Investigação do efeito do laser de baixa potência na recuperação de lesões epidérmicas e musculares em roedores

Tese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas, Programa de Pós-graduação em Bioquímica, Florianópolis, 2013Estudos nos últimos anos tem demonstrado que a terapia com laser de baixa potência (LBP) modula vários processos bioquímicos, principalmente aqueles relacionados ao aumento da respiração mitocondrial e síntese de ATP, com consequente redução de lesões musculares e dérmicas, bem como acelerando o processo de cicatrização. O presente estudo foi desenvolvido com o objetivo de avaliar a influência da terapia com LBP sobre o processo de cicatrização de feridas dérmicas e também em um modelo de lesão muscular traumática. Além disso, foram avaliados os efeitos da irradiação com LBP sobre atividade da cadeia respiratória em mitocôndrias isoladas de músculo esquelético e em parâmetros relacionados com a respiração celular em células musculares da linhagem L6. Primeiramente nossos resultados demonstram que o LBP Hélio Neônio (HeNe) e Arsêneto de Gálio (AsGa) com comprimentos de onda de 632 e 904 nm, respectivamente, e doses de 1 e 3 J/cm2 induziram um maior fechamento da ferida estimulou a produção de colágeno, aumentando o conteúdo de hidroxiprolina e diminuiu os danos oxidativos a lipídeos e proteínas e as atividades das enzimas antioxidantes superóxido dismutase (SOD) e catalase (CAT) em um modelo de lesão epidérmica, quando comparado com o grupo lesão sem laser. No segundo experimento, demonstramos que o laser AsGa com doses de 3 J/cm2 com início de irradiação 2 horas após a lesão muscular traumática induziu uma diminuição em todos os parâmetros de estresse oxidativo analisados, tais como os conteúdos de oxidação de lipídeos e proteínas, atividades das enzimas antioxidantes glutationa peroxidase (GPX), SOD e CAT e na concentração de nitritos no plasma e no músculo induziu uma diminuição na expressão gênica de IL-6, VEGF e BDNF como também um aumento na IL-10 no tecido muscular traumatizado e irradiado até 5 dias após a lesão. Os resultados relacionados a parâmetros comportamentais de déficit motor (campo aberto) também comprovaram que os animais com lesão muscular e tratados com a dose de 3 J/cm2 tiveram um aumento na distância percorrida e no número de levantamentos comparados com o grupo lesão muscular, demonstrando uma melhora no déficit motor induzido pela lesão muscular. Os resultados com mitocôndrias musculares saudáveis demonstraram que a irradiação com LBP HeNe com doses de 30 e 60 J/cm2 induziu um aumento significativo nas atividades do complexo I e IV 5 minutos após a irradiação. Além disso, o LBP com doses de 10, 30 e 60 J/cm2 também aumentou a atividade do complexo IV 1 hora após a irradiação. Estudos in vitro com células saudáveis L6 demonstraram que a irradiação com LBP HeNe e dose de 10 J/cm2 teve uma contribuição maior da ATP sintase ao consumo de oxigênio basal e uma concentração maior de FCCP foi usada para induzir a capacidade máxima respiratória. Neste contexto, o presente estudo evidencia que o LBP pode induzir um aumento na velocidade de cicatrização de feridas e acelerar o processo inflamatório agudo da lesão muscular traumática, provavelmente devido a sua capacidade de aumentar o estado energético da mitocôndria e diminuir parâmetros de inflamação e de estresse oxidativo.Abstract: Many studies performed in the past 40 years have proposed that lowpower laser (LPB) therapy reduces dermal and muscle injuries and accelerates the healing process, by modulating several biochemical processes, including those associated with increased mitochondrial respiration and ATP synthesis. The present study aimed to evaluate the influence of LPB therapy on dermal wound healing and on the recovering from muscle injury. In addition, the effects of LPB irradiation on mitochondrial respiratory chain activity and oxygen consumption in skeletal muscle and muscle L6 cell line, were also evaluated. Irradiation with LPB Helium Neon (HeNe) and Galeo arsenide (GaAs) at wavelengths of 632 and 904 nm, and doses of 1 and 3 J/cm2, respectively, accelerated the wound repair, stimulated the collagen production, by increasing the hydroxyproline content and also by decreasing the concentration of oxidized lipids and proteins. The activities of the antioxidants enzymes, including superoxide dismutase (SOD) and catalase (CAT) were also decreased after the LPB treatment. The second set of experiments demonstrated that the application of the AsGa laser with 3 J/cm2, 2 hours after the traumatic-induced muscle lesion decreased the oxidative stress parameters, including the activity of antioxidant enzymes, oxidative damage markers and nitrite levels in plasma and muscle tissue. In addition, the irradiation decreased the gene expression of IL-6, VEGF e BDNF, and increased the IL-10 content on injured muscle after 5 days of irradiation. Rat spontaneous locomotor activity, measured in the open field arena, was reduced in traumareceiving animals. The LPB irradiation (3 J/cm2) reverted this effect by increasing the distance traveled and the number of withdrawals of the animals. The third group of experiments showed that healthy muscle mitochondria irradiated with LPB HeNe, with doses of 30 and 60 J/cm2, induced a significant increment in the activity of complexes I and IV 5 minutes after the irradiation. Additionally, LPB, with doses of 10, 30 60 J/cm2, also increased complex IV activity one hour after irradiation. In agreement, studies performed in irradiated (LPB HeNe) healthy L6 cells, with dose and 10 J/cm2, provoked increased of ATP synthaselinked oxygen consumption and required higher concentration of FCCP for inducing the maximal mitochondrial respiratory capacity. In this context, the present study demonstrates that LPB induces wound healing process and accelerates the inflammatory phase during acute traumatic muscle injury, probably by increasing the energy state of mitochondria

