Remodeling Factors, Transcription Factors and Angiogenetic Factors in Cholesteatoma in Ontogenetic Aspect

Publisher Copyright: © 2022 Mashhad University of Medical Sciences. All rights reserved.Introduction: The main goal of our study was to describe the transcription factor (NF-κβ), angiogenetic factor (VEGF), and remodeling markers (MMP-9 and TIMP-4) of the cholesteatoma tissue compared to control skin tissue. There are still uncertainties how transcription, angiogenetic and remodeling factors affect the cholesteatoma course. Materials and Methods: Eight cholesteatoma tissue specimens were retrieved from children, seven – from adults, seven skin controls – from cadavers. Obtained material immunohistochemically were stained for NF-κβ, MMP-9, TIMP-4, VEGF. Non-parametric statistic methods were used. Results: A statistically significant higher numbers of NF-κβ and TIMP-4 immunoreactive cells in the cholesteatoma compared to control group. A very strong positive correlation between MMP-9 and TIMP-4 was seen in the patient group. A strong positive correlation - between MMP-9 in matrix and MMP-9, VEGF in perimatrix, between TIMP-4 in matrix and TIMP-4 in perimatrix, NF-κβ in the matrix and VEGF; between TIMP-4 in perimatrix and NF-κβ in the matrix. Conclusions: Correlation between MMP-9 and TIMP-4 suggests that TIMP-4 in cholesteatoma tissue intercorrelates to MMP9. TIMP-4 likely regulates the development of cholesteatoma. Disbalance between MMPs and TIMPs affects NFκβ and causes uncontrolled cell proliferation and immune response in this tumor. There is a lack of VEGF strong expression in cholesteatoma perimatrix.publishersversionPeer reviewe

Complex Evaluation of Tissue Factors in Pediatric Cholesteatoma

Funding Information: Funding: Riga Stradin,š University’s funding is gratefully acknowledged. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.The aim of this study was to describe the appearance and distribution of tissue remodeling markers (MMP-2, MMP-9, TIMP-2, TIMP-4), Sonic hedgehog gene protein (Shh), pro- and anti-inflammatory cytokines (IL–1, IL–10), transcription factor (NF-κβ), proliferation marker (Ki–67), angiogenetic factor (VEGF), tissue defensins (HβD–2, HβD–4) of the pediatric cholesteatoma. Sixteen cholesteatoma samples were obtained from children, eleven skin controls from cadavers. Tissues were stained for MMP-2, MMP-9, TIMP-2, TIMP-4, Shh, IL–1, IL–10, NF-κβ, Ki–67, VEGF, HβD–2, HβD–4. Non-parametric statistic, Mann–Whitney, and Spearman’s coefficient was used. A statistically significant difference was seen between Shh and HβD–2 in perimatrix and control connective tissue, between NF-κβ in cholesteatoma and control skin, and between HβD–4 in matrix and skin epithelium. Complex intercorrelations between MMPs, NF-κβ and VEGF cause the intensification of angiogenesis in cholesteatoma. The persistent increase in Shh gene protein expression in cholesteatoma perimatrix suggests the stimulation of the cholesteatoma growth in children. Similar expression of IL-1 and IL-10 and their intercorrelation, proves there is a balance between pro- and anti-inflammatory cytokines. NF-κβ, and not Ki-67, seems to be the main inducer of cellular proliferation. The main antimicrobial protection is provided by HβD-2.publishersversionPeer reviewe

Histologic description of acquired cholesteatomas: comparison between children and adults

