Efficient Bone Conduction Hearing Device With a Novel Piezoelectric Transducer Using Skin as an Electrode.

OBJECTIVE: Bone conduction hearing aids are the only non-surgical devices used for conductive hearing loss. However, they are impractical for lifelong use since they require close contact of the transducer with the head skin, causing skin erosion and discomfort. Bone conduction hearing implants and active middle ear implants do not present these issues; however, they require surgery and can sometimes cause issues in the skin surrounding the devices. This study aimed to develop a new bone conduction hearing device that does not exert pressure on the skin or require surgery. METHODS: Our device modified a piezoelectric element by using the skin of a pinna as one of the two electrodes of a conventional piezoelectric device. We compared the sound transmission of a speaker, a conventional piezoelectric device, or the new device to the Guinea pig cochlea, a physiological sound transducer to the auditory nerve, in normal and air-conductive hearing loss conditions. RESULTS: The novel device transmitted sound to the cochlea even after causing air-conductive hearing loss. Its bone conduction was more efficient than the speaker and the conventional piezoelectric device. CONCLUSION: We developed a novel type of bone conduction device that efficiently transmits sound to the cochlea by skipping the external auditory canal, tympanic membrane, and middle ear ossicles. This device does not exert pressure on the skin that can result in skin damage, an adverse effect of a conventional bone conduction hearing aid. SIGNIFICANCE: Our novel hearing device can be used as a substitute for current bone-conduction hearing devices.

Neck and superior mediastinal granular cell tumor excised via a combined approach.

Granular cell tumor (GCT) is an uncommon tumor of Schwann cell origin. GCT occurs in various sites throughout the body, but mediastinal GCT is very rare. We present an extremely rare case of GCT of the neck and superior mediastinum. A 36-year-old man with a 3-month history of cough was investigated at our hospital. CT and MRI revealed a spindle-shaped mass in the left neck and superior mediastinum, with features suggesting a neurogenic tumor. The tumor was successfully excised via combined neck incision and video-assisted thoracoscopic surgery. Histopathological examination showed proliferation of polygonal and spindled cells with eosinophilic granule-rich cytoplasm. These cells were S-100 protein positive, and the cytoplasmic granules were periodic acid-Schiff positive. Based on these histopathological and immunohistochemical findings, a diagnosis of GCT was established. The patient developed hoarseness immediately after the operation, and laryngoscopy revealed the left vocal cord palsy in the paramedian position, which resolved after about 3 months. The tumor did not recur during the following 10 months.

[A case of metastatic salivary duct carcinoma successfully treated with trastuzumab-based targeted therapy].

Salivary duct carcinoma is a malignant salivary neoplasm with a poor prognosis. Effective treatment for remote metastases has not been recognized. We report herein on a case of this tumor overexpressing HER2 successfully treated with trastuzumab-based molecular targeted therapy. The patient was a 69-year-old man, who developed remote metastases into the liver and the thoracic vertebra six months after surgery and postoperative irradiation for the primary and nodal lesions. After targeted therapy including paclitaxel and trastuzumab, these metastatic lesions showed rapid and continued regression. After paclitaxel was discontinued due to peripheral neuropathy in the extremities, trastuzumab monotherapy followed without resulting in cardiotoxicity. After three years since development of remote metastases, the patient is doing well without re-progression of the disease.

[Successful treatment of dysphagia due to Wallenberg syndrome using intermittent air stretching method with balloon catheter: a case report].

A 69-year-old woman presented with dysphagia due to Wallenberg syndrome. Videofluorography revealed unilateral dysfunction of the cricopharyngeal muscle, which caused stenosis of the esophageal entrance on the affected side. Pharyngeal fiberscopy indicated that glottal function and the cough reflex were preserved and that the pharyngeal reflex was lost. The stenosis of the esophageal entrance due to dysfunction of the cricopharyngeal muscle on the affected side and the loss of the pharyngeal reflex were considered to cause the patient's dysphagia. Based on above mentioned findings, the intermittent air stretching method with balloon catheter(IASM)was performed. As a result, dysphagia showed rapid improvement without any complications such as aspiration or bleeding. Thus, the IASM may be effective in some cases of dysphagia due to Wallenberg syndrome.

