Cholesteatoma Growth Research Articles

Cholesteatoma Severity Determines the Risk of Recurrent Paediatric Cholesteatoma More Than the Surgical Approach

Objective: To evaluate factors that influence the rate of cholesteatoma recurrence (growth of new retraction cholesteatoma) in children. Methods: Review of children with primary acquired or congenital cholesteatoma. Severity was classified by extent and EAONO-JOS stage, and surgery by SAMEO-ATO. Primary outcome measure was 5-year recurrence rate using Kaplan–Meier or Cox regression analysis. Results: Median age was 10.7 years for 408 cholesteatomas from which 64 recurred. Median follow up was 4.6 years (0–13.5 years) with 5-year recurrence rate of 16% and 10-year of 29%. Congenital cholesteatoma (n = 51) had 15% 5-year recurrence. Of 216 pars tensa cholesteatomas, 5-year recurrence was similar at 14%, whereas recurrence from 100 pars flaccida cholesteatomas was more common at 23% (log-rank, p = 0.001). Sub-division of EAONO-JOS Stage 2 showed more recurrence in those with than without mastoid cholesteatoma (22.1% versus 10%), with more in Stage 3 (31.9%; p = 0.0003). Surgery without mastoidectomy, including totally endoscopic ear surgery, had 11% 5-year recurrence. Canal wall-up tympanomastoidectomy (CWU) and canal wall-down/mastoid obliteration both had 23% 5-year recurrence. Multivariate analysis showed increased recurrence for EAONO-JOS Stage 3 (HR 5.1; CI: 1.4–18.5) at risk syndromes (HR 2.88; 1.1–7.5) and age < 7 years (HR 1.9; 1.1–3.3), but not for surgical category or other factors. Conclusion: Young age and more extensive cholesteatoma increase the risk of recurrent cholesteatoma in children. When controlling for these factors, surgical approach does not have a significant effect on this outcome. Other objectives, such as lower post-operative morbidity and better hearing outcome, may prove to be more appropriate parameters for selecting optimal surgical approach in children.

Study of Angiogenic, Pro-Apoptotic, and Pro-Inflammatory Factors in Congenital and Acquired Cholesteatomas.

Despite recent advances in biomolecular research that have improved our knowledge of cholesteatoma pathogenesis, the reasons behind its highly variable clinical course are still not clarified. It has been proposed that biological signaling between peri-matrix and matrix cells could play a critical role in disease homeostasis. The aim of our study was to analyze the expression of inflammatory (IL-1β), hyper-proliferative (STAT-3, TGF-β), and angiogenic (VEGF-C, PDGFr) factors in congenital and acquired cholesteatomas (both in adults and children), which might correlate with the clinical features observed. We performed an experimental study on 37 patients (29 males and 8 females, ranging from 4 to 66 years of age) who were diagnosed with cholesteatoma between 2020 and 2021 in our institution. All patients underwent clinical, audiologic, and radiologic assessments. Bone erosion grading and staging of cholesteatoma growth were assessed through preoperative evaluation and intraoperative middle ear findings, according to the PTAM System proposed by the Japan Otological Society (2016). Retro-auricular skin specimens were intraoperatively collected in all patients. Skin and cholesteatoma samples were analyzed through histopathological, western blot, and immunohistochemical evaluations. The expression rate was measured to find out the differences between congenital and acquired cholesteatomas as well as between the adult and pediatric populations. Expression of angiogenic, inflammatory, and proliferative biomarkers is significantly increased in acquired cholesteatomas in children as compared to congenital and acquired forms in adults, in accordance with the higher stage of disease shown by imaging, surgical, and histological features. Our data suggest that pathways already supposed to be involved in the pathogenesis of cholesteatomas could be differently activated in more destructive forms, typically found in children. The identification of potential biomarkers of cholesteatoma aggressiveness could lead to more personalized management (timing of intervention, recurrence prevention) and the future identification of anti-growth/anti-proliferative agents as non-surgery therapeutic options.

Expression and Correlation Research of MicroRNA10a-5p and PIK3CA in Middle Ear Cholesteatoma.

