Protective effect of N-acetylcysteine against cisplatin ototoxicity in rats: a study with hearing tests and scanning electron microscopy / Efeito protetor da N-acetilcisteína na ototoxicidade por cisplatina em ratos: um estudo com testes auditivos e microscopia eletrônica de varredura

Abstract Introduction Ototoxicity is a health problem appearing after powerful treatments in serious health conditions. It is sometimes inevitable when treatment of the serious disease is required. Cisplatin is an antineoplastic agent which was investigated previously to reveal increased nitrogen and reactive oxygen radicals that damages hair cells, resulting in ototoxicity. N-acetylcysteine, previously shown to decrease ototoxicity caused by different agents, is known to be a powerful in vitro antioxidant. Probably N-acetylcysteine, in addition to its antioxidant effect, blocks a cascade where reactive oxygen species result in apoptosis in the cochlea. Objectives The possible preventive effect of N-acetylcysteine in cisplatin ototoxicity was studied with auditory brain stem responses, otoacoustic emissions, and histopathological investigation of the cochlea in a scanning electron microscopy. Methods This study was conducted on 21 Wistar Albino rats in four groups. 1 mL/kg/day three times in total intraperitoneal (i.p.) Saline (n = 5), 500 mg/kg/day i.p. three times in total N-acetylcysteine (n = 5), i.p. 15 mg/kg cisplatin alone (single dose) (n = 5) and i.p. 15 mg/kg cisplatin plus 500 mg/kg/day N-acetylcysteine (n = 6) were administered. The rats were anesthetized to study the hearing tests before and after the experiment. The rats were sacrificed to investigate the cochleas by scanning electron microscopy. Results Auditory brain stem responses and otoacoustic emissions values were attenuated in the cisplatin group. The group that received N-acetylcysteine in addition to cisplatin had better auditory brain stem responses thresholds and otoacoustic emissions. The samples obtained from the cisplatin group showed surface irregularities, degeneration areas, and total or partial severe stereocilia losses. The changes were milder in the cisplatin + N-acetylcysteine group. Conclusion Cisplatin ototoxicity can be detected by auditory brain stem responses and otoacoustic emissions testing in rats. N-acetylcysteine may protect the cochlear cells from histopathological changes. We concluded that N-acetylcysteine given 4 h after cisplatin injection has a potential otoprotective effect against cisplatin ototoxicity. which suggests it could be used in clinical trials.

Resumo Introdução A ototoxicidade é um problema que pode ocorrer após certos tipos de tratamentos para condições graves de saúde. Às vezes é inevitável quando o tratamento da doença é necessário. A cisplatina é um agente antineoplásico cujo uso em pesquisas anteriores demonstrou aumentar os radicais livres de nitrogênio e espécies reativas de oxigênio que danificam as células ciliadas e resultam em ototoxicidade. Por outro lado, a N-acetilcisteína, que já demonstrou diminuir a ototoxicidade causada por diferentes agentes, é conhecida por ser um potente antioxidante in vitro. Provavelmente a N-acetilcisteína, além de seu efeito antioxidante, bloqueia uma cascata onde espécies reativas de oxigênio resultam em apoptose na cóclea. Objetivos Estudar o possível efeito preventivo da N-acetilcisteína na ototoxicidade por cisplatina por meio de potencial evocado auditivo de tronco encefálico, emissões otoacústicas e investigação histopatológica da cóclea por microscopia eletrônica de varredura. Método Este estudo foi realizado em 21 ratos albinos Wistar, separados em quatro grupos. Foram administrados: 1 mL/kg/dia intraperitoneal (i.p.) de solução salina (n = 5), três vezes no total; 500 mg/kg/dia i.p. de N-acetilcisteína (n = 5), três vezes no total; 15 mg/kg i.p. (dose única) somente de cisplatina (n = 5) e 15 mg/kg i.p. de cisplatina e 500 mg/kg/dia i.p. de N-acetilcisteína (n = 6). Os ratos foram anestesiados para estudo dos testes auditivos antes e depois do experimento. Os ratos foram sacrificados para investigação da cóclea por microscopia eletrônica de varredura. Resultados Os potenciais evocados auditivos de tronco encefálico e os valores das emissões otoacústicas estavam atenuados no grupo cisplatina. O grupo que recebeu N-acetilcisteína além da cisplatina apresentou melhores limiares de respostas auditivas do tronco encefálico e emissões otoacústicas. As amostras obtidas do grupo cisplatina apresentaram irregularidades de superfície, áreas de degeneração, com perdas graves totais ou parciais de estereocílios. As alterações foram mais leves no grupo cisplatina + N-acetilcisteína. Conclusão A ototoxicidade por cisplatina pode ser detectada por meio de potenciais evocados auditivos de tronco encefálico e pelo teste de emissões otoacústicas em ratos. A N-acetilcisteína pode proteger as células cocleares contra alterações histopatológicas. Concluímos que a N-acetilcisteína administrada 4 horas após a injeção de cisplatina tem potencial efeito otoprotetor contra a ototoxicidade por cisplatina e pode ser utilizada em ensaios clínicos.

