Submitted by Nadir Basilio (nadirsb@uninove.br) on 2015-07-21T14:58:44Z No. of bitstreams: 1 Manoel Carneiro de Oliveira Junior.pdf: 4260915 bytes, checksum: ba3950686530a475258ace49a0675cdd (MD5) / Made available in DSpace on 2015-07-21T14:58:45Z (GMT). No. of bitstreams: 1 Manoel Carneiro de Oliveira Junior.pdf: 4260915 bytes, checksum: ba3950686530a475258ace49a0675cdd (MD5) Previous issue date: 2013-09-30 / Acute respiratory distress syndrome (ARDS) is a syndrome that presents high mortality rates, and the results of both insults pulmonary or extra-pulmonary (pneumonia or septic shock) are high, and is a disease characterized by respiratory insufficiency from the inflammatory response that leads to alteration of alveolar-capillary permeability, pulmonary edema and hypoxemia refractory to high flow oxygen. One of the most important mechanisms that determined the severity of this injury is the magnitude of the injury of alveolar epithelial barrier. The possibility of repairing epithelial at an early stage is the major determinant of recovery. Many of therapeutic modalities based on the attempt to decrease lung inflammation to minimize the initial injury and much of the inflammatory process occurs through activation of local and systemic cytokines such as TNF-α and IL-1β. A growing number of studies report that Low Level Laser Therapy (LLLT) have anti-inflammatory effects in models of LPS-induced pulmonary ARDS, however, so far, only the red spectrum lasers were studied. Therefore, this study aimed to investigate the role of infra red laser (830nm), 3J/cm2, 35mw, 80 seconds per point (03 points per application), in pulmonary inflammation, lung using LPS model (intratracheal) and also extrapulmonary (intraperitoneal) inducing ARDS. The laser application was performed directly in contact with the skin in the chest three points (corresponding to the end of the trachea - Section 01 right lung - point 02 and left lung - point 03), three times, beginning 01 hour after LPS administration. BALB / c mice (n = 40) were divided into control (n = 08; not administered LPS), IT (n = 07; intratracheal administered LPS (10 µg / mouse), IT + LLLT (n = 09; intratracheal LPS administered (10 µg / mouse) + LLLT), IP (n = 07; LPS administered intraperitoneal (100 µg / mouse), IP + LLLT (n = 09; administered intraperitoneal LPS (100 µg / mouse) + LLLT). Twenty-four hours after administration of LPS and Laser, animals were euthanized and the lungs removed for studies of pulmonary inflammation: Total cell count and differential, bronchoalveolar lavage (BAL), cytokines (IL-1beta, IL-6, IL-10, KC and TNF-α), BAL levels were also analyzed quantitatively the number of neutrophils in the lung parenchyma in lung tissue using histomorphometry techniques. Results showed that LLLT significantly reduced pulmonary and extra-pulmonary LPS induced in both configurations Experimental of ARDS, as evidenced by a reduction in the number of total cells and neutrophils in BAL, reduced levels of IL-1β, IL-6, KC, and TNF-α in BAL fluid as well as the number of neutrophils in the lung parenchyma. Therefore, we conclude that the 830nm infrared laser is effective in reducing pulmonary inflammation in both models pulmonary or extrapulmonary LPS-induced experimental ARDS. / A síndrome do desconforto respiratório agudo (SDRA) é uma síndrome que apresenta altas taxas de mortalidade, que pode ser resultante tanto de insultos pulmonares como extrapulmonares. A síndrome é caracterizada pela insuficiência respiratória proveniente da resposta inflamatória que cursa com alteração de permeabilidade alvéolo-capilar, edema e hipoxemia refratária aos altos fluxos de oxigênio. Um dos mais importantes mecanismos que determinam a severidade desta injúria é a magnitude da lesão da barreira epitélio alveolar. A possibilidade de reparação do epitélio em um estágio precoce é o maior determinante da recuperação. Muitas das modalidades terapêuticas baseiam-se na tentativa de diminuição da inflamação pulmonar para minimizar a lesão inicial, a qual se deve em grande parte ao processo inflamatório mediado pela ativação local e sistêmica por citocinas como TNF-α e IL-1β. Um número crescente de estudos relata que o laser de