Endolymphatic hydrops (EH) represents a histopathologic finding in which the structures bounding the endolymphatic space are distended by an enlargement of endolymphatic volume. EH primarily involves the cochlear duct and saccule but can involve the utricle and ampullae of the semicircular canals. EH is a consistent finding in patients with Meniere's disease, however, the reverse is not true. EH may occur as a consequence of a variety of disorders, including DFNA 9, Alport syndrome, serous labyrinthitis, suppurative labyrinthitis, otosyphilis, temporal bone fracture, surgical trauma, neoplasm, immune disorders, otosclerosis, or Paget's disease. The mechanism of development of hydrops is also unclear. This review provides information to understand the recent pathophysiologic mechanism and causal disoders in EH.
Cochlear Duct ; Ear, Inner ; Edema ; Endolymphatic Hydrops ; Humans ; Immune System Diseases ; Labyrinthitis ; Meniere Disease ; Nephritis, Hereditary ; Otosclerosis ; Saccule and Utricle ; Semicircular Canals ; Temporal Bone

BACKGROUND AND OBJECTIVES: Although the role of aquaporin-2 (AQP2) in the kidney has been well defined, its role in the inner ear remains to be determined. The present study was to investigate the effect of water deprivation on the expression of AQP2 in the inner ear. MATERIALS AND METHOD: Healthy male guinea pigs weighing 250 g were used. The experimental group underwent water restriction and the control underwent water loading with sucrose-containing water for 3 days. Concentrations of plasma arginine-vasopressin (AVP) were determined and electrocochleography (ECoG) recordings were made. An RT-PCR, real-time PCR and Westernblotting analysis were used for quantitative analysis of AQP2 mRNA and AQP2 protein expression. Immunohistochemistry was also used to evaluate the distribution of AQP2 water channel proteins in the inner ear. RESULTS: AQP2 was mainly expressed in the epithelium of endolymphatic sac, spiral limbus, spiral ligament and stria vascularis of scala media. The concentrations of plasma AVP were 9.2+/- 0.8 pg/mL in the experimental group and 0.78+/-0.3 pg/mL in the control. The summation potential/ action potential (SP/AP) ratio in ECoG was markedly increased in the experimental group (0.55 in the experimental and 0.29 in the control). RT-PCR and real time PCR as well as Western blot analysis showed that the level of AQP2 mRNA and protein in the cochlea and endolymphactic sac of the water-deprived group was significantly higher than those in the control group. CONCLUSION: These data suggest that AQP2 is one of the important water channels in fluid homeostasis in the inner ear. Moreover, the volume of endolymphatic space can be increased via AVP-AQP2 system in response to water deprivation.
Action Potentials ; Animals ; Aquaporin 2 ; Aquaporins ; Arginine Vasopressin ; Audiometry, Evoked Response ; Blotting, Western ; Cochlea ; Cochlear Duct ; Ear, Inner ; Endolymphatic Hydrops ; Endolymphatic Sac ; Epithelium ; Guinea ; Guinea Pigs ; Homeostasis ; Humans ; Immunohistochemistry ; Kidney ; Male ; Plasma ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; Spiral Ligament of Cochlea ; Stria Vascularis ; Water Deprivation

OBJECTIVES: To investigate the otoprotective effects of mouse nerve growth factor (mNGF) in A/J mice. METHODS: The mice at postnatal day 7 (P7) were randomly separated into a mNGF treated group (mNGF group) and a distilled water (for injection) treated group (control group). The mNGF dissolved in distilled water or distilled water alone was given to the mice once every other day from P7 by intramuscular injection in the hips. The otoprotective effects of mNGF in A/J mice were observed in a time course manner. The thresholds of auditory-evoked brainstem response (ABR) were tested from the age of the 3rd to the 8th week. Sections of the inner ears were stained by hematoxylin and eosin, and spiral ganglion neurons (SGNs) were observed at the age of the 3rd, the 6th,and the 8th week. Counts of whole mount outer hair cells (OHCs) in the cochleae were made at the age of 8 weeks. Expression of apoptosis related genes was determined by quantitative real-time polymerase chain reaction and Western blotting. RESULTS: ABR thresholds of the mNGF group were significantly lower than those of the control group at the age of the 6th and the 8th week. Moreover, the mNGF preserved OHC and SGN in the mouse cochleae in this period. Further experiments showed that the expression of caspase genes (including caspase-3) was inhibited in the mouse inner ears in the mNGF group. CONCLUSION: The mNGF improves hearing in A/J mice by preserving SGN and OHC in the cochleae.
Animals ; Apoptosis ; Blotting, Western ; Brain Stem ; Cochlea ; Ear, Inner ; Eosine Yellowish-(YS) ; Hair Cells, Auditory, Outer ; Hearing* ; Hematoxylin ; Hip ; Injections, Intramuscular ; Mice* ; Nerve Growth Factor* ; Neurons ; Real-Time Polymerase Chain Reaction ; Spiral Ganglion ; Water