Histology of the testis and the epididymal ducts from hoary fox Lycalopex vetulus (Lund, 1842) / Histologia dos testículos e ductos epididimários da raposa-do-campo Lycalopex vetulus (Lund, 1842)

The hoary fox Lycalopex vetulus (Lund, 1842) is a small canid, endemic to Brazil, belonging to the Canidae family, widely distributed in the country, occurring records in different regions and habitats. The objective of this study is to describe morphologically the testicles and epididymal ducts of hoary fox. The animals, coming from the zoo of Federal University of Mato Grosso, Brazil, had died by natural causes. The male reproductive system was dissected to collect the testicles. The samples were fragmented and histologically examined. Microscopically, the testes were coated by the vaginal and albuginea tunic, formed by modeled dense connective tissue with large amount of collagen fibers. Into the organ, convoluted seminiferous tubules were surrounded by a basement membrane characterized by the presence of myoid and Sertoli cells and germinative epithelium composed by Between the seminiferous tubules, interstitial tissue composed of connective tissue, blood and lymph vessels and Leydig cells in polyhedral shape was present. The epididymal ducts showed pseudostratified columnar epithelium with secretory cells, in which stereocilia located on a basement membrane filled by myoid cells were found. The structures observed by us are very similar to those described for other mammals.(AU)

A raposa-do-campo Lycalopex vetulus (Lund, 1842) é um canídeo de pequeno porte, endêmico do Brasil, pertencente a família Canidae, com ampla distribuição no país, ocorrendo registros em várias regiões e habitats diferentes. Com base nessa informação, o objetivo deste trabalho é caracterizar morfologicamente os testículos e ductos epididimários da raposa-do-campo. O animal, oriundo do zoológico da Universidade Federal de Mato Grosso, Brasil, veio a óbito por causas naturais e o sistema reprodutor masculino foi dissecado para coleta dos testículos. As amostras retiradas foram fragmentadas e histológicamente examinadas. A partir das análises microscópicas dos testículos foram identificados: a túnica vaginal e albugínea, formada por tecido conjuntivo denso modelado, com grande quantidade de fibras colágenas; túbulos seminíferos enovelados e revestidos por epitélio germinativo e células de Sertoli, envolvidos por uma membrana basal com presença de células mioides; tecido intersticial entre os túbulos seminíferos constituído de tecido conjuntivo, vasos sanguíneos e linfáticos, e células de Leydig em formato poliédrico. Os ductos epididimários apresentaram epitélio cilíndrico pseudoestratificado com células secretoras dos quais projetam estereocílios, situados sobre uma membrana basal repleta por células mióides. As estruturas por nós observadas possuem muita semelhança com as demais descrições para mamíferos.(AU)

