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Ossos temporais humanos constituem valiosas fontes de estudo das doenças relacionadas à orelha. Apesar dos vários métodos existentes para obtenção de amostras de ossos temporais de cadáveres, geralmente estes métodos não são descritos em detalhes nos artigos envolvendo pesquisa. OBJETIVOS: Descrever um método prático e rápido para obtenção de amostras de meato acústico externo e membrana timpânica, e verificar sua viabilidade para estudo histológico e imunohistoquímico. MATERIAL E MÉTODO: Para este estudo experimental, foram obtidas 31 amostras de meato acústico externo e membrana timpânica de cadáveres com a utilização de furadeira convencional e broca do tipo serra copo. O material foi dissecado para obtenção de amostras em bloco contendo o meato acústico externo e a membrana timpânica. Estas amostras foram analisadas por histologia e imunohistoquímica. RESULTADOS: Foi possível a remoção de amostras de pequeno volume e de boa qualidade, o que facilitou a obtenção de amostras contendo todo o meato acústico externo e membrana timpânica. Em todas as amostras, foi possível realização de estudo histológico e imunohistoquímico. CONCLUSÃO: O método aqui descrito foi facilmente exequível, reprodutível e produziu amostras de boa qualidade, seja para treinamento ou pesquisa experimental. Todas as amostras foram viáveis para a realização de estudo histológico e imunohistoquímico.Temporal bones are valuable resources to study ear diseases. Although there are several methods for removing temporal bones from cadavers, such methods are not usually described in enough details in experimental research papers. OBJECTIVES: To describe a simple and rapid method for ear canal and tympanic membrane removal, and to evaluate its viability for histologic and immunohistochemical studies. MATERIALS AND METHODS: In this experimental study, we obtained 31 ear canal and tympanic membrane samples from cadavers, with a conventional power drill and plug cutter. The material was dissected and samples containing ear canals and tympanic membranes were obtained in blocks. The samples were analyzed by histology and immunohistochemistry. RESULTS: Removal of small and good quality samples containing entire ear canals and tympanic membranes. In all the samples, it was possible to perform both histological and immunohistochemical analyses. CONCLUSION: This method was easily achievable, reproducible and yielded good quality samples, both for training purposes and for experimental research. All the samples were viable for histological and immunohistochemical analyses.

Ossos temporais humanos constituem valiosas fontes de estudo das doencas relacionadas a orelha. Apesar dos varios metodos existentes para obtencao de amostras de ossos temporais de cadaveres, geralmente estes metodos nao sao descritos em detalhes nos artigos envolvendo pesquisa. OBJETIVOS: Descrever um metodo pratico e rapido para obtencao de amostras de meato acustico externo e membrana timpânica, e verificar sua viabilidade para estudo histologico e imunohistoquimico. MATERIAL E METODO: Para este estudo experimental, foram obtidas 31 amostras de meato acustico externo e membrana timpânica de cadaveres com a utilizacao de furadeira convencional e broca do tipo serra copo. O material foi dissecado para obtencao de amostras em bloco contendo o meato acustico externo e a membrana timpânica. Estas amostras foram analisadas por histologia e imunohistoquimica. RESULTADOS: Foi possivel a remocao de amostras de pequeno volume e de boa qualidade, o que facilitou a obtencao de amostras contendo todo o meato acustico externo e membrana timpânica. Em todas as amostras, foi possivel realizacao de estudo histologico e imunohistoquimico. CONCLUSAO: O metodo aqui descrito foi facilmente exequivel, reprodutivel e produziu amostras de boa qualidade, seja para treinamento ou pesquisa experimental. Todas as amostras foram viaveis para a realizacao de estudo histologico e imunohistoquimico.

