Orientador: Maria Alice da Cruz Hofling, Albetiza Lôbo de Araújo / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-17T23:01:57Z (GMT). No. of bitstreams: 1 Barbosa-Souza_Valeria_M.pdf: 7722866 bytes, checksum: 7544e873dc1e9f79280e8045930c4c37 (MD5) Previous issue date: 2011 / Resumo: Os acidentes por serpentes venenosas podem causar alterações locais graves e de rápido desenvolvimento. O veneno de Bothrops lanceolatus (VBL) contém, dentre outras toxinas, metaloproteinases hemorrágicas e pouco hemorrágicas (SVMPI e SVMPIII) e atividade pró-trombina que causam inflamação (vias das ciclooxigenases-COX e lipoxigenases), alterações hemostáticas e degenerativas. O veneno de Bothrops lanceolatus (100 ?g/100 ?l) foi injetado no gastrocnêmio de ratos para investigar a patogênese da mionecrose (1,3,6,18 horas 1, 2 dias) e regeneração muscular (3,7,14 e 21 dias) ocasionada pelo envenenamento através de análise histológica quantitativa (medida do menor diâmetro 1 hora e 21 dias pós- VBL) e imunohistoquímica para a osteopontina (OPN), citocina inflamatória, quimiotática, com sítios de ligação para integrinas de matriz e células. A medida da expressão dos macrófagos residentes (CD68+) e dos macrófagos migrantes (CD163+), de myoD e miogenina, membros da família de fatores transcricionais miogênicos foi feita com o objetivo de correlacionar com a expressão da OPN nos diferentes estágios patológicos (n=6/período) ao longo do período experimental. Os grupos controles foram injetados com PBS (100 ?l). O envenenamento produziu hemorragia local, edema, infiltrado neutrofílico e macrofágico e desorganização das bainhas perimisiais de tecido conjuntivo, após 48 horas. As fibras mionecróticas apresentavam-se em número moderado. Aos 3 dias, havia focos de deposição de colágeno (tipo I) no meio dos quais se viam mioblastos. O número de macrófagos CD68+ atingiu o máximo às 24 horas, e manteve-se significativamente maior que nos grupos controles até aos sete dias. A expressão de OPN foi significativamente maior das 6 horas aos 3 dias e dos 7 aos 14 dias, sendo expressa em fibras musculares, macrófagos, mioblastos, miotubos e fibroblastos. Não foi obtida marcação para anti-myoD; a expressão de miogenina, que tipicamente é nuclear, foi observada também no citoplasma de mioblastos e miotubos até o 70 dia (pico), sendo sua expressão somente nuclear aos 14 dias. A retenção de miogenina no citoplasma tem sido interpretada como mecanismo de retardo na diferenciação, já que a presença no núcleo é necessária para o seu papel regulador transcricional. Aos 21 dias as fibras regeneradas, com núcleo central, não haviam atingido o tamanho das fibras maduras intactas (VBL) e dos controles. Considerando que as SVMPs e a enzima com atividade tipo trombina de VBL, podem aumentar a capacidade de ligação de sítios da OPN a integrinas de matriz e de superfícies de células, sugerimos que durante a patogênese da regeneração muscular post- VBL a OPN estaria criticamente envolvida no processo inflamatório agudo, e como tal atuaria primordialmente como citocina ativadora de células satélites quiescentes, seguida por atividade quimiotática e adesiva, favorecendo migração, proliferação de mioblastos e a diferenciação de miotubos. Segundo a literatura a OPN é profibrótica em certas doenças. Sugerimos que a fibrose intersticial observada, mais a presença tardia de macrófagos no local, mais a expressão citoplasmática da miogenina podem ter sido fatores relevantes que levaram ao atraso da regeneração das fibras musculares, como constatado pelo tamanho menor diâmetro das fibras. Sugere-se para a OPN um papel dual, isto é, tanto próregenerativo, como anti-regenerativo na patogênese das alterações causadas pelo veneno de B. lanceolatus / Abstract: Accident caused by venomous snakes can lead to local changes of serious and rapid development. Bothrops lanceolatus venom (BLV) contains, among other toxins, hemorrhagic metalloproteinases and non-hemorrhagic (SVMPI and SVMPIII) and prothrombin activity. As result, the venom causes hemostatic and inflammatory (via lipoxygenase and cyclooxygenase) and degenerative alterations. In this study, Bothrops lanceolatus venom (100 ?g/100?l) was injected into the gastrocnemius of rats to investigate the pathogenesis of myonecrosis (one, three, six, 18 hours and two days) and regeneration (three, seven, 14 and 21 days) by quantitative histological analysis (measurement of the small diameter one hour and 21 days post-BLV injection) and immunohistochemistry for osteopontin (OPN) protein, an inflammatory and chemotactic cytokines, with integrin binding sites to matrix proteins and cells. The number of macrophages CD68+ (resident macrophages), and CD163 (migrants macrophages), the expression of myoD and myogenin, members of the myogenic, both transcription factors family were correlated (n = 6/period). Control groups were injected with PBS (100?l). The envenomation produced local hemorrhage (initially massive), acute interstitial edema and myofibers, neutrophil and macrophage infiltration, and disruption of the perimysium sheath of connective tissue. These changes were observed up to 48 hours. The number of myonecrotic fibers was in moderate number. At 3 days, there were foci of collagen (type I) deposition surrounding groups of myoblasts. The number of macrophages (CD68 +) peaked at 24 hours, and remained significantly higher for seven days: there was no expression of CD163 macrophages. The OPN expression showed two steps, a significant increase in, from six hours to three days and seven to 14 days, and it was expressed in muscle fibers, macrophages, myoblasts, myotubes and fibroblasts. There was no