ABSTRACT 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.

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.

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.

Os objetivos do estudo foram avaliar as propriedades físicas e a atividade antibacteriana de uma resina composta fotopolimerizável modificada com nanopartículas de prata. Discos foram produzidos com resina não modificada (grupo controle - GC) e resina modificada com nanopartículas de prata em duas concentrações 0,3 e 0,6% em massa (MR03 e MR06, respectivamente). Biofilmes de Streptococcus mutans e Lactobacillus acidophilus foram induzidos in vitro pela incubação dos discos em meio líquido com 20% de sacarose, seguida pela sonicação e contagem de células viáveis após 1, 4 e 7 dias. A rugosidade aritmética de todos os grupos foi avaliada por microscopia de força atômica (MFA). Os dados foram submetidos à ANOVA e ao Teste de Tukey (α=0,05%). Nos três períodos de tempo, o número de células viáveis foi estatisticamente mais baixo para MR03 e MR06, em comparação com GC, para ambas as espécies. MR03 e MR06 não apresentaram diferenças significantes. A análise por meio de MFA demonstrou que não houve diferença significante para a rugosidade entre os três grupos. MR03 apresentouse mais resistente em compressão do que GC, e o desempenho em compressão em MR06 foi estatisticamente pior do que em GC e MR03. Foi possível concluír que as resinas modificadas de MR03 inibiram o crescimento dos biofilmes, sem comprometimento da resistência à compressão e da rugosidade superficial.

The objectives of this study were to evaluate physical properties and antibacterial activity of a light-activated composite modified with silver nanoparticles. Discs were produced with unmodified resin (control group - CG) and modified resin with silver nanoparticles at two concentrations, 0.3% wt (MR03) and 0.6% wt (MR06). Streptococcus mutans and Lactobacillus acidophilus biofilms were induced in vitro by incubation of discs in a 20% sucrose medium, followed by sonication and counting of viable cells after 1, 4 and 7 days (n=9). The arithmetic roughness of all three groups was evaluated by atomic force microscopy (n=9). Compression assay was conducted in all groups to measure the compressive strength at failure and elasticity modulus (n=5). Data were subjected to ANOVA and Tukey's tests (α=0.05%). At all three time points the number of viable cells was statistically lower for MR03 and MR06 compared with CG, for both specimens. MR03 and MR06 showed no significant differences. Microscopic analysis demonstrated no significant differences for roughness among the three groups (p>0.05). The MR03 was stronger to compression than CG, and MR06 was statistically lower than CG and MR03. It was concluded that the MR03 were less conducive to biofilm growth, without compromising the strength in compression and surface roughness.

OBJETIVO: Avaliar a função mitocondrial do fígado remanescente (FR) na fase precoce da regeneração hepática em ratos após hepatectomia parcial (HP) a 70%. MÉTODOS: Sessenta ratos machos Wistar (200 - 250g) submetidos à HP a 70%, 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.

PURPOSE: To evaluate the mitochondrial function of the remnant liver (RL) in the early phase of liver regeneration in rats after 70% partial hepatectomy (PH). METHODS: Sixty male Wistar rats (200-250g) submitted to 70% 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.

O objetivo deste estudo foi avaliar o efeito de parâmetros específicos de irradiação com laser de baixa intensidade sobre biofilmes formados por Streptococcus mutans (S. mutans), Candida albicans (C. albicans) ou associação de ambas as espécies. Biofilmes isolados ou associados destes microrganismos foram irradiados com um dispositivo laser infra-vermelho próximo de diodos InGaAsP (LaserTABLE 780 ±3 nm, 0,04W), utilizando-se para isto o dispositivo LASERTable. Quinze horas após a irradiação, foi demonstrado, por meio da avaliação da viabilidade celular (Teste de MTT), da morfologia das células (MEV) e do crescimento do biofilme (UFC/mL), que esta terapia foi capaz de reduzir o metabolismo celular, número de microrganismos presentes no biofilme, bem como seu crescimento no local. Quanto à viabilidade celular, a resposta à irradiação do biofilme de S. mutans (SSB) foi semelhante para todas as doses de energia, sendo que o crescimento do biofilme foi dose dependente. Porém, quando associado à C. albicans, este microrganismo apresentou resistência à fototerapia. Já a C. albicans associada ao S. mutans apresentou redução de crescimento significativa, sendo este resultado também foi dose dependente. A morfologia dos microrganismos não foi alterada pelas irradiações realizadas quando em biofilmes isolados. A associação entre os microrganismos promoveu redução na formação de hifas pela C. albicans. A laserterapia de baixa intensidade apresentou efeito inibitório sobre microrganismos, sendo que esta capacidade pode ser alterada de acordo com a interação entre diferentes microrganismos.