This study aimed to examine the roles of miR-10a-5p and phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α in the pathogenesis of middle ear cholesteatoma. We enrolled 27 patients with middle ear cholesteatoma and collected samples of intraoperative cholesteatoma and normal posterior ear skin tissues. The mRNA expression levels of miR-10a-5p and PIK3CA were detected using real-time quantitative polymerase chain reaction. PIK3CA protein expression was measured by immunohistochemistry and western blotting. Middle ear cholesteatoma tissues showed significantly lower miR-10a-5p expression levels and significantly higher PIK3CA expression levels than normal posterior ear skin tissues (both P < .05). Furthermore, the miR-10a-5p and PIK3CA expression levels were significantly negatively correlated in middle ear cholesteatoma tissues (r = -0.926, P < .001). Low miR-10a-5p expression levels in middle ear cholesteatoma tissues may inhibit the growth and proliferation of cholesteatoma, whereas high PIK3CA expression level may promote its growth and proliferation. In addition, miR-10a-5p may affect the proliferation and differentiation of cholesteatoma by negatively regulating its target gene, PIK3CA.

Epigenetic Regulation as a New Therapeutic Target for Middle Ear Cholesteatoma.

To evaluate the effectiveness of the menin-MLL inhibitor, MI503, as a conservative treatment of middle ear cholesteatoma (cholesteatoma) in a mouse model and to confirm its safety profile regarding auditory function in vivo. Cholesteatoma is a mass formed by the keratinizing squamous epithelium in the tympanic cavity and/or mastoid and subepithelial connective tissue and by the progressive accumulation of keratin debris with/without a surrounding inflammatory reaction. Although the main treatment is surgical therapy, the techniques to prevent recurrence remain a critical area of research. Recently, the use of MI503 in experiments resulted in the inhibition of the growth of cholesteatoma in vivo under histone modification. After cholesteatoma was induced in ICR mice (n = 7) by keratinocyte growth factor expression vector transfection, MI503 (50 μM) or phosphate-buffered saline was topically injected for 14 days. The effects of MI503 against cholesteatoma were analyzed by micro-computed tomography images. For the in vivo ototoxicity study, a single intratympanic injection of MI503 (50 or 500 μM) or phosphate-buffered saline (n = 4 each) was done in the ICR mice. An auditory brainstem response was performed at days 0, 1, and 14. For morphological analysis, immunostaining for Phalloidin/F-actin and Myo7a was performed. MI503 reduced keratinocyte growth factor-induced cholesteatoma in vivo (4 of 4 [100%]). No difference was found in the mean variation of the average of the auditory brainstem response thresholds between the three groups in the in vivo ototoxicity study, thus confirming its safety profile regarding auditory function. MI503 does not demonstrate any deleterious effects on murine hair cells when assessed by immunostaining. These findings demonstrate an encouraging safety profile for the use of menin-MLL inhibitor for the conservative treatment of cholesteatoma.

Downregulation of Calreticulin and Annexin A2 Expression in Acquired Middle Ear Cholesteatoma by 2-DE Analysis.

Many factors are thought to be associated with the development of cholesteatoma, while the mechanisms of its formation remain unclear. This study aimed to identify the potential mechanisms of the proliferation and growth of cholesteatoma by analysis of the differential expressions of proteins in cholesteatoma and retroauricular skin tissue collected from the same patients. The present study is a retrospective study performed in an academic medical center. Comparative proteomics analyses using two-dimensional gel electrophoresis (2-DE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS), in addition to immunohistochemical analysis, were conducted to identify differentially-expressed proteins in cholesteatoma tissue as compared with retroauricular skin tissue. Western blotting was also employed to verify the expression patterns of the specific proteins identified by 2-DE and to measure the changes in potential modulators related to cholesteatoma proliferation and growth. Calreticulin (CRT) and annexin A2 (AnxA2) were identified as being differentially-expressed in cholesteatoma by 2-DE and LC-MS/MS, the results of which were in agreement with the results of immunohistochemical analysis and western blotting. Downregulation of CRT and AnxA2 were observed in cholesteatoma. Our data suggests that CRT and AnxA2 downregulation are seen in cholesteatoma compared to retroauricular skin. We speculate that the reduced expression of CRT and the persistent inflammatory response play important roles in the epithelial proliferation of cholesteatoma.

Non-coding RNA and cholesteatoma.

