Use of scanning electron microscopy in the cochlea of guinea pigs / Microscopia eletrônica de varredura em cócleas de cobaias

Abstract Introduction: The use of electron microscopy in the study of the inner ear has allowed us to observe minute details of the hair cells, especially in ototoxicity studies; however, the preparation of this material is a difficult and delicate task. In an attempt to simplify the handling of these materials, two agents, toluidine blue and ethylenediamine tetra-acetic acid were tested, in addition to the elimination of osmium tetroxide during the preparation of albino guinea pig cochleae. We also tested the applicability of these methodologies in an ototoxicity protocol. Objective: To verify the quality of the images obtained with and without the use of ethylenediamine tetra-acetic acid, toluidine blue and osmium tetroxide in the preparation of cochleae of albino guinea pigs for the scanning electron microscopy. Methods: Three groups of cochleae were used. In Group 1, 10 cochleae were prepared with the usual methodology, dissecting the optical capsule without decalcification and using osmium tetroxide as a post-fixative agent. In Group 2, we prepared 10 cochleae decalcified with ethylenediamine tetra-acetic acid, injecting toluidine blue in the endolymphatic space to facilitate the identification of the organ of Corti. In Group 3, we used 4 cochleae of guinea pigs that received 3 doses of cisplatin (7.5 mg/kg, D1-D5-D6), two prepared according to the methodology used in Group 1 and two with that used in Group 2. Scanning electron microscopy images were obtained from the organ of Corti region of the basal turn of each cochlea. Results: The organ of Corti was more easily identified with the use of toluidine blue. The dissection of the cochlea was more accurate in the decalcified cochleae. The quality of the images and the preservation of the organ of Corti obtained with the two methodologies were similar. Conclusion: The proposed modifications resulted in images of similar quality as those observed using the traditional methodology.

Resumo Introdução: O emprego da microscopia eletrônica no estudo da orelha interna permitiu observar detalhes minuciosos das células ciliadas especialmente em estudos de ototoxicidade. Entretanto, o preparo desse material é trabalhoso e delicado. Para simplificar a manipulação desses materiais, testou-se o uso de dois agentes, azul de toluidina e ácido etilenodiamino tetra-acético, além da retirada do tetróxido de ósmio na preparação de cócleas de cobaias albinas. Testamos também a aplicabilidade dessas metodologias em um protocolo de ototoxicidade. Objetivo: Verificar a qualidade das imagens obtidas com e sem o uso de ácido etilenodiamino tetra-acético, azul de toluidina e tetróxido de ósmio na preparação de cócleas de cobaias albinas para a microscopia eletrônica de varredura. Método: Foram utilizados três grupos de cócleas. No Grupo 1 preparou-se 10 cócleas com a metodologia usual, dissecando a cápsula ótica sem descalcificac¸ão e utilizando tetróxido de ósmio como pós-fixador. No Grupo 2 preparamos 10 cócleas descalcificadas com ácido etilenodiamino tetra-acético, injetando azul de toluidina no espac¸o endolinfático para facilitar a identificação do órgão de Corti. No Grupo 3 utilizamos 4 cócleas de cobaias que receberam 3 doses de cisplatina (7,5 mg/kg, D1-D5-D6), duas preparadas com a metodologia do Grupo 1 e duas com a do Grupo 2. Foram obtidas imagens da microscopia eletrônica de varredura da região do órgão de Corti do giro basal de cada cóclea. Resultados: O órgão de Corti foi mais facilmente identificado com o azul de touidina. A dissecção da cóclea foi mais precisa nas cócleas descalcificadas A qualidade das imagens e a preservac¸ão do órgão de Corti obtidas com as duas metodologias foi similar. Conclusão: As modificações propostas resultaram em imagens de qualidade similar as observadas com o uso da metodologia tradicional.

Use of scanning electron microscopy in the cochlea of guinea pigs.

