Complex Evaluation of Tissue Factors in Pediatric Cholesteatoma

Abstract

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.

Review of potential medical treatments for middle ear cholesteatoma

“…Furthermore, a significant correlation was found between TLR4 and activated NF-κB [ 37 ] in MEC tissue, and we assume, that the same could be shown for RAGE as well. Further, another study demonstrated a correlation between NF-kB and biomarkers of angiogenesis and osteolysis in MEC tissue isolated from patients [ 84 ]. In addition to this, the DNA-binding protein inhibitor Id1, known to be induced by chronic TLR4 signalling [ 85 ], was proven to be abundantly expressed in MEC epithelium [ 86 ] where it accelerates proliferation and survival of keratinocytes in vitro via NF-κB signalling [ 87 ].…”

“…The most prominent is the vascular endothelial growth factor (VEGF), for which expression is upregulated in compared to normal skin [ 241 , 282 ]. A correlation between NF-κB driven inflammation and VEGF was shown to promote the neovascularisation in MEC tissue [ 84 ]. VEGF signalling can be inhibited by approved drugs on the level of intracellular signalling by drugs like sunitinib [ 288 ] or axitinib [ 289 ], which are approved for treatment of renal cell carcinoma or nintedanib applied against pulmonary fibrosis [ 290 ] or lucitanib used in clinical trials against cancer [ 291 ].…”

“…These MMPs are well known to be crucial for the decomposition of the extracellular matrix making the tissue susceptible for vascular outgrowth. An immunofluorescence correlation study in cholesteatoma tissue demonstrated, that correlations between NF-κB and these MMPs causes the intensification of angiogenesis in MEC [ 84 ] and can be targeted by drugs described in the previous subchapter.…”

Morphopathogenesis of Adult Acquired Cholesteatoma

“…However, additional information on how HβD-2 acts in cholesteatoma tissue is needed. There are limited data about the importance of HβD-4 in cholesteatoma, but our previous study showed that HβD-2 is more heavily expressed in pediatric cholesteatoma tissue than HβD-4 [ 26 ].…”

“…Chiang et al proved that the loss of Shh causes external- and middle -ear pathologies [ 28 ]. However, the exact role of Shh in cholesteatoma tissue is not yet clear, although our previous study showed the upregulation of Shh in cholesteatoma compared to skin tissue [ 26 ].…”