Thesis (M.D.S. 1973) from the Department of Dental Health, University of Adelaide.

No description available.

A German pathologist first described defects in the elastic tissues of human arteries over one hundred years ago. Much evidence now supports the involvement of these elastic tissue defects (ETDs) in the initiation and progression of atherosclerosis, although this association is not well accepted. Recent research has determined that the migration of medial smooth muscle cells into the intima (and therefore the start of the atherosclerotic process) is initiated in an attempt to repair these defects and in addition, that there is a correlation between the extent of intimal thickening and the degree of elastic tissue disruption. The Brown Norway (BN) strain appears to have an increased predilection, having a significantly greater incidence of ETDs within the caudal and renal arteries and the abdominal aorta compared with other rat strains. These defects appear morphologically identical to those observed in the arteries of young humans. The purpose of this study was to determine the magnitude of the genetic and environmental components in the formation of these ETDs in the aorta. Previous studies have demonstrated that the spontaneous formation of elastic tissue defects in the abdominal aorta of the Brown Noway rat is a genetically inherited phenotype, passed from parent to offspring in an autosomal dominant manner. Following crossbreeding of the BN rat with four other strains (two hypertensive and two normotensive) it was determined that, although the inheritance mode of the ETD phenotype followed an autosomal dominant pattern, the expression or penetrance of this phenotype was reduced in F₁ all crossbred groups. Moreover, the early postnatal growth profile of the F₁ pups appeared to be differentially associated with defect formation. To further examine the relationship between aortic ETDs and birthweight, a well-studied model of in utero growth restriction was investigated in the BN rat. On day 18 of a 23-day gestation the uterine arteries were ligated, which resulted in offspring that were 14% smaller than un-operated control pups. This short-term insult resulted in significantly increased numbers of ETDs in growth-restricted animals at 8 weeks of age, an effect that was also observed in 16-week old males. The effect of in utero growth restriction on ETDs in the guinea pig and ApoE knockout mouse was also examined, to determine if ETDs (and subsequent early atherosclerotic events) may be influenced by the exposure to a growth-restricting event in utero. Despite this work leading to the novel characterisation of ETDs in the guinea pig aorta, the growth restricting surgery resulted in poor maternal and pup outcomes, which limited the conclusions that could be drawn from these studies. Furthermore, microarray techniques were employed to examine changes in aortic gene expression following growth restriction, by comparing amplified mRNA extracts from 8-week old growth restricted BN pup aortas with extracts from a group of average birthweight, un-operated BN pups. In combination, these studies propose both genetic inheritance and the in utero environment regulate elastic tissue defect phenotype, which in turn potentially affects the initiation and progression of early atherosclerosis.

The extracellular matrix (ECM) is the material found surrounding the cells in a tissue. One component of the ECM is the elastic fiber, which confers the property of elasticity to its environment. Organs such as the lung, skin and major blood vessels have an abundance of elastic fibers so that they are able to expand and recoil. Elastic fibers are composed of two main components; elastin and microfibrils. Microfibrils are composed primarily of fibrillin-1 and provide a scaffold unto which tropoelastin monomers assemble. Elastic fibers interact with many other proteins in the ECM, one of which is fibulin-5. Based on the severe elastic fiber defects observed in the fibulin-5 null mouse, it was established that fibulin-5 plays an essential role in elastic fiber development. This role may be in the deposition of tropoelastin onto microfibrils and/or in stabilizing the elastic fibers in the extracellular matrix. In the present study, the relationship between fibulin-5 and the elastic fiber was investigated through a number of in vivo and in vitro experiments. To test the hypothesis that fibulin-5 requires the presence of elastin to assemble in the ECM, full-length recombinant fibulin-5 (rF5) was purified from transfected cells and used to make a fibulin-5 antibody. Solid-phase binding assays using rF5 showed that fibulin-5 binds tropoelastin at two sites; the initial portion of the C-terminus and the first calcium-binding epidermal growth factor-like domain at the N-terminus. Immunofluorescence staining of elastin null mouse embryonic fibroblast cultures revealed that fibulin-5 does not require elastin to be present in the ECM in order to assemble. Subsequently, solid-phase binding assays showed that fibulin-5 can bind to the N-terminus of fibrillin-1. To determine if fibulin-5 could exist independent of elastin and/or fibrillin-1 in vivo, an immunohistochemical analysis was conducted on heart, liver, lung, colon, spleen, testis and kidney. All three proteins were co-localized in all organs except in the kidney, where fibrillin-1 was found to independently stain the capillary tufts of the renal corpuscles and renal tubules. Thus, fibulin-5 may be co-regulated with elastin and is not present on elastin-independent microfibrils. Additionally, novel locations of elastic fibers were uncovered in the heart, liver, colon, spleen and testis. Overall, this study provides important insights as to the role of fibulin-5 in elastic fiber structure and assembly and also reveals the complexity in understanding the pathogenesis of diseases involving elastic fiber proteins.