MyoD imunolabeling. The myogenin expression, which is known to be typically nuclear, was also observed in the cytoplasm of myoblasts and myotubes until 7 days (peak). At 14 days, the myogenin expression became nuclear. The cytoplasmic retention of myogenin has been interpreted as a mechanism to delay myotube differentiation, since the presence in the nucleus is required for its transcriptional regulatory role. At 21 days, the regenerated fibers with central nucleus had not reached the size of intact fibers (VBL), nor that of controls. Whereas SVMPs and an enzyme with thrombin-like activity are known to increase the capacity of OPN binding to integrin of matrix and cell surfaces, we suggest that, during the pathogenesis of muscle regeneration post-VBL injection, OPN would be critically involved in an acute inflammatory process. OPN would act primarily as a cytokine activator of quiescent satellite cells and later play chemotactic and adhesive activities, promoting migration, proliferation of myoblasts, and formation and differentiation of myotubes. According to the literature, OPN is profibrotic in certain diseases. We suggest that the foci of interstitial fibrosis and the late presence of macrophages at the site of regeneration, as well as the cytoplasmic expression of myogenin may be relevant factors that lead to regeneration delay. It is suggested a pro-regenerative and anti-regenerative roles for OPN in the pathogenesis of alterations caused by B. lanceolatus venom / Mestrado / Farmacologia / Mestre em Farmacologia

A seleção de linhagens de frangos de corte com altas taxas de crescimento e aumento de rendimento muscular vem contribuindo para o surgimento de perdas significativas na indústria avícola em decorrência de modificações anatômicas. Dentre estas alterações, destaca-se uma miopatia que se caracteriza pelo aparecimento de graus de estrias esbranquiçadas no músculo Pectoralis major, as quais seguem a direção da fibra muscular, referida como white striping. A etiologia da white striping ainda é desconhecida, mas avaliações histológicas demonstraram que esta alteração é usualmente associada à degeneração muscular e alterações miopáticas. Problemas de qualidade da carne podem ser resultantes de mudanças estruturais, morfométricas e bioquímicas do tecido muscular, com consequente alteração nas fibras musculares. Para tanto este trabalho, conduzido através de dois experimentos, teve como objetivos caracterizar histopatologicamente a miopatia white striping em diferentes graus de severidade (NORM, MOD, SEV) no músculo Pectoralis major de frangos de corte e as características morfométricas das fibras musculares, bem como avaliar a incidência desta condição em frangos de corte de linhagem Cobb abatidos aos 42 dias de idade. No primeiro experimento, a avaliação histopatológica demonstrou uma miopatia degenerativa nos graus de severidade classificados como MOD e SEV, entretanto as amostras de grau MOD apresentaram necrose ou necrose multifocal moderada, enquanto as de grau SEV apresentaram necrose difusa e acentuada, sugerindo avanço do processo degenerativo. Não foi observada proliferação de tecido conjuntivo em nenhum dos graus de white striping analisados, caracterizando este caso como de ocorrência aguda. No segundo experimento, foram verificados menor área de tecido muscular, menor diâmentro e maior densidade de fibras nos músculos Pectoralis major de frangos acometidos com white striping (MOD e SEV) quando comparado aos peitos sem esta condição (NORM). Conforme os resultados obtidos nesta pequisa, pode-se concluir que as fibras com os graus MOD e SEV de white striping demonstraram o início de um processo de regeneração muscular, de característica hiperplásica. / Genetic selection of broiler chickens for growth rate and increase of muscle yield has contributed to the appearance of significant losses in the poultry industry caused by anatomical changes. Among these alterations stands out a myophathy characterized by the occurrence of white striations on Pectoralis major muscle, seen parallel to the direction of muscle fibers, referred as white striping. The etiology of white striping is still unknown but histological changes demonstrated that this alteration is usually associated with muscle degeneration and myopatic changes. Meat quality problems may result from structural, morphological and biochemical changes of muscle tissue with consequent alterations in muscle fibers. To this study, two experiments were conducted, and aimed to histopathologically characterize the myopathy white striping at varying degrees of severity (NORM, MOD, SEV) on broilers Pectoralis major muscle and the morphometric muscle fibers characteristics as well as evaluate the incidence of this condition on Cobb 500 broiler breast fillets slaughtered at 42 days of age. The histopathological evaluation on experiment 1 showed a degenerative myopathy in both MOD and SEV degrees, however the samples classified as MOD showed microscopically necrosis or necrosis multifocal moderate, while the classified as SEV showed necrosis diffuse and strong suggesting an advancement in the degenerative process. The proliferation of connective tissue was not observed for either of the degrees of white striping investigated, characterizing this profile as an acute occurrence. In experiment 2, small area and diameter and higher densities of muscle fiber were observed in broilers Pectoralis major muscle with white striping (MOD and SEV) compared to NORM. According to the results, we can conclude that muscle fibers with white striping showed the beginning of a muscle regeneration process with hyperplasic characteristics.

