). The aim of this study was to evaluate whether we could; (1) achieve a similar effect using an inexpensive light-emitting diode (LED) to photoactivate RB and (2) to evaluate whether our PDT regime could be combined with standard antifungal drug therapy and increase its effectiveness. METHODS: We designed and built our own inexpensive green (530 nm) LED source and tested its efficacy as part our RB-PDT regime in vitro against T. rubrum. We also examined the potential benefits of incorporating PDT as part of combination therapy and whether the order in which this was done had an impact. First we subjected spore suspensions to sub-inhibitory concentrations of a number of antifungal agents (CLT, MCZ and TRB) for 72 hours followed by RB-PDT. Secondly we subjected spore suspensions to sub-inhibitory PDT followed by drug treatment and evaluated if there were any changes to the minimum inhibitory concentrations (MICs) of the drugs tested. RESULTS: The optimal conditions for photoinactivation of T. rubrum using RB-PDT alone were 140 µM of RB and 24 J/cm2 of LED (equating to a 30-minute exposure). These parameters also caused a 100% reduction in the viability of the pathogenic yeast Candida albicans and the model fungus Saccharomyces cerevisiae. By combining our RB-PDT regime as an adjunct to antifungal drugs we were able to dramatically reduce the exposure times. Treatment of spore suspensions using a sub-inhibitory dose of clotrimazole (CLT) followed by RB-PDT, this order was critical, significantly reduced the exposure times required to achieve 100% inhibition of T. rubrum to 15 minutes as compared to RB-PDT alone. CONCLUSIONS: The combination of antifungal drug and RB-PDT represents an attractive alternative to the current antifungal therapies used to treat superficial fungal diseases. Our approach has the potential to reduce treatment times and drug dosages which can also reduce drug toxicity and improve patient compliance.

Effects of low-level laser irradiation on the pathogenicity of Candida albicans: in vitro and in vivo study.

OBJECTIVE: The purpose of this study was to evaluate the effects of low-level laser irradiation (LLLI) on the in vitro growth characteristics and in vivo pathogenicity of Candida albicans in a murine model in the absence of a photosensitizer. BACKGROUND DATA: C. albicans is an opportunistic commensal organism that causes a wide variety of diseases in human beings, ranging from superficial infections to life-threatening invasive candidiasis. The incidence of C. albicans infection is increasing, because of the greater frequency of acquired immunodeficiency conditions. A high recurrence rate has been reported for vulvovaginal and oral candidiasis, despite the best available treatments. Therefore, the search for new treatment modalities seems quite rational. METHODS: Candida culture plates were exposed to common clinical energies of LLLI: 3, 5, 10, and 20 J at 685 nm (BTL Laser 5000, Medicinos Projektai, Czech Republic, Prague, max power output 50 mW) and 3, 5, 10, 30, and 50 J at 830 nm (BTL Laser 5000, Medicinos Projektai, Czech Republic, Prague, max power output 400 mW). RESULTS: Following LLLI with energies >10 J at both 685 and 830 nm wavelengths, statistically significant effects were observed in vitro on the turbidimetric growth kinetics of C. albicans and in vivo on the survival rate of infected mice (p value ≤ 0.05). Therefore, this energy could be considered a threshold for clinical investigation. CONCLUSIONS: Translating our data into the clinical setting, it can be proposed that a direct laser-based approach without using a photosensitizing dye can significantly reduce the pathogenicity of Candida albicans. It can also be concluded that laser light at specific wavelengths could be a possible promising novel treatment for superficial and mucocutaneous C. albicans infections.

Inactivation of Candida albicans following exposure to 624-nanometer light from a supraluminous diode array.

OBJECTIVE: To determine the potential for visible light (405 or 624 nm) to produce an inhibitory effect on Candida albicans. In addition, the study sought to evaluate a series of doses in terms of their respective inhibiting capabilities. BACKGROUND DATA: The authors have studied the effect of blue light on Staphylococcus aureus and found that a bactericidal outcome can be obtained with low doses of blue light. METHODS: Candida albicans was tested because of its common appearance in human skin and mucous membrane infections. The organism was treated in vitro with 405-nm (blue) and with 624-nm (red) light emitted from a supraluminous diode array. Doses of 3, 9, 15, 30, and 60 J/cm(2) were used. Colony counts were performed and compared with untreated controls using Student t tests and 1-way analysis of variance with Tukey post hoc analysis. RESULTS: The results revealed no inhibition produced by 405 nm on C albicans (F4,20 = 0.901; P = .482). However, 624 nm did inhibit growth of C albicans at 3, 9, and 30 J/cm(2) (F4,20 = 6.064; P = .002). CONCLUSIONS: Appropriate doses of 624-nm light from a supraluminous diode array can inhibit the growth of C albicans in vitro. Three, 9, and 30 J/cm(2) are all effective dose levels.