Circling mouse (C57BL/6J-cir/cir) deleted the transmembrane inner ear (Tmie) gene is an animal model for human non-syndromic recessive deafness, DFNB6. In circling mouse, hair cells in the cochlea have degenerated and hair bundles have become irregularity as time goes on. Tmie protein carries out a function of the mechanoelectrical transduction channel in cochlear hair cells. Myosin7a (MYO7A) protein has key roles in development of the cochlear hair bundles as well as in the function of cochlear hair cells. To find whether Tmie protein interacts with MYO7A proteins in the cochlea postnatal developmental stage, we investigated expression of the MYO7A proteins in the cochlear hair cells of circling mice by western blot analysis and whole mount immunofluorescence at postnatal day 5 (P5). The expression of MYO7A showed statistically significant increase in the cochlea of C57BL/6J-+/cir and C57BL/6J-cir/cir mice than that of C57BL/6J-+/+ mice. The MYO7A intensity of the cochlear hair cells also increased in C57BL/6J-+/cir and C57BL/6J-cir/cir mice compared with those of C57BL/6J-+/+ mice. Taken together, the results indicate that Tmie protein may have an important role with MYO7A protein in the development and maintenance of the stereociliary bundles during postnatal developmental stage of the cochlea.
Animals ; Blotting, Western ; Cochlea ; Deafness ; Ear, Inner ; Fluorescent Antibody Technique ; Hair ; Hair Cells, Auditory* ; Humans ; Mice* ; Models, Animal

BACKGROUND AND OBJECTIVES: Locacorten Vioform (Novartis UK) is frequently prescribed for otomycosis. Its component, Clioquinol, also has anti-bacterial properties. Up to this point, its ototoxic potential has not been evaluated. Our objective aims to evaluate Locacorten Vioform’s potential ototoxicity when applied directly to the middle ear cavity. MATERIALS AND METHODS: We performed an experimental prospective animal study in our animal research center with 20 Hartley guinea pigs divided into 2 groups. The first group (experimental) was treated with Locacorten Vioform in one ear and with a physiologic saline solution in the other. The second group (positive control) was treated with concentrated gentamycin in one ear and physiologic saline in the other. Auditory brainstem response measurements were obtained before and after three sets of injections. Statistics were analyzed using a variance analysis with repeated measures. The histological state of cochlear outer hair cells was compared between the two groups using scanning electron microscopy. RESULTS: Average hearing loss in ears treated with Locacorten Vioform was 32.1 dB, compared with a 2.5 dB average loss in the saline-treated ears. Ears treated with gentamycin lost an average of 33.0 dB. There were clinically and statistically significant differences between the two ears of the guinea pigs in both groups (p < 0.001). Scanning electron microscopy revealed severe pericochlear and cochlear inflammation and ossification in the Locacorten Vioform-treated ears. Gentamycin caused significant destruction of outer hair cell architecture. CONCLUSIONS: Locacorten Vioform induces a hearing loss similar to that caused by gentamycin when applied directly to the middle ear of a guinea pig model. Electron microscopy indicates a pericochlear and cochlear inflammatory reaction with ossification.
Animal Experimentation ; Animals ; Clioquinol ; Ear ; Ear, Middle ; Evoked Potentials, Auditory, Brain Stem ; Gentamicins ; Guinea Pigs ; Guinea ; Hair ; Hair Cells, Auditory, Inner ; Hair Cells, Auditory, Outer ; Hearing Loss ; Inflammation ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Otomycosis ; Prospective Studies ; Sodium Chloride