ObjectiveCholesteatoma is a challenging chronic pathology of the middle ear for which pharmacologic therapies have not been developed yet. Cholesteatoma occurrence depends on the interplay between genetic and environmental factors while master regulators orchestrating disease progression are still unknown. Therefore, in this review, we will discuss the diagnostic and therapeutic potential of non‐coding RNAs (ncRNA) as a new class of regulatory molecules.MethodsWe have comprehensively reviewed all articles investigating ncRNAs, specifically micro RNAs (miRNAs) and long ncRNAs (lncRNA/circRNA) in cholesteatoma tissue.ResultsCandidate miRNA approaches indicated that miR‐21 and let‐7a are the major miRNAs involved in cholesteatoma growth, migration, proliferation, bone destruction, and apoptosis. Regulatory potential for the same biological processes was also observed for miR‐203a. The NF‐kB/miR‐802/PTEN regulatory network was in relation to observed miR‐21 activity in cholesteatoma as well. High throughput approaches revealed additional ncRNAs implicated in cholesteatoma pathology. Competitive endogenous RNA (ceRNA) analysis highlighted lncRNA/circRNA that could be “endogenous sponge” for miR‐21 and let‐7a based on the hypothesis that RNA transcripts can communicate with and regulate each other by using shared miRNA response elements.ConclusionIn this review, we summarize the discoveries and role of ncRNA in major pathways in cholesteatoma and highlight the potential of miRNA‐based therapeutics in the treatment of cholesteatoma. Level of Evidence: NA.

Complex Evaluation of Tissue Factors in Pediatric Cholesteatoma.

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.

On the hearing effects of a cholesteatoma growing: A biomechanical study.

Chronic otitis media enables the appearance of a benign middle ear tumor, known as a cholesteatoma, that may compromise hearing. To evaluate the influence of a cholesteatoma growth on the hearing function, a computational middle ear model based on the finite element method was used and three different size of cholesteatoma were modeled. The cholesteatoma solidification and the consequent degradation of the ossicles were also simulated as two condition that commonly occurs during cholesteatoma evolution. A sound pressure level of 80 dB SPL was applied in the tympanic membrane and a steady state analysis was performed for frequencies from 100 Hz to 10 kHz. The displacements of both the tympanic membrane and the stapes footplate were measured. The results were compared with a healthy case and it was shown that the cholesteatoma development leads to a decrease in the umbo and stapes displacements. The ossicles degradation simulation showed the higher difference comparing with the cholesteatoma in an initial stage, with lower displacements in the stapes footplate mainly for high frequencies. The observed displacement differences are directly connected to hearing loss, being possible to conclude that cholesteatoma evolution in the middle ear will lead to hearing problems, mainly in an advanced stage.

Hsa_circ_0074491 regulates the malignance of cholesteatoma keratinocytes by modulating the PI3K/Akt pathway by binding to miR-22-3p and miR-125a-5p: An observational study.

Cholesteatoma is a benign cystic lesion that can continue to grow like a tumor. Circular ribonucleic acid (RNA) hsa_circ_0074491 (circ_0074491) has been reported to be down-regulated in cholesteatoma tissues. However, the role and regulatory mechanism of circ_0074491 in the growth of cholesteatoma are unclear.The expression of circ_0074491, microRNA (miR)-22-3p, and miR-125a-5p in cholesteatoma tissues was detected by quantitative real-time polymerase chain reaction. The proliferation, cell cycle, apoptosis, migration, and invasion of cholesteatoma keratinocytes were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, plate clone, flow cytometry, or transwell assays. Several protein levels were examined by western blotting. The targeting relationship between miR-22-3p or miR-125a-5p and circ_0074491 was verified via dual-luciferase reporter and RNA pull-down assays.We observed the downregulation of circ_0074491 in cholesteatoma tissues. Furthermore, circ_0074491 knockdown facilitated cell proliferation, migration, invasion, and repressed cell apoptosis in cholesteatoma keratinocytes. Circ_0074491 was verified as a decoy for miR-22-3p and miR-125a-5p in cholesteatoma keratinocytes. Both miR-22-3p and miR-125a-5p silencing reversed the impacts of circ_0074491 silencing on proliferation, apoptosis, migration, and invasion of cholesteatoma keratinocytes. Also, circ_0074491 knockdown activated the PI3K/Akt pathway in cholesteatoma keratinocytes via miR-22-3p and miR-125a-5p.Circ_0074491 played a suppressive role in cholesteatoma through inactivating the PI3K/Akt pathway via binding to miR-22-3p and miR-125a-5p, which provided a novel evidence for the involvement of circRNA in the development of cholesteatoma.

