Perfil de IL-6 e TNF na Formação de Células Espumosas: Um Método Aprimorado Usando a Sonda de Isotiocianato de Fluoresceína (FITC) / Profile of IL-6 and TNF in Foam Cell Formation: An Improved Method Using Fluorescein Isothiocyanate (FITC) Probe

Resumo Fundamento A formação de células espumosas ocorre devido ao aumento em lipoproteína plasmática de baixa densidade (LDL) e desregulação da inflamação, sendo importante para o desenvolvimento da aterosclerose. Objetivo Avaliar o perfil do fator de necrose tumoral alfa (TNF-α) e da interleucina-6 (IL-6) no método de formação da célula espumosa existente, otimizando esse protocolo. Métodos A LDL foi isolada, oxidada e marcada com sonda de isotiocianato de fluoresceína (FITC). As células espumosas foram geradas de célula derivada de monócitos humanos THP-1 e incubadas na ausência (controle) ou presença de FITC-ox-LDL (10, 50, 100, 150 ou 200 μg/mL), por 12, 24, 48 ou 72 horas. A FITC-ox-LDL na célula foi quantificada por microscopia. O ensaio de imunoabsorção enzimática foi avaliado para quantificar a IL-6 e o TNF-α, com um p <0,05 considerado significativo. Resultados Todas as concentrações de FITC-ox-LDL testadas apresentaram fluorescência mais alta em comparação com o controle, demonstrando maior acúmulo de lipoproteínas nas células. Quanto mais alta a concentração de FITC-ox-LDL, maior a produção de TNF-α e IL-6. A produção de IL-6 pelas células espumosas foi detectada até o valor de 150 µg/mL da LDL máxima de estímulo. Concentrações acima de 50 μg/mL de LDL estimularam maior liberação de TNF-α comparado ao controle. Conclusões Nosso modelo contribui para o entendimento da liberação de IL-6 e TNF-α em resposta a várias concentrações de ox-LDL usando o método otimizado para a formação de células espumosas.

Abstract Background The formation of foam cells occurs due to the increase in low-density plasma lipoprotein (LDL) and dysregulation of inflammation, which is important for the development of atherosclerosis. Objective To evaluate the profile of tumor necrosis factor-alpha (TNF-α) and Interleukin-6 (IL-6) in the existing foam cell formation method, optimizing this protocol. Methods The LDL was isolated, oxidized, and labeled with a Fluorescein isothiocyanate (FITC) probe. Foam cells were generated from THP-1 human monocyte-derived cells and incubated in the absence (control) or presence of FITC-ox-LDL (10, 50, 100, 150, or 200 μg/mL), for 12, 24, 48, or 72 hours. The accumulated FITC-ox-LDL in the cell was quantified by microscopy. The enzyme-linked immunosorbent assay was evaluated to quantify the IL-6 and TNF-α, with p < 0.05 considered significant. Results All the FITC-ox-LDL concentrations tested showed a higher fluorescence when compared to the control, showing a greater accumulation of lipoprotein in cells. The higher the concentration of FITC-ox-LDL, the greater the production of TNF-α and IL-6. The production of IL-6 by foam cells was detected up to the value of 150 µg/mL of the maximum stimulus for LDL. Concentrations above 50 μg/mL LDL stimulated greater release of TNF-α compared to control. Conclusions Our model contributes to the understanding of the release of IL-6 and TNF-α in response to different concentrations of ox-LDL, using an optimized method for the formation of foam cells.

Ultrasound device as a minimally invasive approach for caries dentin removal

Abstract The aim of this study was to evaluate the efficacy of an ultrasound device and the dentin surface morphology after removal of the caries dentin lesions by removal rate and scanning electron microscopy (SEM). The Knoop hardness test on the bovine dentin blocks (n = 20, 4x4x2mm) was performed to standardize the samples and only those with 38 ± 2 KHN were included. The dentin blocks were submitted to induction of artificial caries lesions, using the bacterial model. Strains of Streptococcus mutans and Lactobacillus acidophilus were used for 7 days. The caries dentin lesion was removed for 1 min, according to two methods: G1 - carbide bur under low-speed rotation (control group) and G2 - ultrasound device under refrigeration. For the removal rate, the samples were weighed 3 times: T0 (before induction), T1 (after induction) and T2 (after removal). Morphology evaluation of the residual dentin surface was performed by SEM. Data normality was verified by Shapiro-Wilk test (p ≥ 0.240). T-test for independent samples was applied to evaluate the removal rate. A significance level of 5% was adopted. G2 provided lower removal rate than G1 (G1: 3.68 mg and G2 = 2.26 mg). SEM images showed different morphological characteristics between the groups. G2 showed absent of smear layer, while G1 showed a visible smear layer over the surface. We concluded that ultrasound device provides minimally invasive removal with residual dentin exhibiting open dentin tubules and no smear layer formation and no bacteria, which infer the removal of the infected tissue.

Resumo O objetivo deste estudo foi avaliar a eficácia de um dispositivo ultrassônico e a morfologia da superfície dentinária após a remoção da dentina cariada por meio da taxa de remoção e imagens de microscopia eletrônica de varredura (MEV). Foi realizado o teste de dureza Knoop nas amostras de dentina bovina (n = 20, 4x4x2mm) afim de padronizar as amostras e foram incluídas apenas aquelas com 38 ± 2 KHN. As amostras de dentina foram submetidas à indução de lesões artificiais de cárie, utilizando-se modelo biológico contendo cepas de Streptococcus mutans e Lactobacillus acidophilus durante por 7 dias. As lesões foram removidas por 1 min, de acordo com dois métodos: G1 - fresa esférica sob baixa rotação (grupo controle) e G2 - dispositivo ultrassônico sob refrigeração. Para a taxa de remoção, as amostras foram pesadas em três tempos: T0 (antes da indução), T1 (após a indução) e T2 (após a remoção). A avaliação morfológica da superfície residual da dentina foi realizada por MEV. A normalidade dos dados foi verificada pelo teste de Shapiro-Wilk (p ≥ 0,240). Teste T para amostras independentes foi aplicado para avaliar a taxa de remoção. Foi adotado nível de significância de 5%. G2 apresentou menor taxa de remoção que G1 (G1: 3,68 mg e G2 = 2,26 mg). As imagens de MEV mostraram características morfológicas diferentes entre os grupos. G2 mostrou ausência de smear layer, enquanto G1 mostrou grande quantidade de smear layer sobre a superfície. Com base neste estudo in vitro, o dispositivo de ultrassom promoveu remoção minimamente invasiva e dentina residual exibindo túbulos dentinários abertos e mínima formação de smear layer.