Measurement of the Mechanical Deformation of Organ of Corti in a Model of Acute Endolymphatic Hydrops / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Endolymphatic hydrops has been considered as an important histologic substrate of Meniere's disease. A permanent displacement of basilar membrane (BM) by increased endolymphatic pressure has been thought to be an explanation for hearing change. Direct observation of histological sections of temporal bones, however, suggested that stereocilia and tectorial membrane decoupling is more associated with pressure induced by mechanical deformation of the organ of Corti rather than with the displacement of BM. METERIALS AND METHOD: 26 cochleae from 13 female pigmented ginea pigs were harvested. One cochlea per each animal was injected with artificial perilymph. The other one was used as control. After fixation, followed by embedding and mid-modiolar sectionning, specimens were observed with a microscope. Morphometric parameters of each row and turn of the organ of Corti were measured and quantified. RESULTS: The average area and height of the organ of Corti were significantly smaller in the apical turn of the experimental group (p<0.05). The lengths of outer hair cell and Deiters cell in the apical turn were also significantly reduced in the experimental group (p<0.05). The angle between the outer hair cell and Deiters cell was smaller in the apex and in the 3rd turn of the experimental group (p<0.05). CONCLUSION: Results show that compression and deformation of the organ of Corti, especially in the apical turn, is a prominent feature in the acute endolymphatic hydrops model. We suggest that the deformation of organ of Corti is the primary cause of hydrops that induce the decoupling of tectorial membrane and stereocilia rather than the displacement of BM.

Noise-Induced Hearing Loss / 한양의대학술지

Noise-induced hearing loss (NIHL) is the second most common cause of permanent hearing impairment after age-related hearing loss. NIHL is influenced by environmental and genetic factors and the effects of noise can be exacerbated by the administration of ototoxic drugs or exposure to chemicals. The pathophysiology of NIHL is classified as either mechanical injury or metabolic (or biochemical) injury. Exposure of cochleae to intense sounds has been found to disrupt the stereocilia on the hair cells by separating the tip links and to depolymerize actin filaments, resulting in a disturbance in signal transduction. Major mechanisms of metabolic injuries include accumulation of reactive oxygen species enhanced by oxidative stress, cochlear ischemia followed by reperfusion injury, and excitotoxicity to auditory neuron induced by excessive release of the cochlear afferent neurotransmitter, glutamate. Many studies involving therapeutic or preventive trial with antioxidants, JNK inhibitors, and NMDA antagonists have shown partial effectiveness. However, protection from noise before cochlear injury occurs is very important because damaged hair cells and auditory neurons in the mammalian cochleae are unable to regenerate.

Effect of nicotine on the structure of cochlea of guinea pigs / 대한해부학회지

Smoking has been positively associated with hearing loss in human. However, its effect on the cochlea has not been previously evaluated. Aim of work is to investigate the effect of nicotine, which is the primary pharmacological component of tobacco, on the structure of the cochlea of adult male guinea pigs. Fifteen male guinea pigs were classified into two groups: group I (control) and group II (nicotine treated group). Group II was further subdivided into two subgroups; IIA and IIB according to the dose of nicotine (3 mg/kg and 6 mg/kg, respectively). The cochlea was harvested and processed for light microscopy, transmission electron microscopy and scanning electron microscopy. Nicotine administration induced damage of outer hair cells which were distorted in shape with vacuolated cytoplasm and heterochromatic nuclei. Topography revealed damage of the stereocilia which included disorganization, bent and limp or complete loss and expansion of the surrounding supporting cells. These changes were more pronounced in the basal turn of the cochlea and mainly involved the outer hair cells. High dose induced more damage and resulted in protrusion of the apical poles of hair cells (blebing), particularly the outer two rows. Nicotine is proved to be harmful to the cells of the cochlea, particularly the outer hair cells of the basal turn. High doses induce blebing of hair cells.