Immunomodulation in Cholesteatoma

The etiopathogenesis of chronic otitis media epitympanalis/cholesteatoma and its proliferative destructive course with possible complications such as destruction of bony structures with hearing loss, vestibular dysfunction, facial nerve paralysis and intracranial complications are still unexplained. Surgery is still the way to go. New studies are increasingly looking at the innate immune system. Our studies were carried out in a mouse model in WT mice and immundeficient KO-mice, as well as in cholesteatoma and healthy ear canal skin and middle ear tissue, which was removed during ear surgery. The expression analyses were carried out at the gene and protein level using TNF as the major target for therapy evaluation. By means of TUNEL staining and immunohistochemistry the level of apoptosis was evaluated. The uncontrolled undirected cholesteatoma growth shows an immunomodulatory profile with up and down-regulation of various gene networks, especially those involved in TNF downstream and upstream signaling pathways. TNF in cholesteatoma is modulated both inflammatorily and apoptotically and therefore is suitable as a possible therapeutic approach in various models. Cholesteatoma might be immunomodulatory regulated.

The Use of Serial Non EPI DWI MRI Scans to Determine the Growth of Cholesteatoma

Introduction It is an established practice to use non-EPI DWI MRI scans to detect the presence of cholesteatoma post operatively. In the present era of Covid-19 where routine surgery to remove cholesteatoma has been suspended resulting in potentially unprecedented demands on the service, a review of serial MRI scans performed over a 7 year period was undertaken to determine the rate of growth of cholesteatoma. Materials and Methods A retrospective longitudinal study identified 24 middle ear cholesteatomas in 17 patients with serial non-EPI DWI MRI scans (having excluded those having surgical intervention between scans) for a median period of 33 months (range of 6-91 months). Cholesteatomas were measured by the first author and by the consultant radiologist. Results Of 24 cholesteatomas, 1 resolved completely, 5 reduced, 6 stayed the same size, 4 grew slowly and 8 grew significantly. Conclusion Non-EPI DWI MRI scans to determine cholesteatoma growth in asymptomatic ears is useful in triaging patients in the Covid-19 era.

Menin-MLL inhibitor blocks progression of middle ear cholesteatoma in vivo

ObjectiveCholesteatoma is an epithelial lesion that expands into the middle ear, resulting in bone destruction. The acceleration of the proliferative activity of epithelial stem/progenitor cells is involved in the pathogenesis of cholesteatoma. Recently, the use of a menin-mixed lineage leukemia 1 (MLL1) inhibitor, MI503, in experiments has resulted in inhibition of the growth of tumors under histone modification. In this study, we investigated the effects of the menin-MLL inhibitor against cholesteatoma growth in an in vivo model. MethodsWe first correlated the expression level of histone H3 trimethylation at lysine 4 (H3K4me3) among cholesteatoma cases, chronic otitis media cases and normal skin tissues. Based on the role of keratinocyte growth factor (KGF) in the development of cholesteatoma, KGF-expression vector was transfected into the ear and we analyzed the expression level of H3K4me3. After cholesteatoma was induced, MI503 was administered daily into the ear for 14 days. ResultsWe detected the highest labeling index of H3K4me3 in the cholesteatoma specimens. After KGF-expression vector transfection in the mouse ear, a high expression level of H3K4me3 was observed in the epithelial layers. The use of MI503 reduced cholesteatoma in the in vivo model and decreased the proliferation of epithelial stem/progenitor cells in a dose-dependent manner. ConclusionWe demonstrated that inhibition of the menin-MLL interaction may be a potentially useful strategy in the conservative treatment of cholesteatoma.

Low expression of microRNA-125b enhances the expression of STAT3 and contributes to cholesteatoma growth.