Orientador: Vera Aparecida Madruga / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Educação Física / Made available in DSpace on 2018-08-18T10:46:08Z (GMT). No. of bitstreams: 1 Conceicao_MiguelSoares_M.pdf: 1532956 bytes, checksum: 8b4cb337b2669a48e0b8c68602a72f25 (MD5) Previous issue date: 2011 / Resumo: O presente estudo analisou as alterações nos marcadores indiretos de dano muscular após uma sessão de ações excêntricas de flexores do cotovelo com diferentes velocidades de movimento, em mulheres jovens e na pós-menopausa. Participaram da pesquisa 37 mulheres, voluntárias, com idade compreendida entre 18 a 65 anos. Essas voluntárias foram divididas em quatro grupos. Grupo de mulheres jovens que executou ações excêntricas com velocidade rápida (n=10) e grupo de jovens que executou ações excêntricas com velocidade lenta (n=9). Grupo de mulheres na pós-menopausa que realizou as ações excêntricas com velocidade rápida (n=10) e grupo de mulheres na pós-menopausa que realizou as ações excêntricas com velocidade lenta (n=8). As ações excêntricas foram realizadas no braço não dominante, utilizando-se um dinamômetro isocinético, e consistiram em 30 ações excêntricas máximas divididas em cinco séries de seis repetições. Para analisar a magnitude do dano muscular foram utilizados marcadores indiretos como a contração isométrica voluntária máxima (CIVM), amplitude de movimento (AM), circunferência de braço, dor muscular, creatina quinase (CK), interleucina-6 (IL-6), fator de necrose tumoral (TNF-?) e prostaglandina (PGE2). A CIVM, AM, dor muscular e circunferência de braço foram avaliadas pré, imediatamente após, 24, 48 e 72h após a execução de ações excêntricas. Para a atividade plasmática da CK, IL-6, TNF-? e PGE2, as coletas aconteceram pré, 24, 48 e 72h após as ações excêntricas. Como resultado desta dissertação é apresentado um artigo original. O objetivo do estudo original apresentado foi comparar as alterações dos marcadores de dano muscular e inflamação entre mulheres jovens e na pós-menopausa após ações excêntricas. Diferenças significantes entre grupos foram encontradas para torque excêntrico e TNF-? na qual, mulheres na pós-menopausa apresentaram maiores valores (p<0,05). Uma correlação positiva significante (p<0,05) se estabeleceu entre idade x dor e idade x PGE2. Concluindo, esses resultados não suportam a hipótese que mulheres na pós-menopausa são mais susceptíveis ao dano muscular induzido por ações excêntricas comparado a mulheres jovens / Abstract: The present study have analyzed the changes in muscle damage and inflammatory markers after an acute bout of elbow flexors eccentric exercise at difference velocities, in young and post menopausal women. 37 women aged between 18 and 65 participated of the study. They were divided into four groups, (1) fast velocity, young women (n = 10); (2) slow velocity, young women (n = 9); (3) fast velocity, postmenopausal women (n = 10) and (4) slow velocity, postmenopausal women (n = 8). Eccentric exercise consisted of five sets of six maximal elbow flexors eccentric repetitions performed with the nondominant arm in an isokinetic dynamometer. It was assessed voluntary isometric contraction (MVIC), range of motion (ROM), arm circumference, muscle soreness, creatine kinase (CK), interleukin-6 (IL-6), tumor necrosis factor (TNF-?) and prostaglandin (PGE2). The MVIC, ROM, muscle soreness and arm circumference were assessed before, immediately after, 24, 48 and 72h after eccentric exercise. For the plasma activity of CK, IL-6, TNF-? and PGE2, the blood samples were made before, 24, 48 and 72 hours after the eccentric protocol. As result of the present dissertation it is presented an original article. The aim of the presented original study was to compare changes in indirect markers of muscle damage and inflammation in young and postmenopausal women after maximal eccentric exercise. Significant differences were found between groups for eccentric torque and TNF-?, with greater values for postmenopausal women (p<0.05). A positive significant correlation (p<0.05) was found between age x muscle soreness and between age x PGE2. In conclusion, these results do not support the hypothesis that postmenopausal women were more susceptible to eccentric exercise-induced muscle damage than young women / Mestrado / Atividade Fisica, Adaptação e Saude / Mestre em Educação Física

Desenvolvemos um instrumento capaz de provocar uma lesão muscular aguda por mecanismo de impacto, no músculo gastrocnêmio de ratas, através de um trauma direto e não-invasivo. Após a produção da lesão experimental avaliamos os efeitos do ultra-som terapêutico (UST), adicionado ou não à imobilização gessada, como forma de tratamento imediato à lesão muscular pela análise das propriedades mecânicas deste músculo. Foram utilizadas 68 ratas da raça Wistar, sendo divididas em 7 grupos experimentais da seguinte forma: Grupo 1 – Controle intacto; Grupo 2 – Sem tratamento efetivo, Grupo 3 – Imobilização gessada por 24 horas; Grupo 4 – Imobilização gessada por 72 horas; Grupo 5 – Estimulação com UST sem presença de imobilização; Grupo 6 – Imobilização gessada por 24 horas associada à estimulação com UST; Grupo 7 – Imobilização gessada por 72 horas associada à estimulação com UST tratadas durante 6 dias