baixa intensidade apresenta efeitos antiinflamatórios em modelos de SDRA induzida por LPS e isquemia e reperfusão da artéria pulmonar. No entanto, até o momento, apenas lasers no espectro vermelho (650 – 655 nm) foram estudados. Portanto, o presente estudo tem como objetivo investigar o papel do laser de baixa intensidade (LBI), na faixa do infravermelho (830nm), 3J/cm2, 35mw, 80 segundos por ponto (03 pontos por aplicação), na inflamação pulmonar, usando um modelo de SDRA de origem pulmonar (LPS intratraqueal) e também extrapulmonar (LPS intraperitoneal). A aplicação do laser foi realizada diretamente em contato com a pele, em três pontos do tórax (correspondente ao final da traquéia - ponto 01, pulmão direito - ponto 02 e do pulmão esquerdo ponto 03), por três vezes, 01 hora após a administração de LPS. Camundongos BALB/c (n = 40) machos foram distribuídos em Controle (n = 08; não administrado com LPS), IT 10 (n = 07; LPS intratraqueal; 10 µg/camundongo), IT + Laser (n = 09; LPS intratraqueal; 10µg/camundongo + Laser), IP (n= 07; LPS intraperitoneal; 100µg/ camundongo), IP + Laser (n = 09; LPS intraperitoneal; 100 µg/camundongo + Laser). Os animais foram eutanaziados vinte e quatro horas após a administração de LPS. Foi avaliada a contagem de células totais e diferenciais no lavado bronco alveolar (LBA), os níveis de citocinas (IL-1β, IL-6, IL 10, KC e TNF-α), a densidade de neutrófilos no parênquima pulmonar. Os resultados demonstraram que o LBI significativamente reduziu o número de células totais e de neutrófilos no Lavado Bronco Alveolar (LBA), o número de neutrófilos no parênquima pulmonar, e os níveis de citocinas pró-inflamatórias no LBA tanto no modelo de SDRA pulmonar quanto extrapulmonar. Portanto, concluímos que o laser infravermelho 830nm é eficaz para reduzir a inflamação pulmonar, em ambos os modelos de SDRA intrapulmonar e extrapulmonar induzida por LPS.

The photobiomodulation therapy (PBMT) has many demonstrated applications in the health area including anti-inflammatory and wound healing effects. The main objective of this work is to verify if the PBMT causes measurable changes in the mechanical properties of cells, specifically in red blood cells, epithelial cells and fibroblasts. In addition, to contribute to the knowledge of the action mechanisms of the PBMT, this study intends to support applications of the PBMT during invasive procedures, such as the direct photo-treatment of the blood in surgical procedures with cardiopulmonary bypass, regarding security of the cellular integrity. For this analysis, three experimental techniques were used: optical magnetic twisting cytometry (OMTC), defocusing microscopy and confocal laser-scanning microscopy. Human bronchial epithelial cells were evaluated with OMTC. The epithelial cell culture was either photo-treated or not, with red laser (lambda=660 nm), and fixed power and time (power density of 153 mW/cm2, time 300 s). It was not possible to observe significant differences between photo-treated and control epithelial cells, for the hysteresivity (ratio between the cell loss and elastic shear moduli). The defocusing microscopy, similar to a phase contrast microscopy, was used to study human red blood cells from fresh blood. The red blood cells were either photo-treated or not, with red laser (lambda=660 nm), and different powers and times (power densities from 0 to 510 mW/cm2, times from 0 to 180 s). Some morphological and mechanical characteristics of individual red blood cells were evaluated, such as volume, radial profile of cell thickness, lateral and vertical membrane fluctuations, for the photo-treated and control red blood cells. It was not possible to detect differences between the two groups, for any of the parameters analyzed. For both techniques, the absence of detectable differences might be due to several factors, such as the non-action of the PBMT, with the parameters used, in the epithelial cells and red blood cells or to the small sensitivity of each technique. Confocal laser-scanning microscopy was used to evaluate the actin filaments of mouse fibroblasts. The fibroblast cell culture was either photo-treated or not, with red (lambda=625 nm) or infrared (lambda=808 nm) light