MicroRNA-125b has been found to be down-regulated in many types of malignant tumours and diseases with excessive proliferation of keratinocytes, such as cutaneous squamous cell carcinoma and psoriasis. Cholesteatoma, which is mainly composed of keratinocytes, also has characteristics of abnormal proliferation similar to a malignant tumour. However, the expression and regulatory mechanisms of miR-125b and its downstream genes in cholesteatoma have not been clarified. Real time fluorescence quantitative PCR was applied to detect the expression of miR-125b in the cholesteatoma and corresponding retroauricular skin. Immunohistochemical staining and western blot were used to detect signal transducers and activators of transcription 3 (STAT3) and the downstream gene cyclin D1, survivin, and vascular endothelial growth factor (VEGF) in the cholesteatoma and corresponding retroauricular skin. The targeted regulatory relationship between miR-125b and STAT3 was confirmed by dual luciferase reporter assay. Proliferation and apoptosis of transfected HaCaT cells were detected by MTS, cell cycle, and apoptosis assays. We observed down-regulation of miR-125b and up-regulation of STAT3, cyclin D1, survivin, and VEGF in cholesteatoma tissues. STAT3 was a direct target gene of miR-125b. Inhibition of miR-125b enhanced STAT3 and its downstream genes expression, promoted HaCaT cell proliferation, and inhibited apoptosis. The results of this study demonstrate that miR-125b can influence the growth of cholesteatoma by targeting STAT3 and its downstream genes, including cyclin D1, survivin, and VEGF, thus providing an opportunity to establish new medical therapy strategies and facilitating further study of the pathogenesis of cholesteatoma.

Growth and Late Detection of Post-Operative Cholesteatoma on Long Term Follow-Up With Diffusion Weighted Magnetic Resonance Imaging (DWI MRI): A Retrospective Analysis From a Single UK Centre.

To evaluate the growth rate and late detection of residual cholesteatoma on long-term follow-up with diffusion weighted magnetic resonance imaging (DWI MRI) in clinically stable ears following definitive surgery, in order to define surveillance imaging protocols. Retrospective case review. Tertiary referral center. Patients who underwent DWI MRI at our institution between February 2007 and May 2013 for postoperative cholesteatoma follow-up. Non-echo planar imaging (non-EPI) Half-Fourier Acquisition Single-shot Turbo spin Echo (HASTE) DWI MRI. Intervals between the definitive surgery and the first and subsequent DWI MRI, the maximum coronal dimension of the lesion on DWI and length of follow-up. The study evaluated 152 postoperative DWI studies performed for 88 patients. In 12 cases, DWI was initially negative but became positive on repeat imaging after a mean interval of 3.8 years from the initial cholesteatoma surgery (median 3.7 years, range 1.6-7.9). Of these, 3/12 had more than one negative/indeterminate DWI before disease was eventually detected on imaging; in this subgroup, the mean interval between surgery and positive DWI was 3.2 years (median 2.6 years, range 2.3-4.2). 39 DWI positive foci with serial imaging demonstrated a mean growth rate of 4 mm/year (median 2 mm, range 0-18). After negative initial DWI, it is proposed that interval imaging should be considered for a minimum of 5 years in stable ears following definitive cholesteatoma surgery. In view of the marked variability in growth rate, an additional interval scan between 2 and 3 years postoperatively is indicated.

Measuring growth of residual cholesteatoma in subtotal petrosectomy

Background: Little is known about the growth rate of cholesteatoma in patients.Objective: Investigate the growth of residual cholesteatoma in subtotal petrosectomy based on volume measured in MRI scans.Materials and methods: Retrospective case series in a Tertiary Medical Centre. Thirteen residual cholesteatomas were identified in 10 patients after subtotal petrosectomy for which a wait-and-scan policy was adopted. Volume of the residual cholesteatoma was calculated by manual segmentation as well as the ‘box method’.Results: Mean growth rate was 27.9 mm3/month (SD 22.8), with a large individual variation ranging from 2.2 to 69.8 mm3/month. No complications were reported in 10 patients with a wait-and-scan policy for residual cholesteatoma in subtotal petrosectomy. The box method overestimates growth rate compared to the reference method manual segmentation and a linear increase of this systematic error was seen with increasing size of the cholesteatoma.Conclusions: Residual cholesteatoma growth rate shows a large individual variation. A wait-and-scan policy could be considered in case of a (small) residual in subtotal petrosectomy with ample room to grow before destroying any remaining structures. Furthermore, the clinically more applicable and less time-consuming box method can be used to accurately measure volumes of small cholesteatomasup to a volume of 500 mm3.

