Total mouth photodynamic therapy mediated by red LED and porphyrin in individuals with AIDS.

Due to the immune changes resulting from HIV/AIDS infection, systemic and local infections throughout the body are common. The use of high activity antiretroviral therapy has been widely used during treatment, which, added to the use of antibiotics, antifungals, and the patients' own immunocompromised state, cause important changes in the oral microbiota. The emergence of pathological microorganisms and with high resistance to drug therapies are frequent and cause serious damage to the oral health of these patients. In this sense, antimicrobial photodynamic therapy (aPDT) appears as a promising alternative in the control of these oral infections. The aim of the study was to test the effectiveness of a therapeutic protocol for total oral aPDT mediated by a 660-nm red LED (light-emitting diode) associated with porphyrin in individuals with AIDS. Patients were selected by exclusion criteria and randomly distributed into groups to test the effectiveness of antimicrobial aPDT with 50 µg/ml porphyrin associated with the red LED. Before and after the treatments, saliva samples were collected and processed in duplicate in selective culture media. Colonies were counted and the results obtained in Log10 CFU/ml and tested statistically. It was concluded that aPDT was effective in reducing oral enterobacteria, in addition to reducing Streptococcus spp. and general count of microorganisms, when considering the numbers of TCD4 and TCD8 lymphocytes.

Photodynamic therapry with curcumin in the reduction of enterococcus faecalis biofilm in bone cavity: rMicrobiological and spectral fluorescense analysis.

BACKGROUND: Antimicrobial photodynamic therapy (PDT) has emerged as a therapeutic strategy to conventional procedures using antibiotics. OBJECTIVE: To evaluate the antimicrobial effectiveness of PDT using blue light emitting diode (LED) associated with curcumin on biofilms of Enterococcus faecalis in bovine bone cavities and also to analyze the presence of these biofilms through spectral fluorescence. MATERIALS AND METHODS: Standardized suspensions of E. faecalis (ATCC 29212) were incubated in artificial bone cavities for 14 days at 36 °C ± 1 °C for biofilm formation. The test specimens were distributed among the four experimental groups (n = 10): L-C- (control), L + C- (LED for 5 min), L-C+ (curcumin for 5 min) and L + C+ (PDT). Aliquots were collected from the bone cavities after treatments and seeded on BHI agar for 24 h at 36 °C ± 1 °C for CFU count. Before and after each treatment the specimens were submitted to spectral fluorescence, whose images were compared in the Image J program. The log10 CFU/mL results were submitted to the Kruskal-Wallis test (5%) and the biofilm fluorescence spectroscopy results were submitted to the Wilcoxon test (5%). RESULTS: All treatments presented statistical difference when compared to the control, and PDT was responsible for the largest reduction (1.92 log10 CFU/mL). There was a reduction in the fluorescence emitted after the treatments, with greater statistical difference in the PDT group. CONCLUSION: PDT was efficient in the reduction of E. faecalis biofilms. In all groups post treatment there was a significant reduction of biofilms in the fluorescence spectroscopy images with greater reduction in the PDT group.

Fluorescence spectroscopy of Candida albicans biofilms in bone cavities treated with photodynamic therapy using blue LED (450 nm) and curcumin.