Afferent deprivation elicits a transcriptional response associated with neuronal survival after a critical period in the mouse cochlear nucleus.

The mechanisms underlying enhanced plasticity of synaptic connections and susceptibilities to manipulations of afferent activity in developing sensory systems are not well understood. One example is the rapid and dramatic neuron death that occurs after removal of afferent input to the cochlear nucleus (CN) of young mammals and birds. The molecular basis of this critical period of neuronal vulnerability and the transition to survival independent of afferent input remains to be defined. Here we used microarray analyses, real-time reverse transcription PCR, and immunohistochemistry of the mouse CN to show that deafferentation results in strikingly different sets of regulated genes in vulnerable [postnatal day (P)7] and invulnerable (P21) CN. An unexpectedly large set of immune-related genes was induced by afferent deprivation after the critical period, which corresponded with glial proliferation over the same time frame. Apoptotic gene expression was not highly regulated in the vulnerable CN after afferent deprivation but, surprisingly, did increase after deafferentation at P21, when all neurons ultimately survive. Pharmacological activity blockade in the eighth nerve mimicked afferent deprivation for only a subset of the afferent deprivation regulated genes, indicating the presence of an additional factor not dependent on action potential-mediated signaling that is also responsible for transcriptional changes. Overall, our results suggest that the cell death machinery during this critical period is mainly constitutive, whereas after the critical period neuronal survival could be actively promoted by both constitutive and induced gene expression.

A new method for drug application to the inner ear.

Inner ear sensory cells are very susceptible to injuries and recovery after damage is very difficult. Recently several drugs including neurotrophic factors have been reported to protect against inner ear injury. The purpose of this experimental study is to find new methods for applying drugs to the inner ear that effectively protect against inner ear damage. Biodegradable hydrogel was used as a carrier for application of brain-derived neurotrophic factor (BDNF) into the inner ear of guinea pigs through the round window membrane. After application of BDNF the number of surviving spiral ganglion neurons increased following injury of inner ear hair cells and spiral ganglion neurons by ototoxic treatment. This result indicates that BDNF provides effective protection against inner ear damage and that biodegradable hydrogel is useful for application of drugs to the inner ear.

Novel strategy for treatment of inner ears using a biodegradable gel.

OBJECTIVE: The present study aimed to evaluate the efficacy of a biodegradable hydrogel as a drug-delivery medium for the inner ear. Brain-derived neurotrophic factor (BDNF) was chosen as the agent to be administered. METHOD: First, we used an enzyme-linked immunosorbent assay to measure BDNF concentrations in the cochlear fluid after placing a hydrogel containing this agent onto the round-window membrane of the ear. Second, the functional and histologic protection of the auditory primary neurons (spiral ganglion neurons [SGNs]) by BDNF applied through the hydrogel was examined using an animal model of SGN degeneration. RESULTS: The results revealed sustained delivery of BDNF into the cochlear fluid by way of the hydrogel. Second, the functional and histologic protection of the auditory primary neurons (SGNs) by BDNF applied through the hydrogel was examined using an animal model of SGN degeneration. The measurement of electrically evoked auditory-brainstem responses demonstrated that BDNF delivered by way of the hydrogel significantly reduced the threshold elevation. Immunohistochemistry for neurofilament 200 kD demonstrated increased survival of SGNs because of BDNF application through the hydrogel. CONCLUSION: These findings indicate that biodegradable hydrogels can be used for drug delivery to the inner ear.

Engraftment of embryonic stem cell-derived neurons into the cochlear modiolus.

This study aimed to evaluate the potential of embryonic stem cell-derived neural progenitors for use as transplants for the replacement of the auditory primary neurons, spiral ganglion neurons. Mouse embryonic stem cell-derived neural progenitors were implanted into the base of the cochlear modiolus of normal or deafened guinea pigs, which contains spiral ganglion neurons and cochlear nerve fibers. Histological analysis demonstrated the survival and neural differentiation of transplants in the cochlear modiolus and active neurite outgrowth of transplants toward host peripheral or central auditory systems. Functional assessments indicated the potential of transplanted embryonic stem cell-derived neural progenitors to elicit the functional recovery of damaged cochleae. These findings support the hypothesis that transplantation of embryonic stem cell-derived neural progenitors can contribute to the functional restoration of spiral ganglion neurons.