and fixed power and time (power density from 113 to 158 mW/cm2, time 300 s). The nucleus and cell areas increased slightly when comparing photo-treated and control cells. On the other hand, the total actin, total actin density and the number of filaments decreased. These changes were detected for a short time after treatment, however, after 24 h they are not anymore detectable. The total branch length does not seem to suffer any modifications. In summary, with the data acquired with the three techniques, it was found that the PBMT, in the red range, with the parameters used, could not cause noticeable changes in red blood cells and epithelial cells, in vitro. On the other hand, the PBMT in the red and near-infrared range, with the power and times used, cause changes in actin filaments of fibroblasts, in vitro, in particular the decrease of the total actin density. / A terapia por fotobiomodulação tem muitas aplicações na área de Saúde devido a sua ação anti-inflamatória e de reparação tecidual. O objetivo geral desse trabalho é verificar se a terapia por fotobiomodulação provoca mudanças nas propriedades mecânicas de células, em particular em hemácias, células epiteliais e fibroblastos. Além de contribuir com o conhecimento dos mecanismos de ação da terapia por fotobiomodulação, este estudo pretende subsidiar aplicações da terapia por fotobiomodulação durante procedimentos mais invasivos, como a iluminação direta do sangue em procedimentos cirúrgicos com circulação extracorpórea, sob o ponto de vista da segurança quanto à integridade celular. Para essa análise foram utilizadas três técnicas experimentais: citometria óptica magnética de oscilação (OMTC), microscopia de desfocalização e microscopia confocal. Com a técnica de OMTC foram avaliadas células epiteliais brônquicas humanas em cultura, foto-tratadas com laser vermelho (lambda=660 nm), com potência e tempo fixos (densidade de potência de 153 mW/cm2, tempo 300 s). Não foi possível constatar diferenças significativas entre as células epiteliais foto-tratadas e as células controle, para a histerisividade (razão entre os módulos viscoso e elástico das células). Com a técnica de microscopia de desfocalização, semelhante a uma microscopia de contraste de fase, foram estudadas hemácias humanas de sangue recém coletado. As hemácias foram tratadas com laser vermelho (lambda=660 nm), com potências e tempos variados (densidade de potência de 0 a 510 mW/cm2, tempo de 0 a 180 s). Foram avaliadas algumas características morfológicas e mecânicas das hemácias individualmente, como o volume, perfil radial de espessura, flutuações lateral e vertical da membrana, tanto para hemácias foto-tratadas quanto para hemácias controle. Não foi possível detectar diferenças entre as hemácias foto-tratadas e controle para nenhum dos parâmetros avaliados. Para ambas as técnicas, a falta de mudanças observáveis poderia ser devida a diversos fatores, como a não ação da terapia por fotobiomodulação nas células epiteliais e nas hemácias, com os parâmetros aqui empregados, ou à falta de sensibilidade de cada uma das técnicas usadas. A microscopia confocal foi utilizada para avaliar os filamentos de actina de fibroblastos de camundongo em cultura, os quais foram foto-tratados com luz vermelha (lambda=625 nm) ou infravermelha (lambda=808 nm) e potência e tempo fixos (densidade de potência de 113 a 158 mW/cm2, tempo 300 s). Foi possível constatar ligeiro aumento nas áreas nuclear e celular das células foto-tratadas em relação aos fibroblastos controle. Também foi possível verificar a diminuição da quantidade total de actina, densidade de actina e do número de filamentos de actina nos fibroblastos foto-tratados. Essas mudanças são detectadas para tempos curtos após o tratamento, sendo que depois de 24 h elas desaparecem. O tamanho total dos filamentos parece não sofrer alterações. A partir dos dados coletados com as três técnicas, foi possível constatar que a terapia por fotobiomodulação, com os parâmetros utilizados, não consegue provocar mudanças perceptíveis em hemácias e em células epiteliais, in vitro. Porém, causa mudanças nos filamentos de actina de fibroblastos, in vitro, em particular a diminuição da densidade de actina total.

