Effect of low-power diode laser on infected root canals

Abstract This study evaluated the effect of photodynamic therapy (PDT) on infected root canals. Twenty-one human teeth were selected, and 18 were infected by E. faecalis for 60 days. The antimicrobial strategies tested were: G1. Root canal preparation (RCP) using Niquel-Titanium (NiTi) rotary instruments, 2.5% NaOCl, and final irrigation with 17% EDTA, followed by PDT with methylene blue photosensitizer and laser diode low power; G2. RCP using stainless steel files and the same irrigation and PDT protocols as G1; G3. Same RCP protocol as G1 without PDT; G4. Only irrigation with 2.5% NaOCl; G5. Same PDT protocol as G1 without RCP; G6. Negative control; G7. Positive control. Samples for microbiological tests were collected initially (S1), after RCP (S2), and after PDT (S3). Subsequently, the roots were sectioned and prepared for Scanning Electron Microscopy (SEM) analysis. Bacterial growth was analyzed according to the turbidity of the culture medium, followed by spectrophotometric optical density (nm). The effect of PDT on the dentinal structure was evaluated at magnifications 1,600X and 5,000X and described qualitatively. The Wilcoxon test was used for the comparisons from the same specimens, and the Mann-Whitney test was used to compare groups ((=5%). Bacteria were found in all experimental groups' microbiological samples (S1, S2 and S3). The optical density of culture media was lower in S2 than in S1 of G1, 2, 3, and 4 (p> 0.05). After PDT (S3) in G1 and 2, there was an additional reduction in optical density of the culture medium, respectively (p>0.05). In Group 5, the analysis of culture media at S2 revealed an increase in optical density compared to S1(p>0.05). In SEM images of G1, 2, and 5, dentin with melting and recrystallization areas were evidenced. After preparation of the root canal with the rotary system or manually associated with 2.5% NaOCl, PDT was not able to completely eliminate E. faecalis present in the root canal.

Resumo Este estudo avaliou o efeito da terapia fotodinâmica (PDT) em canais radiculares infectados com E. faecalis. Vinte e um dentes humanos extraídos foram selecionados, e 18 foram infectados por E. faecalis por 60 dias. As estratégias antimicrobianas testadas foram: G1. Preparo do canal radicular (PCR) com instrumentos rotatórios de NiTi, NaOCl 2,5% e irrigação final com EDTA 17%, seguido de PDT com fotossensibilizador azul de metileno e laser diodo de baixa potência; G2. PCR usando limas de aço inoxidável e os mesmos protocolos de irrigação e PDT do G1; G3. Protocolo de PCR similar que G1 sem PDT; G4. Somente irrigação com NaOCl 2,5%; G5. Protocolo similar ao G1, sem PCR; G6. Controle negativo; G7. Controle positivo. Amostras para exames microbiológicos foram coletadas inicialmente (S1), após PCR (S2) e após PDT (S3). Na sequência, as raízes foram seccionadas e preparadas para análise em microscopia eletrônica de varredura (MEV). O crescimento bacteriano foi analisado de acordo com a turbidez do meio de cultura seguida pela densidade óptica espectrofotométrica (nm). O efeito da PDT na estrutura dentinária foi avaliado em aumentos de 1.600X e 5.000X, e descrito qualitativamente. O teste de Wilcoxon foi utilizado para as comparações dos mesmos espécimes e o teste de Mann-Whitney para as comparações entre os grupos ((=5%). Bactérias foram encontradas em todos os grupos experimentais, e em todas as coletas microbiológicas (S1, S2 e S3). A densidade óptica dos meios de cultura foi menor em S2 do que em S1 de G1, 2, 3 e 4 (p>0,05). Após a PDT (S3) em G1 e 2, houve redução adicional na densidade óptica do meio de cultura de 90,0% e 92,0%, respectivamente (p>0,05). No Grupo 5, a análise dos meios de cultura em S2 revelou um aumento de 3,2% na densidade óptica em comparação com S1(p>0,05). Nas imagens de MEV do G1, 2 e 5 foram evidenciadas dentina com áreas de fusão e recristalização. O PDT utilizado após preparo do canal radicular com sistema rotatório ou manual, associado ao NaOCl 2,5%, não foi capaz de eliminar completamente o E. faecalis em biofilme maduro presente no canal radicular.

Evaluation of the Antimicrobial Effect of Photodynamic Therapy and Er:YAG Laser Irradiation on Root Canals Infected with Enterococcus faecalis.

Objective: To carry out a histological and morphometric analysis of the antimicrobial effect of Er:YAG laser irradiation combined with photodynamic therapy (PDT) on root canals infected with Enterococcus faecalis. Background: PDT and Er:YAG laser irradiation may be alternatives for effective endodontic disinfection but there are no data on the combination of these therapies. Materials and methods: Forty single-rooted bovine teeth had their roots contaminated with E. faecalis for 72 h. The teeth were randomly divided into four groups (n = 10): group 1, irrigation with 2.5% sodium hypochlorite (NaOCl); group 2, Er:YAG laser (λ2940 nm, 15 Hz, 100 mJ); group 3, PDT with 0.07% methylene blue as photosensitizer and laser irradiation (λ660 nm, power 40 mW, 5 min); and group 4, Er:YAG laser + PDT. After treatment, the teeth were examined by confocal laser scanning microscopy to verify bacterial viability, and morphometric analysis of the images was performed. Results: The PDT and Er:YAG + PDT treatments promoted the greatest reduction in bacteria among the proposed therapies, whereas 2.5% NaOCl was the least effective in bacterial elimination. A statistically significant difference (p < 0.05) was observed among the groups studied, except between the group combining Er:YAG and PDT and the group treated with PDT alone. Conclusions: PDT combined or not with Er:YAG laser was found to be more effective in root canal disinfection when compared with the other groups.