Effects of aquatic exercise on mental health, functional autonomy and oxidative stress in depressed elderly individuals: A randomized clinical trial

OBJECTIVES: The aim of this study was to investigate the effects of aquatic exercise on mental health, functional autonomy and oxidative stress parameters in depressed elderly individuals. METHODS: Initially, ninety-two elderly individuals were included in the study and were allocated into the depression group (n=16) and nondepression group (n=14). Both groups engaged in the aquatic exercise program for 12 weeks, including two weekly sessions (45 min/session) at a low intensity (between 50% and 60% of maximal heart rate or Borg scale scores of 13 to 14) throughout the intervention. All outcomes were evaluated at baseline and 12 weeks later. RESULTS: The patients were 63.5±8.8 years old. The following scores were decreased after training in the depressed group: depression (53%), anxiety (48%), and Timed Up & Go (33%). The following scores increased: Berg Balance Scale (9%) and flexibility (44%). Regarding the blood-based parameters, there were decreases in protein carbonylation (46%) and nitric oxide (60%) and increases in glutathione (170%) and superoxide dismutase (160%) in the depression group (po0.005). CONCLUSIONS: The aquatic exercise program reduces depression and anxiety, improves functional autonomy and decreases oxidative stress in depressed elderly individuals

Tratamento com plasma rico em plaquetas para fotoenvelhecimento facial: ensaio clínico randomizado duplo-cego

Artigo submetido ao Curso de Medicina da UNESC como requisito parcial para obtenção do Título de Bacharel em Medicina.Fundamentos: o plasma rico em plaquetas (PRP) é definido como um plasma sanguíneo autólogo com concentração de plaquetas acima dos valores de base. O PRP se trata de um procedimento pouco invasivo, sem contraindicações ou risco de rejeições, mínimos efeitos adversos, além de baixo custo para a sua realização. Objetivo: analisar a eficácia de PRP para o tratamento do fotoenvelhecimento facial em mulheres selecionadas no ambulatório de dermatologia de uma universidade no Extremo Sul Catarinense. Métodos: Ensaio clínico randomizado duplo-cego, que foi realizado com duas mulheres adultas. Foram feitas quatro aplicações de PRP em dois meses e acompanhamento por quatro meses após o procedimento. A avaliação foi feita através de biópsia, fotografias e aplicação de questionário elaborado pelas pesquisadoras. Resultados: Após a aplicação do PRP, as duas pacientes perceberam melhora com o procedimento e recomendariam a intervenção a conhecidos. Todas estariam dispostas a continuar a terapia e negaram a existência de efeitos adversos. As participantes, após o estudo, foram mantidas na classificação tipo III da Escala de Rugas de Glogau. As duas tiveram redução do influxo de células inflamatórias (placebo: p=0,102; PRP: p=0,144) e de fibroblastos (placebo: p=0,102, PPR: p=0,0465). Somente a que recebeu PRP teve estímulo de angiogênese (placebo: p=0,564, PRP: p=0,066) e, além disso, nesta participante houve maior expressão de colágeno (p=0,289). Conclusão: o tratamento com PRP mostra-se uma técnica promissora para o rejuvenescimento facial, embora estudos bem delimitados e com amostras maiores sejam realizados para avaliar de maneira mais fiel seu papel na dermatologia

Effect of aquatic exercise on mental health, functional autonomy, and oxidative dysfunction in hypertensive adults