In der vorliegenden Arbeit wurde mit einer Vielzahl an objektiven Messverfahren und subjektiven Einschätzungen, die Wirkungen und Nebenwirkungen von drei unterschiedlichen fraktional ablativen Lasersystemen in der Therapie der gealterten und chronisch lichtgeschädigten Haut einzelner Gesichtsareale, sowie des gesamten Gesichtes dargestellt: CO 2 -Laser; Er:YAG hot Laser; Er:YAG cold Laser. Dabei kamen profilometrische Messungen, Messung von Hautfunktionsparameter (Elastizität, Feuchtigkeit, Sebumgehalt), Bewertungen von Fotografien und Fragebögen zur Erfassungs des Wirkungsprofils einer Lasertherapie zur Anwendung.:Inhaltsverzeichnis 1 EINLEITUNG 1 1.1 EINFÜHRUNG IN DIE THEMATIK 1 1.2 Darstellung der Lasersysteme für die Therapie der gealterten Haut 2 1.3 Darstellung von Verfahren zur Evaluation der Therapieergebnisse 6 1.4 Zielstellung 9 1.5 Fragestellungen 9 2. MATERIAL UND METHODEN 10 2.1 Studiendesign und Studienprotokoll 10 2.2 Probanden 11 2.3 Randomisierung 11 2.4 Versicherung und Ethik 11 2.5 Ablauf der Behandlung 12 2.6 Angewandte Lasersysteme 13 2.7 Messgeräte zur Erfassung von Hautveränderungen 15 2.8 Patientenfragebögen 23 2.9 Digitale Fotografien 24 2.10 STATISTIK 24 3 ERGEBNISSE 26 3.1 Hautveränderungen nach 3 Behandlungen gemessen mit PRIMOS 28 3.2 Veränderung der Hautfunktionsparameter nach 3 Behandlungen gemessen mit MPA 5 37 3.3 Auswertung der digitalen Fotografien: Blinded Evaluation 43 3.4 Auswertung der Patientenfragebögen 47 4 DISKUSSION 71 4.1 Evaluation der mit PRIMOS gewonnenen Messergebnisse 71 4.2 Evaluation der mit MPA 5 gewonnenen Messergebnisse 74 4.3 Evaluation subjektiver Behandlungseindrücke mittels Fragebögen 76 4.4 Vergleich der Ergebnisse der digitalen Fotografien mit ausgewählten objektiven Parametern und subjektiven Probandenbeurteilungen 81 4.5 Beantwortung der Fragestellungen 84 4.6 Schlussfolgerungen 85 5 ZUSAMMENFASSUNG DER ARBEIT 86 6 LITERATURVERZEICHNIS 90 7 ANHANG 97

A thesis submitted to the College of Dental Medicine of Nova Southeastern University of the degree of Master of Science in Dentistry. Introduction: One of the challenges in orthodontics is obtaining desired tooth movement. A recent development in orthodontics offers patients more comfort by minimizing pain, and shortening the treatment time. Low level laser therapy (LLLT) emerged as a technology that may accelerate the velocity of tooth movement and shorten the orthodontic treatment period. LLLT (630-1000nm) has been shown to modulate various biological processes including wound healing and bone remodeling. Bone remodeling is one of the biological processes that ensue during tooth movement. Bone remodeling is a continuous process characterized by bone deposition at sites of tension and bone resorption on the pressure sites. At cellular level bone remodeling is regulated by receptor activator of NF-κB (RANK) and receptor activator of NF-κB ligand (RANKL) and Osteoprotegrin (OPG). RANK and RANKL promotes osteoclastic differentiation and promotes bone resorption. OPG is a soluble decoy receptor that competes with RANK for binding to RANKL and inhibits the osteoclastic activity. The goal of the research is to understand the regulatory effects of LLLT on bone metabolism at the cellular level. Furthermore, the purpose of this study is to evaluate the critical parameters of low level lasers on the early stage of ostegenic regulation of human osteoblast cells. In this study, CRL-1427 cells derived from human osteosarcoma which have an osteoblast phenotype were used as cell model. Methods: Human osteoblast cells CRL1427(ATCC, Manassas, VA) were cultured in minimum essential medium supplemented with 10% fetal bovine serum and 1% antibiotics and incubated in at 37°C with 5% CO2. The monolayer of cells after reaching 70-80% confluency were irradiated with a single dose of Galium Aluminum Arsenide (Ga-Al-As) laser, with a wave length of 830nm and output power of 85mW with 0.6, 1.5, 1.8 Joules/cm2 energy exposure regimes. The mRNA expressions of Alkaline phosphatase (ALP), Osteoprotegerin (OPG), RANKL and RANK were compared after seven days by quantitative real time PCR. Results: We observed that treatment of CRL 1427 cells with LLLT (1.5, 1.8 Joules/cm2) irradiation significantly increased the expression of ALP, OPG, RANKL and RANK mRNAs compared to the control group (P≤0.05). There was no significant difference between the control group and mRNA expression of ALP, OPG, RANK, or RANKL at 0.6J/cm2of irradiation. Conclusion: LLLT irradiation can directly influence the expression of genes associated with bone metabolism and potentially represents a mechanism that facilitates rapid bone remodeling.

