Effectiveness of antimicrobial photodynamic therapy (AmPDT) on Staphylococcus aureus using phenothiazine compound with red laser.

The aim of this study was to evaluate, in vitro, the bactericidal effect of antimicrobial photodynamic therapy (AmPDT) using phenothiazinium dyes (Toluidine Blue O and methylene blue, 1:1) using different concentrations (100, 50, 25, 12.5, and 6.25 µg/mL) associated to red laser with different energy densities (2.4, 4.8, 7.2, 9.6, and 12 J/cm2) on a strain of Staphylococcus aureus (ATCC 23529). On this study, tests were performed in triplicate and the samples were distributed into 36 test groups: Control and bacterial suspensions were irradiated with the different energy densities, respectively, in the absence of photosensitizer, bacterial suspensions were irradiated with the laser in the different concentrations of the photosensitizer, and finally bacterial suspensions only in the presence of phenothiazinium dye. The pre-irradiation time was 5 min. Therefore, we analyzed the potential of the AmPDT by counting colony-forming units. The logarithm of CFU/mL (log10 CFU/mL) was calculated and the data was analyzed statistically (ANOVA, Tukey's test, p < 0.05). The results showed that the association 50 and 100 µg/mL with 12 J/cm2 showed the highest percentage of inhibition (100 %). Based upon the present results, it may be concluded that the AmPDT was able to enhance the antimicrobial effect of phenothiazines and both concentration of the compound and energy density are important factors for greater effectiveness of therapy.

Histological evaluation of skin lesions induced by Leishmania braziliensis treated by PACT using Laser light and 1.9 dimethyl-methylene blue.

This study aimed to perform a histological evaluation in skin lesions caused by Leishmania braziliensis after PACT treatment using Laser associated with 1.9. dimethyl methylene blue BALB/c mouse ear infection model was used. A total of 40 animals were assigned into two groups considering time intervals at 5 and 10 weeks and subdivided into four subgroups: Control, Photosensitizer, Laser and PACT. Two therapeutic interventions were performed after the 5th week of infection at 48 h intervals. 1.9 Dimethyl methylene blue was used as a photosensitizer at the concentration of 7 ng/mL, with a non-invasive topical administration method associated with Laser (λ = 660 nm, 40 mW, 12 J/cm2). Sample collection occurred 5 or 10 weeks after therapeutic interventions. The main histological findings were observed in the laser and PACT groups at the 10-week evaluation. The Laser group showed reduced lymphoplasmacytic inflammation and histiocytes (p = 0.0079). The PACT group showed reductions in lymphoplasmacytic inflammation at 5 and 10 weeks, discrete reduction of histiocytes and a higher percentage of tissue remodeling. PACT with non-invasive topical administration of the photosensitizer was able to reduce lymphoplasmacytic inflammation and increase tissue remodeling in leishmaniosis skin lesions. This protocol may be easily used in humans and clinical trial shall be carried out to confirm the animal's findings.

Enhancement of photodynamic inactivation of planktonic cultures of Staphylococcus aureus by DMMB-AuNPs.

Photodynamic inactivation is a promising method for the treatment of infectious diseases. Nanotechnology through gold nanoparticles, as a tool to improve the delivery of photosensitizer is an attractive approach to enhance photodynamic inactivation of bacteria. Moreover, gold nanoparticles enchance the absorption of light due to their plasmon resonance. The aim of this study was to evaluate in vitro photodynamic inactivation effects of 1.9-Dimethyl-Methylene Blue (DMMB)-AuNPs associated with the red LED (λ630 ηm ± 20 ηm, 125 mW, 12 J / cm², 192 s) on S. aureus strain. Eight experimental groups were studied: Control, LED, AuNPs, AuNPs + LED, DMMB, DMMB + LED, DMMB + AuNPs, DMMB + AuNPs + LED. After incubation, the number of bacteria surviving each treatment was determined and then enumerated by viable counting (CFU / mL). The logarithm of CFU / mL (CFU/mL log10) was calculated. All experiments realized in triplicate. The statistical analyses included one-way ANOVA tests, Tukey's multiple comparisons and nonlinear regression, p values <0.05 were considered statistically significant. According to results, the photodynamic inactivation of S. aureus on groups DMMB + LED and DMMB-AuNPs + LED, showed a significant reduction of the microbial load (p < 0.0001) when compared to the Control group. The decimal reduction (RD) of these groups were 99.96 % (RD = 3) and 99.994 % (RD = 4) respectively. In conclusion, these findings demonstrated that photodynamic inactivation is enhanced by using DMMB-AuNPs on S. aureus.

aPDT using nanoconcentration of 1,9-dimethylmethylene blue associated to red light is efficacious in killing Enterococcus faecalis ATCC 29212 in vitro.