Disruption and restoration of cell-cell junctions in mouse vestibular epithelia following aminoglycoside treatment.

The intracellular junction complexes, which consist of tight junctions (TJ), adherens junctions (AJ), and desmosomes, mediate cell-cell adhesion in epithelial cells. E-cadherin, which is a major component of AJ, plays a role not only in the maintenance of cell-cell junctions, but also in repressing cell proliferation. In this study, we examined changes of E-cadherin expression in mouse vestibular epithelia following local application of neomycin using immunohistochemistry and western blotting, and morphology of cell-cell junctions by transmission electron microscopy (TEM). Immunohistochemistry and western blotting revealed down-expression of E-cadherin and its consecutive recovery. TEM demonstrated temporal disruption of cell-cell junctions. Morphology of cell-cell junctions was more rapidly restored than recovery of E-cadherin expression. Transient disruption of cell-cell junctions and down-expression of E-cadherin is a rational response for the deletion of dying hair cells, and may be associated with a limited capacity for cell proliferations in mammalian vestibular epithelia following their rapid restoration.

Nuclear translocation of beta-catenin in developing auditory epithelia of mice.

Beta-catenin, a protein component of adherens junctions, plays a role in the signalling pathway for cell proliferation. In this study, we examined the cellular distribution of beta-catenin in developing auditory epithelia of mice. Immunohistochemistry for Ki67 and cyclin D indicated active cell proliferation in premature auditory epithelia. In this period, the nuclear localization of beta-catenin in epithelial cells was observed together with expression of the lymphoid enhancer factor, a transcription factor in beta-catenin signalling. Epithelial cells showing nuclear localization of beta-catenin disappeared at the same time, as there was a decrease of cell proliferation. These findings indicate that nuclear translocation of beta-catenin plays a role in cell proliferation in developing auditory epithelia.

Elevation of superoxide dismutase increases acoustic trauma from noise exposure.

The generation of superoxide has been implicated as a cause of cochlear damage from excessive noise. Cu/Zn superoxide dismutase (SOD1) generally will protect against superoxide-mediated tissue injury but protection by this enzyme against noise trauma is controversial. This study assessed auditory function in C57BL/6 mice overexpressing SOD1 or treated with lecithinized SOD1 (PC-SOD1). Noise exposure caused significantly higher threshold shifts in PC-SOD1-treated animals than physiological saline-treated animals. Cochlear tissues of PC-SOD1-treated animals exhibited significant elevation of the levels in the SOD activity, not in the catalase activity, in comparison with those of saline-treated animals. Likewise, transgenic mice overexpressing SOD1 tended to suffer higher threshold shifts than nontransgenic littermates from noise exposure. The findings indicate that increasing SOD1 enhances auditory dysfunction following noise exposure.

Aging effects on vestibulo-ocular responses in C57BL/6 mice: comparison with alteration in auditory function.

Age-related changes in auditory function are well documented in animal models; however, this is not the case as regards vestibular function. In this study, we evaluated age-related changes in vestibulo-ocular responses in C57BL/6 mice that are considered as a model of presbycusis. The functional data were substantiated by the findings of histological analysis of vestibular and auditory peripherals. The gain in vestibulo-ocular reflex, which reflects functionality of the vestibular system, increased in an age-dependent manner until 12 weeks and exhibited limited functional loss due to aging after 24 weeks. By contrast, no alteration in the thresholds of the auditory brainstem response (ABR) was observed from 3 to 12 weeks of age; however, ABR thresholds were significantly elevated from age 24 weeks and onwards. Histological analysis demonstrated that the degeneration of auditory peripherals was closely related with functional loss due to aging. Vestibular peripherals also exhibited age-related degeneration morphologically, although age-related dysfunction was not apparent. Age-related changes in the vestibular function of C57BL/6 mice followed a different time course when compared to changes in auditory function. These findings indicate that mechanisms for age-related changes in vestibular function differ from those of auditory function.

Destiny of autologous bone marrow-derived stromal cells implanted in the vocal fold.