INTRODUCTION: The use of electron microscopy in the study of the inner ear has allowed us to observe minute details of the hair cells, especially in ototoxicity studies; however, the preparation of this material is a difficult and delicate task. In an attempt to simplify the handling of these materials, two agents, toluidine blue and ethylenediamine tetra-acetic acid were tested, in addition to the elimination of osmium tetroxide during the preparation of albino guinea pig cochleae. We also tested the applicability of these methodologies in an ototoxicity protocol. OBJECTIVE: To verify the quality of the images obtained with and without the use of ethylenediamine tetra-acetic acid, toluidine blue and osmium tetroxide in the preparation of cochleae of albino guinea pigs for the scanning electron microscopy. METHODS: Three groups of cochleae were used. In Group 1, 10 cochleae were prepared with the usual methodology, dissecting the optical capsule without decalcification and using osmium tetroxide as a post-fixative agent. In Group 2, we prepared 10 cochleae decalcified with ethylenediamine tetra-acetic acid, injecting toluidine blue in the endolymphatic space to facilitate the identification of the organ of Corti. In Group 3, we used 4 cochleae of guinea pigs that received 3 doses of cisplatin (7.5mg/kg, D1-D5-D6), two prepared according to the methodology used in Group 1 and two with that used in Group 2. Scanning electron microscopy images were obtained from the organ of Corti region of the basal turn of each cochlea. RESULTS: The organ of Corti was more easily identified with the use of toluidine blue. The dissection of the cochlea was more accurate in the decalcified cochleae. The quality of the images and the preservation of the organ of Corti obtained with the two methodologies were similar. CONCLUSION: The proposed modifications resulted in images of similar quality as those observed using the traditional methodology.

Specialized features of the outer hair cell shapes in the cochlear fovea of bats.

In this study, we examined the specialized features of the outer hair cells (OHCs) and the stereocilium bundles of the bat cochlear fovea. Bat cochlea hair cells were observed by scanning and transmission electron microscopy, and the auditory brainstem response thresholds were assessed. The stereocilia bundles of the OHCs were extremely short. The OHC bodies were flask-shaped and cambiform or ball-shape in the cochlear fovea. Digitations in the Deiters cells had exaggerated lengths, and cup formation of the Deiters cell, housed at the bottom of the OHC in the base of the cell, showed a specialized shape. Our results provide the first evidence that different shapes of the OHCs in the cochlea fovea are related to the high-frequency function of auditory response. Echolocating bats have cochlear morphologies that differ from those of non-echolocating animals. Bat cochlear foveae are specialized for analyzing the Doppler-shifted echoes of the first-harmonics of the CF2 component; these are overrepresented in the frequency range around the dominant harmonic of the echolocation calls of bats. However, the OHCs of the bat cochlear fovea have not been fully characterized.

[Lack of protection against gentamicin ototoxicity by auditory conditioning with noise]. / Falta de proteção contra a ototoxicidade da gentamicina pelo condicionamentoauditivo com ruído.

INTRODUCTION: Auditory conditioning consists of the pre-exposure to low levels of a potential harmful agent to protect against a subsequent harmful exposure. OBJECTIVE: To confirm if conditioning with an agent different from that used to cause the trauma can also be effective. METHODS: This was an experimental study with 17 guinea pigs, divided into three groups: an ototoxic control group (Cont) that received intramuscular administration of gentamicin 160 mg/kg/day for ten consecutive days, but no sound exposure; a sound control group (Sound) that was exposed to 85 dB broadband noise centered at 4 kHz, 30 min each day for ten consecutive days, but received no ototoxic medications; and an experimental group (Expt) that received sound exposure identical to the Sound group and after each noise presentation, received gentamicin similarly to Cont group. The animals were evaluated by distortion product otoacoustic emissions (DPOAEs), brainstem auditory evoked potentials (BAEPs), and scanning electron microscopy. RESULTS: The animals that were conditioned with noise did not show any protective effect compared with the ones that received only the ototoxic gentamicin administration. This lack of protection was observed functionally and morphologically. CONCLUSION: Conditioning with 85 dB broadband noises, 30 min a day for ten consecutive days does not protect against an ototoxic gentamicin administration of 160 mg/kg/day for ten consecutive days in the guinea pig.

Lack of protection against gentamicin ototoxicity by auditory conditioning with noise / Falta de proteção contra a ototoxicidade da gentamicina pelo condicionamento auditivo com ruído

INTRODUCTION: Auditory conditioning consists of the pre-exposure to low levels of a potential harmful agent to protect against a subsequent harmful presentation. OBJECTIVE: To confirm if conditioning with an agent different from the used to cause the trauma can also be effective. METHOD: Experimental study with 17 guinea pigs divided as follows: group Som: exposed to 85 dB broadband noise centered at 4 kHz, 30 minutes a day for 10 consecutive days; group Cont: intramuscular administration of gentamicin 160 mg/kg a day for 10 consecutive days; group Expt: conditioned with noise similarly to group Som and, after each noise presentation, received gentamicin similarly to group Cont. The animals were evaluated by distortion product otoacoustic emissions (DPOAEs), brainstem auditory evoked potentials (BAEPs) and scanning electron microscopy. RESULTS: The animals that were conditioned with noise did not show any protective effect compared to the ones that received only the ototoxic gentamicin administration. This lack of protection was observed functionally and morphologically. CONCLUSION: Conditioning with 85 dB broadband noise, 30 min a day for 10 consecutive days does not protect against an ototoxic gentamicin administration of 160 mg/kg a day for 10 consecutive days in the guinea pig. .