The Enterococcus faecalis is a microorganism that causes multiple forms of resistance to a wide range of drugs used clinically. aPDT is a technique in which a visible light activates photosensitizer (PS), resulting in generation of reactive oxygen species that kill bacteria unselectively via an oxidative burst. aPDT is an alternative to antibiotics with the advantage of not causing resistance. The search for an alternative treatment of infections caused by E. faecalis, without using antibiotics, is off great clinical importance. The aim of present investigation was to assess the efficacy of using 3.32 ηg/mL of 1,9-dimethylmethylene blue (DMMB) as photosensitizer associated with the use of either Laser (λ660 nm) or LED (λ632 ±â€¯2 nm) using different energy densities (6, 12 and 18 J/cm2) to kill E. faecalis in vitro. Under different experimental conditions, 14 study groups, in triplicate, were used to compare the efficacy of the aPDT carried out with either the laser or LED lights using different energy densities associated to DMMB. The most probable number method (MPN) was used for quantitative analysis. Photodynamic antimicrobial effectiveness was directly proportional to the energy density used, reaching at 18 J/cm2, 99.999998% reduction of the counts of E. faecalis using both light sources. The results of this study showed that the use of 3.32 ηg/mL of DMMB associated with the use 18 J/cm2 of LED light (λ632 ±â€¯2 nm) reduced >7-log counts of planktonic culture of E. faecalis.

LED antimicrobial photodynamic therapy with phenothiazinium dye against Staphylococcus aureus: An in vitro study.

The objective of this study was to evaluate, in vitro, the bactericidal effect of AmPDT on Staphylococcus aureus (ATCC 25923) using different concentrations (100, 50, 25, 12.5 and 6.25µg/mL) of phenothiazine compound combined with LED light (λ632±2nm) using varied energy densities (12, 9.6, 7.2, 4.8 and 2.4J/cm2). The experiments were carried out in triplicate and the samples were divided into groups: Control, Irradiated (treated only with light at different energy densities), Photosensitizer (treated only in the presence of the dye), AmPDT (treatment with light associated with dye). Counts of the colony forming units and the data obtained were statistically analyzed (ANOVA, Tukey's test, p<0.05). The results showed no difference between irradiated and Control groups. However, using the photosensitizer alone caused significant increased cytotoxicity and consequent reduction on the CFU counts (12.5µg/mL (p<0.001), 25µg/mL, 50µg/mL and 100µg/mL (p<0.0001). When AmPDT was used significant inhibition above 70% were detected for all concentrations of the photosensitize (p<0.0001) except for 6.25µg/mL. The results indicate a dose-response dependent when the photosensitizer is used alone but not for the sole use of the light is used. It is concluded that, a single application of AmPDT, using energy density of 12J/cm2 associated either to 12.5 (81.52%) or 25µg/mL (91.57%) resulted in higher in vitro inhibition of S. aureus.

Does the use of laser photobiomodulation, bone morphogenetic proteins, and guided bone regeneration improve the outcome of autologous bone grafts? An in vivo study in a rodent model.

OBJECTIVE: The aim of the present investigation was to histologically assess the effect of laser photobiomodulation (LBPM) on the repair of autologous bone grafts in a rodent model. BACKGROUND DATA: A major problem in modern dentistry is the recovery of bone defects caused by trauma, surgical procedures, or pathologies. Several types of biomaterials have been used to improve the repair of these defects. These materials are often associated with procedures of guided bone regeneration (GBR). MATERIALS AND METHODS: Twenty four animals were divided into four groups: group I (control); group II (LPBM of the bone graft); group III (bone morphogenetic proteins [BMPs] + bone graft); and group IV (LPBM of the bed and the bone graft + BMPs). When appropriate the bed was filled with lyophilized bovine bone and BMPs used with or without GBR. The animals in the irradiated groups received 10 J/cm(2) per session divided over four points around the defect (4 J/cm(2)), with the first irradiation immediately after surgery, and then repeated seven times every other day. The animals were humanely killed after 40 d. RESULTS: The results showed that in all treatment groups, new bone formation was greater and qualitatively better than the untreated subjects. Control specimens showed a less advanced repair after 40 d, and this was characterized by the presence of medullary tissue, a small amount of bone trabeculi, and some cortical repair. CONCLUSION: We conclude that LPBM has a positive biomodulatory effect on the healing of bone defects, and that this effect was more evident when LPBM was performed on the surgical bed intraoperatively, prior to the placement of the autologous bone graft.