OBJECTIVES: The aim of this study was to investigate the destiny of implanted autologous bone marrow-derived stromal cells (BSCs) containing mesenchymal stem cells. We previously reported the successful regeneration of an injured vocal fold through implantation of BSCs in a canine model. However, the fate of the implanted BSCs was not examined. In this study, implanted BSCs were traced in order to determine the type of tissues resulting at the injected site of the vocal fold. METHODS: After harvest of bone marrow from the femurs of green fluorescent transgenic mice, adherent cells were cultured and selectively amplified. By means of a fluorescence-activated cell sorter, it was confirmed that some cells were strongly positive for mesenchymal stem cell markers, including CD29, CD44, CD49e, and Sca-1. These cells were then injected into the injured vocal fold of a nude rat. Immunohistologic examination of the resected vocal folds was performed 8 weeks after treatment. RESULTS: The implanted cells were alive in the host tissues and showed positive expression for keratin and desmin, markers for epithelial tissue and muscle, respectively. The implanted BSCs differentiated into more than one tissue type in vivo. CONCLUSIONS: Cell-based tissue engineering using BSCs may improve the quality of the healing process in vocal fold injuries.

Mechanisms of apoptosis induced by cisplatin in marginal cells in mouse stria vascularis.

Degeneration of the stria vascularis (SV) is amongst the major causes of cisplatin (CDDP)-induced hearing impairment. The pathways of apoptosis occurring in the SV due to CDDP were examined using a mouse experimental model. Temporal bones of adult C57BL/6 mice were collected on days 3, 7 and 14 after the local application of CDDP. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay and immunostaining for apoptosis-related proteins or reactive radical species were employed for analysis. Local application of CDDP caused apoptotic cell death of marginal cells 3 days after CDDP treatment. Immunohistochemical analyses demonstrated activation of caspase-3 and -9, but not -8, and redistribution of cytochrome c in affected marginal cells, indicating a caspase-dependent, mitochondrion-mediated apoptotic pathway in marginal cells. Temporary expression of hydroxynonenal, nitrotyrosine and inducible nitric oxide synthase in the SV was observed at the induction of apoptosis in marginal cells. CDDP toxicity generates reactive radical species in the SV, which causes mitochondrial membrane permeabilization leading to apoptosis of marginal cells.

Transplantation of bone marrow stromal cells into the cochlea of chinchillas.

This study aimed to evaluate the potential of bone marrow stromal cells for treatment of inner ear diseases. Autologous marrow cells labeled with Dil were implanted into the inner ear of five gentamicin-treated chinchillas. Histological analysis 3 weeks later revealed robust survival of grafted marrow cells in multiple regions within the cochlea. Marrow cells implanted in the basal turn of the cochlea migrated as far as the apical end or into the spiral ligament of the cochlea. Some grafted cells expressed a neuronal or glial cell marker, indicating their ability to differentiate into neuronal or glial cells. Survival, migrational mobility and differentiation of autologous marrow cells in damaged cochlea suggest their potential as transplants for treatment of various degenerative inner ear diseases.

Expression of beta-catenin in developing auditory epithelia of mice.

This study investigated the role of beta-catenin in the development of mouse auditory epithelia. Inner ears obtained from embryonic and newborn mice were used. Expression of beta-catenin was examined together with the expression of Ki-67, a marker for proliferating cells, or myosin VIIa, a marker for differentiated hair cells. In the early phase of development, intense expression of beta-catenin was found in auditory epithelia in which a number of Ki-67-positive cells were identified. Together with a decrease in proliferating cells, the intensity and area of beta-catenin expression were reduced. In addition, during differentiation and maturation of hair cells, the area of beta-catenin expression was further limited. These findings suggest that patterns of expression of beta-catenin are closely linked with the status of auditory epithelia development.

Induction of cell proliferation and beta-catenin expression in rat utricles in vitro.