consecutivos com UST, durante 5 minutos diários. Avaliamos as propriedades mecânicas obtidas através de ensaios mecânicos de tração longitudinal e os resultados foram analisados estatisticamente. O estudo evidenciou que a propriedade de alongamento no limite de proporcionalidade não foi um bom parâmetro de correlação para avaliar a efetividade dos tratamentos empregados junto ao processo de reparação muscular. A utilização isolada do UST como técnica de tratamento apresentou melhores resultados em relação às propriedades mecânicas de carga no limite de proporcionalidade, rigidez, resiliência, carga e alongamento máximo. No entanto, a utilização da imobilização gessada por 72 horas associada ou não a estimulação ultra-sônica foi capaz de proporcionar melhorias somente em relação ao alongamento máximo. Sugere-se ainda, que a utilização da imobilização gessada por períodos de 24 horas associada ou não ao uso do UST não foi capaz de apresentar resultados benéficos para as propriedades mecânicas avaliadas / We developed an instrument capable to provoke a muscular injury acute for impact mechanism, in the muscle gastrocnemius of female rats, through a direct and non-invasive. After the production of experimental injury we evaluate the effects of the therapeutic ultrasound (UST), added or not to the cast immobilization, as form of immediate treatment to the muscular injury, for the analysis of the mechanical properties of this muscle. Sixty eight female Wistar rats were used and being divided in seven experimental groups: Group 1 – Intact Control; Group 2 – Without treatment; Group 3 -Immobilization in spica cast for 24 hours; Group 4 - Immobilization in spica cast for 72 hours; Group 5 – Stimulation with UST without immobilization presence; Group 6 - Immobilization in spica cast for 24 hours associated to the stimulation with UST and Group 7 - Immobilization in spica cast for 72 hours associated to the stimulation with UST, treated during six consecutive days with UST, for five minutes daily. We evaluted properties we carry through assays of logitudinal traction in the universal machine of assays and the gotten results had been analyzed statical test. The study it evidenced that the property of proportional limit strength in the was not a good parameter of correlation to evaluate the effectiveness of the employed treatments together to the process of muscular repairing. The isolated use of the UST as treatment technique presented better resulted in relation to the mechanical load properties in the proportional limits, stiffness, resilience, load and strength maximum. However, the use of the spica cast for periods of 72 hours associated or not ultrasound stimulation was not capable to only provide improvements in relation to the maximum strength. One still suggests, that the use of the spica cast in the immobilization for 24 hours periods associate or not to the use of the UST was not capable to present resulted beneficial for the evaluated mechanical properties

Thesis (PhD (Physiological Sciences))--University of Stellenbosch, 2011. / Includes bibliography. / ENGLISH ABSTRACT: Muscle injuries are associated with changes in skeletal muscle as well as the immune system. All studies investigating possible treatment modalities have found both positive and negative effects on muscle recovery. Since no universally accepted treatment modality exists, this thesis aims to determine whether a plant-derived antioxidant, proanthocyanidolic oligomer (PCO), might prove beneficial as treatment for sports injuries in order for athletes to return to the sports field quicker. The difference in recovery of muscle following both chronic (supplementation started 14 days prior to injury and continued thereafter) and acute supplementation (supplementation started two hours after injury) were also investigated. Both chronic and acute PCO supplementation in a rat hindlimb contusion injury model resulted in earlier muscle recovery, verified by an earlier satellite cell response compared to the placebo group. This effect was most prominent already at the four hour time point following injury, compared to day seven and three after chronic and acute placebo treatment respectively. PCO supplementation also resulted in quicker foetal myosin heavy chain (MHCf) expression compared to placebo treatment. Chronic supplementation specifically resulted in a blunted circulatory pro-inflammatory cytokine response, whilst allowing for a significant increase in IL-10, an anti-inflammatory cytokine, on day three (in the PCO group only). At tissue level, the response of the muscle pro-inflammatory cytokines, TNF- and IL- 6, coincided with the satellite cell response. Macrophage infiltration into the injured muscle also followed a similar pattern to that seen for the pro-inflammatory cytokines. Macrophages invaded the injured area quicker when supplemented with PCO chronically, however, macrophage infiltration could not explain the cytokine response seen with acute supplementation. Both chronic and acute supplementation with PCO was responsible for a severely blunted neutrophil response, a novel finding of this particular antioxidant. The main findings of the in vivo rodent study were that PCO was able to blunt the neutrophil response, whilst allowing