Identification of risk factors for residual cholesteatoma in children and adults: a retrospective study on 110 cases of revision surgery

IntroductionResidual disease after cholesteatoma removal is still a challenge for the otorhinolaryngologist. Scheduled “second-look” surgery and, more recently, radiological screenings are used to identify residual cholesteatoma as early as possible. However, these procedures are cost-intensive and are accompanied by discomfort and risks for the patient. ObjectiveTo identify anamnestic, clinical, and surgery-related risk factors for residual cholesteatoma. MethodsThe charts of 108 patients, including children as well as adults, having undergone a second-look or revision surgery after initial cholesteatoma removal at a tertiary referral hospital, were analyzed retrospectively. ResultsGender, age, mastoid pneumatization, prior ventilation tube insertion, congenital cholesteatoma, erosion of ossicles, atticotomy, resection of chorda tympani, different reconstruction materials, and postoperative otorrhea did not emerge as statistically significant risk factors for residual disease. However, prior adenoid removal, cholesteatoma growth to the sinus tympani and to the antrum and mastoid, canal-wall-up 2 ways approach, and postoperative retraction and perforation were associated with a statistically higher rate of residual disease. A type A tympanogram as well as canal-wall-down plus reconstruction 2 ways approach for extended epitympanic and for extended epitympanic and mesotympanic cholesteatomas were associated with statistically lower rates of residual disease. A score including the postoperative retraction or perforation of the tympanic membrane, the quality of the postoperative tympanogram and the intraoperative extension of the cholesteatoma to the sinus tympani and/or the antrum was elaborated and proved to be suitable for predicting residual cholesteatoma with acceptable sensitivity and high specificity. ConclusionCholesteatoma extension to the sinus tympani, antrum and mastoid makes a residual disease more likely. The canal-wall-down plus reconstruction 2 ways approach seems safe with similar rates of residual cholesteatoma and without the known disadvantages of canal-wall-down surgery. The described score can be useful for identifying patients who need a postoperative radiological control and a second-look surgery.

Cholesteatoma labyrinthine fistula: prevalence and impact

IntroductionLabyrinthine fistula is one of the most common complications associated with cholesteatoma. It represents an erosive loss of the endochondral bone overlying the labyrinth. Reasons for cholesteatoma-induced labyrinthine fistula are still poorly understood. ObjectiveEvaluate patients with cholesteatoma, in order to identify possible risk factors or clinical findings associated with labyrinthine fistula. Secondary objectives were to determine the prevalence of labyrinthine fistula in the study cohort, to analyze the role of computed tomography and to describe the hearing results after surgery. MethodsThis retrospective cohort study included patients with an acquired middle ear cholesteatoma in at least one ear with no prior surgery, who underwent audiometry and tomographic examination of the ears or surgery at our institution. Hearing results after surgery were analyzed according to the labyrinthine fistula classification and the employed technique. ResultsWe analyzed a total of 333 patients, of which 9 (2.7%) had labyrinthine fistula in the lateral semicircular canal. In 8 patients, the fistula was first identified on image studies and confirmed at surgery. In patients with posterior epitympanic and two-route cholesteatomas, the prevalence was 5.0%; and in cases with remaining cholesteatoma growth patterns, the prevalence was 0.6% (p=0.16). In addition, the prevalence ratio for labyrinthine fistula between patients with and without vertigo was 2.1. Of patients without sensorineural hearing loss before surgery, 80.0% remained with the same bone conduction thresholds, whereas 20.0% progressed to profound hearing loss. Of patients with sensorineural hearing loss before surgery, 33.33% remained with the same hearing impairment, whereas 33.33% showed improvement of the bone conduction thresholds’ Pure Tone Average. ConclusionLabyrinthine fistula must be ruled out prior to ear surgery, particularly in cases of posterior epitympanic or two-route cholesteatoma. Computed tomography is a good diagnostic modality for lateral semicircular canal fistula. Sensorineural hearing loss can occur post-surgically, even in previously unaffected patients despite the technique employed.

Downregulation of MiR-203a Disinhibits Bmi1 and Promotes Growth and Proliferation of Keratinocytes in Cholesteatoma.

