Effect of photodynamic antimicrobial chemotherapy on Candida albicans in the presence of glucose.

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/cm2), 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.

The In Situ Effect of Titanium Tetrafluoride Gel on Erosion/Abrasion Progression in Human Dentin

Abstract Erosion incidence is increasing and its control is still a challenge in clinical practice. This study evaluated 4% TiF4-gel effects on eroded human dentin subjected to in situ erosive/abrasive episodes. Seventy-two previously eroded dentin slabs (0.05 M citric acid, pH 2.3, 20 min) were allocated to 6 groups (n=12) according to the treatment to be performed during the in situ phase and number of erosive/abrasive cycles, as follows: 4% TiF4-gel applied once (TiF41), twice (TiF42) or three times (TiF43) followed by 1, 2 and 3 erosive/abrasive cycles, respectively. Gel was applied before the beginning of the next cycle. Control groups were subjected to 1 (C1), 2 (C2) and 3 (C3) erosive/abrasive cycles only. A seventh group (n=12) comprised in vitro uneroded samples (UN) subjected to 3 erosive/abrasive cycles. Each cycle corresponded to 2 days of erosive (citric acid 0.5%, pH 2.6, 6x/day) and abrasive (electric toothbrush, 10 s/sample, 1 x/day) challenges. Samples were evaluated under profilometry and environmental scanning electron microscopy (ESEM). Atomic force microscopy images (AFM) were also made (n=3). Repeated measures 2-way ANOVA and Tukey test (p<0.001) showed that TiF42, which did not differ from TiF41 and TiF43, revealed a significant reduction in surface loss compared to all control groups. TiF41 and TiF43 showed no significant difference from C1, but both groups demonstrated significantly smaller surface loss than C2 and C3. ESEM and AFM micrographs suggested alterations on treated surfaces compared to samples from control groups, showing reduced diameters of dentinal tubules lumens. Therefore, TiF4 was able to reduce the progression of erosive/abrasive lesions.

Resumo A incidência da erosão tem aumentado e o seu controle ainda é um desafio na prática clínica. Este estudo avaliou os efeitos do gel de TiF4 a 4% sobre a dentina humana erodida submetida a episódios erosivos/abrasivos in situ. Setenta e dois fragmentos de dentina previamente erodida (ácido cítrico 0,05 M, pH 2,3, 20 min) foram distribuídas em 6 grupos (n=12) de acordo com o tratamento a ser realizado durante a fase in situ e o número de ciclos erosivos/abrasivos, como descrito a seguir: gel de TiF4 a 4% aplicado uma (TiF41), duas (TiF42) ou três vezes (TiF43) seguido de 1, 2 e 3 ciclos erosivos/abrasivos, respectivamente. As aplicações dos géis foram realizadas antes do início do ciclo erosivo seguinte. Grupos controle foram submetidos a 1 (C1), 2 (C2) e 3 (C3) ciclos erosivos/abrasivos apenas. Um sétimo grupo (n=12) compreendia amostras sem erosão in vitro (UN) submetidas a 3 ciclos erosivos/abrasivos. Cada ciclo correspondia a 2 dias de desafios erosivos (ácido cítrico a 0,5%, pH 2,6, 6x/dia) e abrasivos (escova de dentes elétrica, 10 s/amostra, 1x/dia). As amostras foram avaliadas em perfilômetro e Microscopia Eletrônica de Varredura Ambiental (MEV). Imagens de microscopia de força atômica (AFM) também foram capturadas (n=3). ANOVA a 2-fatores para medidas repetidas e o teste de Tukey (p<0,001) demonstraram que TiF42, que não diferiu do TiF41 e TiF43, revelou redução significativa na perda de superfície quando comparado a todos os grupos controle. TiF41 e TiF43 não apresentaram diferença estatisticamente significativa em relação ao C1, mas ambos os grupos demonstraram perda de superfície significativamente menor que C2 e C3. Micrografias de MEV e AFM sugeriram alterações nas superfícies tratadas quando comparadas a amostras dos grupos controle, apresentando redução no diâmetro das luzes dos túbulos dentinários. Portanto, o TiF4 foi capaz de reduzir a progressão das lesões erosivas/abrasivas.

Glucose modulates antimicrobial photodynamic inactivation of Candida albicans in biofilms.