for earlier macrophage infiltration. To establish possible mechanisms by which PCO might exert these beneficial effects, further analysis included determining macrophage phenotypes and neutrophil numbers in circulation. An in vitro neutrophil migration assay was also employed to further elucidate PCO’s ability to blunt neutrophil infiltration into the injured area. For this study, conditioned plasma were harvested from experimental animals and added together with neutrophils from control rats and granulocyte colony stimulating factor (G-CSF) to the insert of the migration chamber. A chemotactic factor, N-formyl methionine-leucine-phenylalanine (fMLP), was added to the bottom well and neutrophils were allowed to migrate for two hours. Results from this study indicated that neutrophil migration was attenuated in vitro in the presence of conditioned plasma from PCO supplemented rats only. The studies in this thesis on the effect of PCO on parameters of muscle and the immune system led to the following main conclusions: a) PCO supplementation resulted in earlier muscle recovery as a result of earlier satellite cell activation and MHCf synthesis; b) PCO favours an anti-inflammatory cytokine reaction, whilst blunting the pro-inflammatory cytokine response; and c) PCO blunted the neutrophil response whilst facilitating earlier macrophage infiltration into the injured area. The specific mechanism of action of PCO to blunt the neutrophil response specifically, possibly includes the ability to suppress adhesion molecule expression on the neutrophils themselves. However, this warrants further investigation. / AFRIKAANSE OPSOMMING: Spier beserings word geassosiëer met veranderinge in skeletspier sowel as die immuunstelsel. Meeste studies wat moontlike behandelingsopsies ondersoek, het beide positiewe en negatiewe spierherstel gerapporteer. Omrede daar geen universele behandelingsmoontlikheid bestaan nie, is die doel van hierdie tesis om die effek van ‘n plantgebaseerde anti-oksidant, pro-antosianiedoliese oligomeer (PSO), as ‘n voordelige behandelingstrategie vir sportbeserings te toets. Die verskil in spierherstel na beide kroniese (supplementering wat 14 dae voor besering begin is, en volgehou is daarna) en akute supplementering (supplementering het twee uur na besering begin), is ook ondersoek. Beide kroniese en akute PSO supplementering, in ‘n rot agterbeen-kneusbeseringmodel, het gelei tot vroeë spierherstel. Die bevindinge is geverifiëer deur ‘n vroeë satelietselrespons in vergelyking met die plasebo groep. Hierdie effek was reeds opvallend vier uur na besering, in vergelyking met die dag sewe en dag drie tydpunt tydens kroniese en acute plasebo behandeling onderskeidelik. In vergelyking met die kontrole groep, het PSO supplementering ook gelei to vininger uitdrukking van miosienswaarketting (MHCf). Kroniese supplementering het spesifiek gelei to ‘n onderdrukte sirkulatoriese pro-inflammatoriese sitokien response, terwyl ‘n betekenisvolle toename in IL-10 op dag drie (in die PSO groep alleenlik) waargeneem is. Op weefselvlak, het die pro-inflammatoriese sitokiene, IL-6 en TNF- , dieselfde patron gevolg as die van satelietselle. Makrofaaginfiltrasie binne die beseerde spier het ook ‘n soorgelyke patroon gevolg. Makrofage het die beseerde area vinniger geïnfiltreer in die kronies PSO-gesupplementeerde groep, maar kon nie die sitokienrespons, wat waargeneem is met akute supplementasie, verklaar nie. Beide kroniese en akute PSO supplementering was verantwoordelik vir ‘n onderdrukte neutrofiel respons, wat ‘n nuwe bevinding is vir hierdie spesifieke anti-oksidant. Die hoof bevindinge in die in vivo rotstudies, is dat PSO instaat is om die neutrofielrespons te onderdruk, en sodoende vroeë makrofaaginfiltrasie teweeg te bring. Om meganismes waarby PSO hierdie voordelige effekte veroorsaak te ondersoek, is verdere analises gedoen om makrofaagfenotipe en neutrofielgetalle in die sirkulasie te bepaal. ‘n In vitro neutrofielmigrasie studie is ook aangewend om PSO se vermoë om neutrofielinfiltrasie in die beseerde area te onderdruk, te ondersoek. Neutrofiele van kontrole rotte, tesame met gekondisioneerde plasma van eksperimentele diere en granulosiet-kolonie stimulerende faktor (G-KSF), is toegelaat om vir twee ure in die teenwoordigheid van ‘n chemotaktiese faktor, N-formiel metionien-leusien-fenielalanien (fMLP) te migreer. Resultate van hierdie studie het aangetoon dat neutrofielmigrasie, in vitro, alleenlik onderdruk word in die teenwoordigheid van gekondisioneerde plasma van PSO-gesupplementeerde rotte. Die studies in hierdie tesis oor die effek van PSO op parameters van spier en die immuunsisteem, het tot die volgende hoofgevolgtrekkings gelei: a) PSO supplementering het vroeë spierherstel, as gevolge van vroeë satelietselaktivering en MHCf sintese, teweeg gebring; b) PSO verkies ‘n anti-inflammatoriese sitokien reaksie, terwyl dit die proinflammatoriese sitokienrespons onderdruk; en c) PSO onderdruk die neutrofielrespons, terwyl vroeë makrofaaginfiltrasie in die beseerde area gefasiliteer word. Die spesifieke meganisme van aksie van PSO, om die neutrofielrespons te onderdruk, kan moontlik die vermoë van neutrofiele om adhesie molekule uit te druk, insluit. Hierdie aanname moet egter verder ondersoek word.