INTRODUÇÃO: O condicionamento auditivo consiste da pré-exposição de um agente lesivo em baixos níveis para proteger contra uma posterior apresentação lesiva. OBJETIVO: Confirmar se o condicionamento com um agente diferente do utilizado para causar o trauma pode ser efetivo. MÉTODO: Estudo experimental com 17 cobaias albinas divididas como a seguir- grupo Som: exposto a um ruído branco de 85 dB centrado em 4 kHz, 30 minutos por dia, por 10 dias consecutivos; grupo Cont: administração intramuscular de gentamicina 160 mg/kg por dia, por 10 dias consecutivos; grupo Expt: condicionado com ruído como o grupo Som. Após cada exposição ao ruído, recebeu gentamicina similarmente ao grupo Cont. Os animais foram avaliados por emissões otoacústicas produto de distorção (EOAPDs), potencial evocado auditivo de tronco encefálico (PEATE) e microscopia eletrônica de varredura (MEV). RESULTADOS: Os animais que foram condicionados com ruído não mostraram qualquer efeito protetor quando comparados com os que receberam apenas a gentamicina em doses ototóxicas. Esta ausência de proteção foi observada tanto funcionalmente quanto morfologicamente. CONCLUSÃO: Os autores concluíram que o condicionamento com ruído branco a 85 dB por 30 minutos, por dia por 10 dias consecutivos, não protege contra uma administração de gentamicina 160 mg/kg/dia, por 10 dias consecutivos. .

Otoprotection in guinea pigs exposed to pesticides and ginkgo biloba.

UNLABELLED: Pesticides are widely used in agriculture, despite the risk of hearing loss related to the exposure to their chemical components. This study looks into protective drugs to counteract the ototoxicity of pesticides. OBJECTIVE: This study aims to analyze the effect ginkgo biloba extract may have in protecting against possible cochlear damage caused by organophosphate pesticides (methamidophos). Anatomic changes are assessed through surface and electron microscopy. MATERIALS AND METHODS: This is a prospective experimental study. Twenty-one guinea pigs were given saline solution, pesticide, and ginkgo biloba alone or combined for seven consecutive days. Then their cochleas were removed and examined in a scanning electron microscope. RESULTS: Pesticide-exposed guinea pigs had morphological alterations in their cochleas and injuries in the three turns analyzed through electron microscopy. Injury intensity varied according to the dosages of the agents given to the test subjects. Guinea pigs treated with pesticide and ginkgo biloba maintained the architecture of their outer hair cells in all cochlear turns. CONCLUSION: The antioxidant properties found in the ginkgo biloba extract protected guinea pigs from pesticide ototoxicity.

Otoproteção em cobaias expostas a agrotóxico e ginkgo biloba / Otoprotection in guinea pigs exposed to pesticides and ginkgo biloba

Os agrotóxicos são amplamente utilizados na agricultura e, atualmente, fazem parte do grupo de agentes químicos que podem levar à perda auditiva. A identificação de drogas que, associadas aos ototóxicos, possam atuar como otoprotetores é objeto de estudo. OBJETIVO: Analisar a existência de efeito otoprotetor do extrato de Ginkgo biloba aos possíveis danos cocleares causados pelo agrotóxico do grupo dos organofosforados - metamidofós, avaliando-se as alterações anatômicas por meio da microscopia eletrônica de superfície. MATERIAL E MÉTODO: Estudo experimental prospectivo utilizando 21 cobaias, que sofreram ação da administração de soro fisiológico, agrotóxico e ginkgo biloba isoladamente e associadas, durante sete dias consecutivos. Após, as cócleas foram removidas e avaliadas anatomicamente pela microscopia eletrônica de varredura. RESULTADOS: As cobaias submetidas ao agrotóxico apresentaram alterações morfológicas cocleares, com lesões nas três espiras analisadas na microscopia eletrônica, intensificadas de acordo com a dosagem recebida do agente. As cobaias tratadas com agrotóxico e Ginkgo biloba apresentaram uma manutenção da arquitetura ciliar nas células ciliadas externas em todas as espiras da cóclea. CONCLUSÃO: O extrato de Ginkgo biloba, por sua ação antioxidante, atuou como fator otoprotetor à ototoxicidade pelo agrotóxico em cobaias.