Fluorescence spectroscopy may assisst in the diagnosis and control of infectious processes associated with bone lesions of the oral cavity. The aim of this study was to analyze, through fluorescence spectroscopy, Candida albicans biofilms formed in artificial bone cavities treated with photodynamic therapy (PDT) mediated with 450-nm blue light-emitting diode (LED) and curcumin. Another aim of this study was to analyze the existence of a correlation between the effectiveness of the photodynamic treatments and the fluorescence spectroscopy images. Artificial bone lesions (n = 40) were made in bovine bones and inoculated with standard suspensions of Candida albicans (ATCC 18804) for biofilm formation (14 days / 36 °C ± 1 °C). The 40 specimens were distributed among four experimental groups (n = 10): L-C- (control), L + C- (LED for 5 min), L-C+ (curcumin for 5 min), and L + C+ (PDT). Aliquots of 100 µL were collected from the bone cavities after treatments and were seeded in duplicate on Sabouraud dextrose agar for 24 h at 36 °C ± 1 °C and the colony-forming units (CFU/ mL) were counted. Before and after each treatment, the specimens were subjected to spectral fluorescence and the images were compared using the Image J program. The log10 CFU/mL were compared with Kruskal-Wallis and Dunn's Multiple Comparison post-test (significance level at 0.05). The fluorescence histogram values before and after treatment were compared using Wilcoxon test (95%).The correlation between Candida albicans log10 CFU/mL and the number of the fluorescence red pixels spectroscopy was verified using Spearman correlation test. The reduction of Candida albicans log10 CFU/mL in the L + C+ (PDT) group was the most relevant and the fluorescence spectroscopy was correlated to the microbiological result. It was concluded that there was a consistency between the number of Candida albicans log10 CFU/mL and the red pixel data of the fluorescence images, demonstrating that the fluorescence diagnostic device reflects the true microbiological condition of Candida albicans biofilms in the bone cavities during the pre-treatment and post-treatment, thus providing the clinician the ability to dynamically, simply, and instantaneously verify the performance of the treatment used.

Reduced methicillin-resistant Staphylococcus aureus biofilm formation in bone cavities by photodynamic therapy.

Photodynamic Therapy (PDT) is a promising alternative for the treatment of infectious bone lesions in the oral cavity. The objective of this study was to evaluate the antimicrobial effectiveness of PDT using blue LED associated with curcumin in methicillin-resistant Staphylococcus aureus biofilms (MRSA) in bovine bone cavities by fluorescence spectroscopy. Standardized suspensions of MRSA culture were inoculated into bone lesions to form biofilm. Forty bone species were distributed in three distinct groups: L-C- (control); L + C- (LED for 5 min); L-C+ (curcumin incubation for 5 min) and L + C+ (PDT). Aliquots of 100 µL were collected from the bone cavities after the treatments and were cultived in BHI for 24 h at 36 °C ±â€¯1 and bacterial colonies counting were performed. Statistical analysis were performed using the paired t-test and analysis of variance (ANOVA) for the variables studied. RESULTS: The control and PDT groups presented statistically significant differences (p < 0.001). It was possible to reduce 3.666 log10 CFU/mL of MRSA and a reduction in the fluorescence emitted after the treatments was observed. The MRSA reduction in biofilms by PDT was the most efficient treatmnent. There was a significant reduction of biofilms in the L + C- and non-PDT groups by fluorescence spectroscopy images.

Application of photodynamic therapy, laser therapy, and a cellulose membrane for calcaneal pressure ulcer treatment in a diabetic patient: A case report.

Diabetes mellitus is a metabolic disorder in which a person has high blood glucose levels due to inadequate insulin production by the pancreas. Wounds in these individuals cannot heal properly over time due to circulatory changes that hinder and stagnate the healing process. We report the case of an 82-year-old female type 2 diabetes mellitus carrier, presenting to clinical-dermatological examination pressure ulcer (PU) in the right calcaneus region. The patient was treated with photodynamic therapy using curcumin and blue light-emitting diodes (LEDs), laser therapy, and the application of a cellulose membrane in order to promote ulcer decontamination by local action, accelerate wound healing, and maintain favorable conditions of asepsis and moisture, respectively. The ulcer healing occurred after 30days of treatment and total epithelialization was observed. From the results obtained in this case report, we conclude that the combination of photodynamic therapy, laser therapy, and coating with a cellulose membrane is a promising treatment for the healing of PU in diabetic patients.

Photodynamic therapy controls of Staphylococcus aureus intradermal infection in mice.