Candida albicans biofilm is a main cause of infections associated with medical devices such as catheters, contact lens and artificial joint prosthesis. The current treatment comprises antifungal chemotherapy that presents low success rates. Photodynamic inactivation (PDI) involves the combination of a photosensitizing compound (PS) and light to generate oxidative stress that has demonstrated effective antimicrobial activity against a broad-spectrum of pathogens, including C. albicans. This fungus senses glucose inducing an upregulation of membrane transporters that can facilitate PS uptake into the cell. The aim of this study was to evaluate the effects of glucose on methylene blue (MB) uptake and its influence on PDI efficiency when combined to a red LED with central wavelength at λ=660nm. C. albicans biofilms were grown on hydrogel disks. Prior to PDI assays, MB uptake tests were performed with and without glucose-sensitization. In this system, the optimum PS administration was determined as 500µM of MB in contact with the biofilm during 30min before irradiation. Irradiation was performed during 3, 6, 9, 12, 15 and 18min with irradiance of 127.3mW/cm2. Our results showed that glucose was able to increase MB uptake in C. albicans cells. In addition, PDI without glucose showed a higher viability reduction until 6min; after 9min, glucose group demonstrated a significant decrease in cell viability when compared to glucose-free group. Taken together, our data suggest that glucose is capable to enhance MB uptake and modulate photodynamic inactivation of C. albicans biofilm.

The In Situ Effect of Titanium Tetrafluoride Gel on Erosion/Abrasion Progression in Human Dentin.

Erosion incidence is increasing and its control is still a challenge in clinical practice. This study evaluated 4% TiF4-gel effects on eroded human dentin subjected to in situ erosive/abrasive episodes. Seventy-two previously eroded dentin slabs (0.05 M citric acid, pH 2.3, 20 min) were allocated to 6 groups (n=12) according to the treatment to be performed during the in situ phase and number of erosive/abrasive cycles, as follows: 4% TiF4-gel applied once (TiF41), twice (TiF42) or three times (TiF43) followed by 1, 2 and 3 erosive/abrasive cycles, respectively. Gel was applied before the beginning of the next cycle. Control groups were subjected to 1 (C1), 2 (C2) and 3 (C3) erosive/abrasive cycles only. A seventh group (n=12) comprised in vitro uneroded samples (UN) subjected to 3 erosive/abrasive cycles. Each cycle corresponded to 2 days of erosive (citric acid 0.5%, pH 2.6, 6x/day) and abrasive (electric toothbrush, 10 s/sample, 1 x/day) challenges. Samples were evaluated under profilometry and environmental scanning electron microscopy (ESEM). Atomic force microscopy images (AFM) were also made (n=3). Repeated measures 2-way ANOVA and Tukey test (p<0.001) showed that TiF42, which did not differ from TiF41 and TiF43, revealed a significant reduction in surface loss compared to all control groups. TiF41 and TiF43 showed no significant difference from C1, but both groups demonstrated significantly smaller surface loss than C2 and C3. ESEM and AFM micrographs suggested alterations on treated surfaces compared to samples from control groups, showing reduced diameters of dentinal tubules lumens. Therefore, TiF4 was able to reduce the progression of erosive/abrasive lesions.

Aggregatibacter actinomycetemcomitans biofilm can be inactivated by methylene blue-mediated photodynamic therapy.

BACKGROUND: The purpose of this study was to evaluate the antibacterial effects of photodynamic action of methylene blue (MB) against Aggregatibacter actinomycetemcomitans organized on biofilm. METHODS: After the biofilm growth in 96 flat-bottom well plate, the following groups were used: control group, untreated by either laser or photosensitizer (PS); MB group or dark toxicity group, which was exposed to MB alone (100µM) for 1min (pre-irradiation time); laser group, irradiated with laser for 5min in the absence of PS and three antimicrobial photodynamic inactivation (APDI) groups, with three exposure times of 1, 3 and 5min of irradiation, corresponding to fluences of 15, 45, and 75J/cm(2) respectively. The results were compared to the control group for statistical proposes. Scanning electronic microscope analysis was used to access structural changes in biofilm. RESULTS: Red laser alone and MB alone were not able to inactivate bacterial biofilm. APDI groups showed differences when compared to the control group and they were dependent on the exposure time. No statistically significant differences were observed among the APDI groups at 1 and 3min of irradiation. On the other hand, 5min of APDI showed 99.85% of bacterial reduction (p=0.0004). In addition, the biofilm loose its structure following 5min APDI. CONCLUSIONS: The results of this study suggest that A. actinomycetemcomitans biofilm can be inactivated by MB mediated APDI.