Thesis (Ph. D.)--University of Adelaide, Dept. of Pathology, 1985. / Includes bibliographical references (leaves 266-303).

Bibliography: leaves 266-303 / xviii, 303 leaves : ill ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Pathology, 1985

Introdução: A luz intensa pulsada(LIP) tem sido muito utilizada no tratamento do fotoenvelhecimento sem completo conhecimento de seu mecanismo de ação. Métodos: Foram acompanhados 26 pacientes apresentando fotoenvelhecimento grau II-III (GLOGAU, 1994), os quais foram submetidas à avaliação clínica , histológica e imunoistoquímica 6 e 12 meses após o término do tratamento com LIP. Foram realizadas cinco sessões com intervalos de trinta dias. Além da quantificação histomorfométrica das fibras colágenas e elásticas na derme, foram avaliados CD1, CD4, CD8 e ICAM-1. Resultados: Após 6 meses houve melhora clínica moderada e intensa em 76,92% dos casos e a nota média de satisfação foi 8,57 correspondendo à melhora moderada. Após 12 meses do término do tratamento observou-se que 51,52% das pacientes apresentaram uma melhora clínica moderada em relação à clínica inicial. Os efeitos colaterais foram eritema (11/26), edema (10/26), ardência (7/26) e crostas (8/26). A quantificação das fibras colágenas mostrou aumento médio de 51,33% proporção média de fibra colágena na derme após 6 meses de tratamento e o aumento em relação a 12 meses do término do tratamento foi 30,17%; as fibras elásticas mostraram aumento de 44,13% após 6 meses e aumento de 143,19% após 12 meses do término do tratamento. Na análise imunoistoquímica não houve alteração de CD1 e CD8. Em relação ao CD4 houve redução significante após 12 meses do término do tratamento. Quanto ao ICAM-1 houve aumento em 6 meses com retorno aos níveis normais após 12 meses do término do tratamento. Conclusão: A melhora clínica observada foi comprovada pelo estudo histopatológico, que mostrou aumento das fibras colágenas e elásticas na derme. Após 12 meses do término do tratamento observou-se discreta redução do aspecto clínico da pele correlacionado ao histopatológico. Poucos efeitos colaterais foram observados, sendo todos reversíveis. Desta forma, a LIP constitui boa opção de tratamento para o fotoenvelhecimento cutâneo, sendo uma técnica não ablativa, segura e eficaz. / Introduction: The intense pulsed light has been used in the treatment of photoaging without full knowledge of its mechanism of action. Material and Methods: 26 patients were followed-up presenting photoaging grade II-III (GLOGAU, 1994), who were submitted to clinical evaluation, histological and immunohistochemistry 6 and 12 months after the treatment termination with LIP. Five sessions were made with 30-day intervals. In addition to histomorphometric quantification of collagen and elastic fibers in the dermis, CD1, CD4, CD8 and ICAM-1 were evaluated. Results: After 6 months there were moderate and intense clinical improvement on 76.92% of the cases and the mean score of satisfaction was 8.57 corresponding to moderate improvement. After 12 months of the treatment termination, it was observed that 51.52% of the patients presented a moderate clinical improvement in relation to initial clinic. The side effects were erythema (11/26), edema (10/26), burning (7/26) and crusts (8/26). The quantification of collagen fibers has shown mean increase of 51.33% in the dermis after 6 months of treatment and the increase regarding to 12 months of the treatment termination was of 30.17%; the elastic fibers has shown an increase of 44.13% after 6 months and increase of 143.19% after 12 months of the treatment termination. In the immunohistochemistry analysis there was no alteration of CD1 and CD8. In relation to CD4, there was a significant reduction after 12 months of treatment termination. Regarding the ICAM-1, there was an increase in 6 months with return to normal levels after 12 months of treatment termination. Conclusion: The observed clinical improvement was verified by the histopathologic study, which showed increase in the elastic and collagen fibers in the dermis. After 12 months of treatment termination, it was observed a discrete reduction of clinical aspect of skin correlated to histopathology. A few side effects were observed, being all reversible. Thus, LIP constitutes good option of treatment for skin photoageing, being a non-ablative, safe and effective technique.