Infections caused by Staphylococcus aureus lead to skin infections, as well as soft tissues and bone infections. Given the communal resistance to antibiotics developed by strains of this bacterium, photodynamic therapy emerges as a promising alternative treatment to control and cure infections. Females of the Balb/C mice were infected with 108 CFU of methicillin-resistant S. aureus (MRSA) and divided into four distinct groups: P-L- (negative control group), P+L- (group exposed only to curcumin), P-L+ (group exposed only to LED incidence of 450 nm, 75 mW/cm2, and 54 J/cm2 for 10 min), and P+L+ (group exposed to curcumin followed by 10 min of LED irradiation) (n = 24). The mice were euthanized 48 and 72 h after infection, and biologic materials were collected for analysis of the bacterial load, peripheral blood leukocyte counts, and draining lymph nodes cell counts. The normalization of data was checked and the ANOVA test was applied. The bacterial load in the draining lymph node of P+L+ group was lower when compared to the control groups 72 h post infection (p < 0.0001), indicating that the LED incidence associated with curcumin controls of the staphylococci intradermal infection. The number of the total lymph node cells shows to be lower than control groups in the two availed times (p < 0.01). The histological analysis and the counting of white blood cells did not show differences among cells in the blood and in the tissue of infection. This is the first report showing that photodynamic therapy may be effective against MRSA infection in a murine model of intradermal infection.

In vitro effectiveness of 455-nm blue LED to reduce the load of Staphylococcus aureus and Candida albicans biofilms in compact bone tissue.

The aim of this study was to evaluate the effectiveness of a 455-nm blue light-emitting diode (LED), at different application times, to reduce the load of Staphylococcus aureus and Candida albicans biofilms applied to compact bone tissue. The microorganisms S. aureus (ATCC 25923) and C. albicans (ATCC 18804) were used to form biofilms on 160 specimens of compact bones that had been divided into eight experimental groups (n = 10) for each microorganism, according to the times of application of the 455-nm blue LED (1, 2, 3, 4, 5, 7, and 10 min) with an irradiance of 75 mW/cm2. After LED application, decimal dilutions of microorganisms were performed, plated on BHI or Sabouraud agar and incubated for 24 h/35 °C to obtain CFU/mL counts. The findings were statistically analyzed using a ANOVA 5 %. For the group of S. aureus biofilms, all groups of 455-nm LED application differ compared with the control group (p < 0.05), in which no treatment was given. The largest reduction was obtained in the group receiving LED for 10 min (p = 0.00); within this group, a 3.2 log reduction was observed. For the C. albicans biofilms, only those samples receiving 3, 7, and 10 min of LED application presented a significant difference compared with the control group (p < 0.00), indicating that longer application times are required to achieve efficacy. The results of this study show that 455-nm LED light was effective to reduce the load of S. aureus and C. albicans biofilms, especially during 10 min of application.

Antimicrobial photodynamic inactivation of Staphylococcus aureus biofilms in bone specimens using methylene blue, toluidine blue ortho and malachite green: An in vitro study.

OBJECTIVE: To evaluate the in vitro effectiveness of APDI with a 660 nm laser combined with methylene blue (MB), toluidine blue ortho (TBO) and malachite green (MG) dyes to inactivate Staphylococcus aureus (ATCC 25923) biofilms in compact and cancellous bone specimens. METHODS: Eighty specimens of compact and 80 of cancellous bone were contaminated with a standard suspension of the microorganism and incubated for 14 days at 37°C to form biofilms. After this period, the specimens were divided into groups (n=10) according to established treatment: PS-L- (control - no treatment); PSmb+L-, PStbo+L-, PSmg+L- (only MB, TBO or MG for 5 min in the dark); PS-L+ (only laser irradiation for 180 s); and APDImb, APDItbo and APDImg (APDI with MB, TBO or MG for 180 s). The findings were statistically analyzed by ANOVA at 5% significance levels. RESULTS: All experimental treatments showed significant reduction of log CFU/mL S. aureus biofilms when compared with the control group for compact and cancellous bones specimens; the APDI group's treatment was more effective. The APDI carried out for the compact specimens showed better results when compared with cancellous specimens at all times of application. For the group of compact bone, APDImg showed greater reductions in CFU/mL (4.46 log 10). In the group of cancellous bone, the greatest reductions were found in the APDImb group (3.06 log 10). CONCLUSION: APDI with methylene blue, toluidine blue ortho and malachite green dyes and a 660 nm laser proved to be effective in the inactivation of S. aureus biofilms formed in compact and cancellous bone.

