Extensive qPCR analysis reveals altered gene expression in middle ear mucosa from cholesteatoma patients
Cecilia DrakskogNele de KlerkSusanna Kumlien GeorénJohanna WesterbergElina Mäki‐TorkkoLars‐Olaf Cardell
0
Citation
0
Reference
10
Related Paper
Abstract:
Abstract The middle ear is a small and hard to reach compartment, limiting the amount of tissue that can be extracted and the possibilities for studying the molecular mechanisms behind diseases like cholesteatoma. In this paper 14 reference gene candidates were evaluated in the middle ear mucosa of cholesteatoma patients and two different control tissues. ACTB and GAPDH were shown to be the optimal genes for the normalisation of target gene expression when investigating middle ear mucosa in multiplex qPCR analysis. Validation of reference genes using c-MYC expression confirmed the suitability of ACTB and GAPDH as reference genes and showed an upregulation of c-MYC in middle ear mucosa during cholesteatoma. The occurrence of participants of the innate immunity, TLR2 and TLR4, were analysed in order to compare healthy middle ear mucosa to cholesteatoma. Analysis of TLR2 and TLR4 showed variable results depending on control tissue used, highlighting the importance of selecting relevant control tissue when investigating causes for disease. It is our belief that a consensus regarding reference genes and control tissue will contribute to the comparability and reproducibility of studies within the field.The advent of antibiotics has not reduced the incidence of cholesteatoma resulting from chronic otitis media. This incidence remains high due to continued Eustachian tube problems and to other poorly understood mechanisms. Because the otologist will continue to encounter cholesteatoma in clinical practice, he should be prepared to judge the extent and severity of disease and its potential for complications based on the initial office examination. This judgment is best accomplished through an understanding of the anatomy and embryology of the middle ear cleft--the compartments which encourage growth of cholesteatoma and the membranes which limit its growth. This paper reviews the anatomy and embryology of the compartments of the middle ear cleft in an attempt to understand the pathogenesis of cholesteatoma and its potential for complication.
Eustachian tube
Embryology
Pathogenesis
Ear disease
Cite
Citations (11)
Objective To study expression of caspase-8 in human middle ear cholesteatoma and its roles in the pathogenetic mechanisms of human middle ear cholesteatoma. Methods Immunohistochemistry and Western Blot tech-niques were used to examine expression of caspase-8 in 36 specimens of middle ear cholesteatoma and 20 samples of normal external ear canal skin. Results Expression of caspase-8 was extremely higher in middle ear cholesteatoma than in normal external canal skin (P 0.01). Conclusicon Caspase-8 may play an important role in human middle ear cholesteatoma, maybe through deregulation of apoptosis and proliferation in middle ear cholesteatoma.
Cite
Citations (0)
Objective To explore the diagnostic effectiveness of HRCT in middle ear cholesteatoma. Methods To compare the findings on CT scan and during operation in 238 cases with middle ear cholesteatoma proved by mastoidotympanectomy and post-operation pathological examination. Results (1) Sensitivity of CT diagnosis is the highest for attic cholesteatoma(81.25%) and the lowest for mastoid cholesteatoma (50%); (2) The radiological features on the CT scan were: ① a tissue mass (cholesteatoma) in the tympanic and mastoid cavity; ② a low density ring around the mass; ③ sclerosing of the bony borders of the tympanic sinus and mastoid cavity; ④ enlargment of the tympanic and mastoid cavity;⑤ destruction of the auditoy ossicle chain. Conclusions CT scan is helpful in the assessment of and planning the management for the middle ear cholesteatoma before surgery.
Sinus (botany)
Tympanic cavity
Cite
Citations (0)
Objective—To establish the eventual presence of human β-defensins (hBD) in middle ear cholesteatoma and paired retroauricular skin samples obtained during surgical treatment. Material and Methods—hBD were detected using immunohistochemical techniques and reverse transcriptase-polymerase chain reaction (RT-PCR). Results—It is hypothesized that hBD-1 and -2 are upregulated in cholesteatoma in comparison with retroauricular skin. The immunohistochemical study demonstrated stronger hBD-2-positive staining in middle ear cholesteatoma in comparison with retroauricular skin. No significant differences in hBD-1 expression were detected between cholesteatoma and skin samples. RT-PCR established elevated expression of hBD-2 mRNA in middle ear cholesteatomas. HBD-1 mRNA was constitutively expressed in cholesteatoma and retroauricular skin specimens. Conclusion—These preliminary experimental results lead us to believe that hBD may play an important role in the chronic inflammatory state of middle ear cholesteatoma.