Effects of laser photobiomodulation on tgf-ß and vegf expression in burn wound: systematic review and meta-analysis in the animal model / Influencia de la fotobiomodulación por láser en la expresión de tgf-ß y vegf en heridas por quemadura: revisión sistemática y meta-análisis en modelo animal

SUMMARY: Laser photobiomodulation (laser PBM) is known to be able to accelerate burn wound healing in the animal model; however little evidence exists on the action of laser PBM on the expression of important proteins in wound healing in the animal model, such as VEGF and TGF-ß1. The aim of this study was to carry out a systematic review in order to analyse the effect of laser PBM on VEGF and TGF-ß expression during burn wound repair in the animal model. A systematic review was carried out of the EMBASE, PubMed/ MEDLINE and LILACS databases. The studies included were preclinical studies that analysed the action of laser PBM on the expression of VEGF and TGF-ß (1, 2, 3) during burn wound repair in the animal model. The SYRCLE risk of bias tool was used. Random effect models were used to estimate the combined effect. Increased VEGF expression was observed with the use of laser PBM at 4.93 J/cm2 per point in the first two weeks after induction of the burn wound, with greater size of effect in the second week (SDM = 5.72; 95% CI: 3.14 to 8.31, I2 = 0 %; very low certainty of evidence). We also observed that the effect of laser PBM on TGF-ß1 expression was greater than in the control in the first week (SDM = -0.45; 95% CI: -1.91 to 1.02, I2 = 51 %; very low certainty of evidence), but diminished in the third week after induction of the lesion (SDM = -2.50; 95% CI: 3.98 to -1.01, I2 = 0 %; very low certainty of evidence). Laser PBM has an effect on TGF-ß1 and VEGF expression, promoting burn wound repair in the animal model.

RESUMEN: Es sabido que la fotobiomodulación por láser (FBM láser) puede acelerar el proceso de curación de heridas por quemadura en modelo animal, sin embargo aún se carece de mayor evidencia sobre la acción de la FBM láser en la expresión de proteínas importantes en el proceso de curación de heridas en modelo animal, como VEGF y TGF-ß1. Así, el objetivo de este estudio fue realizar una revisión sistemática a fin de analizar el efecto de la FBM láser sobre la expresión de VEGF, TGF-ß durante el proceso de reparación de heridas por quemadura en modelo animal. Se realizó una búsqueda sistemática en las bases de datos EMBASE, PubMed/MEDLINE y LILACS. Se incluyeron estudios preclínicos que analizaron la acción de la FBM láser en la expresión de VEGF, TGF-ß (1, 2, 3) durante el proceso de reparación de heridas por quemadura en modelo animal. Se utilizó la herramienta de riesgo de sesgo SYRCLE. Se utilizaron modelos de efectos aleatorios para estimar el efecto combinado. Observamos aumento de la expresión de VEGF con el uso de FBM láser 4.93 J/cm2 por punto, en las dos primeras semanas tras inducción de la herida por quemadura, con mayor tamaño de efecto en la segunda semana (SDM = 5,72; IC del 95%: 3,14 a 8,31, I2 = 0 %; certeza de la evidencia muy baja). También se observó el efecto de la FBM láser en la expresión del TGF- ß1 que fue mayor que el control en la primera semana (SDM = - 0,45; IC del 95%: -1,91 a 1,02, I2 = 51 %; certeza de la evidencia muy baja), disminuyendo en la tercera semana tras inducción de la lesión (SDM = -2,50; IC del 95%: -3,98 a -1,01; I2 = 0 %; certeza de la evidencia baja). La TFB por láser ejerce influencia en la expresión de TGF-ß1 y VEGF favoreciendo el proceso de reparación de heridas por quemadura en modelo animal.

Fluorescence spectroscopy for clinical transplantation liver grafts monitoring: possibilities offered by 408 nm excitation

Purpose: Fluorescence spectroscopy techniques have been investigated aiming to reduce the invasiveness of methods for investigation of tissue. In transplantation procedures, it may offer the possibility of a complementary technique for the monitoring of liver grafts' conditions prior to and during the transplantation procedure stages involving cold perfusion. The objective of this study was to evaluate fluorescence spectroscopy under violet light excitation (408 nm) for the monitoring of clinical hypothermic liver transplantation procedures. Methods: Organ grafts were monitored from before the removal of the donor's body to 1 h after the implant into the receptor's body. Fluorescence spectroscopy was assessed over five stages within these transplant stages. Results: The study provided evidence of a correlation between fluorescence information collected during liver grafts transplantation and the survival of patients. Conclusions: Fluorescence spectroscopy can become a tool to monitor transplantation grafts, providing objective information for the final decision of surgeons to use organs.