Pesticides are widely used in agriculture, despite the risk of hearing loss related to the exposure to their chemical components. This study looks into protective drugs to counteract the ototoxicity of pesticides. OBJECTIVE: This study aims to analyze the effect ginkgo biloba extract may have in protecting against possible cochlear damage caused by organophosphate pesticides (methamidophos). Anatomic changes are assessed through surface and electron microscopy. MATERIALS AND METHODS: This is a prospective experimental study. Twenty-one guinea pigs were given saline solution, pesticide, and ginkgo biloba alone or combined for seven consecutive days. Then their cochleas were removed and examined in a scanning electron microscope. RESULTS: Pesticide-exposed guinea pigs had morphological alterations in their cochleas and injuries in the three turns analyzed through electron microscopy. Injury intensity varied according to the dosages of the agents given to the test subjects. Guinea pigs treated with pesticide and ginkgo biloba maintained the architecture of their outer hair cells in all cochlear turns. CONCLUSION: The antioxidant properties found in the ginkgo biloba extract protected guinea pigs from pesticide ototoxicity.

Spontaneous reversibility of damage to outer hair cells after sodium salicylate induced ototoxicity.

BACKGROUND: High sodium salicylate doses can cause reversible hearing loss and tinnitus, possibly due to reduced outer hair cell electromotility. Sodium salicylate is known to alter outer hair cell structure and function. This study determined the reversibility and cochlear recovery time after administration of an ototoxic sodium salicylate dose to guinea pigs with normal cochlear function. STUDY DESIGN: Prospective experimental investigation. METHODS: All animals received a single 500 mg sodium salicylate dose, but with different durations of action. Function was evaluated before drug administration and immediately before sacrifice. Cochleae were processed and viewed using scanning electron microscopy. RESULTS: Changes in outer hair cell function were observed to be present 2 hours after drug administration, with recovery of normal anatomy beginning after 24 hours. Subsequently, derangement and distortion of cilia reduced, with effects predominantly in row three. At 168 hours, cilia were near-normal but with mild distortions which interfered with normal cochlear physiology. CONCLUSIONS: Ciliary changes persisted for up to 168 hours after ototoxic sodium salicylate administration.

Effects of hyperbaric oxygen treatment on auditory hair cells after acute noise damage.

Acute acoustic trauma (AAT) is a sudden sensorineural hearing loss caused by exposure of the hearing organ to acoustic overstimulation, typically an intense sound impulse, hyperbaric oxygen therapy (HOT), which favors repair of the microcirculation, can be potentially used to treat it. Hence, this study aimed to assess the effects of HOT on guinea pigs exposed to acoustic trauma. Fifteen guinea pigs were exposed to noise in the 4-kHz range with intensity of 110 dB sound level pressure for 72 h. They were assessed by brainstem auditory evoked potential (BAEP) and by distortion product otoacoustic emission (DPOAE) before and after exposure and after HOT at 2.0 absolute atmospheres for 1 h. The cochleae were then analyzed using scanning electron microscopy (SEM). There was a statistically significant difference in the signal-to-noise ratio of the DPOAE amplitudes for the 1- to 4-kHz frequencies and the SEM findings revealed damaged outer hair cells (OHC) after exposure to noise, with recovery after HOT (p = 0.0159), which did not occur on thresholds and amplitudes to BAEP (p = 0.1593). The electrophysiological BAEP data did not demonstrate effectiveness of HOT against AAT damage. However, there was improvement of the anatomical pattern of damage detected by SEM, with a significant reduction of the number of injured cochlear OHC and their functionality detected by DPOAE.

Organophosphate-related ototoxicity: Description of the vestibulocochlear system ultrastructural aspects of guinea pigs.

