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.

Background and Objectives. The aim of this study was to compare the distribution of proliferation markers (Ki-67, NF-κβ), tissue-remodeling factors (MMP-2, MMP-9, TIMP-2, TIMP-4), vascular endothelial growth factor (VEGF), interleukins (IL-1 and IL-10), human beta defensins (HβD-2 and HβD-4) and Sonic hedgehog gene protein in cholesteatoma and control skin. Methods. Nineteen patient cholesteatoma tissues and seven control skin materials from cadavers were included in the study and stained immunohistochemically. Results. Statistically discernible differences were found between the following: the Ki-67 in the matrix and the Ki-67 in the skin epithelium (p = 0.000); the Ki-67 in the perimatrix and the Ki-67 in the connective tissue (p = 0.010); the NF-κβ in the cholesteatoma matrix and the NF-κβ in the epithelium (p = 0.001); the MMP-9 in the matrix and the MMP-9 in the epithelium (p = 0.008); the HβD-2 in the perimatrix and the HβD-2 in the connective tissue (p = 0.004); and the Shh in the cholesteatoma’s perimatrix and the Shh in the skin’s connective tissue (p = 0.000). Conclusion. The elevation of Ki-67 and NF-κβ suggests the induction of cellular proliferation in the cholesteatoma. Intercorrelations between VEGF, NF-κβ and TIMP-2 induce neo-angiogenesis in adult cholesteatoma. The similarity in the expression of IL-1 and IL-10 suggests the dysregulation of the local immune status in cholesteatoma. The overexpression of the Sonic hedgehog gene protein in the cholesteatoma proves the selective local stimulation of perimatrix development.

Introduction:The complex appearance of proliferation markers, cytokines, remodelling factors, antimicrobial peptides and genes still have not been studied together despite their possible crucial role in the development of cholesteatoma. Therefore, the aim of this study was complex research of appearance and distribution of proliferation, remodeling markers, pro-and anti-inflammatory cytokines, local tissue defensins and genes of the cholesteatoma tissue.Materials and Methods: Four cholesteatoma tissue samples were obtained from children, 5 cholesteatoma specimens were obtained from adults, and 7 deep external meatal skin controls were obtained from cadavers. Tissues were stained for Ki-67, MMP-2, TIMP-2, IL-1, IL-10, HβD-2, HβD-4 and Shh immunohistochemically. Non-parametric statistic, Mann-Whitney and Spearman's coefficient were used.Results: A statistically significant difference was seen between the numbers of Ki-67, TIMP-2 positive cells in matrix and control group epithelium and between numbers of HβD-2, HβD-4 Shh positive cells in perimatrix in comparison to control group connective tissue. A very strong positive correlation was detected in control tissue epithelium between Ki-67 and TIMP-2, IL-10, Shh; between MMP-2 and TIMP-2, IL-1; between IL-10 and TIMP-2, Shh. In the connective tissue, a very strong correlation was detected between Ki-67 and Shh. A strong correlation was detected in epithelium between Ki-67 and MMP-2, and between MMP-2 and IL-10. Conclusion:Cholesteatoma tissue with the low appearance of Ki-67 proves the indistinct proliferation activity in the already developed tumour, but Ki-67 correlations with HβD-2 and HβD-4 in cholesteatoma indicate the direct link between the proliferation and expression of antimicrobial peptides. HβD-2 seems to be the most important factor for antimicrobial defence, but the correlations between HβD-2 and HβD-4 suggest their common action against chronic middle ear infection in case of choleasteatoma. Higher levels of MMP-2 positive cells in cholesteatoma and lower levels of TIMP-2 positive cells suggest the prevalence of enzymatic processes over their suppression, possibly being characteristic for cholesteatoma of different age patients. Shh gene might play a crucial role in the development of cholesteatoma.