Raman spectroscopy and sciatic functional index (SFI) after low-level laser therapy (LLLT) in a rat sciatic nerve crush injury model.

Axonotmesis causes sensorimotor and neurofunctional deficits, and its regeneration can occur slowly or not occur if not treated appropriately. Low-level laser therapy (LLLT) promotes nerve regeneration with the proliferation of myelinating Schwann cells to recover the myelin sheath and the production of glycoproteins for endoneurium reconstruction. This study aimed to evaluate the effects of LLLT on sciatic nerve regeneration after compression injury by means of the sciatic functional index (SFI) and Raman spectroscopy (RS). For this, 64 Wistar rats were divided into two groups according to the length of treatment: 14 days (n = 32) and 21 days (n = 32). These two groups were subdivided into four sub-groups of eight animals each (control 1; control 2; laser 660 nm; laser 808 nm). All animals had surgical exposure to the sciatic nerve, and only control 1 did not suffer nerve damage. To cause the lesion in the sciatic nerve, compression was applied with a Kelly clamp for 6 s. The evaluation of sensory deficit was performed by the painful exteroceptive sensitivity (PES) and neuromotor tests by the SFI. Laser 660 nm and laser 808 nm sub-groups were irradiated daily (100 mW, 40 s, energy density of 133 J/cm2). The sciatic nerve segment was removed for RS analysis. The animals showed accentuated sensory and neurofunctional deficit after injury and their rehabilitation occurred more effectively in the sub-groups treated with 660 nm laser. Control 2 sub-group did not obtain functional recovery of gait. The RS identified sphingolipids (718, 1065, and 1440 cm-1) and collagen (700, 852, 1004, 1270, and 1660 cm-1) as biomolecular characteristics of sciatic nerves. Principal component analysis revealed important differences among sub-groups and a directly proportional correlation with SFI, mainly in the sub-group laser 660 nm treated for 21 days. In the axonotmesis-type lesion model presented herein, the 660 nm laser was more efficient in neurofunctional recovery, and the Raman spectra of lipid and protein properties were attributed to the basic biochemical composition of the sciatic nerve.

Impact of photobiomodulation therapy on the morphological aspects of submandibular gland submitted to excretory duct ligation and hypothyroidism: an animal study.

The aim of this study was to evaluate the impact of photobiomodulation therapy (PBMT) on histomorphological aspects of submandibular gland (SMG) submitted to salivary gland duct obstruction in hypothyroid rats. Fifty-six male Wistar rats (250 to 300 g) were divided into 4 groups (n = 14): euthyroid (EU), EU + PBMT, hypothyroid (HYPO), and HYPO + PBMT. Duct obstruction of the left submandibular gland (LSMG) was performed in all animals by a ligature procedure. For the induction of hypothyroidism, total thyroidectomy was performed. PBMT groups received irradiation with AlGaInP diode laser (808 nm, 0.04 W, 0.04cm2 spot size, 60 s, 2.4 J per point, 60 J/cm2, 1 W/cm2). Irradiation was performed immediately, 24 h, and 48 h after the obstruction of the salivary gland duct, in one point, extra oral and perpendicular to the gland. Animals were sacrificed after 24 h and 72 h after duct ligature. Our results indicated that salivary duct obstruction and hypothyroidism caused negative modifications on the salivary glands' histomorphology, especially acinar atrophy, after 24 h and 72 h. HYPO + PBMT showed a significant reduction of the inflammatory infiltrate, congested blood vessels, and acinar atrophy in the SMG submandibular salivary gland in 72 h compared to 24 h (p < 0.05). In conclusion, obstruction of the salivary gland excretory duct and hypothyroidism causes severe sialoadenitis with expressive atrophy of the glandular parenchyma. However, PBMT was able to modulate the inflammatory process and delaying acinar atrophy. This study provided insights to better understand the role of the PBMT on the altered salivary gland by duct ligation and associate hypothyroidism.

Photobiomodulation Therapy in the Proliferation and Differentiation of Human Umbilical Cord Mesenchymal Stem Cells: An In Vitro Study.