UNLABELLED: Organophosphate toxic agents are used in agriculture and are currently part of the group of toxic agents which can lead to hearing loss, in which we have solvents, metals and asphyxiation agents. AIM: To analyze the acute ototoxic action of a group of organophosphate agents in the vestibulo-cochlear system. This is a prospective experimental study. MATERIALS AND METHODS: We used male albino guinea pigs, broken down into three groups, to which we provided distilled water (group 1 - control), agrotoxic agent - 0.3 mg/Kg/day (group 2), agrotoxic - 3 mg/Kg/day (group 3), during 7 seven consecutive days. The most used agrotoxic agent was Tamaron BR (metamidophos). The anatomical evaluation of the cochlea, saccule and utricle was carried out by means of electronic scanning microscopy after the use of the agrotoxic agent. RESULTS: The guinea pigs submitted to the organophosphate presented cochlear morphological alterations with lesions on the three turns analyzed, as well as cilia alterations in the saccule and utricle, intensified according to the agent dosage. CONCLUSION: The morphological alterations seen in the hair cells exposed to daily doses of organophosphate promote evidences of an acute deleterious effect of agrotoxic agents on the vestibulo-cochlear system.

Ototoxicidade por organofosforados: descrição dos aspectos ultraestruturais do sistema vestibulococlear de cobaias / Organophosphate-related ototoxicity: Description of the vestibulocochlear system ultrastructural aspects of guinea pigs

Os agrotóxicos organofosforados são amplamente utilizados na agricultura, e atualmente fazem parte do grupo de agentes químicos que podem levar à perda auditiva, no qual já estavam incluídos os solventes, os metais e os asfixiantes. OBJETIVO: Analisar a ação ototóxica aguda de um agrotóxico do grupo dos organofosforados na citoarquitetura do sistema vestibulococlear. Trata-se de um estudo experimental prospectivo. MATERIAL E MÉTODO: Foram utilizadas cobaias albinas machos, divididas em três grupos, nos quais se administrou água destilada (grupo 1 - controle), agrotóxico - 0,3mg/Kg/dia (grupo 2), agrotóxico - 3 mg/Kg/dia (grupo 3), durante sete dias consecutivos. O agrotóxico utilizado foi Tamaron BR (metamidofós). A avaliação anatômica da cóclea, sáculo e utrículo foi realizada através da microscopia eletrônica de varredura, após o período de aplicação do agrotóxico. RESULTADOS: As cobaias submetidas ao organofosforado apresentaram alterações morfológicas cocleares, com lesões nas três espiras analisadas, bem como alterações ciliares de sáculo e utrículo, intensificadas de acordo com a dosagem recebida do agente. CONCLUSÃO: As alterações morfológicas observadas nas células ciliadas nos grupos expostos a doses diárias de organofosforado promovem evidências de um efeito agudo degradante dos agrotóxicos no sistema vestibulococlear.

Organophosphate toxic agents are used in agriculture and are currently part of the group of toxic agents which can lead to hearing loss, in which we have solvents, metals and asphyxiation agents. AIM: to analyze the acute ototoxic action of a group of organophosphate agents in the vestibulo-cochlear system. This is a prospective experimental study. MATERIALS AND METHODS: we used male albino guinea pigs, broken down into three groups, to which we provided distilled water (group 1 - control), agrotoxic agent - 0.3mg/Kg/day (group 2), agrotoxic - 3 mg/Kg/day (group 3), during 7 seven consecutive days. The most used agrotoxic agent was Tamaron BR (metamidophos). The anatomical evaluation of the cochlea, saccule and utricle was carried out by means of electronic scanning microscopy after the use of the agrotoxic agent. RESULTS: the guinea pigs submitted to the organophosphate presented cochlear morphological alterations with lesions on the three turns analyzed, as well as cilia alterations in the saccule and utricle, intensified according to the agent dosage. CONCLUSION: the morphological alterations seen in the hair cells exposed to daily doses of organophosphate promote evidences of an acute deleterious effect of agrotoxic agents on the vestibulo-cochlear system.

The use of Maytenus ilicifolia to prevent cisplatin-induced ototoxicity.

