Simple, low-cost, and well-performing method, the outgrowth technique, for the isolation of cells from nasal polyps.

BACKGROUND: Epithelial cells are an important part of the pathomechanism in chronic rhinosinusitis with nasal polyps. It is therefore essential to establish a robust method for the isolation and culture of epithelial cells from nasal polyps to enable further research. In this study, the feasibility of the outgrowth technique for the isolation of the epithelial cells from the nasal polyps was evaluated. RESULTS: Using the outgrowth technique, epithelial cells could be isolated from all tissue samples. Isolated epithelial cells showed a proliferation rate of approximately 7- to 23-fold every 6 days up to the 3rd passage. Over 97% of isolated cells were shown to be cytokeratin- and p63-positive, and over 86% of them were Ki-67-positive in flow cytometry. Interleukin-33 and periostin were detectable in the supernatant. CONCLUSIONS: We introduce a simple, low-cost, and well-performing method for isolating epithelial cells from nasal polyps with the outgrowth technique.

Long-Term Cryopreservation of Nasal Polyp Tissue in a Biobank for the Isolation and Culture of Primary Epithelial Cells.

Epithelial cells may play an important role in the pathologic process of chronic rhinosinusitis with nasal polyps. Therefore, providing epithelial cells from a biobank could greatly contribute to further research. In the present work, the isolation of epithelial cells from long-term cryopreserved tissue is demonstrated. Polyp tissues were cryopreserved in a commercially available freezing medium with dimethyl sulfoxide and stored in liquid nitrogen. The outgrowth and proliferation of epithelial cells from cryopreserved tissue were evaluated and compared to that of fresh tissue. Flow cytometric analysis with anti-cytokeratin, anti-p63, and anti-Ki-67 was performed to identify epithelial cells and determine differentiation and proliferation. A functionality test was performed by determining type 2-relevant proteins, representatively thymic stromal lymphopoietin (TSLP) and periostin, using ELISA. Primary epithelial cells could be isolated from cryopreserved tissues. Cells from cryopreserved tissues showed comparable outgrowth and proliferation to that of fresh tissue. Isolated epithelial cells showed high cytokeratin, p63, and Ki-67 expression and secreted TSLP and periostin. In the present study, a method for long-term cryopreservation of polyp tissue was established, thereby enabling the isolation and cell culture of primary cell culture at a later time. Epithelial cell availability should be greatly improved by including this method in a biobank.

[Syphilis infection in the ENT area]. / Syphilisinfektion im HNO-Bereich.

The number of reported cases of syphilis has been increasing for years. The sexually transmitted disease is caused by the spirochete Treponema pallidum subspecies pallidum and progresses in different stages. Symptoms in the ENT area can occur in all stages. This means that a syphilis infection should always be considered by the ENT doctor as a differential diagnosis if the symptoms are suitable. Thus, with increasing oral sexual intercourse, the primary effect/hard chancre is more often observed in the oral cavity. In addition, symptoms can occur not only in the oral cavity, but also in the ear, nose, larynx, cervical and facial regions. The diagnosis is confirmed by direct pathogen detection or by serological detection. The spirochete cannot be cultivated. The therapeutic gold standard is the administration of benzathine penicillin G or procaine penicillin G. Doxycycline, macrolides or ceftriaxone are available as alternatives. In case of inner ear or cranial nerve involvement, the additional administration of a glucocorticoid is recommended. Before antibiotic therapy is administered, the patient must be informed about the possibility of a Jarisch-Herxheimer reaction. In cases of a syphilis infection, a collegial cooperation with the venereologists is always recommended so that an effective and comprehensive diagnosis and therapy can be carried out.

Incidence of Inner Ear Disorders in Various Forms of Acute Otitis Media in ENT Practices in Germany.