The aim of this study was to evaluate the effect of specific parameters of low-level laser therapy (LLLT) on biofilms formed by Streptococcus mutans, Candida albicans or an association of both species. Single and dual-species biofilms - SSB and DSB - were exposed to laser doses of 5, 10 or 20 J/cm2 from a near infrared InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm, 0.04 W). After irradiation, the analysis of biobilm viability (MTT assay), biofilm growth (cfu/mL) and cell morphology (SEM) showed that LLLT reduced cell viability as well as the growth of biofilms. The response of S. mutans (SSB) to irradiation was similar for all laser doses and the biofilm growth was dose dependent. However, when associated with C. albicans (DSB), S. mutans was resistant to LLLT. For C. albicans, the association with S. mutans (DSB) caused a significant decrease in biofilm growth in a dose-dependent fashion. The morphology of the microorganisms in the SSB was not altered by LLLT, while the association of microbial species (DSB) promoted a reduction in the formation of C. albicans hyphae. LLLT had an inhibitory effect on the microorganisms, and this capacity can be altered according to the interactions between different microbial species.

Tubos de ensaio foram preenchidos com a solução Fricke dopada com Fotogem® em concentrações crescentes; essa hemotoporfirina é utilizada na terapia fotodinâmica. Esses tubos foram irradiados com doses de 5 a 20 Gy. A solução Fricke convencional também foi irradiada com as mesmas doses. As concentrações de íons férricos nas soluções Fricke convencional e dopadas irradiadas foram medidas num espectrofotômetro com comprimento de onda entre 220 e 340 nm. Os resultados mostraram que quando comparado o Fricke convencional com o Fricke dopado irradiado, as amostras dopadas demonstraram um aumento na resposta da dose absorvida que é proporcional a concentração do Photogem® na solução Fricke. Concluímos que esse procedimento pode ser utilizado para propósitos de dosimetria na terapia com radiossensibilizadores.

The vials filled with Fricke solutions were doped with increasing concentrations of Photogem®, used in photodynamic therapy. These vials were then irradiated with low-energy X-rays with doses ranging from 5 to 20 Gy. The conventional Fricke solution was also irradiated with the same doses. The concentration of ferric ions for the Fricke and doped-Fricke irradiated solutions were measured in a spectrophotometer at 220 to 340 nm. The results showed that there was an enhancement in the response of the doped-Fricke solution, which was proportional to the concentration of the photosensitizer. The use of such procedure for studying the radiosensitizing property of photosensitizers based on the production of free radicals is also discussed.

Recentes avanços em técnicas ópticas têm propiciado vasto campo de possibilidades tanto para o diagnóstico quanto para a terapêutica de doenças hepáticas.Com o uso de eficientes fontes de luz como o laser e Light emitting diodes (LED) é possível utilizar a interação luz-tecido para promover a regeneração hepática após hepatectomias parciais,detectar hepatocarcinoma, esteatose e outras alterações do fígado pelo uso da fluorescência óptica,para avaliar o metabolismo hepático durante o transplante de fígado e na abordagem diagnóstica e terapêutica de alterações hepatocelulares. Os autores apresentam uma ampla revisão de técnicas atualmente em desenvolvimento na Divisão de Gastroenterologia Cirúrgica da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo num trabalho cooperativo com o Instituto de Física de São Carlos da USP. Os resultados obtidos até agora têm sido motivo de lista de publicações que são aqui apresentados em forma de revisão. Uma nova perspectiva de moderna aplicação de técnicas ópticas em várias situações clínico-cirúrgicas relacionadas com o fígado é apresentada e amplamente discutida.