Introduction: Since photobiomodulation therapy (PBMT) favors in vitro mesenchymal stem cell (MSC) preconditioning before MSC transplantation, increasing the proliferation of these cells without molecular injuries by conserving their characteristics, in the present in vitro study we analyzed the effect of PBMT on the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells (hUCMSCs). Methods: Irradiation with an InGaAIP Laser (660 nm, 10 mW, 2.5 J/cm2 , 0.08 cm2 spot size, and 10 s) was carried out. The cells were divided into four groups: CONTROL [cells grown in Dulbecco's Modified Eagle Medium (DMEM)], OSTEO (cells grown in an osteogenic medium); PBMT (cells grown in DMEM+PBMT), and OSTEO+PBMT (cells grown in an osteogenic medium plus PBMT). The cell proliferation curve was obtained over periods of 24, 48 and 72 hours using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Osteogenic differentiation was analyzed by the formation of calcium nodules over periods of 7, 14 and 21 days. Morphometric analysis was performed to quantify the total area of nodular calcification. Results: The highest cell proliferation and cell differentiation occurred in the OSTEO+PBMT group, followed by the PBMT, OSTEO and CONTROL groups respectively, at the observed times (P <0.05). Conclusion: PBMT enhanced the osteogenic proliferation and the differentiation of hUCMSCs during the periods tested, without causing damage to the cells and preserving their specific characteristics, a fact that may represent an innovative pretreatment in the application of stem cells.

Anti-Trypanosoma cruzi effect of the photodynamic antiparasitic chemotherapy using phenothiazine derivatives as photosensitizers.

Chagas disease is endemic in Latin America and increasingly found in non-endemic countries. Its treatment is limited due to the variable efficacy and several side effects of benznidazole. Photodynamic antimicrobial chemotherapy (PACT) may be an attractive approach for treating Chagas disease. Here, the trypanocidal activity of PACT was investigated in vitro using phenothiazine derivatives. The cytotoxicity of both, methylene blue (MB) and toluidine blue (TBO), was determined on macrophages cultures using AlamarBlue method. The trypanocidal activity of the two photosensitizers was initially evaluated by determining their IC50 values against trypomastigote forms. After this, the trypanocidal effect was evaluated in cultures of infected macrophages using an automatized image analysis protocol. All experiments were performed in the dark and in the clear phase (after a photodynamic exposure). The compounds showed no cytotoxicity in both phases at the tested concentrations. The IC50 values for the sole use of MB and TBO were 2.6 and 1.2 µM, respectively. The photoactivation of the compounds using a fixed energy density (J/cm2) caused a reduction of the IC50 values to 1.0 and 0.9 µM, respectively. It was found that, on infected macrophage, the use of TBO significantly reduced the number of infected cells and parasitic load, and this effect was increased in the presence of light. The results of the present study are indicative that PACT may be considered as both selective and effective therapeutic intervention for treating Chagas disease.

Photobiomodulation Therapy in Bone Repair Associated with Bone Morphogenetic Proteins and Guided Bone Regeneration: A Histomorphometric Study.

OBJECTIVE: To evaluate the efficacy of photobiomodulation for bone repair of critical surgical wounds with implants of bone morphogenetic proteins (BMPs) and bovine biological membranes, using histological and histomorphometric analysis. BACKGROUND: Tissue engineering has been developing rapidly through the use of various biomaterials for the treatment of bone defects, such as mechanical barriers consisting of biological membranes and implants of biomaterials for bone supply. MATERIALS AND METHODS: Thirty-two male rats were divided into four groups (n = 8): group I-C: control; group II-PT: photobiomodulation therapy; group III-PM: Gen-Pro® BMPs+Gen-Derm® membrane; and group IV-PMPT: Gen-Pro® BMPs+Gen-Derm® membrane+photobiomodulation therapy. A 3 mm bone cavity was performed in the upper third of the lateral surface of the right rat femur to obtain a bone defect considered to be critical. The irradiated groups received seven applications of AlGaAs diode laser 830 nm, P = 40 mW, continuous wave (CW) emission mode, f ∼ 0.6 mm, 4 J/cm2 per point (north, south, east, and west) at 48 h intervals, for a total of 16 J/cm2 per session (final dose: 112 J/cm2). Bone repair was evaluated at sacrifice 15 and 30 days after the procedure. The specimens were embedded in paraffin and stained with hematoxylin and eosin and Picrosirius for analysis by light microscopy and by the Leica interactive measurement module software. Statistical analysis was performed (p < 0.05%). RESULTS: Histological analysis confirmed the histomorphometric results, with the experimental groups showing bone neoformation of significantly higher quality and quantity at the end of 30 days compared with the control group. CONCLUSIONS: Photobiomodulation therapy was effective for bone repair mainly when associated with BMPs and a biological membrane. The results of this study are promising and stimulate further scientific and clinical research.

Does laser phototherapy influence the proliferation of myoepithelial cells in the salivary gland of hypothyroid rats?