SummaryCholesteatoma is constituted of matrix, perimatrix and cystic content. Some authors affirm that, in children, its clinical behavior is more aggressive of the than in adults.AimsHistologic compared cholesteatomas of children and adults.Methodology74 cholesteatomas been analyzed, being 35 of pediatrics patients (<18 years). The average number of cellular layers and hyperplasia in the matrix had been evaluated; thickness, delimitante epithelium, fibrosis, inflammation and granuloma in the perimatrix. The analysis statistics was carried through with program SPSS 10,0, using the coefficients of Pearson and Spearman, test of qui-square and t test. The number of cellular layers in the matrix was of 8,2±4,2. The hyperplasia appears in 17%, fibrosis in 65%, granuloma in 12% and the delimitante epithelium in 21%. The perimatrix presented a medium one of 80 micrometers (37 the 232), minimum value zero and maximum value 1.926. The histological degree of inflammation was considered of moderate the accented one in 60%. When applying the coefficient of Spearman enters the inflammation degree and average of cellular layers of the matrix with the variables of the measure of thickness of the perimatrix we find correlations, significant, with moderate magnitudes of the great ones (rs=0,5 and P<0,0001).ConclusionAdults colesteatomas of and child had not been identified to morphologic differences between. We find correlation enters the intensity of the inflammation and of the average of cellular layers of the matrix with the thickness of the perimatrix, what it can predict its aggressiveness, more studies are necessary to define the paper of this finding in pathogenesis of cholesteatoma

The structure of acquired aural cholesteatoma as revealed by scanning electron microscopy

The structural features of cells, their surfaces and the extracellular matrix were investigated in acquired aural cholesteatoma. Cholesteatomas surgically removed from 30 patients were examined by a scanning electron microscope (SEM). The predominant part of a cholesteatoma was composed of stratified squamous epithelium, showing extensive chaotic desquamation. The surface sculpture of the keratinocytes and corneocytes varied from parallel ridges, irregular microplicae and mirovilli, to flat grooves and pits and a completely smooth surface. Sheetlike lamellar structures, probably representing an intercellular lipid-forming permeability barrier, were also observed. Small crystals located in the perimatrix were observed in one case. According to the SEM observations, cholesteatoma epithelium is characterised by abnormal and uncoordinated keratinisation, with a predominance of the advanced stages of the process. Folia Morphol 2008; 67: 8&#8211;12

Analysis of histopathological aspects in acquired middle ear cholesteatoma

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.O colesteatoma da orelha média é caracterizado pela presença de epitélio estratificado pavimentoso queratinizado neste local, com alto poder invasivo, causando destruição óssea e podendo levar a complicações. OBJETIVO: Estudar os padrões histopatológicos no colesteatoma adquirido da orelha média. Correlacionar esses dados com a idade do paciente. FORMA DE ESTUDO: Clínico e experimental do tipo transversal. MATERIAL E MÉTODO: Foram colhidas amostras de colesteatoma de 50 pacientes submetidos à cirurgia otológica, sendo 34 adultos e 16 crianças, no período de 2006 a 2007. Essas amostras foram submetidas à análise histológica. RESULTADOS: A presença de atrofia foi encontrada em 78% dos casos, acantose em 88%, hiperplasia da camada basal em 88% e cones epiteliais em 62%. As correlações entre acantose e hiperplasia da camada basal, acantose e formação de cones epiteliais, hiperplasia da camada basal foram positivas e significativas. Não houve diferença estatisticamente significativa em relação aos padrões histopatológicos entre os dois grupos etários (p>0,05). CONCLUSÃO: O colesteatoma tem características hiperproliferativas, com acantose, hiperplasia da camada basal e presença de cones epiteliais na sua matriz.SCSP FCM Depto. MorfologiaUNIFESP-EPMSCSP FCM Depto. de OtorrinolaringologiaFundação Antônio PrudenteUNIFESP, EPMSciEL

Proliferation activity and apoptosis in granulation tissue and cholesteatoma in middle ear reoperations