Protective Effect of Hexane and Ethanol Extract of Piper Longum L. on Gentamicin-Induced Hair Cell Loss in Neonatal Cultures

OBJECTIVES: Gentamicin (GM) is a commonly used aminoglycoside antibiotic that generates free oxygen radicals within the inner ear, which can cause vestibulo-cochlear toxicity and permanent damage to the sensory hair cells and neurons. Piper longum L. (PL) is a well-known spice and traditional medicine in Asia and Pacific islands, which has been reported to exhibit a wide spectrum of activity, including antioxidant activity. In this study, we evaluated the effect of hexane:ethanol (2:8) PL extract (subfraction of PL [SPL] extract) on GM-induced hair cell loss in basal, middle and apical regions in a neonatal cochlea cultures. METHODS: The protective effects of SPL extract were measured by phalloidin staining of cultures from postnatal day 2-3 mice with GM-induced hair cell loss. The anti-apoptosis activity of SPL extract was measured using double labeling by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and myosin-7a staining. The radical-scavenging activity of SPL extract was assessed using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. RESULTS: SPL extract at a concentration of 1 microg/mL significantly inhibited GM-induced hair cell loss at basal and middle region of cochlea, while 5 microg/mL was effective against apical region hair cell loss. The protective effect of SPL extract was concentration dependent and hair cells retained their stereocilia in explants treated with SPL extract prior to treatment with 0.3 mM GM. SPL extract decreased GM-induced apoptosis of hair cells as assessed by TUNEL staining. The outer hair and inner hair counts were not decreased in SPL extract treated groups in compare to GM treated explants. Additionally, SPL extract showed concentration dependent radical scavenging activity in a DPPH assay. CONCLUSION: An anti-apoptosis effect and potent radical scavenger activity of SPL extract protects from GM-induced hair cell loss at basal, middle and apical regions in neonatal cochlea cultures.

The Effect of Pretreatment of CoCl2 on the Prevention of Noise-Induced Hearing Loss / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: The study was designed to investigate the changes in the expression of hypoxia inducible factor-1alpha (HIF-1alpha) according to time after being exposed to noise trauma and find out the effect of HIF-1 alpha in the prevention of noiseinduced hearing loss by pre-treatment with cobalt chloride (CoCl2). SUBJECTS AND METHOD: BALB/c hybrid mice with 25 dB HL or less ABR were used in this study. In the study group, subjects were exposed to 120 dB SPL broad white band noise for 3 hours per day for 3 days. The changes in their hearing were documented before and after 1, 2, 3, 4, and 9 days of the first noise exposure. CoCl2 was injected into the peritoneum 2 hours prior to each noise exposure to see the effect of induced HIF-1alpha on noise-induced hearing loss. For the control, injection with distilled water was performed and hearing thresholds were measured in the same manner. Cochlea from each group was obtained in order to observe the morphological changes in the inner ear and the expression of the HIF-1alpha using immunohistochemial staining and immuno-fluorescein staining along with quantification of the hair cell loss. RESULTS: Mice exposed to the noise for 3 days, showed permanent threshold shift and the expression of HIF-1alpha was increased. When HIF-1alpha was induced by pre-treatment of CoCl2 prior to the noise exposure, however, hearing recovery was observed to some degree. And hair cell survival rate was also higher when treated with CoCl2 compared to the distilled water treated group. CONCLUSION: When pre-treated with CoCl2, inducing HIF-1alpha before the noise trauma, it allowed for a less stereocilia loss in the hair cells in the organ of Corti. HIF-1alpha may play an important role in the prevention of noise-induced hearing loss.

Morphological Change of Mouse Inner Ear Hair Cells after Noise Exposure / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Mechanism of inner ear hair cell distortion after noise exposure has been well described. The present study was designed to determine the response to the auditory system of a genetically well-defined laboratory mouse in preparation for examining the effect of noise on mice with specific genetic mutations. So it is important to recognize the relationship between noise exposure duration and hair cell morphological changes. We try to reveal the hearing loss and inner ear hair cell morphological changes after applying the noise protocol. SUBJECTS AND METHOD: The mice were BALB/c hybrids and aged 8 weeks. Six mice served as non-noise-exposed controls and 8 mice were exposed for 3 hours per day to white band noise with a center frequency from 0.2 kHz to 70 kHz and a sound pressure level of 120 dB. And we divided the noise exposure group into 3 subgroups(1 day, 3 day, 5 day noise exposure group). We checked the photographs of FITC phalloidin stain and scanning electron microscopy of cochlea after noise exposure. RESULTS: The hearing level of mice decreased after noise exposure. We could see the stereocilia damage in cochlea after FITC phalloidin stain in cochlea and sterocilia loss was more severe in basal turn. In scanning electron microscopy, morphological changes of stereocilia were observed to be more severe in the cochlear basal turn than other area. Significant hair cell loss in the cochlear basal turn could be calculated using cochleocytogram. CONCLUSION: 120dB broad white band noise can damage the hair cell of cochlea in mice. These changes were especially severe in the cochlear basal turn. Noise exposure duration is the other important factor in damaging cochlear hair cells. Therefore, we can guess that harmful noise level and noise exposure duration are the main risk factors that injure the inner ear hair cell.