Thyroid hormones influence both development and growth of organs and tissues and guarantee metabolic demands that interfere with the quality of digestive secretions, including those of the salivary glands. Laser phototherapy - LPT can modulate various biological phenomena and its diverse effects permit the action on different cell types. The aim of this study was to evaluate the influence of laser phototherapy on myoepithelial cells of salivary glands of hypothyroid rats. Forty-two albino Wistar rats were divided into two main groups: euthyroid (EU) and hypothyroid (HYPO). Hypothyroidism was induced using propylthiouracil (PTU) for 4weeks. Each group was divided into subgroups: control (without laser) and laser groups (Red/infrared - IR). LPT was used on the submandibular gland and was carried out using a diode laser (λ660 or λ780nm, 40mW, spot size 0.04cm2, irradiation area 1cm2, 300s, 6J/cm2 per gland, 12J/cm2 per session) and started two weeks after PTU treatment. LPT was repeated every other day for two weeks. After animal death, the glands were removed, dissected and processed for immunohistochemical analysis. It was observed an increase in the number of myoepithelial cells of hypothyroid control rats in comparison to euthyroid controls (p=0.001). Visible LPT (λ660nm) caused significant higher proliferation of myoepithelial cells in EU rats when compared to IR LPT (λ 780nm)(p≤0.001).It is concluded that, despite the LPT protocol used did not influence myoepithelial proliferation on hypothyroid rats it significantly increased the proliferation on euthyroid animals.

Effectiveness of photodynamic therapy on Candida species isolated from oral samples of children exposed and not exposed to HIV / Efetividade da terapia fotodinâmica sobre espécies de Candida isoladas de amostras bucais de crianças expostas e não expostas ao HIV

ABSTRACT Objective: Identify yeast species isolated from unexposed, exposed and HIV-carrier children, and verify the effectiveness of low power laser photodynamic therapy (PDT) on the yeasts species belonging to the Candida genus. Methods: Fifty children assisted by the Public Health Program of the city of Vitória da Conquista, Bahia, were selected and divided into three groups: unexposed to HIV, exposed to HIV during pregnancy, and HIV-carrier. Saliva samples were collected in a disposable sterile universal container and were plated to Sabouraud dextrose agar supplemented with 0.1 mg/mL chloramphenicol. The plates were incubated at 37°C for 48 h. Three strains of each patient were identified by using an API 20 C AUX system. The strains were submitted to photodynamic therapy (PDT) with a 660 nm low power laser and methylene blue dye at different times of irradiation (90, 180 and 282 sec.). Results: The results showed that the most prevalent species was Candida albicans followed by Candida famata (second most prevalent in unexposed to HIV and HIV carriers)) and Candida parapsilosis (second most prevalent in exposed to HIV group). The CFU/mL of Candida spp. decreased significantly (p<0,05) in all groups treated with PDT compared to the controls. Photodynamic therapy treatments at different exposure times (e.g., PS+L90+, PS+L180+, PS+L282+) revealed that the exposure time of 282 sec. gave the highest reduction of the mean logarithmic CFU/mL. Conclusion: Candida albicans was the most prevalent Candida species in these three groups and Candida non-albicans species, when combined, amounted to a significant percentage of Candida isolates. Photodynamic therapy was effective in inactivating the Candida spp. isolated from the oral cavity of children not exposed to HIV, exposed to HIV and HIV-carriers, with the best photodynamic therapy irradiation time being 282 sec.

RESUMO Objetivo: Identificar espécies de leveduras isoladas de crianças não expostas, expostas ao HIV e portadoras de AIDS, e verificar a eficácia da terapia fotodinâmica com laser de baixa potência sobre as espécies de leveduras pertencentes ao gênero Candida. Métodos: Cinquenta crianças atendidas pelo Programa de Saúde Pública da cidade de Vitória da Conquista, Bahia, foram selecionadas e divididas em três grupos: não expostas ao HIV, expostas ao HIV durante a gravidez e portadoras da AIDS. Amostras de saliva foram coletadas em um recipiente universal descartável estéril e foram semeadas em ágar Sabouraud dextrose suplementado com 0,1 mg / mL de cloranfenicol. As placas foram incubadas a 37°C durante 48h. Três cepas de cada paciente foram identificadas utilizando o sistema API 20 C AUX. As cepas foram submetidas à Terapia Fotodinâmica com laser de baixa potência de 660 nm e corante azul de metileno em diferentes tempos de irradiação (90, 180 e 282 segundos). Resultados: Os resultados mostraram que a espécie isolada mais prevalente nos grupos estuddos foi Candida albicans, seguida de Candida famata (segunda mais prevalene nos grupos não expostos ao HIV e com AIDS) e Candida parapsilosis (segunda mais prevalente no grupo exposto ao HIV). Houve diminuição significante de CFU/ml de Candida spp. (p <0,05) em todos os grupos tratados com terapia fotodinâmica, em comparação com os controles. A terapia fotodinâmica, nos diferentes tempos (e.g., PS+L90+, PS+L180+, PS+L282+) mostrou que o tempo de 282 seg. apresentou a maior redução em media de logarítmo de UFC/mL. Conclusão: Candida albicans foi a espécie de Candida mais prevalente isolada nos três grupos e as espécies de Candida não-albicans, quando combinadas, contribuíram com porcentagem significativa dos isolados de Candida. A PDT foi eficaz na inativação de Candida spp. isoladas a partir da cavidade oral de crianças não expostas, expostas ao HIV e portadoras da AIDS, com o melhor tempo de irradiação sendo o de 282 seg.

Photodynamic Antimicrobial Chemotherapy (PACT) in osteomyelitis induced by Staphylococcus aureus: Microbiological and histological study.