UNLABELLED: Maytenus ilicifolia is a native plant from South America, with several medicinal properties including antioxidant effects. AIM: using an original cisplatin induced ototoxicity model, we evaluated a possible otoprotection caused by Maytenus ilicifolia extract. MATERIALS AND METHODS: clinical and experimental study design with female albino guinea pigs divided in groups as follows: 9 animals receiving cisplatin only (three doses of 7.5mg/kg/day), 4 animals receiving the plant extract only, 10 animals receiving the cisplatin protocol and 1g/kg/day of extract for 8 days, 5 animals with cisplatin and 3g/kg/day of extract for 8 days, and 5 animals receiving extract for 3 weeks and cisplatin in the last week. The tests were distortion product otoacoustic emissions, brainstem auditory response, before and after medication and scanning electron microscopy. RESULTS: the animals receiving cisplatin plus plant extract, had alterations in all the tests, showing lesions on the basal cochlear region under electron microscopy. CONCLUSIONS: Despite of the plant extract's antioxidant effect, it was not sufficient to protect the cochlea against cisplatin ototoxicity.

O uso de Maytenus ilicifolia na prevenção da ototoxicidade induzida pela cisplatina / The use of Maytenus ilicifolia to prevent cisplatin-induced ototoxicity

Maytenus ilicifolia é uma planta sul americana apresenta várias propriedades medicinais, entre elas, a ação antioxidante. OBJETIVO: Por meio de um modelo original de ototoxicidade induzida pela cisplatina, verificar uma possível ação otoprotetora do extrato aquoso desta planta. MATERIAL E MÉTODO: Estudo clínico e experimental com cobaias fêmeas, albinas divididas em 5 grupos: 9 animais recebendo somente 3 doses de 7,5mg/kg/d do protocolo de cisplatina, 4 animais somente com o extrato, 10 animais com cisplatina e 1g/kg/d de extrato por 8 dias, 5 animais com cisplatina e 3g/kg/d do extrato por 8 dias e 5 animais recebendo extrato por 3 semanas e cisplatina na última semana. Os exames foram emissões otoacústica por produtos de distorção, potencial de tronco encefálico pré e após administração de cisplatina e, microscopia eletrônica de varredura. RESULTADOS: Os animais que receberam a cisplatina com o extrato, independente da dose, obtiveram alterações em todos os testes, com lesões na região basal na microscopia eletrônica. CONCLUSÃO: Apesar do efeito antioxidante da Maytenus ilicifolia, ela não foi suficiente para bloquear o efeito ototóxico da cisplatina.

Maytenus ilicifolia is a native plant from South America, with several medicinal properties including antioxidant effects. AIM: using an original cisplatin induced ototoxicity model, we evaluated a possible otoprotection caused by Maytenus ilicifolia extract. MATERIALS AND METHODS: clinical and experimental study design with female albino guinea pigs divided in groups as follows: 9 animals receiving cisplatin only (three doses of 7.5mg/kg/day), 4 animals receiving the plant extract only, 10 animals receiving the cisplatin protocol and 1g/kg/day of extract for 8 days, 5 animals with cisplatin and 3g/kg/day of extract for 8 days, and 5 animals receiving extract for 3 weeks and cisplatin in the last week. The tests were distortion product otoacoustic emissions, brainstem auditory response, before and after medication and scanning electron microscopy. RESULTS: the animals receiving cisplatin plus plant extract, had alterations in all the tests, showing lesions on the basal cochlear region under electron microscopy. CONCLUSIONS: Despite of the plant extract's antioxidant effect, it was not sufficient to protect the cochlea against cisplatin ototoxicity.

Gentamicin attenuates gentamicin-induced ototoxicity - self-protection.

Aminoglycoside antibiotics cause considerable toxicity to the inner ear. A progressive hearing loss at high frequencies resulted from the loss of hair cells in the base of the cochlea and a constant preoccupation with finding a treatment that protects against their toxic effects. A self-protection phenomenon to high ototoxic doses of gentamicin is proposed in this paper. Thirty-eight adult guinea pigs with normal hearing were tested using Preyer's reflex and the distortion product otoacoustic emission (DPOAE) test, and their cochleae were analyzed by scanning electron microscopy. To the four groups investigated, group I (control) and group II (low dose, 10 mg/kg/day for 30 days) showed a normal DPOEA and normal outer hair cells; group III (high dose, 160 mg/kg/day for 10 days) showed the absence of DPOEA and damage to the outer hair cells; and group IV (low dose, 10 mg/kg/day for 30 days followed by a high dose of 160 mg/kg/day for 10 days) showed a normal DPOEA and normal outer hair cells. These results demonstrate that there was a considerable self-protection phenomenon by gentamicin.

Aminoglycoside antibiotic derivatives: preparation and evaluation of toxicity on cochlea and vestibular tissues and antimicrobial activity.