Effect of Melatonin on the Cisplatin Induced Ototoxicity in Rats / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Cisplatin (CP), an antitumor agent widely used in the treatment of head and neck cancers, has side effects such as ototoxicity and nephrotoxicity. These side effects are closely related to oxidative stress. In the present study, we attempted to suppress CP-induced ototoxicity in rats by administering melatonin, an antioxidant. MATERIALS AND METHOD: Male Sprague-Dawley rats were divided into different groups and were treated as follows: 1) saline control, 2) CP (16 mg/kg, i.p.), 3) CP plus melatonin (10 mg/kg, i.p.). The rats were sacrificed at the 6th day after CP treatment. RESULTS: CP-treated rats showed increase in cochlear malondialdehyde, hydrogen peroxide, glutathione peroxidase and glutathione reductase levels, and the decrease in cochlear superoxide dismutase (SOD) and catalase levels. CP-treated rats showed markedly decreased in the number of stereocilia on the inner hair cells and mildly decreased in the number of outer hair cells in organ of Corti under the light and scanning electron microscopic examination. Light and electron microscopic findings, and cochlear hydrogen peroxide, malondialdehyde, SOD, catalase, glutathione peroxidase and glutathione reductase levels were restored in the rats injected with CP plus melatonin than those with CP alone. CONCLUSION: These results suggest that melatonin suppresses CP-induced ototoxicity via the suppression of the increased production of reactive oxygen species.

The Morphological Changes of the Cochlea in the Guinea Pig after Single Blast Wave Stimulation / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: The mechanism of the noise damage to the cochlea is not fully understood in spite of many studies. The morphological changes in a noise stimulated model show various features relating to the duration of the exposure and sound intensity. Until now there had been a lot of the morphological studies on noise induced hearing loss models, but only a few reports of single blast injury of more than 150 dB. MATERIALS AND METHOD: We observed with a transmission electron microscope by using the microslicing technique for minimizing the tissue injury and successful hair cell counting in resin embedded cochlea of the guinea pig, stimulated by a 178 dB single blast wave. RESULTS: The area of hair cell damage in L.M. showed stereocilia bending at 24 hours, proliferation of Hensen's body, subsurface cistern in outer hair cell and mitochondria proliferation in inner hair cell within 2wks. The area of no hair cell damage in L.M. showed minimal changes in E.M., as well. CONCLUSION: These results suggest that the 178 dB single blast wave injury may not cause severe distortion of the organ of Corti in the guinea pig and thus the compensation process can be advanced gradually.