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Laser therapies can provide a minimally invasive treatment alternative to surgical resection of tumors. However, therapy effectiveness is limited due to nonspecific heating of target tissue, leading to healthy tissue injury and extended treatment durations. These therapies can be further compromised due to heat shock protein (HSP) induction in tumor regions where non-lethal temperature elevation occurs, thereby imparting enhanced tumor cell viability and resistance to subsequent therapy treatments. Introducing nanoparticles (NPs), such as multi-walled nanotubes (MWNTs) or carbon nanohorns (CNHs), into target tissue prior to laser irradiation increases heating selectivity permitting more precise thermal energy delivery to the tumor region and enhances thermal deposition thereby increasing tumor injury and reducing HSP expression induction. This research investigates the impact of MWNTs and CNHs in untreated and laser-irradiated monolayer cell culture, tissue phantoms, and/or tumor tissue from both thermal and biological standpoints. Cell viability remained high for all unheated NP-containing samples, demonstrating the non-toxic nature of both the nanoparticle and the alginate phantom. Up-regulation of HSP27, 70 and 90 was witnessed in samples that achieved sub-lethal temperature elevations. Tuning of laser parameters permitted dramatic temperature elevations, decreased cell viability, and limited HSP induction in NP-containing samples compared to those lacking NPs. Preliminary work showed MWNT internalization by cells, which presents imaging and multi-modal therapy options for NT use. The lethal combination of NPs and laser light and NP internalization reveals these particles as being viable options for enhancing the thermal deposition and specificity of hyperthermia treatments to eliminate cancer. / Master of Science

Introdução: A cicatrização e a manutenção da integridade tissular da úlcera venosa (UVe) consiste em uma cascata de eventos celulares e moleculares que interagem entre si para que ocorra a regeneração tecidual. O seu tratamento depende da associação de diferentes mecanismos por meio da aplicação de produtos tópicos e do uso de terapia compressiva. Apesar dos avanços tecnológicos relativos à oferta e ao desenvolvimento de coberturas tecnológicas ainda não há impacto efetivo na incidência dessas lesões, o que repercute em elevados custos financeiros e em prejuízo aos pacientes. Atualmente, a Terapia a Laser de Baixa Potência (TLBP) vem sendo utilizada para acelerar o processo cicatricial em diferentes lesões graças à sua ação bioestimulatória e anti-inflamatória, o que possibilita o seu uso em UVe. Para direcionar a escolha do tratamento mais adequado, entretanto, é fundamental uma avaliação acurada da lesão. Diante disso, a Nursing Outcomes Classification (NOC) possui os resultados de enfermagem, que permitem avaliar o efeito do tratamento por meio de indicadores clínicos. Objetivo: Comparar o efeito do tratamento adjuvante de Terapia a Laser de Baixa Potência (TLBP) com o tratamento convencional na reparação tecidual de úlcera venosa em pacientes ambulatoriais. Método: Ensaio clínico randomizado realizado em um hospital universitário brasileiro. A amostra consistiu de 40 pacientes com UVe, randomizados em número igual para o Grupo Controle (GC) e Grupo Intervenção (GI), acompanhados no segmento de dezesseis semanas ou até a cicatrização da lesão. O GC recebeu tratamento convencional com curativos tópicos e terapia compressiva, enquanto que ao GI foi adicionado a TLBP como adjuvante. Foi utilizada uma dose de energia de 1 a 3 J/cm2 com um laser AsGalaser, com comprimento de onda de 660 nm. A aplicação ocorreu de forma pontual nas bordas e método de varredura no leito da lesão. O processo de cicatrização foi avaliado, em ambos os grupos, por oito indicadores clínicos do resultado Cicatrização de feridas: segunda intenção (1103): Tamanho da ferida diminuído, Formação