A single session of antimicrobial photodynamic therapy does not influence the alveolar repair process in rats

Abstract: The aim of this study was to use microscopic and molecular techniques to evaluate the effects of a single session of antimicrobial photodynamic therapy (aPDT) on the alveolar repair process after tooth extraction in rats. The study sample included 84 rats divided into four groups, as follows: a) Control - untreated socket; b) Laser - socket treated using photobiomodulation; c) TBO - socket treated with topic application of the photosensitizer agent, toluidine blue O (TBO); and d) aPDT - socket treated with TBO and laser irradiation. An additional rat was used for thermal mapping during socket irradiation. The animals were euthanatized at 6, 15, and 28 days after unilateral extraction of the upper incisor. Quantitative and qualitative analyses of the connective and bone tissues, blood clot, blood vessel, and inflammatory infiltrate were performed, and real-time polymerase chain reaction was used to study the expression of genes (collagen type I, osteocalcin, alkaline phosphatase [ALP], runt-related transcription factor 2 [RUNX2], and vascular endothelial growth factor [VEGF]) involved in the bone healing process. No statistically significant differences in microscopic and molecular outcomes were observed between the groups (p > 0.05). A positive correlation was seen to exist between blood clot and VEGF (p = 0.000), and a negative correlation was observed between bone tissue and ALP (p = 0.028) and blood vessel and VEGF (p = 0.018). A single session of aPDT in the dental extraction site did not influence the alveolar repair process in rats.

Use of wide-field optical fluorescence for visualization of oral biofilm in a patient with peri-implant mucositis: a new approach / Uso da fluorescência óptica de campo amplo para visualização de biofilme oral em paciente com mucosite peri-implantar: uma nova abordagem

ABSTRACT Peri-implant diseases, caused by bacteria from biofilm related to dental implants, are one of the main causes of late loss of implants. In this sense, peri-implant diseases are divided into peri-implant mucositis, when it affects only the soft tissues, and peri-implantitis, when there is a bone involvement, which can lead to the failure of dental implant therapy. Thus, biofilm removal is essential for peri-implant health, allowing long-term success in implant therapy. To improve the visualization of oral biofilm, which is usually transparent or colorless, disclosing agents have been routinely used. However, disclosing agents have allergenic potential and can cause staining extrinsically in restorative and prosthetic materials, leading to aesthetic impairment. Thus, the use of fluorescence has been studied as an alternative for visualization of oral biofilm. Therefore, this report describes the use of wide-field optical fluorescence for visualization of oral biofilm associated with implants and teeth, in a routine appointment and follow-up of a partially edentulous patient with peri-implant mucositis. In addition, this report showed wide-field optical fluorescence can be used in a clinical routine of care of patients with dental implants. In this sense, wide-field optical fluorescence allowed easy and immediate visualization of the mature oral biofilm for its adequate removal, evaluation of the quality of restoration to sealing of screw access-hole of implant and identification of cariogenic lesions, without risk of allergic reactions or staining of prostheses and restorations.

RESUMO Doenças peri-implantares, causadas por bactérias de biofilme relacionadas a implantes dentários, são uma das principais causas de perda tardia de implantes. Nesse sentido, as doenças peri-implantares são divididas em mucosite peri-implantar, quando afeta apenas tecidos moles, e peri-implantite, quando há comprometimento ósseo, o que pode levar ao fracasso da terapia com implantes dentários. Assim, a remoção do biofilme é essencial para a saúde peri-implantar, permitindo sucesso a longo prazo na terapia com implantes. A fim de melhorar a visualização do biofilme oral, que geralmente é transparente ou incolor, agentes reveladores têm sido rotineiramente utilizados. No entanto, esses agentes têm potencial alergênico e podem causar manchas extrinsecamente em materiais restauradores e protéticos, levando a prejuízo estético. Assim, o uso da fluorescência tem sido estudado como alternativa para visualização do biofilme oral. Este relato descreve o uso da fluorescência óptica de campo amplo para visualização do biofilme oral associado a implantes e dentes em uma consulta de acompanhamento de rotina de uma paciente parcialmente edêntula com mucosite peri-implantar. Além disso, este relato evidenciou que a fluorescência óptica de campo amplo pode ser utilizada dentro da rotina clínica de atendimento de pacientes com implantes dentários. Nesse sentido, a fluorescência óptica de campo amplo permitiu a visualização fácil e imediata do biofilme oral maduro para sua remoção adequada, a avaliação da qualidade da restauração do selamento do orifício de acesso do parafuso do implante e a identificação de lesões cariogênicas, sem risco de reações alérgicas ou manchamento de próteses e restaurações.

Photodynamic therapy of extrahepatic cholangiocarcinoma using digital cholangioscopy / Terapia fotodinâmica em colangiocarcinoma extra-hepático utilizando colangioscopia digital

ABSTRACT Background: Cholangiocarcinoma is an aggressive neoplasm that usually requires palliative biliary drainage. Photodynamic therapy (PDT) has been described as a successful adjunct treatment to malignant biliary obstruction. Aim: To describe the use of digital cholangioscope to help provide laser light during biliary PDT session using locally developed light source. Method: Patient receives intravenous photosensitizer 24 h before the procedure. It starts with a regular duodenoscopy. After identification of the major papilla and retrograde cannulation, the digital cholangioscope is introduced into the common bile duct. Then, the cholangioscopic examination helps to identify the neoplastic stricture. Under direct visualization lighting catheter is advanced through the cholangioscope. Repositioning is recommended every centimeter to cover all strictured area. At the end of the procedure, a final cholangioscopy assesses the bile duct for the immediate result and adverse events. Result: This procedure was applied in one 82-year-old male due to obstructive jaundice in the last two months. EUS and ERCP revealed a severe dilation of the common bile duct associated with choledocholithiasis. Besides, was revealed dilation of hepatic duct up to a well-circumscribed hypoechoic solid mass measuring 1.8x2 cm compressing the common hepatic duct. The mass was deemed unresectable and the patient was referred for palliative treatment with PDT. He remained asymptomatic for three months. He perished due to complications 15 months after the PDT session. Conclusion: Digital cholangioscopy-guided biliary PDT is feasible and seems safe and effective as an adjunct modality in the palliation of extrahepatic cholangiocarcinoma.