The aquatic exercise is an effective non-pharmacological therapy for prevention and control of hypertension. The objective of the present study was to investigate the effect of aquatic exercise on mental health, functional autonomy, and oxidative dysfunction in hypertensive adults. Methodologically 29 adults (mean age 53 ± 7.5 years) were included in the study, and were randomly grouped as hypertensive (n = 16) and non-hypertensive (n = 13). Both groups underwent low-intensity aquatic exercise program for 12 weeks. Outcomes were evaluated at week 0 and 12. The values for the following parameters decreased in the hypertensive group post training: anxiety (−6.2 ± 2 score; 60%), Timed Up and Go test (−7.4 ± 0.3 sec; 30%), protein carbonylation (−0.15 ± 0.03 nmol/mg protein; 50%), nitric oxide (12.4 ± 6 nmol/mg protein; 62%), interleukin-6 (−27.6 ± 5.7 pg/mg protein; 46%), and tissue necrosis factor-alpha (−52.4 ± 3.8 pg/mg protein; 40%); however, the values of the following parameters increased before training: Berg score (56 ± 2; 7.8%), flexibility (27 ± 1 cm; 71%); glutathione (3.1 ± 1.3 nmol/mg protein; 138%), and superoxide dismutase (1.6 ± 0.4 nmol/mg; 166%). In conclusion, we suggest that low-intensity aquatic exercise program improved anxiety, functional autonomy, and oxidative dysfunction in hypertensive adults

Cigarette smoke exposure induces ROS-mediated autophagy by regulating sestrin, AMPK, and mTOR level in mice

Many pathological conditions linked to cigarette smoking are caused by the production of reactive oxygen species (ROS). The present study was conducted to analyze the effect of ROS on the lungs of Swiss mice exposed to cigarette smoking, focusing on autophagy-mediated mechanisms, and investigate the involvement of SESN2, AMPK, and mTOR signaling. Mice were exposed to cigarette smoke (CS) for 7, 15, 30, 45, and 60 days; the control group was not exposed to CS. Only mice exposed to CS for 45 days were selected for subsequent N-acetylcysteine (NAC) supplementation and smoke cessation analyses. Exposure to CS increased the production of ROS and induced molecular changes in the autophagy pathway, including an increase in phosphorylated AMPK and ULK1, reduction in phosphorylated mTOR, and increases in SESN2, ATG12, and LC3B levels. NAC supplementation reduced ROS levels and reversed all molecular changes observed upon CS treatment, suggesting the involvement of oxidative stress in inducing autophagy upon CS exposure. When exposure to CS was stopped, there were decreases in the levels of oxidative stress, AMPK and ULK1 phosphorylation, and autophagy-initiating molecules and increase in mTOR phosphorylation. In conclusion, these results suggest the involvement of ROS, SESN2, AMPK, and mTOR in the CS-induced autophagic process in the lung

Response of two swimming session on oxidative stress parameters in swimmers

This study investigated the acute effect of two swimming sessions on oxidative stress markers in trained individuals. Twelve male volunteers, students from UNESC (Universidade do Extremo Sul Catarinense, Criciuma, Santa Catarina, Brazil), with a mean age of 28 ± 7 years, initial weight of 72.9 ± 9 kg and height of 1.75 ± 0.08 cm, participated in the study. Blood samples were collected 24 hours before the sessions, immediately after the first and second session, and 24 hours after the end of the sessions. Aliquots were washed, red blood cells were lysed and plasma samples were stored at -80oC until the time of the biochemical assays. Creatine kinase (CK) activity, lipid peroxidation level, protein carbonylation, thiol content and catalase activity were determined. The results showed a significant increase in CK, lipoperoxidation and protein carbonylation and a decrease in thiol content after the second swimming session (p < 0.05) compared to pre-swimming levels. Catalase activity increased after the first and second swimming sessions. The main finding of the present study was that only the second swimming session resulted in oxidative stress

Green Synthesis of Gold Nanoparticles with Curcumin or Açai in the Tissue Repair of Palatal Wounds

This study aimed to evaluate and compare the effects of treatment with gold nanoparticles (GNPs) reduced with Curcumin (Curcuma longa L.) or Açai (Euterpe oleracea) to a standard commercial treatment of the pharmacological type (Omcilon®) and an electrophysical agent (photobiomodulation) in the palatal wounds of rats. As for the in vitro assay, a cell viability test was performed to assess the toxicity of the synthesized nanoparticles. In vivo assay: 60 Wistar rats were divided into five groups (n = 12): I. Palatal Wound (PW); II. PW + Photobiomodulation (PBM); III. PW + Omcilon®; IV. PW + GNPs-Cur (0.025 mg/mL); V. PW + GNPs-Açai (0.025 mg/mL). Animals were first anesthetized, and circular lesions in the palatine mucosa were induced using a 4 mm-diameter punch. The first treatment session started 24 h after the injury and occurred daily for 5 days. The animals were euthanized, and the palatal mucosa tissue was removed for histological, biochemical, and molecular analysis. GNPs-Açai were able to significantly reduce pro-inflammatory cytokines and increase anti-inflammatory ones, reduce oxidant markers, and reduce inflammatory infiltrate while increasing the collagen area and contraction rate of the wound, along with an improved visual qualification. The present study demonstrated that the proposed therapies of GNPs synthesized greenly, thus associating their effects with those of plants, favor the tissue repair process in palatal wounds