OBJECTIVE: The aim of this study was to determine the incidence of inner ear involvement in various forms of acute otitis media (AOM) in ear, nose, and throat (ENT) practices in Germany. METHODS: Patients who had been diagnosed with various forms of AOM in the years 2010 to 2017 were enrolled in the study from a nationwide, representative practice database (Disease Analyzer, IQVIA). In these patients, the incidence of simultaneous or subsequent inner ear disorders (IED) was determined within 7 days and within 12 months from the date of an AOM diagnosis. RESULTS: A total of 286 186 patients with AOM were enrolled. The most frequent diagnoses were "nonsuppurative otitis media, unspecified" (47.6%) and "otitis media, unspecified" (39.0%). The diagnoses of hemorrhagic bullous myringitis (BM) or influenza-induced AOM were very rarely found in the database. The highest incidence of IED after 7 days and 12 months was found in "nonsuppurative otitis media, unspecified" (7.7% and 15.9%, respectively), followed by "otitis media, unspecified" (5.6% and 13.5%, respectively). The incidences of the most frequent IED "hearing loss, unspecified" and "sensorineural hearing loss, unspecified" increased proportionally with increasing patient age, while the rare diagnoses of "labyrinthitis" and "ototoxic hearing loss" were evenly distributed among the age groups. CONCLUSION: In ENT practices in Germany, both various forms of AOM, as well as simultaneous or subsequent IED, are mostly being coded in an unspecific way, while specific forms such as hemorrhagic BM, influenza-induced AOM, and labyrinthitis are coded very rarely. Older patients have a higher risk of IED in AOM. A visit due to AOM seems to be a regular occasion for the initial diagnosis of hearing impairment in the elderly individuals. The highest risk of IED was found in nonsuppurative AOM.

Chronic rhinosinusitis: assessment of changes in nociceptive neurons.

BACKGROUND: Pain is a major symptom of chronic rhinosinusitis (CRS). It is mainly associated with CRS without nasal polyps (CRSsNP) and has a major impact in the decision to move on to surgery. Patients with CRS with nasal polyps (CRSwNP) are characterized by trigeminal hypoesthesia and suffer from less pain. The aim of this study was to investigate whether CRS induces alterations in the peripheral nociceptive neurons, mainly focusing on quantitative changes. METHODS: Sinus mucosa and inferior turbinate (IT) samples were obtained from patients with CRS, and IT tissue of healthy patients served as controls. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was performed for neuronal markers including CNTNAP2, FAM19A1, GFRA2, NEFH, NTRK1, PLXNC1, RET, SCN10A, SCN11A, TRPV1, and PGP 9.5; enzyme-linked immunosorbent assay (ELISA) was performed for KCNK18, SCN10A, MRGPRD, and MAP2. For PGP 9.5, immunohistochemistry was additionally used to analyze tissue slides. RESULTS: We included 35 patients with CRSsNP, 47 patients with CRSwNP, and 18 control patients. No differences in expression of the neuronal markers were observed between CRSsNP, CRSwNP, and controls. SCN10A was the only marker exclusively expressed on nociceptive neurons in sinus tissue. No histological difference in nerve fibers was observed between sinus mucosa of both phenotypes. CONCLUSION: Our results indicate that the nociceptive nerve density in CRSwNP is not lower than in CRSsNP, as was assumed previously. The nociceptive neurons in sinonasal mucosa cannot be classified into subtypes due to the lack of specificity of the respective marker genes. Our findings question the generally accepted claim that nasal polyp tissue does not contain any nerves.

Incidence and Risk of Various Disorders of the External Ear in Patients With Hearing Aids Treated in ENT Practices in Germany.

OBJECTIVE: The aim of the study was to investigate the relationship between the presence of a hearing aid (HA) and the occurrence of various disorders of the external ear, using data from ear, nose, and throat (ENT) practices in Germany from a nationwide, representative practice database. METHODS: In the period between 2012 and 2016, the incidences of disorders of the external ear (ICD-10 groups H60 and H61) in patients with HA prescriptions were compared retrospectively with a control group without HA prescriptions (1:1 matching). RESULTS: A total of 20,127 patients with HA prescriptions were compared with 20,127 controls. The highest 12-month incidences (HA vs. controls) were determined for "Impacted cerumen" (H61.2) (16.5% vs. 4.2%), "Unspecified otitis externa" (H60.9) (2.6% vs. 1.2%) and 'Acute noninfective otitis externa' (H60.5) (2.3% vs. 0.7%). The most significant risk increases for HA wearers were found for "Abscess of external ear' (H60.0, OR 10.03), "Other otitis externa' (H60.8, OR 6.00), and "Impacted cerumen' (H61.2, OR 4.55). A smaller risk increase was found for "Cholesteatoma of external ear' (H60.4, OR 2.26), among others. CONCLUSION: In HA wearers, the risk of developing almost all of the diagnoses reviewed is significantly increased, especially for external auditory canal (EAC) furuncle, chronic otitis externa, and impacted cerumen. This study provides the first epidemiological evidence for HA as a risk factor for the rare EAC cholesteatoma.