RESUMO Racional: Colangiocarcinoma é neoplasia agressiva que geralmente exige drenagem biliar paliativa. A terapia fotodinâmica (TFD) tem sido descrita como tratamento adjunto bem-sucedido para tratar obstrução biliar maligna. Objetivo: Descrever o emprego do colangioscópio digital para ajudar a fornecer luz de laser durante sessão de TFD biliar usando fonte de luz desenvolvida localmente. Método: Paciente recebe fotossensibilizador intravenoso 24 h antes do procedimento que começa com duodenoscopia regular. Após a identificação da papila principal e da canulação retrógrada, o colangioscópio digital é introduzido no ducto biliar comum. Em seguida, o exame colangioscópico ajuda a identificar a estenose neoplásica. Sob visualização direta, o cateter de iluminação avança através do colangioscópio. Reposicionamento é feito a cada centímetro. Ao final colangioscopia avalia o ducto biliar quanto ao resultado imediato e a eventos adversos. Resultado: Este procedimento foi aplicado em um homem de 82 anos devido à icterícia obstrutiva nos últimos dois meses. EUS e CPRE revelaram dilatação grave do ducto biliar comum associada à coledocolitíase. Além disso, havia dilatação do ducto hepático até massa sólida hipoecóica bem circunscrita, medindo 1,8x2 cm, comprimindo o ducto hepático comum. Ela foi considerada irressecável e paciente encaminhado para tratamento paliativo com TFD que permaneceu assintomático por três meses. Morreu devido a complicações 15 meses após a sessão de TFD. Conclusão: A TFD biliar guiada por colangioscopia digital é viável e parece segura e eficaz como modalidade auxiliar na paliação de colangiocarcinoma extra-hepático.

Effects of low-level laser on the repair of orthodontically induced inflammatory root resorption: a systematic review of studies in rats / Efectos del láser de bajo nivel en la reparación de la reabsorción radicular inflamatoria inducida por ortodoncia: una revisión sistemática de estudios en ratas

Orthodontically induced inflammatory root resorption (OIIRR) is a complication of dental treatment which consists of the degradation of local tissue due to an inflammatory reaction provoked by inappropriate orthodontic stimulus. The aim of the present systematic review was to assess the effectiveness of low-level laser therapy (LLLT) in reducing orthodontically induced inflammatory root resorption (OIIRR) in animal models. A systematic review was carried out in the MEDLINE, EMBASE and LILACS databases. Studies of interventions in animals were selected which analysed the effect of LLLT on OIIRR repair. The risk of bias was analysed through the 10 domains of the SYRCLE RoB tool for animal studies. Seventy-one studies were found; 27 were eliminated as duplicates and 44 titles/abstracts were analysed. Of these, 38 were excluded, and five studies were included for qualitative analysis. In 66.6 % of the studies included, the authors state that LLLT was effective in the inhibition/repair of OIIRR. In histological analysis it was observed that root resorption was significantly less in animals treated with laser as compared to the control. Furthermore, LLLT accelerated cicatrization after OIIRR. Laser proved effective in reducing root resorption lacunae and shortening the inflammatory process induced by the application of orthodontic force.

La resorción radicular inflamatoria inducida por tratamiento ortodontico (RRIITO) es una complicación del tratamiento odontológico que consiste en la degradación del tejido local debido a una reacción inflamación provocada por un estímulo ortodóntico inadecuado. El objetivo fue analizar la efectividad de la terapia láser de baja intensidad (LBI) en la disminución de RRIITO en ratas. Se realizó una revisión sistemática en las bases MEDLINE, EMBASE y LILACS. Fueron utilizados los términos 'resorción radicular', 'láser de baja intensidad', 'fototerapia', 'tratamiento ortodóntico', 'movimiento dental'. Fueron seleccionados estudios de intervención en animales, que analizaron el efecto del LBI en la reparación de la RRIITO. Los riesgos de sesgos fueron analizados mediante los 10 domínios de la herramienta SYRCLE RoB para estudios en animales. 71 estudios fueron encontrados, siendo eliminados 27 duplicados, y analizados 44 títulos/abstracts; de estos, fueron incluídos 5 estudios para análisis cualitativa. El 66,6 % de los estudios incluidos afirman que el LBI fue efectivo em reparar la RRIITO. En el análisis histológico se observó que la RRIITO fue significativamente menor en animales tratados con láser en comparación con el control. Además de eso, el LBI aceleró el proceso de cicatrización de la RRIITO. El láser se mostró efectivo en reducir las lagunas de resorción radicular y acortar el proceso inflamatorio inducido por la aplicación de fuerzas ortodonticas.

Mucosal vitiligo in angles of the mouth: clinical and fluorescence aspects

SUMMARY Vitiligo is the most common depigmenting, chronic acquired disease of the skin and mucosa. However, vitiligo of an unclassified type and mucosal subtype affecting only one area of the mucosa is considered quite uncommon. The diagnosis of vitiligo, regardless of its type, is clinical. Nonetheless, a device that allows the visualization of the tissue fluorescence may be useful for confirming the diagnosis. We present the use of wide-field optical fluorescence device for complementary examination and diagnosis of unusual cases of mucosal vitiligo located only in angles of the mouth.

RESUMO O vitiligo é a doença crônica adquirida despigmentante mais comum da pele e/ou da mucosa. Entretanto, o vitiligo do tipo não classificado e subtipo de mucosa afetando apenas uma área da mucosa é considerado bastante incomum. O diagnóstico de vitiligo, independentemente do seu tipo, é clínico. No entanto, o uso de um dispositivo que permite a visualização da fluorescência tecidual pode ser útil para a confirmação do diagnóstico de vitiligo. Apresentamos o uso do dispositivo de exame complementar de fluorescência óptica de campo amplo para o diagnóstico de um caso incomum de vitiligo de mucosa localizado apenas em ângulos da boca.

