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Changes in the Testicular Damage Caused by Indium Arsenide and Indium Phosphide in Hamsters During Two Years After Intratracheal Instillations

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Changes in the Testicular Damage Caused by Indium Arsenide and Indium Phosphide in Hamsters During Two Years After Intratracheal Instillations Changes in the Testicular Damage Caused by Indium Arsenide and Indium Phosphide in Hamsters during Two Years after Intratracheal Instillations Minoru OMURA1,Koji YAMAZAKI1,2,Akiyo TANAKA1,Miyuki HIRATA1, Yuji MAKITA1and Naohide INOUE1 1 Department of Hygiene, 2Department of GraduateGeneral SchoolSurgery of Medical Sciences, Kyushu University , Graduate School of Medical Sciences, Kyushu University Abstract: Changes in the Testicular Damage semiconductor materials is apparent in human subjects. Caused by Indium Arsenide and Indium Phosphide (J Occup Health 2000; 42: 196-204) in Hamsters during Two Years after Intratracheal Instillations: Minoru OMURA,et al. Department of Key words: Indium arsenide, Indium phosphide, Hygiene, Graduate School of Medical Sciences, Indium, Intratracheal instillation,Testicular toxicity,Long Kyushu University-Change in the testicular damage term observation, Sertoli cell, Spermatogonia caused by indium arsenide (InAs) and indium phosphide (InP) was examined during two yr after III-V compound semiconductor materials, such as repetitive intratracheal instillations in hamsters. In this gallium arsenide (GaAs), are now extensively used for study, 4.0 mg/kg body weight/day of InAs or 3.0 mg/kg manufacturing light-emitting diodes, semiconductor body weight/day of InP was instilled intratracheally lasers and microcircuits''. This means that the health risk twice weekly for eight wk. A single instillation dose of due to these materials needs to be properly assessed. indium was 2.4 mg/kg body weight in both groups. Testicular damage was evaluated 0, 8, 16, 40, 64 and Indium is an element in the IIIa column of the periodic 88 wk after the last instillation. Both InAs and InP were table and is incorporated into III-V compound proved to be definite testicular toxicants. Both materials semiconductor materials, such as indium arsenide (InAs) decreased reproductive organ weight and caudal sperm and indium phosphide (InP). Only a few data are count, and caused severe histopathologic changes in available to assess the health risk of indium-containing the testes. InAs-induced testicular damage was always compound semiconductor materials'-", especially to more serious than InP-induced testicular damage. The assess the risk to the reproductive system' ". We serum indium concentration in the InAs group was previously examined the testicular toxicity of GaAs and always higher than that in the InP group, and indium InAs immediately after eight-week repetitive intratracheal was probably a toxic element in both materials. In the instillation (7.7 mg/kg body weight/day, twice a week) histopathologic examination, vacuolization of in rats' and hamsters". GaAs showed a definite testicular seminiferous epithelium was frequently observed as an early histopathologic change and spermatogonia toxicity in both species. InAs showed only a weak remained in general even in the seminiferous tubules testicular toxicity in rats and did not show any sign of with severe histopathologic changes in both groups. It testicular toxicity in hamsters. But our study on hamsters is therefore estimated that Sertoli cells, not stem cell was inadequate to evaluate the testicular toxicity of InAs. spermatogonia, were the target cells of these indium In the study, eight-wk intratracheal instillation could not containing compound semiconductor materials. The be completed because of emaciation of the animals". We threat of InAs and InP to male reproduction was proved also thought that our study on rats might have in this study. We concluded that male reproductive underestimated the testicular toxicity of InAs because disorders should not be overlooked when severe high concentrations of indium and arsenic were still exposure to indium-containing compound detected in the serum of hamsters a long time after the Received March 22, 2000; Accepted April 24, 2000 intratracheal instillations of InAs''. Chemical-induced Correspondence to: M. Omura, Department of Hygiene, Graduate adverse effects on the male reproductive system are School of Medical Sciences, Kyushu University, Fukuoka 812-8582, drawing attention in the field of industrial hygiene in Japans Japan because of 2-bromopropane-induced reproductive hazards in a Korean electronic company'-"' and group and 3.0 mg/kg body weight in the InP group. The widespread anxiety about endocrine-disrupting chemicals. single instillation dose of indium was 2.4 mg/kg body We therefore decided to re-examined the testicular weight in both groups. Hamsters in the control group toxicity of InAs in hamsters. Testicular toxicities of InP were given phosphate buffer solution only. Hamsters were also examined and were compared with those of were thus treated twice weekly for eight weeks. No InAs in order to clarify the contribution of indium to the animals died during the administration