A traumatic skeletal muscle injury that involves the loss of a substantial portion of tissue will not regenerate on its own. Little is understood about the ability of the muscle to recover function after such a defect injury, and few research models exist to further elucidate the repair and regeneration processes of defected skeletal muscle. In the current research, a model of muscle injury was developed in the lateral gastrocnemius (LGAS) of the rat. In this model, the muscle gradually remodels but functional recovery does not occur over 42 days. Repair of the defect with muscle-derived extracellular matrix (ECM), improves the morphology of the LGAS. Blood vessels and myofibers grow into the ECM implant in vivo, but functional recovery does not occur. Addition of bone marrow-derived mesenchymal stem cells (MSCs) to the implanted ECM in the LGAS increases the number of blood vessels and regenerating myofibers within the ECM. Following 42 days of recovery, the cell-seeded ECM implanted LGAS produces significantly higher isometric force than the non-repaired and non-cell seeded ECM muscles. These results suggest that the LGAS muscle defect is a suitable model for the study of traumatic skeletal muscle injury with tissue loss. Additionally, MSCs seeded on an implanted ECM lead to functional restoration of the defected LGAS. / text

O objetivo deste estudo foi investigar o efeito da sobrecarga de leucina no músculo sóleo de ratos submetidos à DGO (Distração Gradual Osteogênica). Vinte e cinco ratos Wistar foram divididos em 5 grupos: controle, sham-lengthening, leucina, DGO e DGO+leucina. A DGO de 2mm/dia foi realizada na tíbia até um ganho de 20% e a sobrecarga de leucina (114mM) foi administrada oralmente. Avaliou-se: estrutura histológica muscular, número de sarcômeros em série, área dos diferentes tipos de fibras musculares e análise do tecido conjuntivo. Os dados obtidos dos cortes histológicos mostram que a DGO promoveu intensa lesão muscular e aumento do tecido conjuntivo, o que foi minimizado pela leucina. Houve uma adição no número de sarcômeros em série (26%) durante a DGO, que foi intensificada pela leucina (33%). Observou-se uma diminuição da área de todos os tipos de fibras com a DGO e um aumento da área das fibras do tipo I com a leucina. Portanto, a leucina minimizou os efeitos causados pela DGO, podendo esta ser utilizada como um agente terapêutico durante a DGO no futuro. / The goal of the present study was to investigate the effect of leucine overload in the rat soleus muscle before and along DGO (Gradual Distraction Osteogenesis). Twenty five male Wistar rats were divided into 5 groups: control, sham-lengthening, leucine, DGO and DGO+leucine. The DGO was 2mm/day and the leucine overload (114mM) was administrated orally. Subsequently analyzed: histological cross-section, sarcomere serial number and cross-sectional area of the types fibers and connective tissue. The histological data showed that the DGO promoted intense injury and increase of connective tissue, which can be minimized by leucine. The sarcomere serial number increased by ~26% and leucine overload increased this gain to 33%. The fibers types showed a decrease of the cross-section area of the all type fibers with the DGO and increase of the type I fibers area with leucine overload. We conclude that leucine overload is a potential therapeutical device in DGO, seeking improvement of skeletal muscle longitudinal growth.