Introdução: A luz intensa pulsada(LIP) tem sido muito utilizada no tratamento do fotoenvelhecimento sem completo conhecimento de seu mecanismo de ação. Métodos: Foram acompanhados 26 pacientes apresentando fotoenvelhecimento grau II-III (GLOGAU, 1994), os quais foram submetidas à avaliação clínica , histológica e imunoistoquímica 6 e 12 meses após o término do tratamento com LIP. Foram realizadas cinco sessões com intervalos de trinta dias. Além da quantificação histomorfométrica das fibras colágenas e elásticas na derme, foram avaliados CD1, CD4, CD8 e ICAM-1. Resultados: Após 6 meses houve melhora clínica moderada e intensa em 76,92% dos casos e a nota média de satisfação foi 8,57 correspondendo à melhora moderada. Após 12 meses do término do tratamento observou-se que 51,52% das pacientes apresentaram uma melhora clínica moderada em relação à clínica inicial. Os efeitos colaterais foram eritema (11/26), edema (10/26), ardência (7/26) e crostas (8/26). A quantificação das fibras colágenas mostrou aumento médio de 51,33% proporção média de fibra colágena na derme após 6 meses de tratamento e o aumento em relação a 12 meses do término do tratamento foi 30,17%; as fibras elásticas mostraram aumento de 44,13% após 6 meses e aumento de 143,19% após 12 meses do término do tratamento. Na análise imunoistoquímica não houve alteração de CD1 e CD8. Em relação ao CD4 houve redução significante após 12 meses do término do tratamento. Quanto ao ICAM-1 houve aumento em 6 meses com retorno aos níveis normais após 12 meses do término do tratamento. Conclusão: A melhora clínica observada foi comprovada pelo estudo histopatológico, que mostrou aumento das fibras colágenas e elásticas na derme. Após 12 meses do término do tratamento observou-se discreta redução do aspecto clínico da pele correlacionado ao histopatológico. Poucos efeitos colaterais foram observados, sendo todos reversíveis. Desta forma, a LIP constitui boa opção de tratamento para o fotoenvelhecimento cutâneo, sendo uma técnica não ablativa, segura e eficaz. / Introduction: The intense pulsed light has been used in the treatment of photoaging without full knowledge of its mechanism of action. Material and Methods: 26 patients were followed-up presenting photoaging grade II-III (GLOGAU, 1994), who were submitted to clinical evaluation, histological and immunohistochemistry 6 and 12 months after the treatment termination with LIP. Five sessions were made with 30-day intervals. In addition to histomorphometric quantification of collagen and elastic fibers in the dermis, CD1, CD4, CD8 and ICAM-1 were evaluated. Results: After 6 months there were moderate and intense clinical improvement on 76.92% of the cases and the mean score of satisfaction was 8.57 corresponding to moderate improvement. After 12 months of the treatment termination, it was observed that 51.52% of the patients presented a moderate clinical improvement in relation to initial clinic. The side effects were erythema (11/26), edema (10/26), burning (7/26) and crusts (8/26). The quantification of collagen fibers has shown mean increase of 51.33% in the dermis after 6 months of treatment and the increase regarding to 12 months of the treatment termination was of 30.17%; the elastic fibers has shown an increase of 44.13% after 6 months and increase of 143.19% after 12 months of the treatment termination. In the immunohistochemistry analysis there was no alteration of CD1 and CD8. In relation to CD4, there was a significant reduction after 12 months of treatment termination. Regarding the ICAM-1, there was an increase in 6 months with return to normal levels after 12 months of treatment termination. Conclusion: The observed clinical improvement was verified by the histopathologic study, which showed increase in the elastic and collagen fibers in the dermis. After 12 months of treatment termination, it was observed a discrete reduction of clinical aspect of skin correlated to histopathology. A few side effects were observed, being all reversible. Thus, LIP constitutes good option of treatment for skin photoageing, being a non-ablative, safe and effective technique.