In vitro effectiveness of antimicrobial photodynamic therapy (APDT) using a 660 nm laser and malachite green dye in Staphylococcus aureus biofilms arranged on compact and cancellous bone specimens.

The aim of this study was to evaluate the in vitro effectiveness of antimicrobial photodynamic therapy (APDT) using a 660 nm visible laser combined with malachite green (MG) dye in the inactivation of Staphylococcus aureus (ATCC 25923) biofilms formed within compact and cancellous bone specimens. Specimens of 80 compact bones and 80 cancellous bones were contaminated with a standard suspension of S. aureus and incubated for 14 days at 37 °C to allow for the formation of biofilms. The specimens were divided into the following groups (n = 10) according to the treatment conditions: PS-L - (control - no treatment), PS+L - (only MG for 5 min), PS-L + 90 (only laser irradiation for 90 s), PS-L + 180 (only laser irradiation for 180 s), PS-L + 300 (only laser irradiation for 300 s), APDT90 (APDT for 90 s), APDT180 (APDT for 180 s), and APDT300 (APDT for 300 s). The findings were statistically analyzed using an ANOVA 5%. All of the experimental groups were significantly different from the control group for both the compact and cancellous bone specimens. The compact bone specimens that received APDT treatment (for either 90, 180, or 300 s) showed reductions in the log10 CFU/ml of S. aureus by a magnitude of 4 log10. Cancellous bone specimens treated with 300 s of APDT showed the highest efficacy, and these specimens had a reduction in S. aureus CFU/ml by a factor of 3 log10. APDT treatment using these proposed parameters in combination with MG was effective at inactivating S. aureus biofilms in compact and cancellous bone specimens.

Effectiveness of six different disinfectants on removing five microbial species and effects on the topographic characteristics of acrylic resin.

PURPOSE: The aim of this study was to evaluate the effectiveness of disinfectant solutions (1% sodium hypochlorite, 2% chlorhexidine digluconate, 2% glutaraldehyde, 100% vinegar, tabs of sodium perborate-based denture cleanser, and 3.8% sodium perborate) in the disinfection of acrylic resin specimens (n = 10/group) contaminated in vitro by Candida albicans, Streptococcus mutans, S. aureus, Escherichia coli, or Bacillus subtilis as measured by residual colony-forming unit (CFU). In a separate experiment, acrylic resin was treated with disinfectants to monitor potential effects on surface roughness, Ra (microm), which might facilitate microbial adherence. MATERIALS AND METHODS: Three hundred fifty acrylic resin specimens contaminated in vitro with 1 x 10(6) cells/ml suspensions of standard strains of the cited microorganisms were immersed in the disinfectants for 10 minutes; the control group was not submitted to any disinfection process. Final counts of microorganisms per ml were performed by plating method for the evaluation of microbial level reduction. Results were compared statistically by ANOVA and Tukey's test (p< or = 0.05). In a parallel study aiming to evaluate the effect of the tested disinfectant on resin surface, 60 specimens were analyzed in a digital rugosimeter before and after ten cycles of 10-minute immersion in the disinfectants. Measurements of superficial roughness, Ra (mum), were compared statistically by paired t-test (p< or = 0.05). RESULTS: The results showed that 1% sodium hypochlorite, 2% glutaraldehyde, and 2% chlorhexidine digluconate were most effective against the analyzed microorganisms, followed by 100% vinegar, 3.8% sodium perborate, and tabs of sodium perborate-based denture cleanser. Superficial roughness of the specimens was higher after disinfection cycles with 3.8% sodium perborate (p= 0.03) and lower after the cycles with 2% chlorhexidine digluconate (p= 0.04). CONCLUSION: Within the limits of this experiment, it could be concluded that 1% sodium hypochlorite, 2% glutaraldehyde, 2% chlorhexidine, 100% vinegar, and 3.8% sodium perborate are valid alternatives for the disinfection of acrylic resin.