Submitted by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2018-09-27T17:10:28Z No. of bitstreams: 1 WILLIAN DA SILVA.pdf: 1484606 bytes, checksum: b09c4375bb0dae5662922161c28677b5 (MD5) / Approved for entry into archive by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2018-09-27T17:10:44Z (GMT) No. of bitstreams: 1 WILLIAN DA SILVA.pdf: 1484606 bytes, checksum: b09c4375bb0dae5662922161c28677b5 (MD5) / Made available in DSpace on 2018-09-27T17:10:44Z (GMT). No. of bitstreams: 1 WILLIAN DA SILVA.pdf: 1484606 bytes, checksum: b09c4375bb0dae5662922161c28677b5 (MD5) Previous issue date: 2017 / O extrato de chá verde possui potencial antioxidante e anti-inflamatório, que pode contribuir para recuperar o equilíbrio metabólico e estresse oxidativo pós-exercício intenso, colaborando assim para diminuir episódios de dor muscular de início tardio (DMIT). Neste estudo, buscamos determinar os efeitos da suplementação com extrato de chá verde sobre a DMIT, dano muscular e oxidativo e status antioxidante. Em um ensaio randomizado, triplo cego, placebo controle, 20 homens não treinados com idade média de 24 (±5) anos realizaram exercício para indução da DMIT no tríceps sural antes e depois de 15 dias de suplementação com extrato de chá verde (500 mg/dia) (GT, n=10) ou placebo (PL, n=10). A DMIT foi determinada pela escala visual analógica de dor (pré e pós-suplementação) antes, imediatamente após e 48 horas após o exercício. Amostras sanguíneas foram tomadas para determinar marcadores bioquímicos de dano muscular como creatina quinase (CK), lactato desidrogenase (LDH) e marcadores de estresse oxidativo como níveis de malondialdeido (TBARS), produção de espécies reativas de oxigênio (DCFH-DA), juntamente com a carbonilação de proteínas e status antioxidante como capacidade antioxidante total do plasma (FRAP) e atividade da glutationa (GSH) além da atividade da acetilcolinesterase (AChe). O exercício induziu DMIT e a suplementação com GT reduziu um marcador de dano estrutural (CK), mas não atenuou a DMIT e marcadores de estresse oxidativo e status antioxidante. Este efeito protetivo ao dano estrutural interessa especialmente ao uso desta suplementação como possível fator adjunto para a prevenção de dano muscular e auxílio na recuperação de esforços intensos. / Green tea extracts supplementation (GT) is recognized as a source of antioxidants and anti-inflammatory substances that may contribute to the recovery after intense exercise when muscle soreness (MS) occurs. In this study we determine the effects of GT on MS, muscle damage, oxidative stress and antioxidant status. Here we present a randomized trial triple blind placebo control including 20 adults aged 25 (5) years and physically active that underwent sessions exercise for induction of muscle damage in the triceps sural before and after 15 days of supplementation with GT at dose of 500 mg/day (GT, n=10) or placebo (PL, n=10). MS was determined using a visual analogic scale. Blood samples were collected to determination of biochemical markers of damage considering activity of creatine kinase (CK) and lactate dehydrogenase (LDH), oxidative stress considering levels of malondialdehyde (TBARS), reactive species of oxygen (DCFH), protein carbonylation, and antioxidant status considering total antioxidant capacity (FRAP), glutathione activity (GSH), and acetylcholinesterase (AChE) activity. Exercise induced MS and GT reduces a marker of structural muscle damage without effects on oxidative stress and antioxidant status. This protective effect may be support the use of this supplementation as a possible correlate strategy to prevent muscle damage and therefore contribute to the recovery of intense exercise.

O objetivo deste estudo foi avaliar os efeitos do ultrassom pulsado (USp) no reparo muscular em um modelo experimental de lesão lacerativa do músculo gastrocnêmio medial em ratos Wistar. Vinte ratos foram tratados diariamente com USp (50%), 1MHz, 0,57W/cm² de intensidade por 5 min. formando os grupos tratados (GT), e 20 animais constituíram os grupos controle (CG). A análise histológica, morfométrica (usando o método estereológico) e biomecânica (teste de tensão) foi realizada aos 4, 7, 14 e 24 dias após a lesão. As lesões apresentaram um padrão de reparo similar tanto nos GT como nos GC. Os volumes absolutos da lesão (VL) e das zonas central e de regeneração (VZC e VZR) diminuíram progressivamente ao longo do processo de reparo tanto nos GT como nos GC. No GT, o VL diminuiu significativamente em todos os dias experimentais, sendo que VZC uma diminuição significante aos 4 e 7 dias pós-lesão e o VZR aos 14 dias pós-lesão. A fração de volume de vasos sanguíneos e a fração de superfície de vasos sanguíneos foi maior nos GT aos 4 e 7 dias pós-lesão em relação aos respectivos controles. Apesar de haver uma tendência a um maior volume absoluto de vasos sanguíneos nos GT, a análise estatística mostrou que existe uma maior volume de vasos somente aos 4 dias pós-lesão. Não houve diferença significante na área de superfície total de vasos sanguíneos na lesão quando se comparam os grupos entre si. Houve um aumento significante na fração de volume de fibras de colágeno na lesão nos GT aos 4,7 e 14 dias pós-lesão. Houve um aumento significante na tensão máxima e na rigidez nos GT aos 4 e 24 dias após a lesão. Não houve diferença significante na deformação relativa entre GC e GT. Os resultados sugerem que o USp otimiza a fase inflamatória e estimula as fases proliferativa e de