Osteomyelitis is an inflammation either of medullar spaces or of the surface of cortical bones, which represents a bacterial infection. Photodynamic Antimicrobial Chemotherapy (PACT) is a treatment based on a cytotoxic photochemical reaction that induces a series of metabolic reactions and culminates in bacterial suppression. Such effect led to the idea that it could be used as a treatment of osteomyelitis. Following approval by the Animal Experimentation Committee of the School of Dentistry of the Federal University of Bahia, the present randomized study used eighty Wistar rats with the aim to evaluate, by microbiological and histological analysis, the effects of Photodynamic Antimicrobial Chemotherapy - PACT on tibial surgical bone defects in rats infected by Staphylococcus aureus. The animals were divided in groups: Control (non-infected); Control Osteomyelitis Induction; Saline solution; Photosensitizer; Red Laser and PACT - on this group, a diode laser (40mW; λ660nm ∅=0.04cm(2), CW, 10J/cm(2)) was used in combination with 5µg/ml of toluidine blue as the photosensitizer. On the microbiological study, immediately after treatment, the PACT group presented a bacterial reduction of 97.4% (p<0.001). Thirty days after treatment, there was a bacterial reduction of more than 99.9% (p<0.001). In the histological study, it was observed that the PACT group demonstrated an intense presence of osteocytes and absence of bone sequestration and micro-abscesses. The PACT using toluidine blue was effective in reducing the number of S. aureus enabling a better quality bone repair.

Assessing the biochemical changes of tendons of rats in an experimental model of tenotomy under therapeutic ultrasound and LEDs (625 and 945 nm) by near-infrared Raman spectroscopy.

Given the complexity of the tendon repair process, where the neoformed scar tissue tends to prevent or hamper the tendon from performing its normal functions, ultrasound (US) and light-emitting diode (LED) devices have been applied to modulate the tendon repair process. This study aimed to compare the anti-inflammatory and healing effects of therapeutic US and LED in an experimental model of tenotomy using histomorphometry and Raman spectroscopy. Forty Wistar rats (235 ± 8.3 g) were separated into four groups: tenotomy (TC; control), tenotomy associated with ultrasound (US; 1 MHz, 0.5 W/cm(2)), tenotomy associated with LED at 625 nm (LR; 4 J/cm(2)), and tenotomy associated with LED at 945 nm (LIR; 4 J/cm(2)), in two experimental periods of 7 and 14 days. Histomorphometry showed a decrease of inflammatory cells (macrophages and polymorphonuclear leukocytes) for the group treated with US compared to the TC group (ANOVA, p < 0.0001) and the LIR group compared to the TC group (p < 0.05) at the seventh day, indicating an anti-inflammatory effect. In relation to the LR and TC groups, there was no significant difference. Raman spectroscopy revealed that the LR group presented an increased synthesis of collagen I compared to the other treatments and the TC groups on the 14th day (p < 0.01). US therapy showed an adjuvant effect with anti-inflammatory action, and the LED therapy at 625 nm showed an increase in collagen synthesis, contributing to the process of tendon healing.

Influence of laser photobiomodulation (GaAlAs) on salivary flow rate and histomorphometry of the submandibular glands of hypothyroid rats.

The aim of this study was to analyze the influence of laser photobiomodulation in salivary flow, weight, and histomorphometry of the submandibular glands of hypothyroid rats. Fifty-six male Wistar albino rats were divided in euthyroid group and hypothyroid group, treated with propylthiouracil (PTU) to induce hypothyroidism. Each group was divided into control (without laser) and laser groups (GaAlAs): λ660 nm (40 mW), λ780 nm (40 mW), and λ780 nm (70 mW). The laser application on the submandibular gland occurred after 2 weeks of PTU treatment and repeatedly during 2 weeks every 48 h. The rats were anesthetized, tracheostomized, and the evaluation of the salivary flow rate (µL/min/100 g body weight) was made by the weight of the saliva collected for 15 min from the first drop. After the animals' death, the glands were dissected and processed for histological analysis. There was an evident reduction of the salivary flow of hypothyroid rats in all groups in comparison to euthyroid group (Mann-Whitney test, p < 0.05). No significant difference was found in the salivary flow of rats that received laser photobiomodulation compared with their control groups. Histological analysis revealed a decrease in the parenchyma of the salivary glands of hypothyroid rats, but the laser was not able to reverse this process. Hypothyroid rats irradiated or not with laser showed acini and acinar cells with significantly smaller areas than euthyroid groups. The laser photobiomodulation protocol used was not able to change salivary flow or reverse the acinar atrophy process in the submandibular glands of hypothyroid rats.

Effect of laser (λ 660 nm) and LED (λ 630 nm) photobiomodulation on formocresol-induced oral ulcers: a clinical and histological study on rodents.

