Photodynamic inactivation (PDI) has been reported to be effective to eradicate a wide variety of pathogens, including antimicrobial-resistant microorganisms. However, there are conflicting reports in the literature about the effect of growth phase on the susceptibility to PDI. The aim of this study was to identify the potential molecular targets of PDI on Candida albicans in exponential growth phase after PDI mediated by methylene blue (50μM) and exposure to a 660nm-LED (P=360mW). For this task, scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FT-IR) techniques were employed. Pre-irradiation time was set at 10min and exposure time was 15 min delivering a radiant exposure of 162 J/cm2 on a 24-well plate of about 2 cm2. Morphological analysis revealed cell damage after PDI. FT-IR predominantly showed degradation of functional groups related to C-O of deoxyribose; C-C of DNA; C-O stretching vibration of C-OH group of ribose-RNA; P-O stretching modes from the phosphodiester groups of nucleic acids; C=C, C=N, C=O, N=H proteins and amides. Previous studies from our group had demonstrated different targets on the same cells but in stationary growth phase. Therefore, we can conclude that PDI promoted damage to intracellular structures in fungal cells at exponential-phase growth and information on the susceptibility of different growth phases to PDI can be of great importance for the development of treatment strategies that would lead to inactivation of fungal cells in all possible phases of growth in a way that would turn the clinical PDI treatment effective and predictable.
Antimicrobial Photodynamic Therapy (aPDT) for periodontal disease (PD) treatment has been studied for more than three decades, but there is no consensus among researchers about its role on PD control. PD is an oral infection linked with a variate of systemic diseases affecting mainly immunocompromised and elderly patients. The aim of this study is to evaluate aPDT for PD control and to analyze the periodontal microbiological profile regarding healthcare-associated infections (HAI). Six patients were enrolled in this study and the microbiological profile of 36 sites were evaluated in different moments: initial, after scaling and root planing, after aPDT and one-week post-treatment. aPDT was performed with 100μM water solution of methylene blue (MB), and a diode laser (100mW, 660nm, 90s per point, 9J). After periodontal therapy the pockets were irrigated with hydrogen peroxide (3%), washed with water, and MB was applied when no bleeding was detected. To collect the periodontal pocket content, a sterilized paper point was placed inside the pocket for 30s. The material was transferred to a vial with transport culture medium and sent to a microbiological facility to be analyzed blindly. The results showed the presence of pathogens linked with HAI as Acinetobacter baumannii, Candida albicans e Serratia marcescens immediately after therapy, but one week after treatment, none of them were detected and the periodontal index of all patients improved. This preliminary result point out that aPDT may not only improve periodontal clinical conditions, but also can diminish the systemic-health threat represented by this disease.
Ricardo Scarparo Navarro, Andréa Dias Neves Lago, Clarissa Calil Bonifácio, Fausto Medeiros Mendes, Patrícia Moreira de Freitas, Alessandra Baptista, Silvia Cristina Nunez, Adriana Bona Matos, José Carlos Imparato
The aim of this study was to evaluate the enamel demineralization around cavities prepared by Er,Cr:YSGG laser (2780 nm) and restored with different materials after an acid challenge. The human dental enamel samples were randomly divided in 12 groups (n=10): G1- high-speed drill (HD); G2- Er,Cr:YSGG laser L (3 W, 20 Hz, 53.05 J/cm2)(air 65% - water 55%); G3– L (4 W, 20 Hz, 70.74 J/cm2); G4– L (5 W, 20 Hz, 88.43 J/cm2). Each group was divided in subgroups: 1- glass ionomer cement (GIC), 2- resin modified GIC (RMGIC), 3- composite resin (C). Samples were submitted to an acid challenge (4.8 pH) for7 days. The calcium ion contend (ppm/mm2) from demineralizing solutions were analyzed by atomic emission spectrometry. ANOVA and LSD tests were performed (α=5%). The significant lower average values of calcium loss were observed on G2 + GIC, G2 + RMGIC, G1 + RMGIC (p<0.05); the significant higher values were observed on G1 + C, G4 +GIC, G4 + C (p<0.05). The composite resin showed higher calcium loss than RMGIC and GIC (p<0.05). The lased cavities using lower fluence (53.05J/cm2) showed significant reduced demineralization than higher fluences (70.74 and 88.43J/cm2) (p<0.05). Neither the techniques nor the restorative materials used were able to avoid the enamel demineralization. The findings of this in vitro study suggest that the Er,Cr:YSGG lased cavities restored with GIC or RMGIC or conventional drill cavities with RMGIC were effective on reducing the demineralization around restorations, showing an important potential in preventing secondary caries.