Cite
Citations (10)
We have undertaken the analysis of the specific clinical manifestations of acquired cholesteatoma of the middle ear in 437 chronic patients suffering from this pathology. 96.1% of them presented with primarily acquired cholesteatoma of the middle ear (including 53.3% having attic cholesteatoma, 22.8% with sinus cholesteatoma, and 19.9% of tensa retraction cholesteatoma). 3.9% of the patients exhibited a different mechanism of development of cholesteatoma. The secondary acquired mesotympanic cholesteatoma formed in association with the long-term chronic inflammation of the middle ear, concomitant perforation of the tympanic membrane and epithelial invasion from the edge of the tympanic membrane perforation and middle ear cavity. Typmanosclerosis of different degree and localization played an important role in the enhancement of the prevalence of this condition. All types of acquired cholesteatoma were found to extend beyond the point of origin of the disorder. The maximum destruction of the ossicular chain was documented in the patients presenting with sinus cholesteatoma. Those with secondary acquired cholesteatoma showed the worst functional capacity as a result of rigid fixation of the auditory ossicles. The overall cholesteatoma relapse rate (including both residual and recurrent cholesteatoma) was estimated to be 15.6%. It is concluded that the surgical strategy should be chosen on an individual basis for each concrete patient. The long-term observation of the treated patients with the application of the up-to-date radiological techniques is believed to be the indispensable prerequisite for the successful management of the complicated pathology under consideration.Проведен анализ клинических проявлений у 437 больных с приобретенной холестеатомой среднего уха. Установлено, что в 96,1% случаев наблюдается первичная приобретенная холестеатома, в том числе в 53,3% - холестеатома расслабленной части барабанной перепонки, в 22,8% - холестеатома синуса, в 19,9% - холестеатома собственно натянутой части. У 3,9% больных выявлен иной механизм формирования холестеатомы - вторичная мезотимпанальная холестеатома на фоне длительного хронического воспаления среднего уха с перфорацией барабанной перепонки и тимпаносклерозом различной локализации и степени. Отмечено распространение холестеатомы за пределы точки первоначального роста при всех ее типах. Выявлено, что максимальная степень деструкции оссикулярной цепи наблюдается при холестеатоме синуса, при этом наихудший функциональный результат получен при вторичной холестеатоме из-за фиксации слуховых косточек тимпаносклерозом. Частота формирования рецидивирующей холестеатомы, включая обе ее формы - резидуальную и рекуррентную, составила 15,6%. Индивидуальный выбор хирургической тактики в каждом конкретном случае, длительное послеоперационное наблюдение пациентов с использованием современных радиологических методов обследования являются залогом успешного лечения этой сложной патологии.
Perforation
Tympanic Membrane Perforation
Sinus (botany)
Ossicles
Cite
Citations (6)
Malleus
Conductive hearing loss
Cite
Citations (1)
Background: Middle ear cholesteatoma is an ectopic squamous stratified epithelium in the middle ear.It is a actually risky disease as it leads to destruction of ossicles and/or walls of middle ear that causes severe complications as hearing loss and brain abscess.Pre-operative assessment of cholesteatoma by Multi-Detectors Computed Tomography (MDCT) is essential for accurate diagnosis and appropriate management to prevent recurrence after surgery. Aim of Study:To assess the diagnostic accuracy of preoperative MDCT of middle ear cholesteatoma with surgical correlation.Patients and Methods: A prospective study was conducted in Tanta University Hospitals from July 2015 to December 2016 on 30 patients with middle ear cholesteatoma.All patients had a pre-operative MDCT scans.The pre-operative MDCT findings were recorded and correlated with surgical and histopathological results.Sensitivity and specificity of preoperative MDCT findings were calculated considering operative findings as gold standard.Results: The study included 30 patients, 17 were females and 13 were males with a mean age of 35 years.The diagnostic accuracy of the pre-operative MDCT findings when correlated to intraoperative ones was 100% in detecting soft tissue mass inside the middle ear, 100% in detecting the location of cholesteatoma, 100% in detecting ossicular erosion, 93.3% in detecting scutum erosion and 100% in confirming the diagnosis of cholesteatoma proved by post-operative histopathological examination.The mean sensitivity of MDCT in preoperative diagnosis and evaluation of middle ear cholesteatoma was 97.4% while the mean specificity was 95.8%. Conclusion:Middle ear cholesteatoma is a distinctive clinical entity with characteristic findings on MDCT scan, that guides in the surgical approach and management plan.
Ossicles
Cite
Citations (0)
The aim of this study is to present various morphologic aspects of the middle ear cholesteatoma, concerning both container (tympanic cavity) and content (cholesteatoma). There are two different aspects of the study: a mezoscopic study of the tympanic walls and the elements within the middle ear (ossicular chain, folds, ligaments, middle ear clefts) in order to evidentiate the critical areas (sinus tympani, facial sinus, anterior attic) and the pathways of spread for the cholesteatomas; a classical histological study of the middle ear cholesteatomas, in order to present the structural parts and the modifications in the neighboring structures.
Sinus (botany)
Tympanic cavity
Cite
Citations (5)
Purpose To evaluate the role of MMP-2 in the absorption of the temporal bone in human middle ear cholesteatoma. Methods Immunohistochemestry technique (ABC) method was used to detect the expression and location of matrix metalloproteinase-2 (MMP-2) in 30 cases of human middle ear cholesteatoma, and 5 eases of middle ear granulation tissue were served as control tissue. Results The expression of MMP-2 were positive in 26 middle ear cholesteatoma. No positive signal could be detected in the inflammatory granulation tissue. Conclusion The results indicate that MMP-2 could play an active role in the molecular mechanisms of cholesteatoma invasion into the temproal bone.
Granulation tissue
Cite
Citations (0)
Computed tomography is today the best method of imaging to study chronic middle ear diseases. The aim of this study was to assess the diagnostic value of CT scan in cases of middle ear cholesteatoma in children. The preoperative CT scans were compared with the operative findings in retrospective analysis of 60 ears operated between 1998-2001. Our results show good radiosurgical correlation in cholesteatoma for most middle ear structures except for the integrity of long process of incus. The disadvantage of CT scans is inability to distinguish between cholesteatoma, granulation tissue and effusion. CT scans are an important investigative tool prior to cholesteatoma surgery.
Incus
Cite
Citations (3)