Demographic data and seasonal variation in peripheral vestibular disorders in ENT practices in Germany.

AIM: The aim of this study was to analyze the demographic data pertaining to and seasonal variation in specific vestibular disorders in ear, nose, and throat (ENT) practices in Germany based on data from a representative nationwide practice database. METHOD: The study sample included patients from 116 ENT practices in Germany who received an initial diagnosis (ICD-10 code) of Meniere's disease (MD, H81.0), benign paroxysmal positioning vertigo (BPPV, H81.1), or vestibular neuronitis (VN, H81.2) between January 2014 and December 2016. Collected parameters included age, sex, and month of diagnosis. Seasonal variation was analyzed for younger vs. older patients (≤50 vs. >50 years of age). Two univariate Poisson regression models were fitted to estimate the association between the month of diagnosis and the number of diagnosed patients per practice. RESULTS: A total of 20,720 patients were available for analysis. The average case numbers for MD, BPPV, and VN were 0.8 patients, 2.7 patients, and 1.5 patients per practice per month, respectively. The mean ages of female vs. male patients were 55 and 56 years (MD), 59 and 60 years (BPPV), and 58 and 57 years (VN), respectively. The proportions of female patients with these diagnoses were 62%, 70%, and 61%, respectively. All diagnoses were evenly distributed throughout the years in all age groups. No seasonal variation was observed. CONCLUSIONS: The demographic data of MD and BPPV patients are comparable to those found by previous large-scale epidemiologic studies. However, no seasonal variation was demonstrated for any vestibular disorder in this large sample.

Diagnoses of Dizziness- and Vertigo-related Disorders in ENT Practices in Germany.

AIM: The aim of this study was to analyze the prevalence of dizziness- and vertigo-related diagnoses in ear, nose, and throat (ENT) practices in Germany and the associated demographic characteristics based on data from a representative nationwide practice database. METHOD: The study sample included patients from 138 ENT practices in Germany who received dizziness- and vertigorelated diagnoses (ICD-10 code) between January 2012 and December 2015. Collected parameters included age, sex, insurance status, prescribed medication (anatomical therapeutic chemical [ATC] class), and referrals to other specialists and hospitals. RESULTS: A total of 107,458 patients were available for analysis. Most common diagnoses were "dizziness and giddiness" (67.7%), "benign paroxysmal vertigo" (10.2%) and "disorder of vestibular function, unspecified" (7.2%). Referrals and admissions were made in 12.8%, mostly to radiologists (7.7%), followed by neurologists (3.7%), and hospitals (1.4%). Most referrals were made for unspecific diagnoses and for "vestibular neuronitis." The rate of medical prescriptions was 37.3%, with the most common prescription being for antivertigo preparations. CONCLUSIONS: Dizziness- and vertigo-related disorders are frequently diagnosed in ENT practices in Germany. The majority of these diagnoses are unspecific and lead to an increased rate of referrals and hospital admissions. The medical prescription rate, especially of antivertigo preparations, was high, even among patients with benign paroxysmal positioning vertigo. This study reflects a mostly pragmatic approach to a complex diagnostic and therapeutic challenge in daily ENT practice as well as the limited ability of the ICD-10 system to classify the underlying etiology.

Current healthcare pathways in the treatment of rhinosinusitis in Germany.

BACKGROUND: In the outpatient setting in Germany, patients with rhinosinusitis usually present at general practices (GP) or ear, nose, and throat practices (ENT) for initial diagnosis and treatment. OBJECTIVES: The aim of this study was to analyze the referral patterns of rhinosinusitis patients in GPs and ENT practices in Germany, with respect to existing recommendations. MATERIAL AND METHODS: The study sample included patients from 940 GP and 106 ENT practices from Disease Analyzer database (IQVIA) who received an acute sinusitis (AS) or chronic sinusitis (CS), or nasal polyp (NP) in 2015. RESULTS: The total numbers of patients in GP versus ENT practices were 24,648 versus 12,095 (AS), 26,768 versus 19,826 (CS), and 516 versus 1773 patients (NP). Referrals to ENT practices were made by GP in 12.3% (AS), 14.8% (CS), and 40.5% (NP). The percentages of patients in GP versus ENT practices with subsequent hospital admissions were 6.9 versus 3.3% (AS), 6.3 versus 6.5% (CS), and 9.5 versus 13.8% (NP), respectively. CONCLUSIONS: Although 40% of patients with NP who consult GPs are referred to ENT practices, it remains unclear how the other 60% are being treated. The hospital admission rates of patients with CS as well as of patients with NP were found to be surprisingly low.