Diabetes Mellitus is a chronic disease that can lead to lower-limb ulceration. The photodynamic therapy (PDT) is based on light interaction with a photosensitizer capable to promote bacterial death and tissue repair acceleration. This study analyzed the effects of PDT in the repair of human diabetic ulcers, by means of microbiological assessment. The clinical study was composed of 12 patients of both sexes with diabetic ulcers in lower limbs that were divided into two groups, control group (n=6) and PDT group (n=6). All patients were treated with collagenase/chloramphenicol during the experimental period, in which 6 of them have received PDT with methylene blue dye (0.01%) associated with laser therapy (660 nm), dose of 6 J/cm2¨ and 30 mW laser power. PDT group received ten treatment sessions. Wounds were evaluated for micro-organisms analysis. It was found a reduction in the occurrence of Staphylococcus aureus in both groups, being that reduction more pronounced in the PDT group. Microbial count was performed on PDT group, showing a statistical difference reduction (p<0.05) when compared before and after the treatment. It is concluded that PDT seems to be effective in microbial reduction of human diabetic wounds, promoting acceleration and improvement of tissue repair quality.ty.
An endotracheal tube (ETT) is required for the management of critically ill, mechanically ventilated patients. Ventilatorassociated pneumonia (VAP) affects patients hospitalized in intensive care units; its risk of occurrence is 1% to up 3% for each day of mechanical ventilation. The polymicrobial nature of VAP is established with mixed bacterial-fungal biofilms colonizing the ETT. The microbial interaction enhances the microbial pathogenesis contributing to high indexes of morbidity/mortality. Antimicrobial Photodynamic Therapy (aPDT) could be a suitable therapy for decontamination of oral cavity and ETT at the same time, but the use of a fiber optics inside the ETT seems to not be appropriated since a cannula for secretion aspiration has to be introduced into the ETT to keep it´s lumen. The aim of this study is to proof the concept that an external light source from a LED is capable of reach all areas of the ETT. We use a commercial ETT, 60μM methylene blue (MB), and a 660nm diode laser and calculated the transmission coefficient of light in different situations as only tube, tube with biofilm and biofilm+MB. The results prove that is possible to transmit light through the tube even in the presence of MB and biofilm although a high attenuation of about 60% was measured depending on the tested condition.
The reduction of pathogenic microorganisms in supragingival plaque is one of the principal factors in caries prevention
and control. A large number of microorganisms have been reported to be inactivated in vitro by photodynamic therapy
(PDT). The purpose of this study was to develop a rat model to investigate the effects of PDT on bacterial reduction in
induced dental caries. Twenty four rats were orally inoculated with Streptococcus mutans cells (ATCC 25175) for three
consecutive days. The animals were fed with a cariogenic diet and water with 10% of sucrose ad libitum, during all
experimental period. Caries lesion formation was confirmed by Optical Coherence Tomography (OCT) 5 days after the
beginning of the experiment. Then, the animals were randomly divided into two groups: Control Group: twelve animals
were untreated by either light or photosensitizer; and PDT Group: twelve animals were treated with 100μM of methylene
blue for 5min and irradiated by a Light Emitting Diode (LED) at λ = 640±30nm, fluence of 172J/cm2, output power of
240mW, and exposure time of 3min. Microbiological samples were collected before, immediately after, 3, 7 and 10 days
after treatment and the number of total microaerophiles was counted. OCT images showed areas of enamel
demineralization on rat molars. Microbiological analysis showed a significant bacterial reduction after PDT.
Furthermore, the number of total microaerophiles in PDT group remained lower than control group until 10 days posttreatment.
These findings suggest that PDT could be an alternative approach to reduce bacteria in dental caries.
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