de cicatriz, Granulação, Exsudato, Odor desagradável da ferida, Pele macerada, Eritema na pele ao redor da lesão e Edema perilesão. A avaliação também ocorreu a partir de seis indicadores do resultado Integridade tissular: pele e mucosas (1101): Espessura, Hidratação/Descamação, Pigmentação anormal, Prurido, Dor e Necrose. A análise levou em consideração a escala Likert de 5 pontos, em que 1 corresponde ao pior escore e 5 ao mais desejável, utilizando o Generalized Estimating Equation. O estudo foi aprovado em Comitê de Ética (15-0634). Resultados: A média de idade foi de 64,55±11,69 anos para o GC e 63±12,25 para o GI. No GC, 17(85%) pacientes eram do sexo masculino, enquanto que no GI prevaleceu o sexo feminino com 11(55%) pacientes, gerando a única diferença estatística significativa (p=0,019) entre os grupos. A cor branca (16-80%) e a escolaridade no nível de analfabeto funcional (15-75% no GC e 12- 63,2% no GI) prevaleceram nos grupos. Em relação ao tempo de ferida, 40% dos pacientes de ambos os grupos a possuem de um a cinco anos. Foram avaliadas 82 feridas, sendo 39 do GC e 43 do GI, resultando em 1066 consultas, das quais 551 ocorreram no GC e 515 no GI. Quanto ao resultado Cicatrização de feridas: segunda intenção (1103), o GC iniciou com um escore médio de 2,87±0,35 e, ao final do estudo, apresentou 4,21±0,60, enquanto que o GI iniciou com 2,67±0,46 e alcançou 4,46±0,47 (p=0,025). Houve diferença estatisticamente significativa na comparação entre os grupos em quatro dos oito indicadores: Granulação 9 (p=0,010), Tamanho reduzido (p=0,010), Formação de cicatriz (p=0,034) e Exsudato (p=0,011). Em relação ao Resultado Integridade tissular: pele e mucosas (1101) o GC iniciou com um escore médio de 3,74±0,38 e atingiu 4,27±0,46 na última consulta, enquanto que o GI iniciou com 3,43±0,42 e alcançou 4,29±0,44. Na análise dos seis indicadores do resultado NOC observou-se diferença estatisticamente significativa em três deles: Pigmentação anormal (p=0,008), Espessura (p=0,010) e Hidratação/Descamação (p=0,015) na comparação entre os grupos. Quanto ao tratamento convencional, agrupados em classe de ação, no GC prevaleceu o desbridamento enzimático (17,8%), curativos para o controle da infecção (52,8%) e agentes tópicos para manutenção do meio úmido (52,7%). Já no GI despontaram os curativos para o controle do exsudato (9,0%), agentes tópicos para proteção dos bordos (31,3%) e para pele perilesional (104,2%). Na TLBP foi utilizado o laser vermelho, com o modo de irradiação varredura predominantemente no leito da úlcera (97,7%), com um tempo médio de um minuto, na potência de 30mW. O modo de irradiação pontual foi usado nos bordos da lesão, com maior predomínio (61,8%) de 10 pontos, a uma distância de um cm cada, com energia de 1J/cm2. Esse modo também foi usado na área perilesional, porém, com uma energia de 21J/cm2. Conclusão: A TLBP é um tratamento adjuvante eficaz para a regeneração tecidual de UVe, confirmado pelos indicadores clínicos dos resultados da NOC. / Introduction: The healing and maintenance of tissue integrity of the venous ulcer (VU) consists of a cascade of cellular and molecular events that interact with each other for tissue regeneration to occur. Its treatment depends on the association of different mechanisms through the application of topical products and the use of compressive therapy. Despite technological advances related to the supply and development of technological coverage, there is still no effective impact on the incidence of these injuries, which has high financial costs and damages to patients. Currently, Low-level Laser Therapy (LLLT) has been used to accelerate the cicatricial process in different lesions thanks to its biostimulatory and anti-inflammatory action, which allows its use in VU. In order to guide the choice of the most appropriate treatment, however, an accurate evaluation of the lesion is essential. Nursing Outcomes Classification (NOC) has the results which allow the