Oral squamous papilloma: a view under clinical, fluorescence and histopathological aspects / Papiloma escamoso oral: uma visão sob aspectos clínicos, de fluorescência e histopatológicos

ABSTRACT Oral squamous papilloma is a benign tumor whose pathogenesis has been associated with human papillomavirus infection. Thus, it is noteworthy that human papillomavirus infection is one of the risk factors associated with the development of cervical, anogenital, pharynx, larynx and oral cavity carcinomas. Oral squamous papilloma can affect any region of the oral cavity, and transmission of human papillomavirus can occur by direct contact, sexual intercourse or from mother to child during delivery. The diagnosis is clinical and histopathological, with surgical removal representing the treatment of choice. Recently, widefield optical fluorescence has been used as a complementary examination to the conventional clinical examination in the screening of oral pathological lesions and for the delimitation of surgical margins. We report a case of oral squamous papilloma with its clinical, histopathological features and, in addition, from the perspective of wide field optical fluorescence.

RESUMO O papiloma escamoso oral é um tumor benigno, cuja patogênese tem sido associada à infecção pelo papilomavírus humano. A infecção pelo papilomavírus humano é um dos fatores de risco associado ao desenvolvimento dos carcinomas cervicais, anogenitais, faríngeos, laríngeos e da cavidade oral. O papiloma escamoso oral pode acometer qualquer região da cavidade oral, e a transmissão do papilomavírus humano ocorre por contato direto, relação sexual ou de mãe para filho durante o parto. O diagnóstico é clínico e histopatológico, e a remoção cirúrgica representa o tratamento de escolha. Recentemente, a fluorescência óptica de campo amplo tem sido empregada como exame complementar ao exame clínico convencional, no rastreio de lesões patológicas orais e para delimitação de margens cirúrgicas. Relatamos um caso de papiloma escamoso oral com suas características clínicas e histopatológicas, sob a perspectiva da fluorescência óptica de campo amplo.

Prophylactic application of laser light restores L-FABP expression in the livers of rats submitted to partial ischemia

OBJECTIVES: The objective of the present study was to evaluate the protective effect of pre-conditioning treatment with laser light on hepatic injury in rats submitted to partial ischemia using mitochondrial function and liver fatty acid binding protein as markers. METHODS: Rats were divided into four groups (n=5): 1) Control, 2) Control + Laser, 3) Partial Ischemia and 4) Partial Ischemia + Laser. Ischemia was induced by clamping the hepatic pedicle of the left and middle lobes of the liver for 60 minutes. Laser light at 660 nm was applied to the liver immediately prior to the induction of ischemia at 22.5 J/cm2, with 30 seconds of illumination at five individual points. The animals were sacrificed after 30 minutes of reperfusion. Blood and liver tissues were collected for analysis of mitochondrial function, determination of malondialdehyde and analysis of fatty acid binding protein expression by Western blot. RESULTS: Mitochondrial function decreased in the Partial Ischemia group, especially during adenosine diphosphate-activated respiration (state 3), and the expression of fatty acid binding protein was also reduced. The application of laser light prevented bioenergetic changes and restored the expression of fatty acid binding protein. CONCLUSION: Prophylactic application of laser light to the livers of rats submitted to partial ischemia was found to have a protective effect in the liver, with normalization of both mitochondrial function and fatty acid binding protein tissue expression.

Resultados preliminares do tratamento de insuficiência venosa grave com termoablação da veia safena magna por técnica endovascular com laser de diodo 980nm desenvolvido no Brasil, associado à escleroterapia com polidocanol / Preliminary results of severe venous insufficiency treatment with thermal ablation of the great saphenous vein by endovascular technique with laser diode 980nm developed in Brazil, associated with sclerotherapy with polidocanol

RESUMO A termoablação endovascular das veias safenas insuficientes com laser é descrita como uma técnica menos invasiva, com resultados semelhantes à cirurgia convencional, porém, com efeitos adversos menos frequentes. A técnica de escleroterapia com espuma de polidocanol ecoguiada vem sendo empregada com a mesma finalidade. A combinação de técnicas pode representar uma alternativa para pacientes mais graves, como os portadores de úlcera varicosa. Um equipamento de laser (denominado VELAS) foi desenvolvido no Centro de Pesquisas em Ótica e Fotônica da USP-São Carlos em convênio com a FMB-UNESP para termoablação endoluminal da veia safena insuficiente. Neste estudo apresentamos os resultados preliminares do uso do aparelho de laser VELAS (diodo MMO 980nm) na termoablação endovascular de veias safenas insuficientes, em portadores de úlcera venosa crônica, associado à complementação com espuma de polidocanol para o tratamento de varicosidades, após uma semana. Os desfechos analisados foram o tempo de cicatrização da úlcera venosa, oclusão das veias tratadas e eventos adversos relacionados aos tratamentos. Foram incluídos 12 pacientes portadores de insuficiência de veia safena e úlcera venosa crônica que aceitaram participar do projeto. Todos foram tratados em regime ambulatorial, com anestesia local e termoablação da veia safena insuficiente (VELAS). Após uma semana da cirurgia, as varicosidades foram esclerosadas com polidocanol espuma (técnica de Tessari). O equipamento laser VELAS nacional apresentou fácil manuseio, oclusão venosa total em 83,3% dos pacientes (em sete dias) e a associação das técnicas foi responsável por uma taxa de cicatrização de feridas de 83,3%, sem ocorrência de eventos adversos.

ABSTRACT The endovenous laser ablation (EVLA) of the insufficient saphenous vein has similar results to open conventional surgery, but less morbidity. The echo-guided polidocanol foam sclerotherapy technique has been used for the same purpose. The combined techniques may play a role for more severe diseases, such as those with varicose ulcers. An EVLA device (called VELAS) has been developed in the Optics and Photonics Research Center of USP-São Carlos in agreement with FMB-UNESP. In this study, we present the preliminary results of the VELAS device (MMO 980nm diode) in patients with chronic venous ulcer, associated with echo-guided polidocanol foam sclerotherapy for the treatment of varicosities. Primary outcomes were healing time of the venous ulcer, occlusion of the treated veins and treatment-related adverse events. We included 12 patients with insufficient saphenous vein and chronic venous ulcer. Initially, we treated all of them with thermoablation of the insufficient saphenous vein (VELAS), on an outpatient basis, with local anesthesia. After one week of the procedure, we sclerosed the varicosities with polidocanol foam (Tessari technique). The national VELAS device was easily handled. Total venous occlusion occurred in 83.3% of the patients (in seven days) and the association of the techniques was responsible for a wound healing rate of 83.3%, with no adverse events.