A fisiopatologia do edema de Reinke ainda permanece desconhecida e poucos estudos abordam sobre as alterações das proteínas fibrosas, colágeno e elastina, na matriz extracelular da prega vocal com edema de Reinke. Por isso, este estudo foi idealizado para descrever a distribuição das fibras colágenas e do sistema de fibras elásticas do espaço de Reinke com edema de Reinke comparando com a prega vocal normal e com a severidade do edema. Foram obtidas 20 amostras de pregas vocais de indivíduos com edema de Reinke, sendo nove casos com edema de Reinke grau II e 11 casos grau III. Dezessete indivíduos eram do sexo feminino e três do sexo masculino, com idade variando de 47 a 62 anos (55 ± 4,4 anos). Quinze pacientes eram tabagistas e cinco ex-tabagistas, média e desvio padrão de 22 ± 10,7 maço/ano. O tempo de disfonia apresentou média e desvio padrão de 36 ± 16,6 meses. Dez pregas vocais normais de laringes humanas excisadas foram utilizadas como controles. Os métodos da Picrossírius-polarização e da Resorcina-fucsina de Weigert com oxidação prévia pelo oxone 10% foram empregados para visualização das fibras colágenas e do sistema de fibras elásticas, respectivamente. Uma avaliação semiquantitativa foi utilizada para categorizar os resultados histológicos que foram correlacionados com a idade, consumo de cigarro, tempo de disfonia e com a severidade do edema. Evidenciou-se que o arranjo entrelaçado das fibras colágenas semelhante a uma cesta de vime observado em pregas vocais normais estava desestruturado no edema de Reinke. A desestruturação foi caracterizada por um afastamento das fibras, fragmentação, formando áreas esparsas e tomadas por um estroma mixóide de quantidade variável. Todos os casos de edema de Reinke mostraram uma maior preservação do arranjo das fibras colágenas próximo ao epitélio da prega comparada com as fibras da camada mais profunda do espaço de Reinke. O arranjo do sistema de fibras elásticas formado por fibras finas e onduladas paralelas à membrana basal do epitélio e uma rede de fibras mais finas, logo abaixo da membrana basal do epitélio observado em pregas vocais normais estava desestruturado no edema de Reinke. Na avaliação semiquantitativa observou-se que moderada e intensa desestruturação ocorreram em 90% dos casos com relação às fibras colágenas e em todos os casos para o sistema de fibras elásticas. Houve uma correlação estatisticamente significante entre a idade e o grau de desestruturação das fibras colágenas (r=0,47, p=0,037). Houve diferenças estatisticamente significantes entre os edemas grau II e III quanto à desestruturação das fibras colágenas (p=0,007) e quanto à idade (p=0,036). Observamos com este estudo que as alterações nas proteínas fibrosas presentes no edema de Reinke podem contribuir com a deformidade da prega vocal / The physiopathological mechanisms underlying Reinkes edema are still unknown and few studies addressed alterations in the fibrillar proteins, collagen and elastin, in extracellular matrix with Reinkes edema. This study was idealized to describe the distribution of collagen fibers and elastic system fibers in Reinkes space with Reinkes edema, comparing with normal vocal fold and with the severity of Reinkes edema. Twenty surgical vocal fold specimens were obtained from patients with Reinkes edema, nine cases presented grade II severity and 11 cases grade III. Seventeen subjects were females and three were males, ranging in age from 47 to 62 years (mean±SD 55 ± 4.4 years). Fifteen patients were smokers and five ex-smokers, mean±SD of 22 ± 10.7 pack-years. The duration of dysphonia ranged from 6 to 60 months (mean of 36 ± 16.6 months). Ten vocal folds from human larynges of nonsmokers cadavers were used as normal controls. The Picrosirius polarization method and the Weigerts resorcin-fuchsin stain after oxidation with 10% aqueous oxone were used for visualization of collagen fibers and the elastic system fibers, respectively. Findings were categorized semiquantitatively and correlated with age, cigarette smoking, duration of dysphonia and Reinkes edema severity. The intertwined network of collagen fibers resembling a wicker-basket found in normal vocal folds was disarranged in Reinkes edema. Disarrangement of collagen fibers was characterized by loosely arranged and fragmented fibers intermixed with varying amounts of myxoid stroma. All cases showed a better preservation of collagen fibers arrangement closer to the epithelium compared to fibers of the deeper of the Reinkes space. The elastic system fibers arrangement formed by a delicate network of thin and undulated fibers arranged in parallel to the epithelial basement membrane and a network of thinner fibers immediately below the basement membrane was disarranged. The semiquantitatively analysis showed that moderate and large areas of disarrangement of collagen fibers were found in 90% of cases and in all cases for elastic fibers. Age was significantly correlated with collagen fiber disarrangement (r=0.47, p=0.037). There was a statistical difference in collagen disarrangement (p = 0,007) and age (p=0,036) between grade II and grade III severity. In our study, the alterations in the fibrillar proteins observed in Reinkes edema may contribute to the vocal fold deformity