The aim of this study was to evaluate, clinically and histologically, the influence of laser and LED photobiomodulation in the healing of formocresol-induced oral mucosa ulcers of rats. We used 60 Wistar rats in which oral ulcers were induced on the gingiva of the lower incisors. Forty-eight hours after inducing the ulcers, the animals were divided into three groups: laser, LED, and untreated. Animals from the laser group received irradiation with GaAlAs, 660 nm, CW, 40 mW, φ 4 mm(2), 4.8 J/cm(2). Animals from the LED group received irradiation with InGaAIP, 630 nm, 150 mW, 4.8 J/cm(2), 0.8 cm spot. Forty-eight hours after oral ulcer induction, both irradiations were applied in a punctuate manner in the center of the ulcer at 48-h interval until the end of the experimental period. The animals were killed at 3, 5, 7, and 11 days after day 0. The results of the clinical evaluation showed that the laser and LED phototherapies were able to accelerate the healing of formocresol-induced oral ulcers, which occurred first in the laser group (ANOVA, p < 0.05). Histologically, there was a slight variation between LED and laser therapy; therefore, the laser group proved to be effective in accelerating wound healing, especially at 5 days, whereas the LED group was more effective at the end of the experimental period. It was concluded that laser and LED photobiomodulation were effective in accelerating the healing of formocresol-induced oral ulcers in both clinical and histological aspects.

Análise instrumental da cor através de fotografias digitais após clareamento dentário / Instrumental color analysis using digital photography after tooth bleaching

O propósito deste estudo foi avaliar a efetividade do clareamento dentário através da análise instrumental da cor utilizando fotografias digitais. 54 pacientes foram divididos em três grupos: GI (peróxido de hidrogênio a 35%+ LED), GII (peróxido de hidrogênio a 35%+cálcio) e GIII (peróxido de hidrogênio a 15%+ LED). A cor foi aferida de forma instrumental, através de um espectrofotômetro (VITA Easyshade) e de fotografias digitais (programa Adobe Photoshop) e subjetiva, com as escalas VITAPAN Classical e VITA 3D Master. Foram realizadas duas sessões de clareamento e todos os protocolos seguiram as recomendações dos fabricantes dos produtos. Foi aplicado um questionário visual de avaliação de sensibilidade dentária como forma de validar a segurança dos tratamentos. Os grupos foram comparados pelo teste t de Student, com nível de confiança de 95% (p < 0,05), ANOVA e teste de Tukey. Os resultados dos valores de CIELab* não apresentaram diferenças estatisticamente significativas para L*(p = 0,68) e b*(p = 0,09), mas sim para a*(p < 0,01) na comparação entre a análise da cor aferida pelas fotografias digitais e o espectrofotômetro. Em todos os grupos houve uma redução significativa no tom amarelado (∆b) e aumento da luminosidade (∆L) demonstrando a efetividade do clareamento. Houve maior sensibilidade no Grupo I (p = 0,002). A aferição da cor através de fotografias digitais se constituiu em um instrumento adequado para avaliar a efetividade do clareamento dentário quando comparada com o espectrofotômetro. No entanto deve ser utilizada em conjunto com outros métodos de mensuração já consolidados na rotina clínica...

The purpose of this study was to evaluate the effectiveness of tooth bleaching by instrumental color analysis using digital photographs. 54 patients were divided into three groups: GI (hydrogen peroxide 35% + LED) GII (hydrogen peroxide 35% + calcium) and GIII (hydrogen peroxide 15% + LED). The color was assessed in instrumental form, through a spectrophotometer (VITA Easyshade) and digital photos (Adobe Photoshop program) and subjective, with the scales VITAPAN Classical and VITA 3D Master. There were two sessions of bleaching and all protocols followed the recommendations of the manufacturers of the products. A visual evaluation questionnaire of tooth sensitivity as a way to validate the safety of the treatments was applied. The groups were compared using the Student t test, with a confidence level of 95% (p <0.05), ANOVA and Tukey test. The results of the CIELab* values showed no statistically significant differences in L* (p = 0.68) and b * (p = 0.09), but to a * (p <0.01) when comparing the analysis of color measured by digital photographs and the spectrophotometer. In all groups there was a significant reduction in yellowness (∆b) and increased brightness (∆L) demonstrating the effectiveness of whitening. There was more sensitive in Group I (p= 0.002). Gauging the color using digital photography became an appropriate tool to evaluate the effectiveness of tooth whitening as compared with the spectrophotometer. However it should be used in conjunction with other measurement methods already established in clinical routine...

Cellular effect of low-level laser therapy on the rate and quality of bone formation in mandibular distraction osteogenesis.

OBJECTIVE: Therapeutic lasers have been shown to influence bone physiology and repair. The aim of the present investigation was to evaluate the use of a GaAlAs (λ:810 nm) laser in distraction osteogenesis. BACKGROUND DATA: To reduce problems associated with distraction osteogenesis and shorten the time required for treatment, it is desirable to accelerate the process of bone formation. MATERIALS AND METHODS: Eighteen male rabbits underwent corticotomy of mandibular body, and customized distraction devices were inserted. After a 5-day latency period, the mandibles were lengthened by 0.5 mm/day for 10 days. The rabbits were divided into two groups. A GaAlAs (λ: 810 nm) laser beam with the parameters power (P), 200 mW; energy density (ED), 3 J/cm(2); time (T), 7.5 sec; power density (PD) 400 mW/cm(2); energy (E) 1.5 J and spot diameter, 0.8 mm was directed medially and laterally in the study group; the control group received no laser treatment. The exposure continued with six more doses every other day. Three rabbits from each of the two groups were euthanized on the 10th, 20th, and 40th days post-distraction (consolidation) period. RESULTS: Both light microscopy and scanning electron microscopic (SEM) analysis showed significant improvement in new bone formation in the study group at the 10th and 20th days compared with the control group, but the difference was more prominent on the 10th day. By the 40th day, there were no significant differences between the two groups. CONCLUSIONS: This study shows that a low-level GaAlAs (λ:810 nm; P, 200 mW) laser hastens new bone formation only in the early stages of the consolidation period in distraction osteogenesis, and has no significant effect in later stages.