Limited regenerative response occurs in mammalian vestibular epithelia, although vestibular hair cell regeneration in mammals has been demonstrated under a variety of experimental conditions. Beta-catenin is known to play an important role in both cell-cell adhesion and signal transduction associated with cell proliferation. This study evaluated cell proliferation activity in mammalian vestibular epithelia in organotypic culture and examined the involvement of beta-catenin in cell proliferation in vestibular epithelia. After 72 h of culture, utricles with or without induction of cell proliferation were examined. Cell proliferation was induced by brief exposure to forskolin and supplementation with fetal bovine serum. Cell proliferation activity was assessed by bromodeoxyuridine (BrdU) labeling. Immunohistochemistry was employed for analysis of cellular distribution of beta-catenin. In utricles cultured without induction of cell proliferation. BrdU labeling was not found in vestibular epithelia. Expression of beta-catenin was found in the area corresponding to the distribution of adherens junctions in vestibular epithelia. However, BrdU labeling was identified in sensory epithelia of utricles following induction of cell proliferation, although the number of BrdU-positive cells in sensory epithelia was very limited. Accumulation of beta-catenin was occasionally found in proliferating cells in sensory epithelia; however, it was not always found in BrdU-positive cells. The present findings suggest that beta-catenin may play a role in the induction of cell proliferation in mammal vestibular epithelia.

Surgical techniques for cell transplantation into the mouse cochlea.

This study investigated surgical procedures for cell transplantation into the mouse inner ear. Female C57BL/6 mice were used as recipient animals. Fetal mouse neural stem cells expressing green fluorescence were used as donor cells. Two methods, an injection of transplants from the lateral semicircular canal (LSCC) and from the cochlear lateral wall (CLW), were examined. Two weeks after transplantation, the distribution of transplant-derived cells in the cochlea was examined. Effects on auditory function were assessed by measurement of auditory brain stem responses (ABRs). Cochleae receiving cell transplantation from the LSCC exhibited robust survival of transplant-derived cells mainly in the scala vestibuli and scala tympani. Transplantation from the LSCC caused elevation of ABR thresholds by less than 10 dB SPL. However, transplantation from the CLW resulted in considerable hearing loss, even though transplant-derived cells settled in the scala media. These findings demonstrate that an approach from the LSCC can be utilized for cell transplantation into the perilymph without causing apparent auditory disorder, while an approach from the CLW delivers cells to the endolymph but appears to cause auditory dysfunction.

Fates of mouse embryonic stem cells transplanted into the inner ears of adult mice and embryonic chickens.

The potential of embryonic stem (ES) cells to differentiate into inner ear hair cells was examined in this study. Undifferentiated mouse ES cells transplanted into neomycin-damaged mouse inner ears were evaluated by immunohistochemistry 4 weeks after transplantation. Some ES cells were positive for E-cadherin or NCAM, and most transplanted cells were positive for SSEA3 and Ki67. None were positive for Myosin VIIa or MF20. These results indicate that the damaged inner ear may have some activity inducing ES cells to develop into ectoderm cells, but the effect was insufficient to induce inner ear hair cells. Next, SDIA/BMP-treated ES cells were transplanted into embryonic chicken inner ear rudiments. Embryonic chickens were expected to share the same developmental systems as mice. SDIA/BMP treatment drove ES cells to the population including neural crest cells and probably placode cells ES colonies were found next to or in the otic vesicles but were not a part of vesicle walls, indicating that transplanted ES cells could not be expected to be the same kind of cells as chicken otic vesicle cells Some ES colonies were found at the vestibulo-cochlear ganglions. To induce inner ear hair cells in this system, the competency of ES cells and otic induction signals should be defined further.

Transplantation of neural stem cells into the modiolus of mouse cochleae injured by cisplatin.

This study aimed to examine the possibility of restoration of spiral ganglion neurons, which transmit sound stimulation to the brain, by transplantation of fetal neural stem cells (NSCs) into the modiolus of cochleae. Fetal mouse NSCs expressing green fluorescence were injected into the modiolus of cisplatin-treated cochleae of mice. The temporal bones were collected 14 days after transplantation, and provided histological examination. The cell fate of transplants was determined by immunohistochemistry for a neural or glial cell-marker. Histological analysis 2 weeks after transplantation revealed robust survival of transplant-derived cells in the modiolus of the cochlea. NSCs injected in the basal portion of cochleae migrated as far as the apical end of the modiolus Grafted NSCs expressing a neural cell marker were identified, but the majority of grafted NSCs differentiated into glial cells. These findings suggest the possible use of NSCs in cell therapy for restoration of spiral ganglion neurons. However, further treatments are required to increase the number of NSC-derived neurons in the modiolus to realize functional recovery.