Effect of irradiation with different laser wavelengths on oxidative stress of non-hepatectomized rats

PURPOSE: To assess the effect of two laser wavelengths, either separate or combined, on intact rat livers. METHOD: Nineteen male Wistar rats (200-300 g) were submitted to laser irradiation at 5 different sites on the liver surface.Wavelengths 660 and 780 nm were used, with a dose of irradiation of 60 J/cm2/site.The animals were divided into the groups:control (C) and animals irradiated with 660 nm laser (L1), with 780 nm laser (L2) or withboth wavelengths (L3).Mitochondrial function, mitochondrial swelling, and hepatocellular malondialdehyde (MDA) levels were determined.Data were analyzed by the Mann-Whitney test, with the level of significance set at 5%. RESULTS: There was a reduction of ADP-activated respiration (state 3) in group L1 compared to group C (p=0.0016), whereas the values of group L2 were similar to control.Group L3 also showed a reduction of state 3 (p=0.0159).There was a reduction of RCR in group L1 compared to control (p=0.0001) and to group L2 (p=0.0040).Mitochondrial swelling only differed between group L3 and control (p=0.0286).There was a increase in MDA levels in group L3 compared to control (p=0.0476) and to group L2 (p=0.0286) and in group L1 compared to group L2 (p=0.0132). CONCLUSION: Although laser irradiation reduced mitochondrial function,it did not interfere with the hepatocellular energy status.

Effect of different light-curing techniques on hardness of a microhybrid dental composite resin / Efeito de diferentes técnicas de fotoativação na dureza de uma resina composta dental microhíbrida

Objective: This study assessed the Vickers hardness (VHN) provided by two LCUs when using (1) direct and indirect light-curing techniques, (2) 40 and 60 s and (3) top and bottom surfaces. Material & Methods: One halogen Curing Light 2500 (3M Espe) and one LED (MM Optics) were used by direct and indirect (0,1.0, 2.0 and 3.0 mm of dental structure) techniques during 40 and 60 s. The samples were made with FiltekTM Z250 in a metallic mould with a central orifice (4 mm in diameter, 2 mm in thickness). The samples were stored in dry mean by ± 24 h and the hardness measurements were performed in a testing machine (Buehler MMT-3 digital microhardness tester Lake Bluff, Illinois USA). A 50 gf load was used and the indenter with a dwell time of 30 s. The data were submitted to multiple ANOVA and Newman-Keuls‘s test (p < 0.05). Results: Halogen LCU exhibited higher Vickers hardness values than LED mainly because of the power density used. Hardness values were influenced by LCUs, lightcuring techniques, irradiation times and surfaces. For both LCUs, hardness values were found to decrease with indirect light-curing technique, mainly for the bottom surface. Samples irradiated for 60 s exhibited higher hardness values when the halogen LCU was used. For 60 s, the VHN values were statistically significant greater than 40 s. Significant differences in top and bottom surfaces Vickers hardness number (VHN) values were observed among different LCUs used 40 and 60 s. Conclusion: The LCUs, lightcuring techniques, variations of irradiation times, and surfaces (top and bottom) influence the composite resin hardness.

Objetivo: Este estudo avaliou a dureza Vickers (VHN) em função de duas fontes de luz quando utilizadas diferentes (1) técnicas de fotoativação, direta e indireta, (2) 40 e 60 s, e (3) superfícies de topo e base. Material e Métodos: Uma fonte de luz halógena Curing Light 2500 (3M Espe) e um LED (MM Optics) foram utilizados nas técnicas de fotoativação direta e indireta (0 , 1,0, 2,0 e 3,0 milímetros de estrutura dental), durante 40 e 60 s. As amostras foram feitas utilizando-se FiltekTM Z250 em matriz metálica com orifício central (4 mm de diâmetro, com 2 mm de espessura). As amostras foram armazenadas em meio seco por ± 24 h e as medidas de dureza foram realizadas em microdurômetro (Buehler MMT-3 digital microhardness tester Lake Bluff, Illinois USA). Uma carga de 50 gf durante 30 s foi utilizada. Os dados foram submetidos à análise de variância múltipla e teste de Newman-Keuls (p < 0,05). Resultados: A fonte de luz halógena promoveu os maiores valores de dureza Vickers, principalmente, em função da densidade de potência utilizada. Os valores de dureza foram influenciados pelas fontes de luz, técnicas de fotoativação, tempos de irradiação e superfícies, topo e base. Para ambas as fontes de luz, os valores de dureza diminuíram com a técnica de fotoativação indireta, principalmente para a superfície de base. Amostras irradiadas por 60 s apresentaram valores de dureza maiores quando a fonte de luz halógena foi utilizada. Durante 60 s , os valores de VHN foram estatisticamente significativos maior do que 40 s. Diferenças significativas foram observadas nos valores dureza Vickers (VHN) para as superfícies de topo e base utilizando 40 e 60 s. Conclusão: As fontes de luz, técnicas de fotoativação direta e indireta, tempos de irradiação e superfícies (topo e base) influenciam na dureza da resina composta.