remodelamento, promovendo uma diminuição mais acentuada no volume da lesão, estimulando a angiogênese, assim como, a deposição e a organização do colágeno fibrilar. Os achados histológicos corroboram com os achados biomecânicos, que mostram que os músculos tratados pelo USp tiveram propriedades biomecânicas mais parecidas com as do músculo íntegro / The aim of this study was to evaluate the effect of the pulsed ultrasound therapy (PUT) in stimulating muscle repair in an experimental model of lacerative gastrocnemius medialis muscle lesion in 40 Wistar rats. Twenty rats were treated (TG) daily with 1MHz pulsed ultrasound (50%) at 0.57W/cm² for 5 min, and 20 were control animals (CG). Muscle samples were harvested up on postoperative days 4, 7, 14 and 24 for stereological, histological, and biomechanical analyses. The lesions presented similar repair pattern in both TG and CG. The lesion volume (VL) and the central and regenerative zones volumes (VCZ and VRZ) had a progressive deacrease through the post lesion period both in the TG and CG. The VL decrease was significantly greater in the TG in all experimental days, the VCZ decrease was significant in the TG at 4 and 7 days post lesion, and the VRZ decrease was significant at 14 days post lesion in the TG. Statistically significant increase was found in the blood vessels volume fraction and in the surface fraction of blood vessels in the TG at 4 and 7 days post lesion compared to respective CG. Although there was a tendency to have a greater blood vessels absolute volume within lesion in the TG, the statistical analysis showed that it was only larger at 4 days after surgery in US treated group. No statistically significant increase was found in the surface total area within lesion in all experimental days between CG and TG. There was a significant increase in the volume fraction of fibrilar collagen within the lesion in the TG at 4, 7 and 14 days post lesion. The biomechanical data showed a significant increase in the maximal stress and stiffness in the TG at 4 and 24 days after lesion, although there was a progressive increase of these variables both in the CG and TG. There was no significant difference in the maximal elongation, between CG and TG. Our data suggest that the PUT acts as an inflammatory optimizer and stimulates the proliferative and remodeling phases, promoting a greater decrease in the VL and in the VCZ, stimulating angiogenesis and controlling fibrilar collagen deposition and organization in this experimental model of lacerative gastrocnemius muscle lesion. The histological data are in accordance to the biomechanical data, which shows that the muscles treated by USp have biomechanical properties similar to the noninjuried muscles

Tourniquet mediated Ischemia Reperfusion (I/R) injury causes damage to skeletal muscle, often resulting in prolonged functional impairment. The current study utilizes immunohistochemistry (IHC) to determine whether the controlled release of the anabolic factor, insulin-like growth factor-I (IGF-I), from the biodegradable PEGylated fibrin gel matrix can facilitate the recovery of skeletal muscle from I/R. Treatment groups following a 2-hour tourniquet applied to the limb of 6-9 month rats, included intramuscular injections of saline, PEGylated fibrin gel (PEG-Fib) only and IGF-I conjugated to PEGylated fibrin gel (PEG-Fib-IGF). Expression of the myogenic regulatory factors MyoD and myogenin detected via IHC in the PEG-Fib-IGF group was significantly lower compared to the saline group, showing a 1.4±0.8% nuclear co-localization for MyoD and a 2.0±0.8% nuclear co-localization for myogenin at 14 days of recovery. The saline group showed higher values, 31.4±4.4% and 44.1±7.3% for MyoD and myogenin nuclear co-localization respectively. A significantly greater percentage, 88.8±3.7% of Desmin positive myofibers was seen at 14 days of recovery, while a lower percentage of fibers expressing neonatal myosin, 7.7±2.7% was seen in the PEG-Fib-IGF group compared to the saline treatment group. These results indicate that IGF-I delivered intramuscularly via PEGylated fibrin gel, functions therapeutically in skeletal muscle recovery, from I/R mediated damage. In a separate injury model that deals with volumetric muscle loss, IHC analyses were performed to test the efficacy of a novel tissue engineering strategy utilizing extracellular matrix (ECM) as a scaffold. In this model, also called the defect model, a 1.0 X 1.0 cm piece of the lateral gastrocnemius was removed and replaced with a muscle-derived ECM. The constructs were then seeded with bone marrow derived cells (BMSCs), adipose derived stem cells (ADSCs) or the peroneal nerve was relocated to the area of the ECM implant. 42 days post recovery IHC analysis was performed on the ECM implants. The quantification of desmin-positive regenerating myofibers bearing centrally located nuclei, showed significantly greater values in the top, middle and bottom region of the ECM implants that received peroneal nerve relocation, when compared to the experimental group that received the ECM implant alone. Blood vessel density increases were seen within the middle region of the ECM implant groups that received BMSC+Nerve treatment and the bottom region of the ECM implant groups that received ADSC+Nerve treatment. Thus, these results corroborate the therapeutic effect of peroneal nerve relocation, which stimulated an increase in myofiber regeneration and vascular maintenance within the construct. / text