Outward Rectifying Current in Isolated Deiters' Cells from Guinea Pig Cochlea / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: The Deiters' cell is one of the supporting cells in the organ of Corti and is known to possibly regulate the signal transduction pathway in the organ of Corti. The signal transduction process can be modulated by ATP and acetylcholine, the so-called neurotransmitters, in Deiters' cells. Intracellular Ca2+ concentration can be also increased by these neurotransmitters and the control mechanism on the organ of Corti is highly suggested in Deiters' cells. Potassium ion (K+) is known to be important both in hair cells and supporting cells. Through K+ channel, the membrane potential may be controlled and the signal transduction pathway can be regulated. Furthermore, the motility of outer hair cell and the signal transduction from the apical stereocilia are considered to be regulated by this channel. The aim of this study is to record the K+ current in the isolated Deiters' cells from guinea pig cochlea. MATERIALS AND METHODS: Deiters' cells were isolated from the organ of Corti of guinea pig by using collagenase and a pipet. A whole cell patch clamp was performed under the inverted microscope and the current was measured with List-7 amplifier and pClamp 8.0.2 software. RESULTS: The resting membrane potential was -15.02+/-2.66 mV (n=6). When the cell membrane was hyperpolarized into -110 mV from the -40 mV holding potential, the peak current was -227+/-39.9 pA (n=15). After having depolarized to the maximum, (50 mV), the peak current was 7123+/-737 pA, and the reversal potentials of different external K+ concentration changed in the K+-dependent manner. About 80% of this current was inhibited by TEA. When K+ was substituted by Cs+, the peak current was 1788+/-231 pA at 50 mV step pulse. Activation curve of this outward current showed two different Vh (half activation voltage) and K (slope factor). CONCLUSION: Outward rectifying K+ channels exist in Deiters' cells and they can be inhibited by TEA and permeable to Cs+. More than two types of K+ current can exist and they may play a role in the recovery of membrane potential after depolarization,

The Ototoxicity of Cisplatin is Mediated by NO: A Study using L-NAME and MK-801 in Guinea pigs / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Nitric oxide has been suggested to play an important role in the pathogenesis of cisplatin ototoxicity. L-NAME (NG-Nitroarginine Methyl Ester) is an inhibitor of nitric oxide synthase. MK-801 (Dizocilpine Maleate) is a NMDA receptor antagonist. To evaluate a role of nitric oxide in cisplatin ototoxicity, we investigated whether L-NAME and MK-801 can block the cisplatin ototoxicity in guinea pigs. MATERIALS AND METHOD: In the Group 1, normal saline was injected intraperitoneally as a control group. Group 2, 3, 4, and 5 were injected intraperitoneally as described in the following: Group 2, cisplatin only; Group 3, L-NAME+isplatin; Group 4, MK-801+cisplatin; Group 5, L-NAME+K-801+cisplatin. Using an auditory brainstem response, hearing threshold was tested before cisplatin administration and 5 days after cisplatin injection in each group. The morphological changes of the cochlea were observed by scanning electron microscopy. RESULTS: In the Group 2, a significant hearing loss was observed comparing to Group 1. In contrast , Group 3, 4, and 5 did not demonstrate any significant hearing loss compared to Group 1. In the scanning electron microscopy, the Group 2 showed distorsion and loss of stereocilia of the hair cells. However, the Group 1, 3, 4, and 5 demonstrated well preserved cochlear hair cell morphology. CONCLUSION: Hearing loss induced by ototoxicity of cisplatin was prevented by L-NAME and MK-801. This study suggests that NO may mediate cisplatin ototoxicity.

Cochlear Pathology in Experimental Otitis Media with Effusion ofGuinea Pig Caused by Endotoxin / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Injection of endotoxin into the middle ear of experimental animals results in otitis media with effusion and transient reversible hearing loss, which occurs in the higher concentration of endotoxin than the concentration found in human middle ear effusion. The purpose of this study was to investigate the histopathologic changes of the cochlea in guinea pigs with experimental otitis media with effusion induced by E.coli endotoxin. MATERIALS AND METHODS: Concentrations of lipopolysaccharides (LPS) from E.coli, such as 0.1 mg/ml, 1 mg/ml, and 10 mg/ml, were instillated into the middle ear cavity of 30 normal guinea pigs. Histopathologic changes of the cochlea were observed at 1 week after endotoxin instillation, using both light and electron microscope. Six normal guinea pigs were used as a control group whose middle ears were instillated with the physiologically sterile saline. RESULTS: There were deformation in the organ of Corti, partial loss of stereocilia in the hair cells, and vacuolation in stria vascularis at a concentration of 10 mg/ml lipopolysaccharides from E. coli. Mild morphologic abnormality of the cochlea was observed at the concentrations of 1 mg/ml and 0.1 mg/ml. CONCLUSION: A higher concentration of LPS in the middle ear cavity may produce severe inner ear damage, possibly resulting in the sensorineural hearing loss.