Ampliando a visão bucal com fluorescência óptica / Improving the oral screening with optical fluorescence

A fluorescência óptica pode ser utilizada como um coadjuvante ao exame clínico bucal, uma vez que permite, por autofluorescência, a detecção de inúmeras alterações na cavidade bucal que poderiam passar despercebidas pelo cirurgião-dentista ou até mesmo de difícil percepção apenas pelo método visual. O objetivo do presente estudo foi demonstrar o uso do sistema de fluorescência óptica por imagem no diagnóstico de diferentes lesões da cavidade bucal, seja em tecido duro ou mole, de forma a familiarizar o cirurgião-dentista com o uso do equipamento. Para este estudo foi utilizado um equipamento constituído de um diodo emissor de luz (LEDl, com emissão na região do violeta e um conjunto de filtros ópticos (Evince - MMoptics, São Carlos, SP, Brasil). Para a aquisição das imagens foi acoplado ao equipamento uma câmera fotográfica (Nikon, 090, Bangkok, Tailândia) com o auxílio de um adaptador. O sistema de fluorescência bucal possibilitou observar alterações nos tecidos duros dentais como manchas, presença de placa e cá Icu 10 denta I, lesões incipientes e infiltrações marginais, além de facilitar a diferenciação entre materiais restauradores como resina composta e cerâmica. Em tecidos moles foi possível detectar lesões potencialmente malignas e lesões tumorais. Portanto, pode-se concluir que o sistema de fluorescência óptica permite ao cirurgião-dentista diagnosticar e identificar estruturas e alterações na cavidade bucal de forma simples, não invasiva e em tempo real revelando lesões que não seriam facilmente detectados com a iluminação convencional.

Fluorescence techniques can be used as an adjunct to clinical examination of the mouth, detecting tissue changes in oral mucosa or hard dental tissues, which might be unnoticed by the dentist or even difficult to detect under white light examination. The aim of this study was to demonstrate the use of wide-field fluorescence imaging in the diagnosis of various lesions of the oral cavity, either in hard or soft tissues, in order to familiarize the dentist with the use of the equipment. For this study we used an optical fluorescence system with emission in the violet region (Evince, MMOptics, São Carlos, SP, Brazil). For image acquisition, the fluorescence system was coupled to a digital camera (Nikon 090, Nikon, Bangkok, Tailândia). With the fluorescence system was possible to observe changes in hard dental tissues such as bright spots, dental plaque and calculus, incipient carious and marginal microleakage lesions. The system also facilitated the differentiation between restorative materiais such as composite resin and ceramic. The fluorescence optical was also helpful in screening and detection of potentially malignant lesions and tumors could be observed. Therefore, we conclude that the fluorescence optical system allows the dentist to identify structures and alterations in the oral cavity in a simple, noninvasive, and in real-time procedure, revealing injuries that would not be easily detected with conventional illumination.

Degree of conversion of nanofilled and microhybrid composite resins photo-activated by different generations of LEDs

OBJECTIVE:This study aimed at evaluating the degree of conversion (DC) of four composite resins, being one nanofilled and 3 microhybrid resins, photo-activated with second- and third-generation light-emitting diodes (LEDs). MATERIAL AND METHODS: Filtek TM Z350 nanofilled composite resins and Amelogen® Plus, Vit-l-escenceTM and Opallis microhybrid resins were photo-activated with two second-generation LEDs (Radii-cal and Elipar Free LightTM 2) and one third-generation LED (Ultra-Lume LED 5) by continuous light mode, and a quartz halogen-tungsten bulb (QHT, control). After 24 h of storage, the samples were pulverized into fine powder and 5 mg of each material were mixed with 100 mg of potassium bromide (KBr). After homogenization, they were pressed, which resulted in a pellet that was evaluated using an infrared spectromer (Nexus 470, Thermo Nicolet) equipped with TGS detector using diffuse reflectance (32 scans, resolution of 4 cm-1) coupled to a computer. The percentage of unreacted carbon-carbon double bonds (% C=C) was determined from the ratio of absorbance intensities of aliphatic C=C (peak at 1637 cm-1) against internal standard before and after curing of the specimen: aromatic C-C (peak at 1610 cm-1). RESULTS: The ANOVA showed a significant effect on the interaction between the light-curing units (LCUs) and the composite resins (p<0.001). The Tukey’s test showed that the nanofilled resin (FiltekTM Z350) and Opallis when photo-activated by the halogen lamp (QTH) had the lowest DC compared with the other microhybrid composite resins. The DC of the nanofilled resin (FiltekTM Z350) was also lower using LEDs. The highest degrees of conversion were obtained using the third-generation LED and one of second-generation LEDs (Elipar Free LightTM 2). CONCLUSIONS: The nanofilled resin showed the lowest DC, and the Vit-l-escenceTM microhybrid composite resin showed the highest DC. Among the LCUs, it was not possible to establish an order, even though the second-generation LED Radii-cal provided the lowest DC.

Effect of laser on the remnant liver after the first 24 hours following 70 percent hepatectomy in rats / Efeito do laser no fígado remanescente nas primeiras 24 horas após hepatectomia a 70 por cento em ratos

PURPOSE: To evaluate the mitochondrial function of the remnant liver (RL) in the early phase of liver regeneration in rats after 70 percent partial hepatectomy (PH). METHODS: Sixty male Wistar rats (200-250g) submitted to 70 percent PH were divided into five groups according to the time of euthanasia and application or not of laser light: C = Control, time zero; 2 minutes, 4, 6 and 24 hours after PH. The dose of laser radiation was 22.5 J/cm², wavelength of 660 nm (visible/red), in the remnant liver. We studied the respiration activated by ADP (state 3), basal mitochondrial respiration (state 4), respiratory control ratio (RCR) and mitochondrial membrane potential (MMP). RESULTS: The mitochondrial function of RL changed at 4 and 6 hours after PH, with a significant increase in state 3 and a concomitant increase in state 4 and with maintenance of RCR. MMP differed significantly between the groups biostimulated with laser radiation and the control group 4 hours after HP, with a substantial reduction in the non-laser groups. CONCLUSION: The laser light at the dose used in this study did not induce additional damage to the RL and seems to have delayed the hepatocellular metabolic overload of the remnant liver.