Protocolo de regressão de parestesia após cirurgia de lateralização de nervo alveolar inferior / Surgical protocol for regression of paresthesia after inferior alveolar nerve lateralization

Objetivo: A cirurgia de lateralização do nervo alveolar inferior é pouco difundida devido ao dano neurossensor denominado pa restesia , causado pelo trauma ao feixe nervoso ocasionado pela manipulação cirúrgica. Este trabalho objetiva descrever um protocolo clínico com finalidade de regressão mais rápida da parestesia em pacientes atendidos no curso de especialização em Implantodontia da Unicastelo submetidos à cirurgia de lateralização do nervo alveolar inferior para a instalação de implantes odontológicos, aumentando a previsibilidade clínica da técnica. Materiais e métodos: neste estudo foi utilizado o aparelho de piezocirurgia para o acesso cirúrgico, soft laser para a bioestimulação e um composto polivitamínico para a regeneração tecidual. Resultados: Com a utilização do protocolo obteve-se uma expressiva redução da parestesia local, com retorno da sensibilidade nervosa em média de 20 dias. Dentro deste estudo, o melhor resultado de recuperação sensitiva foi obtido em oito dias pós-cirurgia, enquanto o mais lento ocorreu após seis meses e 21 dias devido a intercorrência de rompimento do feixe nervoso. Como dado comparativo, há 12 anos os pacientes submetidos à técnica cirúrgica apresentavam um quadro estatístico de retorno sensitivo com dois anos de espera. Conclusões: O protocolo de lateralização do nervo alveolar inferior adotado tem reduzido consideravelmente a incidência de parestesia. A utilização de piezocirurgia, "softlaser" e do composto polivitamínico associados a um bom conhecimento de anatomia e domínio da técnica cirúrgica são determinantes para o êxito do tratamento

Objective: The surgery of the inferior alveolar nerve lateralization has a restricted use because of the neurosensitive damage to the nerve bundle called paresthesia, caused by trauma during manipulation of the nerve for implant insertion. This paper aims to describe a clinical protocol with the purpose of the regression of paresthesia in patients enrolled in the specialization course of Implantology at University Unicastelo undergoing surgery of lateralization of the inferior alveolar nerve for dental implant place- ment, increasing the predictability of the clinical technique. Materiais and methods: In this work the piezoelectric device for the surgical approach, soft laser for biostimulation and a multivitamin compound for tissue regeneration were used to obtain a faster recovery of sensation. Results: Using the protocol we obtained a reduction of paresthesia, with consequent return of sensitivity to an average of 20 days, compared 12 years ago when the group began using the technique, where the average incidence of paresthesia was two vears The faster recovery of the area paresthesia in the study group was 8 days and longe r was 6 months and 21 days with the complication of rupture of the nerve bundle. Conclusions: The protocol adopted for inferior alveolar nerve lateralization has considerably reduced the incidence of paresthesia. The use of piezosurgery, softlaser and multivitamin compound associated with a good kno- wledge of anatomy and masterv of surgical technique are essential to the success of treatment

Bactericidal effect of malachite green and red laser on Actinobacillus actinomycetemcomitans.

The aim of this study was to investigate the ability of malachite green (MG) combined with a low-power red laser to kill Actinobacillus actinomycetemcomitans and to investigate MG photodegradation after photodynamic therapy (PDT) by optical absorption spectroscopy. The etiology of periodontal disease is that microorganisms form a bacterial biofilm on the surface of the teeth. It is an infectious disease and A. actinomycetemcomitans is considered an important agent in biofilm ecology. Instead of using antibiotics, PDT is an alternative approach to eradicate bacteria. Cultures of A. actinomycetemcomitans were exposed to a 30 mW diode red laser, in the presence or absence of MG. A group of cultures was treated in dark conditions in the presence of MG (0.01% w/v) for 5 min. In the presence of MG, two exposure times for laser irradiation were used: t=3 min (energy dose=5.4 J/cm(2)), and t=5 min (energy dose=9 J/cm(2)). The samples were diluted and bacterial colonies were counted and converted into colony forming units. Absorption spectra of the bacterial suspensions, MG, MG-stained bacterial suspensions, and photosensitized bacterial suspensions were obtained. A. actinomycetemcomitans can be photoinactivated by a red laser in the presence of MG. Significant differences were observed between the two energy doses used (p<0.05). Red laser alone and MG alone were not able to kill bacteria. Optical absorption showed that MG is photobleached after irradiation. These results indicate that A. actinomycetemcomitans can be photosensitized by red laser combined with MG and that the dye is photodegraded following irradiation.