PURPOSE: To evaluate the CT and MRI findings of the large endolymphatic duct or sac syndrome (LEDS) and its associated anomalies, with clinical features. MATERIALS AND METHODS: We retrospectively reviewed the MR and CT images of 52 ears obtained from 26 patients with LEDS. We reviewed the clinical findings, audiology testing, and treatment results. The degree of hearing loss was classified from normal to profound, based on pure tone audiometry. The largest areas were measured at each endolymphatic duct and analyzed to determine whether a correlation exists with the degree of hearing loss. We also analyzed the differences in measurements between CT and MRI findings. RESULTS: All 26 patients had some degree of sensorineural hearing loss, which resulted in 18 ears to undergo a cochlear implantation. One patient was diagnosed with Cornelia de Lange syndrome. Five patients had a sudden hearing loss onset. Ten ears had incomplete cochlear partitions, whereas 28 ears had enlarged vestibules. All patients had severe to profound hearing loss. We found no statistical correlation between the size of the largest area of the endolymphatic duct and the degree of hearing loss. The mean area of the endolymphatic ducts, as per an MRI examination, revealed slightly greater areas than the CT findings, although the differences were not significant. CONCLUSION: Enlarged vestibules and incomplete partitions of the cochlea were common anomalies associated with LEDS. We found no statistical correlation between the largest area of the endolymphatic duct or sac with the degree of hearing loss.
Audiology ; Audiometry ; Cochlea ; Cochlear Implantation ; Cochlear Implants ; De Lange Syndrome ; Ear ; Ear, Inner ; Endolymphatic Duct ; Hearing Loss ; Hearing Loss, Sensorineural ; Hearing Loss, Sudden ; Humans ; Retrospective Studies

BACKGROUND AND OBJECTIVES: The antioxidant ebselen will be able to limit or prevent the ototoxicity arising from 2-hydroxypropyl-β-cyclodextrin (HPβCD). Niemann-Pick Type C (NPC) disease is a disorder of lysosomal storage manifested in sphingolipidosis. Recently, it was noted that experimental use of HPβCD could partially resolve the symptoms in both animals and human patients. Despite its desirable effect, HPβCD can induce hearing loss, which is the only major side effect noted to date. Understanding of the pathophysiology of hearing impairment after administration of HPβCD and further development of preventive methods are essential to reduce the ototoxic side effect. The mechanisms of HPβCD-induced ototoxicity remain unknown, but the resulting pathology bears some resemblance to other ototoxic agents, which involves oxidative stress pathways. To indirectly determine the involvement of oxidative stress in HPβCD-induced ototoxicity, we tested the efficacy of an antioxidant reagent, ebselen, on the extent of inner ear side effects caused by HPβCD. MATERIALS AND METHODS: Ebselen was applied prior to administration of HPβCD in mice. Auditory brainstem response thresholds and otopathology were assessed one week later. Bilateral effects of the drug treatments also were examined. RESULTS: HPβCD-alone resulted in bilateral, severe, and selective loss of outer hair cells from base to apex with an abrupt transition between lesions and intact areas. Ebselen co-treatment did not ameliorate HPβCD-induced hearing loss or alter the resulting histopathology. CONCLUSIONS: The results indirectly suggest that cochlear damage by HPβCD is unrelated to reactive oxygen species formation. However, further research into the mechanism(s) of HPβCD otopathology is necessary.
Animals ; Ear, Inner ; Evoked Potentials, Auditory, Brain Stem ; Hair Cells, Auditory, Outer ; Hearing Loss ; Hearing ; Humans ; Mice ; Oxidative Stress ; Pathology ; Reactive Oxygen Species ; Sphingolipidoses ; Tight Junctions

Hearing loss has become increasingly prevalent and causes considerable disability, thus gravely burdening the global economy. Irreversible loss of hair cells is a main cause of sensorineural hearing loss, and currently, the only relatively effective clinical treatments are limited to digital hearing equipment like cochlear implants and hearing aids, but these are of limited benefit in patients. It is therefore urgent to understand the mechanisms of damage repair in order to develop new neuroprotective strategies. At present, how to promote the regeneration of functional hair cells is a key scientific question in the field of hearing research. Multiple signaling pathways and transcriptional factors trigger the activation of hair cell progenitors and ensure the maturation of newborn hair cells, and in this article, we first review the principal mechanisms underlying hair cell reproduction. We then further discuss therapeutic strategies involving the co-regulation of multiple signaling pathways in order to induce effective functional hair cell regeneration after degeneration, and we summarize current achievements in hair cell regeneration. Lastly, we discuss potential future approaches, such as small molecule drugs and gene therapy, which might be applied for regenerating functional hair cells in the clinic.
Infant, Newborn ; Humans ; Hair Cells, Auditory, Inner/physiology* ; Ear, Inner/physiology* ; Hair Cells, Auditory/physiology* ; Regeneration/genetics* ; Stem Cells