Aminoglycoside antibiotic derivatives such as neamine, methyl neobiosaminide B, 2-deoxystreptamine, tetra-azidoneamine, tetra-N-acetylneamine, tetra-N-carboxy-benzylneamine, tetra-N-carboxy-methylneamine and tetra-p-methoxy-benzyliminoneamine were prepared and evaluated as to their cochlear and vestibular toxicity. Methyl neobiosaminide B, the most promising derivative in the series showed selective, cochlea-dissociated vestibulotoxic activity and was considered to be a potential lead compound for the treatment of Ménière's disease. Antimicrobial properties of the compounds, qualitatively evaluated against a group of pathogenic bacteria, indicated that neomycin B sulfate, neamine as a free base and methyl-neobiosaminide B dihydrochloride show a broader range of activity when compared to the other derivatives.

Cisplatin ototoxicity and otoprotection with sodium salicylate.

Cisplatin is a potent antineoplastic drug widely used for the treatment of cancer in both adults and children. One of its most important side effects is ototoxicity, which leads to irreversible bilateral hearing loss for high frequencies (4-8 kHz). Several studies have tried to identify drugs that, when combined with cisplatin, may act as otoprotectors. The mechanism of ototoxicity of cisplatin is known to be related to changes in the antioxidant mechanisms of hair cells, especially the outer hair cells of the cochlea. Our proposal was to assess the action of sodium salicylate, which has a known antioxidant property, as a possible otoprotector of outer hair cells against the action of cisplatin, using distortion product otoacoustic emissions (DPOAEs) and scanning electron microscopy. The study was conducted on albino guinea pigs divided into two groups: group 1 (n = 9, 18 cochleae) receiving a cisplatin dose of 8.0 mg/kg/day by the intraperitoneal (ip) route for 3 days, group 2 (n = 10, 20 cochleae) receiving 100 mg/kg sodium salicylate by the subcutaneous route followed 90 min later by cisplatin, 8.0 mg/kg/day ip for 3 days, and group 3 (n = 3, six cochleae) treated with 100 mg/kg day sodium salicylate for 3 days. In group 1, there was damage with the absence of cilia in all three rows of outer hair cells in the basal turn, followed by turns 2 and 3. In group 2, hair cells were present in all cochlear turns, but exhibited disarrangement of the ciliary structure, especially in row 1, and the DPOAEs were absent after 3 days of treatment. We conclude that drugs such as sodium salicylate, because of their antioxidant properties, may protect, at least partially, the outer hair cells against cisplatin ototoxicity.

Anatomia microscópica do canal carótico e suas relações com a cóclea e a cavidade timpânica / Microscopic anatomy of the carotid canal and its relations with cochlea and middle ear

As relações entre as diversas estruturas nobres e vitais que se apresentam na intimidade do osso temporal constituem ainda hoje um grande desafio para o cirurgião otológico. Os estudos micro-anatômicos do mesmo se encontram entre as grandes armas na busca deste entendimento. OBJETIVO: Estudar as correlações anatômicas entre o canal carótico e a cóclea, e a ocorrência de deiscências do mesmo junto à cavidade timpânica. MATERIAL E MÉTODO: Estudo microscópico de 122 ossos temporais humanos. RESULTADOS As distâncias médias entre o canal carótico e os giros cocleares foram: no local de menor distância 1,05mm; no giro basal, 2,04mm; no giro médio, 2,32mm; e no giro apical, 5,7mm. A ocorrência de deiscências do canal carótico na cavidade timpânica foi de 35,2 por cento. CONCLUSÃO: A pequena distância entre estruturas cocleares e o canal carótico, e a alta prevalência de deiscências do mesmo na cavidade timpânica nos relembram o desafio com o qual o cirurgião otológico se depara ao atuar sobre o osso temporal.

Otoproteção da amifostina aos efeitos ototóxicos da cisplatina: estudo em cobaias albinas por emissões otoacústicas produtos de distorção e microscopia eletrônica de varredura

