An in vitro rapid evaluation of drug-induced ototoxicity and of reductive effect of calcium on aminoglycoside ototoxicity using organ culture

Abstract

The ototoxicity of aminoglycoside and anticancer platinum drug was analysed using an organ culture system. The effect of calcium antagonism on aminoglycoside ototoxicity was investigated by the same system. The inner ears of mice, 16-day embryos, were cultured for 5 days with or without gentamicin (GM) or kanamycin (KM) or streptomycin (SM), or ribostamycin (RSM), including 1, 10, 100, or 1000 μg/ml respectively. The 21th gestational day-inner ear was cultured in vitro during 4 days with or without Cisplatin (CDDP) or platinum analog DWA2114R (DWA), including 0.1, 1, or 10 μg/ml, respectively. The 16th gestational day-inner ear was cultured in vitro for 5 days with 10 μg/ml KM, adding 5mM Ca2+or lOmM Ca2+to the culture medium. The damages of crista ampullaris and macula utriculi of cultured inner ear were estimated according to the ototoxicity score based on morphological changes by a light microscopic observation of serial sections of the materials. We difined four grades for the damages according to the following criteria; grade 1: damage of apical surface of the hair cells, grade 2: the existence of debris in the emdolymph space, grade 3: disappearance of the hair cells, grade 4: degeneration of the supporting cells. Uning this system following results were obtained: 1) the effect of aminoglycoside was dose dependent, 2) the order of ototoxicity was following; GM >KM >SM >RSM, 3) the drug concentration of 1000 μg/ml is sufficient to study its ototoxic potential in this system, 4) the effect of both CDDP and DWA was obvious at a concentration of 0.1 μg/ml, 5) DWA showed almost the same ototoxicity as CDDP at the same concentration, 6) Adding 5mM Ca2+or 10mM Ca2+to the culture medium, the ototoxic damage induced by 10 μg/ml of KM was not noticed. © 1991, The Oto-Rhino-Laryngological Society of Japan, Inc. All rights reserved.