OBJETIVO: Avaliar a função mitocondrial do fígado remanescente (FR) na fase precoce da regeneração hepática em ratos após hepatectomia parcial (HP) a 70 por cento. MÉTODOS: Sessenta ratos machos Wistar (200 - 250g) submetidos à HP a 70 por cento, foram distribuídos em cinco grupos de acordo com o tempo de eutanásia e com aplicação ou não de luz laser: C= Controle,tempo zero; 2 minutos, 4, 6 e 24 horas após HP. O laser foi utilizado na dose 22.5 J/cm², 660 nm, no FR.Estudou-se o estado 3 (respiração ativada por ADP), estado 4 (respiração mitocondrial basal), razão de controle respiratório,estado 3/estado 4 (RCR) e o potencial de membrana mitocondrial(PMM). RESULTADOS: A função mitocondrial do FR alterou-se no período de 4 e 6 horas após a HP com aumento significativo do estado 3 e aumento concomitante do estado 4, com manutenção da RCR. O PMM apresentou diferença significativa entre os grupos bioestimulados com laser e o controle a partir de 4 horas pós HP, com queda importante do grupo sem laser e tendência a equiparação dos valores após 24 horas. CONCLUSÃO: A luz laser, na dose utilizada no presente estudo, não induziu lesão adicional ao FR e parece ter retardado a sobrecarga hepatocelular do fígado remanescente.

Photodynamic inactivation of four Candida species induced by photogem®

This study evaluated the in vitro susceptibility of C. albicans, C. dubliniensis, C. tropicalis and C. krusei to photodynamic therapy (PDT) induced by Photogem® and light emitting diode (LED). Suspensions of each Candida strain were treated with three photosensitizer (PS) concentrations (10, 25 and 50 mg/L) and exposed to 18.0, 25.5 and 37.5 J/cm² LED light fluences (λ ~ 455 nm). Control suspensions were treated only with PS concentrations, only exposed to the LED light fluences or not exposed to LED light or PS. Sixteen experimental conditions were obtained and each condition was repeated three times. From each sample, serial dilutions were obtained and aliquots were plated on Sabouraud Dextrose Agar. After incubation of plates (37 ºC for 48 hours), colonies were counted (cfu/mL) and the data were statistically analyzed by ANOVA and the Tukey test (α=0.05). Complete killing of C. albicans was observed after 18.0 J/cm² in association with 50 mg/L of PS. C. dubliniensis were inactivated after 18.0 J/cm² using 25 mg/L of PS. The inactivation of C. tropicalis was observed after photosensitization with 25 mg/L and subsequent illumination at 25.5 J/cm². For C. krusei, none of the associations between PS and light resulted in complete killing of this species. PDT proved to be effective for the inactivation of C. albicans, C. dubliniensis and C. tropicalis. In addition, reduction in the viability of C. krusei was achieved with some of the PS and light associations.

Desenvolvimento e resultados preliminares de um sistema cromático de iluminação para oftalmoscópios indiretos / Development and preliminary results of a chromatic illumination system for indirect ophthalmoscopes

OBJETIVOS: Aplicação da técnica de contraste cromático em exames de fundo de olho com oftalmoscópios indiretos. Apresentamos neste trabalho testes preliminares de um novo sistema de iluminação para oftalmoscópios indiretos, elaborado a partir de técnicas de composição cromática, denominado Sistema de Iluminação Cromático, que permite a composição de cores de luz situadas dentro do espectro visível. A finalidade deste sistema é permitir o uso da técnica de cromaticidade, já estudada em outras áreas da medicina como odontologia e dermatologia, na visualização de fundo de olho em exames de oftalmoscopia indireta. MÉTODOS: Para desenvolvimento deste trabalho foi adquirido um oftalmoscópio comercial comum e de uso geral. O equipamento teve seu sistema de iluminação original de fábrica, composto de uma fonte de luz branca alógena e filtros verde e azul, substituído pelo novo sistema de iluminação cromático implementado neste projeto. RESULTADOS: Foram realizados testes preliminares em um olho mecânico que simula as mesmas características de um olho humano biológico in vivo. Os resultados confirmam a possibilidade de obter-se luz em diferenciados comprimentos de ondas através do uso de fontes de luz independentes nas cores vermelho, verde e azul, para uso em instrumentação oftálmica. Embora exames in vivo com pacientes que possuam doenças de retina ainda estejam em fase de testes em nosso laboratório no IFSC-USP & UNIFESP, os resultados preliminares obtidos aqui demonstram que a técnica de contraste cromático para exames de fundo de olho pode futuramente representar um diferencial na qualidade dos exames de oftalmoscopia indireta. CONCLUSÃO: Foi apresentado neste trabalho o desenvolvimento de um sistema de iluminação totalmente inovador para aplicação em oftalmoscopia. Através dos diferentes contrastes e brilhos nas imagens de fundo de olho proporcionados pelas diferentes cores do sistema de iluminação cromático, acreditamos que em uma próxima...

PURPOSE: Chromatic contrast is a technique used in some areas of medicine to provide better visualization of biological tissues. Based on principles of color composition, a new illumination system was constructed using colored emitting diodes to reproduce the spectral range of visible light. This technique was devised to be used in indirect ophthalmoscopes to improve the visualization of the posterior segment of the eye. METHODS: The original illumination system of a general purpose indirect ophthalmoscope was substituted by a system of color-emitting diodes. RESULTS: Using an electronic interface it was possible to control the intensity of the color lights and therefore generate different wavelengths in the visible spectrum of the light. Preliminary tests undertaken in a mechanical model of the human eye generated very clear and homogenous colors. However in vivo examinations with patients were performed in our laboratory at the IFSC-USP and UNIFESP, and obtained the preliminary results show the possibilities of the chromatic contrast technique, and may represent in the future a differential in the analyses of the posterior segment of the eye. CONCLUSION: The use of color-emitting diodes to reproduce the spectral range of the visible light in indirect ophthalmoscopes seems to be a promising technological advance in the fundoscopy of the eye. This is an innovation that can yield better quality examinations with indirect ophthalmoscopes.