Dose-response Relationship of Morphologic Changes by Cisplatin and Radiation in Guinea Pig Cochleas / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Cisplatin or radiation have been widely used in the treatment of patients with head and neck cancers. However, their adverse effects such as ototoxicity and nephrotoxicity are serious considerations. This study was aimed to establish a dose-dependent response in an animal model, with the hope of contributing to research for prevention of ototoxicity in cancer patients treated with radiation or chemotherapy. MATERIAL AND METHODS: Three different doses (4, 8, 12 mg/kg) of intraperitoneal cisplatin and radiation (17, 23, 33 Gy) were administered to guinea pigs. RESULTS: After 8 days, the temporal bones were prepared for scanning electron microscopic examination. The cochlear hair cells, especially outer hair cells in the third row, were severely damaged in the cisplatin groups and irradiation groups and these damages were related to the dose administered. The main abnormalities were the deletion or distortion of stereocilia of the hair cells and the loss of stiffness. CONCLUSION: A dose-dependent response relationship established in this study will provide as a useful reference for other researches for the prevention of ototoxicity by cisplatin and radiation.

Organotypic Culture of Organ of Corti with Floating Drop Method from Newborn Rat / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Organotypic culture of organ of Corti maintains the basic organization of the spiral lamina and can conserve several factors responsible for the neuronal growth of the nervous components. The explant culture technique has been widely used in organ culture system, however, the floating drop method using collagen gel was also developed as a simple and reliable method. In order to study the effect of growth factors on the regenerative and protective ability of cochlear hair cells, we first had to establish an in vitro model of the inner ear. MATERIALS AND METHODS: Organ of Corti was obtained from newborn rats and cultured with the floating drop method using collagen gel. Immunohistochemical staining was used to visualize the stereocilia and scanning electron microscopic study was also carried out. RESULTS: Explants were maintained up to 10 days without contamination. Morphologically, immunofluorescent staining with phalloidin showed well preserved outer and inner hair cells with stereocilia on the second day of culture. On the tenth day of culture, the staining result showed inner and outer hair cells, although the stereocilia were poorly stained. In scanning electron microscopic examination, an explant on the tenth day of culture showed preserved outer and inner hair cells and stereocilia, although damaged hair cells and stereocilia were also observed. CONCLUSION: The floating drop method was an appropriate method for maintaining the organ of Corti in vitro with the advantage being the easiness in its manual manipulation.

Light and Scaning Electromicroscopic Study on the Experimental on the Experimental Endolymphatic Hydrops in Guinea Pigs / 대한이비인후과학회지

BACKGROUND: Despite numerous investigations, the pathogenesis of Meniere's disease is uncertain. Endolymphatic hydrops (ELH) has been the main histologic finding of Meniere's disease. Experimentally induced ELH in animal model has been developed in order to understand better the consequences of this morphopathology on inner ear structure and function. OBJECTIVES: The aim of this study is to develope the animal model of ELH. MATERIAL AND METHODS: We made an animal endolymphatic hydrops model on giunea pigs by mechanically obliterating the right endolympatic sac and duct. The left ears were used as a controls. RESULTS: After twelve weeks, the temporal bones were prepared for light and electron microscopic examination. The light microscopic study showed the distension and folding of Reissner's membrane, however other pathologic changes were uncertain in hydropic aminal. Under scanning electron microscopic examination, the cellular population of Reissner membrane remained normal but the microvilli of Reissner membrane were significantly decreased in hydrops group. The damage pattern of stereocilia were most severe in the apical turns, followed by the middle and the basal turns, in the order of severeity. The outer hair cell third row was most severely damaged in the entire cochlea, whereas the outer hair cell first row was relatively spared. On the single hair cell examination, abnormal distensions of the tip of stereocilia started from longer stereocilia. The number of microvilli on marginal cell was reduced in stria vascularis, but atrophic change was not observed.