Establishment and Characterization of an In Vitro Model for Cholesteatoma

Abstract

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.

Morgellons Disease: A Chemical and Light Microscopic Study

“…BDD in cattle is associated with hyperkeratosis, keratin filament formation and spirochetal infection [12][13][14][15][16][17][18][19][20]. Hyperkeratosis and excessive keratin production associated with chronic inflammation has been demonstrated in humans with cholesteatoma [23,24], and alterations in keratinocyte expression of HLA markers and tissue enzymes have been reported in association with Bb infection [25,26]. These observations suggest that hyperkeratosis and keratin filament production associated with spirochetal infection is a plausible explanation for the clinical and microscopic findings in MD.…”

Etiopathogenesis of acquired cholesteatoma: Prominent theories and recent advances in biomolecular research

“…In contrast to neoplastic transformation, several studies have revealed an association between the progression of cholesteatoma and host immune response to inflammation, such as that observed in the process of wound healing . In particular, paracrine and autocrine interactions between matrix keratinocytes and perimatrix fibroblasts play an important role in homeostasis and tissue regeneration within cholesteatomas …”

“…For example, TGF‐α and TGF‐β are constitutively expressed in hyperproliferative epithelium, regulating keratinocyte proliferation and differentiation in a process similar to that of the autocrine system (Fig. ) . Additionally, infiltrating inflammatory cells also secrete cytokines to stimulate the induction of hyperproliferative cells in all layers of the cholesteatoma epidermis …”

Characterization and evolution of dermal filaments from patients with Morgellons disease

“…MD lesions are histopathologically distinct, however, from both hairy leukoplakia and myofibroblastic tumors. Increased keratin expression caused by upregulation of keratinocyte growth factor/fibroblast growth factor-7 has been reported in cholesteatoma, a hyperproliferative epithelial disease 22. As a consequence of B. burgdorferi infection and an inflammatory environment, dermal fibroblasts may become activated, modulating epithelial growth and differentiation through increased keratinocyte growth factor/fibroblast growth factor-7 expression 23.…”