period. testicular toxicity of indium-containing compound semiconductor materials. Examination of the effects on the male reproductive system Materials and Methods Eight hamsters died of emaciation (five in the InAs Test materials group and three in the InP group), seven were cannibalized InAs and InP were provided by Mitsuwa Chemicals (four in the control group and three in the InAs group), (Osaka, Japan) and had a purity of more than 99.99%. and four were killed accidentally before examination in These materials were finely pulverized in an agate mortar. the control group during the observation period. These The mean count diameter for InAs and InP was 1.58 ,um animals were excluded from all evaluations. Four to eight [6g (geometric standard deviation): 2.15] and 1.06 ym hamsters in each group were weighed and killed with an [6g: 1.80], respectively. Both powders were analyzed in overdose of ether 0, 8, 16, 40, 64 and 88 wk after the last an energy dispersive X-ray fluorescence element analyzer instillation. The testes and epididymes were removed (MESA-500, Horiba, Ltd., Kyoto, Japan). InAs powder and weighed. The right cauda epididymidis was contained 0.01% (wt%) of zirconium and 0.01% of homogenized in saline containing 0.05% (v/v) Triton X yttrium, and InP powder contained 0.01 % of zirconium 100 in a blender and homogenization-resistant sperm were and a trace amount of yttrium. Test materials were counted in a hemocytometer. The right testis was fixed suspended in particle form in pathogen-free phosphate in Bouin's solution, embedded in paraffin wax, thinly buffer solution (0.025 M, pH 6.86) just before instillation. sectioned and stained with periodic acid Schiff reagent The concentrations of InAs and InP in the suspension (PAS) and hematoxylin. Histopathologic changes in the were 2.0 mg/ml and 1.5 mg/ml, respectively. testis were examined under an optical microscope. All cross-sections of seminiferous tubules in one transverse Animal section of the testis (about 200-400 cross-sections of the Male Syrian golden hamsters were purchased at six tubule) were examined and the seminiferous tubules with weeks of age from Japan SLC, Inc., (Shizuoka, Japan). histopathologic changes were counted. Degeneration of The animals were housed four per stainless cage and were one or two germ cells was not regarded as a maintained in a specific pathogen-free laboratory room histopathologic change. of the Laboratory of Animal Experiments, Faculty of The results of this study concerning the pulmonary Medicine. Kyushu University. The light cycle was 12 h: toxicity and serum concentrations of indium and arsenic 12 h (light/dark), the temperature was 22-25°C, and the have already been reported by Yamazaki et al.". air humidity was 50-60%. The animals were provided with CE-2 feed (Clea Japan Inc., Tokyo, Japan) and tap Statistical analysis water ad libitum. This experiment was reviewed by the Data were analyzed for the mean and standard deviation Committee of Ethics on Animal Experiments in the and F test was performed to evaluate equality of variance. Faculty of Medicine, Kyushu University and was carried If a significant difference was found in a variance, t test out under the Guidelines for Animal Experiment of the with Welch's correction was used for the statistical Faculty of Medicine, Kyushu University and the Law analysis, and otherwise a t test without a correction was (No. 105) and Notification (No. 6) of the Government of used. The differences were interpreted as significant at Japan. p<0.05. There were only two hamsters adequate for evaluation in the control group 8 wk after the last Treatment instillation, (four were killed accidentally before After a two-week acclimation period, 144 hamsters, examination and two had marked unilateral atrophy of eight weeks of age, were randomized into three groups the testis and the epididymis). Therefore, statistical by weight: a control group, an InAs group and an InP analysis was not performed at this point. group. The mean body weights of hamsters in the control Results group, InAs group and InP group were 109.5 g, 111.5 g, and 111.6 g, respectively. Hamsters in each group were Body weight, reproductive organs weights, and sperm intratracheally instilled with a 2.0 ml suspension/kg body count in the cauda epididymidis weight under ether anesthesia. A single instillation dose Body weight in the InAs group had already decreased of test material was 4.0 mg/kg body weight in the InAs significantly immediately (0 wk) after the last instillation and was always approximately 70% of the control value the InP group were compatible with those in the control thereafter (Fig. 1). Body weight in the InP group was group from 0 wk to 8 wk after the last instillation. compatible with that in the control group immediately Reproductive organ weights in this group decreased after the last instillation. Body weight in this group significantly from 16 wk to 64 wk after the last instillation decreased significantly from 16 wk to 64 wk after the and were 60-70% of the control values in this period. In last instillation and was 80-90% of the control value in the InP group, reproductive organ weights became this period.
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