OBJECTIVE: To explore whether the Ad5-atoh1/EGFP could transdifferent the supporting cells into the new hair cells in young adult guinea pigs cochlea in vivo. METHOD: Twelve healthy pigmented guinea pigs weighted 200-250 g were included in this experiment. 5 ul of Ad5-E1/E3 defected-atoh1/EGFP were infused into the scala media through a hole made on the lateral wall of the cochlea. Six of the 12 animal were killed 2 weeks after the infusion operation. The others were killed 4 weeks after the operation. The whole mount of the basal membranes were directly observed under the fluorescence microscope for the expression of the EGFP (enhance green fluorescent protein) or for the expression of the hair cellspecific marker and nuclear after staining with myosin VIIa rabbit polyclonal antibody and Dapi dye. RESULT: New cells with big nuclear, ellipse body and expressed with EGFP were found in the region near to the outmost row of the outer hair cells in 2 animal 2 weeks after the infusion. Moreover there were 3 animals with specific morphologic new cells in the location where ever been located by the outer hair cells and the region as 2 weeks animals 4 weeks after the infusion. Those cells were stained by myosin VIIa antibody. CONCLUSION Atoh1 gene could transdifferent some supporting cells in the basal membrane into hair cell like cells in young adult guinea pigs in vivo. These supporting cells locate in the region of outer hair cells and the basal membrane which do not belong to the region of outer hair cells.
Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; metabolism ; Cell Transdifferentiation ; genetics ; Cochlea ; cytology ; Ear, Inner ; Green Fluorescent Proteins ; genetics ; metabolism ; Guinea Pigs ; Hair Cells, Auditory ; cytology ; Labyrinth Supporting Cells ; cytology

BACKGROUND AND OBJECTIVES: Vacuolar type H+-ATPase (V-H+-ATPase) has a role in the regulation of endolymphatic pH and certain cells (including strial marginal cells, inner hair cells and epithelial cells of the endolymphatic sac) may be specialized for this regulation. Bafilomycin is a specific V-H+-ATPase inhibitor affecting inner ear function by controlling the intracytolic pH decrease. We designed the study to analyze the effect of bafilomycin delivered to the inner ear on the hearing threshold measured by auditory brainstem response (ABR) and endolymphatic potential (EP). MATERIALS AND METHOD: For measuring the hearing threshold change, 13 guinea pigs with normal Preyer's reflex and normal ABR were used. Guinea pigs were randomly divided into control group (n=3, 6 ears) and study groups which were subdivided into the following ;1 mM bafilomycin group (n=3, 6 ears), 5 mM bafilomycin group (n=3, 6 ears) and 10 mM bafilomycin group (n=4, 8 ears). The mastoid cavity was opened to expose the round window and HBSS buffer (300 osm) was applied for the control group and bafilomycin with different concentrations were also applied to the round windows of studied guinea pigs. The hearing threshold was measured using ABR before and after the application of appropriate solutions. For measuring of EP, the cochlea helix and round window of guinea pig with normal hearing were defined and a tungsten micro needle was inserted into the endolymphatic space at 2nd turn of guinea pig's cochlea. EP was measured after application of HBSS buffer as control, 1 mM, 5 mM, and 10 mM bafilomycin. RESULTS: In the control group, the hearing threshold was 21.6+/-2.8 dB (mean+/-SD) initially both before and after mastoidectomy and stayed that way all throughout the study. The hearing threshold increased as bafilomycin was applied. For 1 mM of bafilomycin application, the threshold changed from initial 30.0+/-5 dB to 33.3+/-5.7 dB after 2 hours. For 5 mM of bafilomycin application, the threshold changed after 2 hours from initial 30.0+/-5 dB to 50.0+/-0 dB. With 10 mM of bafilomycin application, the threshold changed after 2 hours from initial 27.5+/-6.4 dB to 52.5+/-8.6 dB. For 5 mM and 10 mM bafilomycin group, there was a significant statistical change of hearing threshold (p<0.05). However, there was no meaningful difference between 5 mM and 10 mM group (p=0.88). Initial EP was 85+/-10 mV and was significantly decreased in 5 mM (35 mV) and 10 mM (19.8 mV) bafilomycin groups, but such was not observed in 1 mM bafilomycin and control group. CONCLUSION: We could observe the elevation of hearing threshold and decrease EP after applying bafilomycin on the round windows of guinea pigs and also observed that this change reached a critical point when the concentration of bafilomycin was 5 mM or higher. From these results, we can conclude that bafilomycin does affect the hearing threshold through its mechanism and the degree of damage has a critical point dependent on the concentration of bafilomycin.
Animals ; Auditory Threshold ; Cochlea ; Ear, Inner ; Epithelial Cells ; Evoked Potentials ; Evoked Potentials, Auditory, Brain Stem ; Guinea Pigs* ; Guinea* ; Hair ; Hearing* ; Hydrogen-Ion Concentration ; Mastoid ; Needles ; Reflex ; Tungsten