Do laser/LED phototherapies influence the outcome of the repair of surgical bone defects grafted with biphasic synthetic microgranular HA + ß-tricalcium phosphate? A Raman spectroscopy study.

The treatment of bone loss is difficult. Many techniques are proposed to improve repair, including biomaterials and, recently, phototherapies. This work studied bone mineralization by Raman spectroscopy assessing intensities of Raman peaks of both inorganic (∼ 960, ∼ 1,070 cm(-1)) and organic (∼ 1,454 cm(-1)) contents in animal model. Six groups were studied: clot, laser, light-emitting diode (LED), biomaterial (HA + ß-tricalcium phosphate), laser + biomaterial, and LED + biomaterial. Defects at right tibia were performed with a drill. When indicated, defects were further irradiated at a 48-h interval during 2 weeks. At the 15th and 30th days, the tibias were withdrawn and analyzed. The ∼ 960-cm(-1) peak was significantly affected by phototherapy on both clot- and biomaterial-filled defects. The ∼ 1,070-cm(-1) peak was affected by both time and the use of the LED light on clot-filled defects. On biomaterial-filled defects, only the use of the laser light significantly influenced the outcome. No significant influence of either the time or the use of the light was detected on clot-filled defects as regards the ∼ 1,454-cm(-1) peak. Raman intensities of both mineral and matrix components indicated that the use of laser and LED phototherapies improved the repair of bone defects grafted or not with biphasic synthetic microgranular HA + ß-tricalcium phosphate.

Effect of low-level laser therapy (λ660 nm) on angiogenesis in wound healing: a immunohistochemical study in a rodent model.

The aim of the present investigation was to evaluate the angiogenesis on dorsal cutaneous wounds in a rodent model treated with λ660 nm laser light. New vessel formation is a multistep process involving vessel sprouting, endothelial cell migration, proliferation and tube formation. Although several in vivo studies have shown that laser phototherapy influences tissue repair, a fully understanding of angiogenesis mechanisms are not yet known. Twenty-four young adult male Wistar rats weighing between 200 and 250 g were used. Under general anesthesia, one excisional wound was created on the dorsum of each animal and they were randomly distributed into two groups: one control and one treated with laser (λ660 nm, 16 mW, 10 J/cm2). Each group was subdivided into three subgroups according to the animal death timing (2, 4 and 6 days). Laser irradiation started immediately after surgery and was repeated every other day during the experiment and marked with Sirius Red, specific for collagen, and immunomarked with anti-TGF-ß and anti-von Willebrand factor. Marked sections underwent histological analysis by light microscopy and the mean area of the wound of each animal was calculated and analyzed by ANOVA and Tukey's test (α=0.05). Although at some death periods, collagen expression and number of blood vessels on irradiated animals were higher than in the control ones, no significant differences were found at any time in relation to TGF-ß expression (p>0.05). It was concluded that laser treatment (λ660 nm) contributed to increase angiogenesis.

Effect of Low-Level Laser Therapy (660 nm) on Angiogenesis in Wound Healing: A Immunohistochemical Study in a Rodent Model

The aim of the present investigation was to evaluate the angiogenesis on dorsal cutaneous wounds in a rodent model treated with λ660 nm laser light. New vessel formation is a multistep process involving vessel sprouting, endothelial cell migration, proliferation and tube formation. Although several in vivo studies have shown that laser phototherapy influences tissue repair, a fully understanding of angiogenesis mechanisms are not yet known. Twenty-four young adult male Wistar rats weighing between 200 and 250 g were used. Under general anesthesia, one excisional wound was created on the dorsum of each animal and they were randomly distributed into two groups: one control and one treated with laser (λ660 nm, 16 mW, 10 J/cm2). Each group was subdivided into three subgroups according to the animal death timing (2, 4 and 6 days). Laser irradiation started immediately after surgery and was repeated every other day during the experiment and marked with Sirius Red, specific for collagen, and immunomarked with anti-TGF-β and anti-von Willebrand factor. Marked sections underwent histological analysis by light microscopy and the mean area of the wound of each animal was calculated and analyzed by ANOVA and Tukey's test (α=0.05). Although at some death periods, collagen expression and number of blood vessels on irradiated animals were higher than in the control ones, no significant differences were found at any time in relation to TGF-β expression (p>0.05). It was concluded that laser treatment (λ660 nm) contributed to increase angiogenesis.