evaluation of the effect of treatment through clinical indicators. Objective: To compare the effect of adjunctive treatment of LLLT with the conventional treatment in tissue repair of venous ulcer in patients at the Outpatient Nursing clinic. Method: Randomized clinical trial performed at a Brazilian university hospital. The sample consisted of 40 patients with VU, equally randomized to the Control Group (CG) and Intervention Group (GI). Patients were followed up at a weekly nursing visit for 16 weeks or until the lesion was healed. GC received conventional treatment with topical dressings and compressive therapy, while GI was added to TLBP as an adjuvant. An energy dose of 1 to 3 J / cm 2 was used with an AsGalaser laser, with a wavelength of 660 nm. The application occurred punctually at the edges and scanning method in the lesion bed. The wound healing process was evaluated in both groups by eight clinical indicators of Wound healing: second intention (1103): Decreased wound size, Scar formation, granulation, Exudate, Fow wound odor, Macerated skin, Surrounding skin erythema and Periwound edema. The evaluation also occurred from six outcome indicators Tissue integrity: skin and mucous membranes (1101): Thickness, Hydration / Skin flaking, Abnormal Pigmentation, Pruritus, Pain and Necrosis. The analysis took into account the 5-point Likert scale, in which 1 corresponds to the worst score and 5 to the most desirable, using the Generalized Estimating Equation. The study was approved by the Ethics Committee (15-0634). Results: The mean age was 64.55 ± 11.69 years for GC and 63 ± 12.25 for GI. In the CG, 17 (85%) patients were male, whereas in GI female patients prevailed with 11 (55%) patients, generating the only statistically significant difference (p = 0.019) between the groups. The white color (16-80%) and schooling at the level of functional illiterate (15-75% in CG and 12- 63,2% in IG) prevailed in the groups. Regarding wound time, 40% of patients in both groups have one to five years. Eighty-two wounds were evaluated, 39 of GC and 43 of GI, resulting in 1066 consultations, of which 551 occurred in GC and 515 in GI. Regarding wound healing: second intention (1103), GC started with a mean score of 2.87 ± 0.35 and, at the end of the study, presented 4.21 ± 0.60, while GI started with 2.67 ± 0.46 and reached 4.46 ± 0.47 (p = 0.025). There was a statistically significant difference in the comparison between the groups in four of the eight indicators: Granulation (p = 0.010), Decreased wound size (p = 0.010), Scar formation (p = 0.034) and Exudate (p = 0.011). Regarding the Outcome Tissue integrity: skin and mucous membranes (1101), GC started with a mean score of 3.74 ± 0.38 and reached 4.27 ± 0.46 in the last visit, while the GI started with 3.43 ± 0.42 and reached 4.29 ± 0.44. In 11 the analysis of the six indicators of the NOC result, a statistically significant difference was observed in three of them: Abnormal pigmentation (p = 0.008), Thickness (p = 0.010) and Hydration / Skin flaking (p = 0.015) in the comparison between groups. Regarding the conventional treatment, grouped in action class, the CG prevailed enzymatic debridement (17.8%), dressings for infection control (52.8%) and topical agents for maintenance of the humid environment (52.7%). In the GI, dressings for exudate control (9.0%), topical agents for edge protection (31.3%) and perlesional skin (104.2%) emerged. In LLLT the red laser was used, with the mode of irradiation sweeping predominantly in the ulcer bed (97.7%), with an average time of one minute, in the power of 30mW. The mode of point irradiation was used at the edges of the lesion, with a predominance (61.8%) of 10 points, at a distance of one cm each, with energy of 1J/cm2. This mode was also used in the perilesional area, however, with an energy of 21J / cm2. Conclusion: LLLT is an effective adjuvant treatment for tissue regeneration of VU and confirms the clinical indicators of NOC outcome.