The aim of the study was to find out the potential prognostic value of proliferation activity and apoptosis in cholesteatoma and granulation tissue removed during middle ear reoperation in recurrent middle ear inflammation. Granulation tissues and recurrent cholesteatoma were analysed after being surgically removed from the middle ear in a group of 25 patients qualified for middle ear reoperation procedure. Paraffin sections were stained with haematoxylin and eosin according to Mallory&#8217;s method. Immunohistochemical reaction Anti-PCNA was performed. Apoptosis was evaluated using the TUNEL method. The percentage of PCNA-positive cells was 42&#8211;95% in the matrix of the cholesteatoma and 29&#8211;81% in the perimatrix. In the granulation tissue it was 35&#8211;75%. The percentage of apoptotic cells was 12&#8211;73% in the matrix of the cholesteatoma, 5&#8211;72% in the perimatrix and 1&#8211;65 % in the granulation tissue. The prognostic value of the parameters studied in the recurrent middle ear inflammatory process is questionable, probably due to the small number of cases under examination

Chronic suppurative otitis media with cholesteatoma and chronic otitis media with granulation investigated by scanning electron microscope based on an analysis of 140 patients

Introduction: Chronic purulent cholesteatoma of the middle ear, as well as to a lesser extent chronic granulomatous otitis media, lead to destruction of bone structures within the middle ear space. The above process is controlled by the OPG/RANKL/RANK system. Aim: The aim of the study is a comparative analysis of surgical material obtained from patients with diagnosed chronić cholesteatoma and chronic otitis media with inflammatory granulation in the assessment of the Scanning Electron Microscope (SEM). Material and methods: An analysis of 140 patients operated on due to chronic otitis media was performed. Forty patients who had been diagnosed with chronic cholesteatoma of the middle ear and chronic granulomatous otitis media were selected for a detailed analysis in the SEM. The final study under SEM included 20 patients. Results: The regular structure of cholesteatoma depicted in the SEM concerned 5 patients. In the remaining 7 patients, the system was irregular and even chaotic. The lack of regularity can also be observed in the case of granulation tissue, which in the SEM image presented itself as an irregular tissue mass without detectable regularities. Conclusions: (1) The regular pattern of the cholesteatoma matrix cells observed in some patients with chronic cholesteatoma of the middle ear reduces the molecular permeability of inflammatory cytokines, concurrently limiting the destructive activity on bone structures; (2) the presence of inflammatory granulation tissue in the middle ear is accompanied by an influx of leukocytes: neutrophils and lymphocytes, which are the source of pro-inflammatory cytokines, the growth of which activates the processes leading to the damage of bone tissue and the development of inflammation; (3) no specimen of acquired cholesteatoma revealed the presence of commensal organisms from Demodex species on the surface of the exfoliated human epithelium

Establishment and Characterization of an In Vitro Model for Cholesteatoma

ObjectivesExperimental models are of importance to study the pathogenesis of middle ear cholesteatoma, however, they were not established until now. We aimed to develop in vitro model of middle ear cholesteatoma using primary keratinocytes and fibroblasts isolated from cholesteatoma tissue. HaCaT cell line was used as a "skin equivalent" and to compare the grade of homogeneity between cholesteatoma keratinocytes and HaCaT cells.MethodsPrimary keratinocytes were isolated from cholesteatoma tissue, co-cultured with preliminary prepared feeder layer from cholesteatoma fibroblasts and subsequently air-exposed. The protein profile of cholesteatoma keratinocytes and HaCaT cells was evaluated by means of immunoblot using monoclonal antibody against cytokeratin (CK) 13 and 16. Tissue localization of CK 13 and 16 was accomplished with immunohistochemistry.ResultsDifferent protein profile and stronger expression of CK 13 and 16 were demonstrated in cholesteatoma keratinocytes in comparison with HaCaT cells. Bigger stratification was observed in the 3D-in vitro systems when both cholesteatoma keratinocytes and HaCaT cells were respectively co-cultured with fibroblasts in comparison with the corresponding control groups without fibroblasts.Conclusion3D-model demonstrates the significance of intercellular interaction between components of cholesteatoma tissue