Adding denture cleanser to microwave disinfection regimen to reduce the irradiation time and the exposure of dentures to high temperatures.

BACKGROUND: The microwave energy is an efficient disinfection method; however, it can generate high temperatures that can result in distortion of the dentures. OBJECTIVES: To evaluate whether the addition of an enzymatic cleanser to microwave disinfection regimen would disinfect dentures with shorter irradiation time. MATERIALS AND METHODS: Seven resin discs colonized with Candida albicans biofilm were placed on the palatal surface of sterile dentures to be randomly assigned to the following treatments: immersion in distilled water for 3 min with 0 (DW), 1 (DW + M1), 2 (DW + M2), or 3 min (DW + M3) of microwave irradiation; or immersion in denture cleanser for 3 min with 0 (DC), 1 (DC + M1), 2 (DC + M2) or 3 min (DC + M3) of irradiation. After the treatments, the viable cells were counted by a blinded examiner. The temperature was measured immediately after irradiation. The data were analyzed by ANOVA and Tukey post hoc tests (α = 0.05). RESULTS: No viable cells were found after DC + M2, DC + M3, and DW + M3 treatments, of which DC + M2 achieved the lowest temperature. No significant difference was found between the effectiveness of DW, DW + M1 and DC treatments (p > 0.05). CONCLUSION: Within the limits of this study, the association of a denture cleanser and microwave energy is efficient to disinfect dentures in lower irradiation time and temperature.

Evaluation of UVB light efficacy for inducing apoptosis in Candida albicans cultures.

BACKGROUND: Candida albicans is known as part of human's skin normal flora; but simultaneously, is the most common opportunistic fungal pathogen of human which can cause a variety of infections including cutaneous candidiasis. Because of the importance of superficial infections like cutaneous candidiasis, we tried to use Ultraviolet B light as a method of phototherapy for inducing apoptosis in irradiated colonies of Candida albicans. MATERIALS AND METHODS: The Ultraviolet B with the wavelength of 302 nanometer was used for irradiating the colonies of Candida albicans. For detecting the eventual apoptotic reactions, the DNA of irradiated colonies as well as control colonies were extracted and then were run in 1% agarose gel electrophoresis containing ethidium bromide to observe luminescent DNA bands. RESULTS: Despite irradiating of Ultraviolet B to yeast cells, no abnormalities including DNA laddering bands or smears were detected in bands formed by total genomic DNA. DISCUSSION: The applied irradiation protocol in this investigation was not successful to induce apoptotic reactions in Ultraviolet-exposed colonies. Maybe, Heat shock proteins as the important part of fungal protein pool, inhibit the inducing pathway of apoptosis in irradiated colonies of Candida albicans.

Denture disinfection by microwave energy: influence of Candida albicans biofilm.

OBJECTIVE: This study evaluated the influence of the area of Candida albicans biofilm on denture disinfection by microwave energy. MATERIALS AND METHODS: Candida albicans biofilm was allowed to form for 72 h on resin discs, and three small coverage or seven large coverage discs were placed onto the palatal surface of sterile maxillary dentures. Each denture was immersed in 200 ml distilled water and individually irradiated at a power of 450, 630 or 900 W for different time intervals (1, 2 or 3 min) (n = 6). The effectiveness of disinfection was evaluated by counting the residual cells. The data were analysed by anova and Tukey's HSD test (α = 0.05). Pearson's correlation test was performed to determine the correlation between effectiveness of sterilisation and temperature. RESULTS: Dentures with a larger area of biofilm demanded a longer irradiation exposure to achieve disinfection (p < 0.001), irrespective of power setting, and in this time no yeast growth was detected. Dentures with small areas of biofilm were disinfected after 1 min at 900 W and 2 min at 450 or 630 W. A positive correlation was found between water temperature and effectiveness of disinfection (r = 0.6170; p < 0.001). CONCLUSION: The C. albicans biofilm area influenced disinfection by microwave energy; therefore dentures with larger biofilm areas required longer irradiation exposure to be disinfected.