Hearing threshold estimation by auditory steady-state responses with narrow-band chirps and adaptive stimulus patterns: implementation in clinical routine.

Objective hearing threshold estimation by auditory steady-state responses (ASSR) can be accelerated by the use of narrow-band chirps and adaptive stimulus patterns. This modification has been examined in only a few clinical studies. In this study, clinical data is validated and extended, and the applicability of the method in audiological diagnostics routine is examined. In 60 patients (normal hearing and hearing impaired), ASSR and pure tone audiometry (PTA) thresholds were compared. ASSR were evoked by binaural multi-frequent narrow-band chirps with adaptive stimulus patterns. The precision and required testing time for hearing threshold estimation were determined. The average differences between ASSR and PTA thresholds were 18, 12, 17 and 19 dB for normal hearing (PTA ≤ 20 dB) and 5, 9, 9 and 11 dB for hearing impaired (PTA > 20 dB) at the frequencies of 500, 1,000, 2,000 and 4,000 Hz, respectively, and the differences were significant in all frequencies with the exception of 1 kHz. Correlation coefficients between ASSR and PTA thresholds were 0.36, 0.47, 0.54 and 0.51 for normal hearing and 0.73, 0.74, 0.72 and 0.71 for hearing impaired at 500, 1,000, 2,000 and 4,000 Hz, respectively. Mean ASSR testing time was 33 ± 8 min. In conclusion, auditory steady-state responses with narrow-band-chirps and adaptive stimulus patterns is an efficient method for objective frequency-specific hearing threshold estimation. Precision of threshold estimation is most limited for slighter hearing loss at 500 Hz. The required testing time is acceptable for the application in everyday clinical routine.

Impact of endolymphatic mastoid shunt surgery on saccule and lateral semicircular canal function.

Endolymphatic mastoid shunt surgery (EMSS) is widely performed in patients with medically intractable Meniere's disease. Although many patients report an improvement of symptoms after surgery, the mechanisms which are responsible for the relief of complaints are not known. To date, only few studies exist which studied the influence of EMSS on vestibular function. The present study examines the effect of EMSS on saccule function by measuring vestibular evoked myogenic potentials and the effect on lateral semicircular canal function by sinusoidal harmonic acceleration (SHA) testing. No changes in vestibulo-collic reflexes were found after surgery compared to before surgery. SHA testing resulted in comparable phase lag and gain pre- and postoperatively. Although central compensation is clinically evident no effect in specific vestibular diagnostic testing is seen. Modulations of canal-otolith interaction might suggest a change of symptoms. The only method so far to evaluate the success of EMSS is the patient's subjective assessment.

Meniere's disease and middle ear pressure: vestibular function after transtympanic tube placement.

CONCLUSION: Selected patients with Meniere's disease (MD) show an improvement of disability from vertigo after transtympanic ventilation tube insertion, although an effect on vestibular function is not seen if ipsilateral middle ear pressure lower than -50 daPa is used as a selection criterion. OBJECTIVES: Transtympanic ventilation tube insertion as a treatment option for MD has been reported but its results have been controversial. So far, no investigations on vestibular function in patients with MD after tube insertion have been carried out. SUBJECTS AND METHODS: Twenty-two patients with unilateral MD who were intractable to medical treatment and who had an ipsilateral middle ear pressure lower than -50 daPa received a transtympanic ventilation tube. Vestibular evoked myogenic potentials (VEMPs) and sinusoidal harmonic acceleration (SHA) testing were recorded pre- and postoperatively and were compared. RESULTS: Most patients (68.2%) reported an improvement of vertigo. Before surgery 63.6% of patients did not show VEMPs, whereas vestibulo-collic reflexes were measured in 36.4% of all cases compared with 68.2% of patients without and 31.8% with recorded VEMPs after surgery. No statistically different findings in gain and phase lag of SHA testing were seen postoperatively compared to preoperative findings.