Acquired middle ear cholesteatoma is a disease which promotes bone erosion resulting in potentially serious complications. The tumor necrosis factor alpha (TNF-α) is present in cholesteatoma and it is related to bone erosion, as shown by different authors. To understand the aggressiveness characteristics of cholesteatoma is necessary, however, to better address the presence and distribution of their receptors. OBJECTIVE: To evaluate the expression of type 2 TNF-α receptor (TNF-R2) in fragments of cholesteatoma and correlate it to the degree of inflammation present. MATERIAL AND METHODS: observational cross-sectional study, which analyzed 33 fragments of cholesteatomas through histological analysis and immunohistochemistry (using as primary antibody to TNF-R2 LabVision® brand). The evaluation was performed by means of a qualitative and semi-quantitative agreement with the observed intensity. For statistical analysis we used the Fisher exact test and Spearman´s correlation coefficient (considered statistically significant when p < 0. 05). RESULTS: The expression of TNF-R2 was present in all fragments, however a statistical analysis showed no correlation or association between inflammation and the expression of TNF-R2. CONCLUSIONS: TNF-R2 is present in cholesteatoma of the middle ear, however, its expression is not directly related to the degree of inflammation observed in patients with this diseaseO colesteatoma adquirido da orelha média promove erosão óssea, ocasionando complicações potencialmente graves. O fator de necrose tumoral alfa (TNF-α) está presente no colesteatoma adquirido da orelha média e relaciona-se com a erosão óssea, como demonstraram diferentes autores. Para que se compreenda as características de agressividade do colesteatoma, é necessário que se estude a presença e a distribuição seus receptores. OBJETIVO: Avaliar a expressão do receptor tipo dois do TNF-α (TNF-R2) em fragmentos de colesteatoma e relacioná-lo com o grau de inflamação. MATERIAL E MÉTODOS: Estudo observacional do tipo transversal. Foram analisados 33 fragmentos de colesteatomas, submetidos à análise histológica e imunoistoquímica (utilizando o TNF-R2 da marca Labvision®). A avaliação foi realizada de forma qualitativa e semiquantitativa, de acordo com a intensidade observada. Para a análise estatística, foram utilizados o teste exato de Fischer e o coeficiente de correlação de Spearman (estatisticamente significativo quando p < 0,05). RESULTADOS: A expressão do TNF-R2 estava presente em todos os fragmentos, entretanto a estatística não evidenciou correlação, nem associação entre o processo inflamatório e a expressão do TNF-R2. CONCLUSÕES: O TNF-R2 está presente no colesteatoma adquirido da orelha média. Entretanto, a sua expressão não está relacionada ao grau de inflamação

Acquired middle ear cholesteatoma is a disease which promotes bone erosion resulting in potentially serious complications. The tumor necrosis factor alpha (TNF-α) is present in cholesteatoma and it is related to bone erosion, as shown by different authors. To understand the aggressiveness characteristics of cholesteatoma is necessary, however, to better address the presence and distribution of their receptors. OBJECTIVE: To evaluate the expression of type 2 TNF-α receptor (TNF-R2) in fragments of cholesteatoma and correlate it to the degree of inflammation present. MATERIAL AND METHODS: observational cross-sectional study, which analyzed 33 fragments of cholesteatomas through histological analysis and immunohistochemistry (using as primary antibody to TNF-R2 LabVision® brand). The evaluation was performed by means of a qualitative and semi-quantitative agreement with the observed intensity. For statistical analysis we used the Fisher exact test and Spearman´s correlation coefficient (considered statistically significant when p < 0. 05). RESULTS: The expression of TNF-R2 was present in all fragments, however a statistical analysis showed no correlation or association between inflammation and the expression of TNF-R2. CONCLUSIONS: TNF-R2 is present in cholesteatoma of the middle ear, however, its expression is not directly related to the degree of inflammation observed in patients with this disease O colesteatoma adquirido da orelha média promove erosão óssea, ocasionando complicações potencialmente graves. O fator de necrose tumoral alfa (TNF-α) está presente no colesteatoma adquirido da orelha média e relaciona-se com a erosão óssea, como demonstraram diferentes autores. Para que se compreenda as características de agressividade do colesteatoma, é necessário que se estude a presença e a distribuição seus receptores. OBJETIVO: Avaliar a expressão do receptor tipo dois do TNF-α (TNF-R2) em fragmentos de colesteatoma e relacioná-lo com o grau de inflamação. MATERIAL E MÉTODOS: Estudo observacional do tipo transversal. Foram analisados 33 fragmentos de colesteatomas, submetidos à análise histológica e imunoistoquímica (utilizando o TNF-R2 da marca Labvision®). A avaliação foi realizada de forma qualitativa e semiquantitativa, de acordo com a intensidade observada. Para a análise estatística, foram utilizados o teste exato de Fischer e o coeficiente de correlação de Spearman (estatisticamente significativo quando p < 0,05). RESULTADOS: A expressão do TNF-R2 estava presente em todos os fragmentos, entretanto a estatística não evidenciou correlação, nem associação entre o processo inflamatório e a expressão do TNF-R2. CONCLUSÕES: O TNF-R2 está presente no colesteatoma adquirido da orelha média. Entretanto, a sua expressão não está relacionada ao grau de inflamação

Summary: Middle ear cholesteatomas are characterized by the presence of stratified squamous epithelium in this cavity with highly invasive properties causing bone destruction and it may lead to complications. Aim: To study the histopathological features in acquired cholesteatomas of the middle ear, correlating this data with patient age. Study design:clinical and experimental cross-sectional study. Material and Methods: Samples were obtained from 50 patients submitted to otologic surgery for the removal of middle ear cholesteatomas from 2006 to 2007. Thirty four patients were adults and 16 were children. Samples were studied by histological analysis. Results: we found the presence of epithelial atrophy (78%), epithelial acanthosis (88%), hyperplasia of the basal layer (88%) and formation of epithelial cones (62%). There was a positive and significant correlation between histopathological variables (such as epithelial acanthosis, hyperplasia of the basal layer and formation of epithelial cones). Histopathological variables presented no statistical significant difference in both age groups (p>0,05). Conclusion: Cholesteatomas have hyperproliferating characteristics with epithelial acanthosis, hyperplasia of the basal layer and the presence of epithelial cones in the matrix. Keywords: cholesteatoma, histopathology, middle ear