O objetivo do trabalho foi avaliar a angiogênese em feridas cutâneas no dorso de ratos tratados com o laser de λ660 nm. A neovascularização é um processo que envolve o aparecimento vascular, a migração das células endoteliais, a proliferação e a formação tubular. Embora diversos estudos in vivo demonstrem que a fototerapia laser influencia no reparo tecidual, uma compreensão completa dos mecanismos da angiogênese ainda não é conhecida. Foram utilizados 24 ratos Wistar novos, machos e adultos pesando entre 200 e 250 g. Uma ferida excisional foi criada no dorso de cada animal sob anestesia geral e os animais foram distribuídos aleatoriamente em dois grupos: G0 (controle) e G1 (laser λ660 nm, 16 mW, 10 J/cm2). Cada grupo foi subdividido em três subgrupos de acordo com o sincronismo da morte dos animais (2, 4 e 6 dias). A irradiação laser foi iniciada imediatamente após a cirurgia, sendo repetida diariamente durante a experiência, avaliada por meio de vermelho de Sirius, específico para o colágeno e avaliação imunológica com anti-TGF-β e o Fator anti-von Willebrand. As seções marcadas foram submetidas à análise histológica no foto-microscópio, onde a área média de cada subgrupo foi calculada e analisada usando o teste ANOVA e de Tukey (α=0,05). Os valores dos animais irradiados foram maiores em certos períodos da morte, na expressão do colágeno e no número de vasos em comparação com os grupos controles. Nenhuma diferença significativa foi encontrada na expressão do TGF-β entre os grupos nos períodos. Conclui-se que o tratamento com laser λ660 nm contribuiu para o aumento da angiogênese.

Effects of imiquimod and low-intensity laser (λ660 nm) in chemically induced oral carcinomas in hamster buccal pouch mucosa.

Squamous cell carcinoma (SCC) is the most common neoplasm of the oral cavity. It is aggressive, highly proliferative, and metastatic. This study aimed to evaluate the effect of LLLT and imiquimod on DMBA chemically induced lesions on the oral mucosa of hamsters. SCCs were induced on 25 hamsters. Animals of G1 (control 1) were killed and the presence of tumors confirmed; G2 (control 2) suffered no interventions for additional 4 weeks; animals of G3 (laser treatment) were irradiated (λ660 nm, 50 mW, CW, Ø=3 mm, 0.07 cm(2), 714.2 mW/cm(2), 133 s, 95 J/cm(2), 6.65 J) at every other day for 4 weeks; animals of G4 (imiquimod treatment) received 5 % imiquimod three times a week for 4 weeks; and animals of G5 (imiquimod and laser treatment) received both treatments for the same period. Samples were taken and underwent histological analysis by light microscopy and were investigated using immunohistochemistry for S-100(+) dendritic cells. In G1, G2, and G3, the evaluations showed malignant tumors and the absence of S-100(+) dendritic cells in the tumor stroma. In G4, 60 % of the animals had no malignant tumors, and S-100(+) dendritic cells were present in the stroma of the tumors as well as dysplasia. In G5, 40 % of the animals presented SCC, with scarce or no S-100(+) dendritic cells. The imiquimod treatment played a direct effect on SCC, demonstrated by the increased number of S-100(+) dendritic cells, which could suggest an important role of immune surveillance against neoplastic proliferation. Furthermore, its association with laser needs to be further investigated.

The efficacy of the use of IR laser phototherapy associated to biphasic ceramic graft and guided bone regeneration on surgical fractures treated with wire osteosynthesis: a comparative laser fluorescence and Raman spectral study on rabbits.

The aim of the present study was to assess, by Raman spectroscopy and laser fluorescence, the repair of surgical fractures fixed with wire osteosynthesis treated or not with infrared laser (λ780 nm, 50 mW, 4 × 4 J/cm(2) =16 J/cm(2), ϕ=0.5 cm(2), CW) associated or not to the use of hydroxyapatite and guided bone regeneration. Surgical tibial fractures were created under general anesthesia on 15 rabbits that were divided into five groups, maintained on individual cages, at day/night cycle, fed with solid laboratory pelted diet, and had water ad libitum. The fractures in groups II, III, IV, and V were fixed with wires. Animals in groups III and V were grafted with hydroxyapatite (HA) and guided bone regeneration (GBR) technique used. Animals in groups IV and V were irradiated at every other day during 2 weeks (4 × 4 J/cm(2), 16 J/cm(2) =112 J/cm(2)). Observation time was that of 30 days. After animal death, specimens were taken and kept in liquid nitrogen and used for Raman spectroscopy. The Raman results showed basal readings of 1,234.38 ± 220. Groups WO+B+L showed higher readings (1,680.22 ± 822) and group WO+B the lowest (501.425 ± 328). Fluorescence data showed basal readings of 5.83333 ± 0.7. Groups WO showed higher readings (6.91667 ± 0.9) and group WO+B+L the lowest (1.66667 ± 0.5). There were significant differences between groups on both cases (p<0.05). Pearson correlation was negative and significant (R (2) = -0.60; p<0.001), and it was indicative that, when the Raman peaks of calcium hydroxyapatite (CHA) are increased, the level of fluorescence is reduced. It is concluded that the use of near-infrared lasertherapy associated to HA graft and GBR was effective in improving bone healing on fractured bones as a result of the increasing deposition of CHA measured by Raman spectroscopy and decrease of the organic components as shown by the fluorescence readings.