Thesis (MSc (Physiological Sciences)--University of Stellenbosch, 2009. / ENGLISH ABSTRACT: Although muscle tissue demonstrates a remarkable capacity for regeneration following injury, this process is slow and often accompanied by the formation of scar tissue and a subsequent decrease in contractile capacity following regeneration. Treatment options are few and mostly supportive in nature. This regeneration process involves muscle stem cells (satellite cells) which ultimately give rise to the regenerated muscle. The contentious field of low level laser therapy (LLLT) has made remarkable claims in facilitating wound healing in soft tissue injuries of various types. Yet, the mechanism(s) invoked in these beneficial effects are poorly understood. We have investigated the effects of LLLT using a 638 nm laser on satellite cells in culture and in-vivo. Using an array of techniques we have measured, amongst other things, metabolic responses to laser irradiation, signaling pathways activated/altered and antioxidant status. In response to laser irradiation satellite cells in culture showed an increase in MTT values (a measure of metabolic activity) and a decrease in antioxidant status (measured using the ORAC assay). In addition laser irradiation also altered the expression and phosphorylation state of several signaling pathways, including Akt and STAT-3. Following on from this the effects of laser irradiation on satellite cells in-vivo was assessed in a rat model of contusion injury. No significant differences in satellite cell number was found following laser irradiation, changes were seen in tissue antioxidant status and blood antioxidant status (measured using the ORAC assay). In the course of this study several standard techniques were used to investigate the effects of laser irradiation on satellite cells both in-vitro and invivo. It has become apparent that several of these techniques have problems associated with them that possibly make them inappropriate for vi further use in studies involving laser irradiation. However the results indicate that laser therapy is induces satellite cell behavior and further study is warranted in this field. / AFRIKAANSE OPSOMMING: Alhoewel spierweefsel merkwaardige regenerasie kapasiteit vertoon ten opsigte van besering, is hierdie proses stadig en word soms vergesel met die vorming van letselweefsel asook ‘n gevolglike afname in kontaktiele kapasiteit na afloop van regenerasie. Behandelingsmoontlikhede is skaars en meesal ondersteunend van aard. Hierdie proses sluit spierstamselle (satelietselle), wat uiteindelik die ontstaan van die regenerasie van spier tot gevolg het, in. Die kontroversiële veld van lae vlak laserterapie (Engels: Low level laser therapy (LLLT)) het merkwaardige aansprake in die fasilitering met verskeie sagteweefsel wondgenesing. Nietemin, die meganisme(s) wat voordelige effekte induseer, word nog nie goed begryp nie. Ons het die effek van LLLT, deur gebruik te maak van ‘n 638 nm laser op kultuur in vitro satelietselle sowel in-vivo, ondersoek. Deur gebruik te maak van verskeie tegnieke is onder meer die metaboliese, sowel die seinstransduksie weë en antioksidantstatus na laserbestraling, gemeet. In reaksie op die laserbestraling het satelietselle (in kultuur) ‘n toename in MTT waardes getoon (‘n maatstaf van die metaboliese aktiwiteit) en ‘n afname in die antioksidantstatus (gemeet deur van die ORAC toets). Addisioneel het laserbestraling ook uitdrukking en fosforilering van verskeie proteïene betrokke in seintransduksieweë beïnvloed, insluitend Akt, STAT-3). Na afloop van hierdie effekte op satelietselle na laserbestraling, is daar gebruik gemaak van ‘n kneusbeseringsrotmodel om hierdie effekte in vivo te ondersoek. Geen betekenisvolle verskille in die aantal satelietselle na laserbestraling is opgemerk nie, maar veranderings is wel opgemerk in weefsel- en bloed-antioksidantstatus (gemeet deur van die ORAC toets gebruik te maak). Gedurende die verloop van die studie is van verskeie standaardtegnieke gebruik gemaak om die effekte van laserbestraling op beide satelietselle in vitro en in vivo te ondersoek. iv Dit het duidelik na vore gekom dat daar wel gepaardgaande probleme met van hierdie tegnieke voorgekom het, en dat van hierdie tegnieke nie gepas is vir ondersoek in laserbestralingsstudies nie. Nietemin, die resultate toon wel dat laserbehandeling. satelietselgedrag induseer wat verdere studie in hierdie veld noodsaak