Background: Keratinocytes are the predominant cell type in a cholesteatoma, and microRNA (miR)-203a has been shown to be essential for the growth and differentiation of keratinocytes. The regulatory mechanisms of miR-203a and Bmi1—the predicted target of miR-203a that is associated with cholesteatoma—have not been clarified.Methods: Real-time PCR and western blot were carried out for the detection of miRNAs, mRNAs, and proteins, including miR-203a, Bmi1, and phosphorylated (p-)Akt. Immunohistochemical staining was applied to observe the expression and distribution of Bmi1 and of p-Akt in cholesteatoma and in control retroauricular skin. The dual luciferase reporter assay was used to analyze the relationship between miR-203a and Bmi1. Ectopic miR-203a and Bmi1 were transfected into an immortalized line of human keratinocytes (HaCaT cells), and the roles of these molecules in cell proliferation, apoptosis, and migration were explored.Results: Cholesteatoma tissues were characterized by downregulation of miR-203a and concomitant upregulation of Bmi1. Results of the dual-luciferase reporter assay indicated that Bmi1 was a direct target gene of miR-203a. Silencing of miR-203a increased Bmi1 expression; promoted proliferation, colony formation, and migration of HaCaT cells; and inhibited apoptosis. Moreover, p-Akt was significantly increased in cholesteatoma tissues and was positively correlated with Bmi1. Suppression of Bmi1 reduced p-Akt expression in HaCaT cells; subsequent inhibition of miR-203a reversed this phenomenon.Conclusions: Our results reveal that miR-203a may regulate cholesteatoma growth and proliferation by targeting Bmi1. These findings provide insight for the development of novel nonsurgical options for cholesteatoma.

The Role of Tympanic Membrane Retractions in Cholesteatoma Pathogenesis.

Objective To analyze the contralateral ear (CLE) of patients with cholesteatoma and to correlate the cholesteatoma growth pattern in the affected ear with the findings in the CLE. Methods Videotoscopy of both ears in 432 patients with cholesteatomas classified as posterior epitympanic (PEC), posterior mesotympanic (PMC), two routes, or undetermined. Tympanic membrane (TM) retractions were classified by location and severity and TM perforations according to signs of previous TM retraction. Results TM retraction was the most prevalent alteration in the CLE (42.6%). Cholesteatoma was observed in 17.4%. In patients with PEC, the retraction in the CLE was more frequent in the PF (66.7%) than in the PT (1.4%), and in those with two-route cholesteatoma, the retraction in the CLE most frequently involved both the PT and PF (65.6%; p < 0.0001). Conclusion Our results confirm the essential role of TM retraction at least in the earlier phases of cholesteatoma pathogenesis.

Monitoring Progression of 12 Cases of Non-Operated Middle Ear Cholesteatoma With Non-Echoplanar Diffusion Weighted Magnetic Resonance Imaging: Our Experience.

The aim of this study is to gain insight into the disease progression and behavior of primary cholesteatoma in a cohort of patients who did not have surgery using non-echoplanar diffusion-weighted magnetic resonance imaging (DW MRI) serial monitoring. Retrospective longitudinal observational study of 12 cases of middle ear cleft cholesteatoma diagnosed between 2009 and 2014 where surgery was not performed for various reasons. All cases were monitored radiologically with non-echoplanar half-Fourier acquisition single-shot turbo spin-echo diffusion weighted imaging annually for a median period of 23 months (between 11 and 45 mo) to evaluate for changes in disease volume and direction of growth. Of the 12 cases, there was one outlier where the cholesteatoma growth was disproportionately high compared with the rest of the cases outside the standard deviation range. A third of the cases had radiological evidence of cholesteatoma growth. The mean growth was about 11.9% of the initial disease volume per year. Seven out of the 12 cases had radiological evidence of cholesteatoma regression in terms of size, with three cases having negative follow-up DW-MRI scans as early as 17 months. The mean regression rate was much higher than the mean growth rate at 54.3% of the initial disease volume per year. The direction of greatest growth is craniocaudally. Within the limits of our longitudinal study, we have shown that by monitoring with non-echoplanar diffusion weighted imaging, cholesteatoma can progress or regress when left untreated by surgery. The greatest progression was recorded in the craniocaudal direction.