INTRODUÇÃO: A cicatriz em prega vocal que ocorre após injúria constitui causa importante de disfonia. A fonocirurgia ideal é aquela em que a lesão efetivamente removida com o menor dano possível ao tecido adjacente, a fim de reestabelecer a função da prega vocal ao mesmo tempo em que a sua ultraestrutura sua ultraestrutura é preservada. Tanto o bisturi frio quanto lasers em geral são aceitos como instrumentos para fonocirurgia. O laser de diodo tem sido usado no tratamento de doenças laríngeas. Entretanto, não há consenso quanto os parâmetros ideais desse aparelho para tal finalidade. Inexistem trabalhos que mostrem a forma ou a extensão da lesão provocada com laser de diodo em laringe e tampouco as reações histológicas e cicatriciais provocadas por ele. OBJETIVO: O objetivo desta pesquisa é estudar as alterações morfométricas e histopatológicas observadas nas pregas vocais de coelhos, sete dias após a lesão provocada com o laser de diodo, comparando diferentes configurações do dispositivo. MÉTODO: Vinte e um coelhos machos albinos da raça New Zealand foram distribuídos aleatoriamente em três grupos com sete animais por grupo. Foi realizada uma lesão única durante 20 segundos em cada prega vocal com a ponta da fibra em contato superficial com o tecido. Duas frequências de pulso foram comparadas no Grupo I (10Hz versus 1000Hz), diferentes potências no Grupo II (3W versus 5W) e modos de radiação distintos no Grupo III (pulsado versus contínuo). Após sete dias, as laringes foram excisadas e submetidas à coloração com hematoxilina-eosina, além de coloração histoquímica para colágeno e elastina. Foi realizada análise histológica quantitativa e subjetiva. RESULTADOS: o laser de diodo provocou: exocitose de células inflamatórias; edema de mucosa e submucosa; infiltrado celular extenso em torno da úlcera, composto por polimorfonucleares (especialmente eosinófilos), linfócitos e histiócitos; tecido de granulação com a presença de fibroblastos e vasos neoformados e áreas de necrose do tipo coagulativa. A extensão do processo inflamatório e a extensão linear da úlcera apresentaram diferença significativa entre as duas potências, e foi maior com uso do laser ajustado para 5W. A extensão do processo inflamatório, extensão linear da úlcera, profundidade da úlcera e profundidade do processo inflamatório apresentaram diferença significativa entre os dois modos de emissão do laser, isto é, maior no modo contínuo. A densidade das fibras colágenas apresentaram-se elevadas apenas ao uso do laser no modo contínuo, quando comparado ao modo pulsado. Não houve diferença estatística quanto à densidade de fibras elásticas. CONCLUSÃO: O uso do laser de diodo ajustado em potência de 5W ao invés 3W e o uso do modo contínuo ao invés do pulsado são capazes de aumentar significativamente a injúria térmica nas pregas vocais de coelhos / INTRODUCTION. Scarring of the vocal folds is a relevant cause of dysphonia after injury. The ideal phonomicrosurgery would be the one that removes the vocal fold disease in order to restore the biomechanical function, while providing minimal disruption to the surrounding vocal fold layered structure. Steel scalpel and laser systems are widely accepted tools for vocal fold surgical procedures. The diode laser technique has been used in the treatment of laryngeal diseases. However, there is great variability among surgeons with regard to the use of the diode laser for laryngeal surgery and the ideal parameters for this procedure remain unclear. No description of the lesion extent or the vocal fold healing after injury with this device have been reported to date. OBJECTIVE: The aim of this research was to study the morphometric and histopathological changes seen in the vocal fold seven days after injury with the diode laser in a rabbit model, comparing different settings of the device. METHODS: Twenty-one male New Zealand white rabbits were randomized into three groups with seven animals per group. A single spot injury during 20 seconds was performed in each vocal fold with the fiber tip in superficially contact with the tissue. Two pulse frequency were compared in group I (10Hz versus 1000Hz), different powers in group II (3W versus 5W) and distinct wave mode of radiation in group III (pulsed versus continuous). After seven days, the larynges were harvested and subjected to H&E staining, histochemical staining for collagen and elastin, with quantitative and subjective histological analysis. RESULTS: the diode laser provoked exocytosis of inflammatory cells; edema of mucosa and submucosa; extensive cell infiltrate around the ulcer with polymorphonuclear cells (especially eosinophils), lymphocytes and histiocytes; granulation tissue with the presence of fibroblasts and neoformed vessels and areas of coagulative necrosis. The extent of the inflammatory infiltrate and the extent of the ablation crater showed to be greater with the 5W power use. The analyzes of extension of the extent of the inflammatory infiltrate, the extent of the ablation crater, the depth of the ablation crater and depth of the inflammatory process presented greater measurements with the continuous mode. The density of collagen fibers was higher when the laser was used in continuous wave mode. There was no statistically significant difference in elastic fiber density. CONCLUSION: Increasing power from 3W to 5W and using continuous wave rather than pulsed wave mode of the diode laser significantly increased the extent of thermal injury in the rabbit vocal folds