A Cisplatina é uma potente droga antineoplásica, largamente utilizada para o tratamento do câncer, tanto em adultos quanto em crianças. Dentre seus efeitos colaterais, a ototoxicidade se apresenta como um dos mais importantes e leva à perda auditiva irreversível, bilateral, para as altas freqüências (4KHz -8KHz). Estudos têm tentado identificar drogas que, associadas à cisplatina, possam atuar como otoprotetores. Sabe-se que o mecanismo da ototoxicidade pela cisplatina está relacionado a alterações nos mecanismos antioxidantes das células ciliadas, principalmente as células ciliadas externas da cóclea. A amifostina tem conhecida ação antioxidante, com conhecido efeito otoprotetor aos efeitos lesivos da radioterapia. OBJETIVO: Nossa proposta foi avaliar através de emissões otoacústicas, por produtos de distorção (EOAPD) e por microscopia eletrônica de varredura (MEV), a existência de possível efeito otoprotetor da amifostina no tratamento com cisplatina. FORMA DE ESTUDO: Experimental. MATERIAL E MÉTODO: O estudo foi realizado em cobaias albinas, que foram divididas em três grupos: Grupo 1: 6 animais -12 orelhas - cisplatina 8,0 mg/Kg/dia (via intraperitoneal) por três dias; Grupo 2: 6 animais - 12 orelhas - amifostina 100 mg/Kg/ dia (via intraperitoneal) e 90 minutos após, cisplatina 8,0 mg/Kg/dia (via intraperitoneal) por três dias; Grupo 3: 03 animais - 06 orelhas - amifostina 100 mg/Kg/dia (via intraperitoneal) por três dias. RESULTADO: Encontramos EOAPD presentes e células ciliadas externas presentes, sem lesão anatômica a MEV, nos grupos 2 e 3. Concluímos que a amifostina, por sua ação antioxidante, atua como otoprotetor a ototoxicidade pela cisplatina. No entanto, seu uso não é recomendável nos casos de tumores potencialmente curáveis, por não se saber exatamente a influência da cisplatina na eficácia da quimioterapia.

[Amifostine otoprotection to cisplatin ototoxicity: a guinea pig study using otoacoustic emission distortion products (DPOEA) and scanning electron microscopy]. / Otoproteção da amifostina aos efeitos ototóxicos da cisplatina: estudo em cobaias albinas por emissões otoacústicas produtos de distorção e microscopia eletrônica de varredura.

UNLABELLED: Cisplatin is an antineoplastic drug for cancer treatment in children and adults. The side effects of cisplatin ototoxicity are significant: irreversible bilateral hearing damage to high frequencies (4 kHz - 8 kHz). Reports recognize some drugs that are associated with cisplatin to obtain an otoprotector effect. The ototoxicity mechanisms of cisplatin are related to injury of hair cell oxidation mechanism, especially of outer hair cells. AIM: Using otoacoustic emissions distortion products (DPOEA) and scanning electron microscopy we intended to verify the action of amifostine, a radioprotective drug that has well known antioxidant characteristics and otoprotector effects to cisplatin injury. STUDY DESIGN: Experimental. MATERIAL AND METHODS: We used an experimental guinea pig model. The study was performed as follows: group 1: 6 animals, 12 ears, cisplatin 8.0 mg/Kg/day (IP), 3 days. Group 2: 6 animals, 12 ears, amifostine 100 mg/Kg/day (IP) and after 90 minutes, cisplatin 8.0 mg/Kg/day (IP), 3 days and group 3: 3 animals, 6 ears, amifostine 100 mg/Kg/day (IP), 3 days. RESULTS: DPOEA were present before and after treatment in groups 2 and 3. The normal cilium architecture of outer hair cells was supported in all cochlear turns in groups 2 and 3. We concluded that amifostine has a potential otoprotector effect against cisplatin ototoxicity and could be used in clinical trials.

Microscopic anatomy of the carotid canal and its relations with cochlea and middle ear.

UNLABELLED: The knowledge of the relations between the noble and vital structures of temporal bone is still a great challenge for the otologic surgeon. The microscopic anatomic studies of the temporal bone are one of the greatest help to prevent lesions during surgical intervention. AIM: To study the anatomic correlations between the carotid canal and the cochlea, and the occurrence of dehiscence of the carotid canal in the middle ear tympanic cavity. MATERIAL AND METHODS: Microscopic study of 122 human temporal bones. RESULTS: The average distance between the carotid canal and the cochlea were: the shortest distance, 1.05 mm; basal turn, 2.04 mm; middle turn, 2.32 mm; and apical turn, 5.70 mm. The occurrence of dehiscence of the carotid canal inside the tympanic cavity was 35.2%. CONCLUSION: The small distances between the cochlea and carotid canal, and the high incidence of dehiscence in the tympanic cavity remind us that anatomical knowledge of the temporal bone is required for the best qualification of otologists.