Temporal bones are valuable resources to study ear diseases. Although there are several methods for removing temporal bones from cadavers, such methods are not usually described in enough details in experimental research papers.To describe a simple and rapid method for ear canal and tympanic membrane removal, and to evaluate its viability for histologic and immunohistochemical studies.In this experimental study, we obtained 31 ear canal and tympanic membrane samples from cadavers, with a conventional power drill and plug cutter. The material was dissected and samples containing ear canals and tympanic membranes were obtained in blocks. The samples were analyzed by histology and immunohistochemistry.Removal of small and good quality samples containing entire ear canals and tympanic membranes. In all the samples, it was possible to perform both histological and immunohistochemical analyses.This method was easily achievable, reproducible and yielded good quality samples, both for training purposes and for experimental research. All the samples were viable for histological and immunohistochemical analyses.

Several studies involving immunohistochemical methods to assess external auditory canal epidermis have been performed with different objectives. With this method it is possible to assess the expression of various antigens such as cytokeratins, cytokines, and hyperproliferation markers among others.to revise, describe and analyze the knowledge generated by identifiable papers published on the worldwide literature about immunohistochemical hyperproliferation markers in normal external auditory canal epidermis.systematic review of the papers published until 2009, in indexed international journals.Various antigens related to hyperproliferation were investigated by immunohistochemical methods among the included papers. The most studied ones were cytokeratin 16, Ki-67 and PCNA.most of the studies utilized external auditory canal epidermis as control sample to study external ear or middle ear cholesteatoma with immunohistochemical methods. There is a hyperproliferative antigen concentration, such as CK16, Ki-67 and PCNA, in the annulus tympanicus, adjacent meatus and tympanic regions, mainly in the lower areas.

Summary Cartilage grafting is an interesting option for rinoplasties refinements. Aim to compare butyl-2-cyanocrylate, compound gelatin-resorcin-formaldeyide (GRF) and suture control to determine the efficacy of these tissue glue preparations in securing grafted cartilage. Study Design Experimental. Methods Fifteen male adult New Zealand rabbits were submitted to a surgical procedure to harvest 6 auricular cartilage grafts from each animal. 2 of these grafts in each animal were glued together with butyl-2-cyanocrylate, 2 with compound gelatin-resorcin-formaldehyde and 2 sewn together with nylon suture. These sandwich grafts were then glued or sewn to the periosteum of the calvaria. After 2, 6 and 12 weeks, groups of 5 animals were sacrificed and histological analysis for inflammation was performed. Cartilage graft migration, adhesion and deformities of the grafts were also evaluated. Results there was less migration of the cartilages glued with GRF than with cyanoacrylate and suture. There was no statistical difference between the 3 materials of graft stabilization in relation to the inflammatory reaction in all evaluated periods. There wasn’t detachment neither deformity in the cartilage sandwiches sewed with suture. The majority of detachment and deformed cartilages were found among those glued with cyanoacrylate. The number of deformed cartilages was directly related to the number of detached cartilages. The data were statistically significant (p Conclusion the compound gelatin-resorcin-formaldehyde revealed to be a stabilization material for cartilage grafts in rabbits better than butyl-2-cyanoacrylate. The compound gelatin-resorcin-formaldeyide was also better than the suture when comparing it's fixation to the periosteum.

Metodos objetivos de avaliacao sao frequentemente cobrados em estudos cientificos. Exames histologicos com coloracao imuno-histoquimica podem ser avaliados por meio de fotometria. OBJETIVO: Comparar este metodo objetivo com a avaliacao subjetiva realizada por tres observadores independentes, utilizando lâminas de colesteatoma adquirido da orelha media. METODO: Foram selecionadas um total de 54 imagens de colesteatomas imuno-histoquimicamente coradas pelos anticorpos anti-TNF-R2 (32 lâminas) e anti-TGF-α; (22 lâminas). O anticorpo secundario utilizado nos dois grupos foi o Max Polimer Detection System (Kit Novo Link, Novocastra®, UK). As amostras foram processadas por um scanner digital de lâminas (modelo ScanScope - Aperio). As areas selecionadas foram submetidas a analise por fotometria. RESULTADOS: A avaliacao objetiva por fotometria foi comparada com a avaliacao subjetiva por tres observadores e submetidas a analise estatistica. A analise estatistica revelou reprodutibilidade moderada (K valores entre 0,41 e 0,60) para os dois grupos. CONCLUSAO: O presente estudo demonstrou que as caracteristicas irregulares das lâminas de colesteatoma da orelha media coradas pela imuno-histoquimica impossibilita a sua adequada avaliacao objetiva, enquanto a avaliacao subjetiva por observadores experientes se mostrou mais confiavel.