Recent advances in optical techniques have created a great range of possibilities for diagnosis and therapeutics in liver related diseases. With the uses of efficient light sources like lasers and LEDs (Light Emitting Diodes) it is possible to employ the light-tissue interaction to promote hepatic tissue regeneration after partial hepatectomy, to detect hepatocarcinoma and steatosis by utilizing optical fluorescence, to evaluate the metabolism of the liver during hepatic transplantation as well as to treat liver tumors. We present here an overview of the technique presently in development at the Ribeirâo Preto Faculty of Medicine - USP in cooperation with the Physics Institute of São Carlos -USP. The results obtained so far have been the subject of a list of publications and are here presented as an overview. A new perspective for modern application of optical techniques in different medical practices related to the liver is presented.

O objetivo deste estudo foi estudar a influência da fonte de luz, LED e lâmpada halógena (LH), na efetividade de polimerização do cimento resinoso dual Enforce fotoativado sob um disco de porcelana. Três tempos de exposição (60, 80 e 120 segundos) foram também avaliados. Dois grupos experimentais, na qual a polimerização do cimento resinoso foi feita através de um disco cerâmico, e dois controles, polimerização direta do cimento, sem presença do disco cerâmico foram subdivididos em três subgrupos, com cinco amostras em cada: G1A- LH 60s; G1B- LH 80s; G1C- LH 120s; G2A- LED 60s; G2B- LED 80s; G2C- LED 120s; e grupos controles: G3A- LH 60s; G3B- LH 80s; G3C- LH 120s; G4A- LED 60s; G4B- LED 80s e G4C- LED 120s. O cimento resinoso foi inserido em uma matriz de aço (4 mm de diâmetro e 1,2 mm de espessura). Nos grupos experimentais um disco de porcelana (6 mm de diâmetro por 2 mm de espessura) foi colocado sobre o cimento. Este foi fotoativado através da porcelana pela lâmpada halógena e pelo LED, por outro lado nos grupos controles a luz foi aplicada sem o disco de porcelana. As amostras foram armazenadas em um frasco que impedia a passagem de luz a 37(0)C por 24 horas, para que depois a dureza Vickers fosse determinada. Os dados foram submetidos à análise de variância a quatro critérios seguido pelo teste Tukey (p£ 0,05). Todas as amostras polimerizadas pelo LED por 60s mostraram valores inferiores quando comparadas com as do grupo da lâmpada halógena. Em geral, a fotoativação realizada por 80 e 120 s com o LED foi estatisticamente similar à fotoativação com a lâmpada halógena (60 e 80 s). A tecnologia LED pode ser viável para a fotoativação de restaurações indiretas de porcelana quando o tempo de polimerização é aumentado.

The aim of this study was to investigate the influence of light source, LED unit and halogen lamp (HL), on the effectiveness of Enforce dual-cured cement cured under a ceramic disc. Three exposure times (60, 80 and 120 s) were also evaluated. Two experimental groups, in which the polymerization of the dual-cured cement was performed through a ceramic disc, and two control groups, in which the polymerization of the dual-cured cement was performed directly without presence of ceramic disc were subdivided into three subgroups (three different exposure times), with five specimens each: G1A- HL 60s; G1B- HL 80s; G1C- HL 120s; G2A- LED 60s; G2B- LED 80s; G2C- LED 120s; and control groups: G3A- HL 60s; G3B- HL 80s; G3C- HL 120s; G4A- LED 60s; G4B- LED 80s and G4C- LED 120s. Cement was applied in a steel matrix (4mm diameter, 1.2mm thickness). In the experimental groups, a ceramic disc was placed on top. The cement was light-cured through the ceramic by a HL and LED, however, the control groups were cured without the ceramic disc. The specimens were stored in a light-proof container at 37ºC for 24 hours, then Vickers hardness was determined. A four-way ANOVA and Tukey test (p£ 0.05) were performed. All specimens cured by LED for 60s showed inferior values compared with the halogen groups. In general, light-curing by LED for 80s and 120s was comparable to halogen groups (60s and 80s) and their control groups. LED technology can be viable for light-curing through conventional ceramic indirect restorations, when curing time is increased in relation to HL curing time.