Minocycline attenuates ototoxicity and enhances antitumor activity of cisplatin treatment in vitro.

OBJECTIVE: Some agents have been shown to prevent cisplatin-induced ototoxicity. The objective is to show that the agent minocycline protects the cochlea against cisplatin damage and enhances the cytotoxicity of anticancer therapies. STUDY DESIGN: In vitro chemotherapeutic assessments of minocycline. SETTING: Research laboratory. SUBJECTS AND METHODS: Hep-2 cells were cultured with and without 100 µM cisplatin, and cell growth inhibition was assessed. Autophagy in the samples was visually evaluated by electron microscopy and by beclin-1 expression using Western blotting. In another experiment, cochlear basilar membranes of 3-day-old rats were isolated and cultured. The cultures were treated with the same concentration of cisplatin or cisplatin combined with minocycline. Immunofluorescence staining was used to identify changes in spiral ganglions. RESULTS: Cell growth was inhibited in a dose-dependent manner following minocycline treatment. Furthermore, the combination of cisplatin and minocycline effectively increased tumor cell death (P < .01). Autophagosomes were also evident in cells treated with minocycline. Beclin-l protein expression was increased after minocycline treatment in Hep-2 cells. In an experiment evaluating cochlear spiral ganglion neuron survival, it was found that the number of surviving cochlear neurons significantly increased in the minocycline pretreatment group compared with the group treated with cisplatin alone (P < .01). CONCLUSION: This study shows that minocycline alone, or in combination with chemotherapeutic drugs, inhibits the growth of tumor cells and attenuates ototoxicity. It is also shown that minocycline activates cell autophagy via the beclin-1 signaling pathway, which may be an additional underlying cause of Hep- 2 cell death.

Functional and morphological effects of fotemustine on the auditory system of the rat.

OBJECTIVE: This study aimed to elucidate the potential inner-ear effects of fotemustine, a chemotherapeutic agent which crosses the blood-brain barrier and is used in the treatment of primary and metastatic brain tumours and metastatic melanoma. METHODS: This study utilised distortion product otoacoustic emissions and transmission electron microscopy in order to conduct electrophysiological and morphological assessments, using a rat experimental model. Twelve ears of six male rats were examined two months following intraperitoneal slow infusion of fotemustine (100 mg/m2 or 7.4 mg/kg). Pre- and post-treatment measurements were compared. Finally, electron microscopy was performed on three rat temporal bones. RESULTS: After infusion of fotemustine, distortion product otoacoustic emissions revealed a significant reduction in signal-to-noise ratios only at 3600 Hz (from 11.95 +/- 7.52 to -0.26 +/- 9.45 dB) and at 3961 Hz (from 18.09 +/- 7.49 to 6.74 +/- 12.11 dB) (referenced to 2f1 - f2). Transmission electron microscopy of the temporal bone revealed ultrastructural changes in the outer hair cells, stria vascularis and cochlear ganglion at the cochlear basal turn. The ganglion cell perikarya were unaffected. CONCLUSIONS: Fotemustine was administered via intraperitoneal slow infusion in a rat experimental model. Twelve ears of six survivors, from 10 rats, were evaluated at the second month. Fotemustine was determined to have a potential for ototoxicity at 3600 and 3961 Hz. Three randomly chosen rats underwent electron microscopy for morphological analysis. Morphological effects in the cochlear basal turn were observed. Oedematous intracytoplasmic spaces and perivascular areas of the stria vascularis, as well as distorted chromatin content, were detected, thereby suggesting potential ototoxic effects for this agent. Further experimental and clinical studies are required in order to determine whether the effect seen in this pilot study is reversible, and to analyse effects in humans.

Morphological evidence of ototoxicity of the iron chelator deferoxamine.

Recent reports of the role of iron-catalyzed free radical formation in gentamicin ototoxicity and the successful attenuation of gentamicin ototoxicity by iron chelators led us to re-examine experimental material from a previously unpublished study of deferoxamine. Deferoxamine was injected i.m. into adult Japanese quail at either 300 or 750 mg/kg body weight for 30 days. Examination of sections from the basilar papilla at the light microscope level indicated that supporting cells were damaged after the lower drug dose, and that both supporting cells and hair cells were damaged after the higher drug dose. High, prolonged exposure to deferoxamine produced pathological changes similar to those seen in the basilar papilla after much lower, shorter doses of gentamicin. These results demonstrate that deferoxamine damages the quail inner ear and are consistent with the idea that the ototoxic actions of gentamicin may be mediated by iron chelation.

Cadmium-metallothionein nephrotoxicity in the rat: transient calcuria and proteinuria.

After a s.c. injection of 0.4 mg Cd/kg as cadmium-metallothionein (CdMT) in rats, a marked increase in urinary protein concentration appeared at 16-40 h. There was a peak of urinary Cd content during the first 4 h after the treatment. Urinary Ca was increased at 8 h after the CdMT injection and returned to normal level at 32 h. Luminal and basolateral renal membrane vesicles were isolated from both control group and CdMT (0.4 mg Cd/kg) group at 24 h after the injection. Calcium uptake and binding of both fractions were decreased in the group treated with CdMT. Cd, Zn and MT concentrations in the kidney cortex were increased, but Ca concentration was not significantly changed. Since injected CdMT is probably only partly reabsorbed by tubular cells at the dose level of 0.4 mg Cd/kg as CdMT, excessive plasma CdMT is rapidly excreted in urine, explaining the increased Cd excretion during the first few hours observed in the present experiment. Decreased Ca binding in the luminal membranes as observed in vitro could be one of the mechanisms of production of calcuria if occurring in vivo. Another possible explanation of calcuria is that Cd ions released from CdMT into the cytoplasm of the tubular cell, may exert ionic interference with Ca transport across the luminal membranes and produce decreased Ca reabsorption. It is known that a disturbance of Ca metabolism could influence the membrane stability and such a change may contribute to explaining the proteinuria characteristic of CdMT nephrotoxicity. The reversibility of the proteinuria observed after a single dose of CdMT may be related to the induction of metallothionein synthesis in the renal cells.