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Chronic Pulmonary Toxicity Study of Indium-Tin Oxide and Indium Oxide Following Intratracheal Instillations Into the Lungs of Hamsters

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Chronic Pulmonary Toxicity Study of Indium-Tin Oxide and Indium Oxide Following Intratracheal Instillations Into the Lungs of Hamsters Journal of J Occup Health 2010; 52: 14–22 Occupational Health Chronic Pulmonary Toxicity Study of Indium-Tin Oxide and Indium Oxide Following Intratracheal Instillations into the Lungs of Hamsters Akiyo TANAKA1, Miyuki HIRATA1, Toshiaki HOMMA2 and Yutaka KIYOHARA1 1Department of Environmental Medicine, Graduate School of Medical Sciences, Kyushu University and 2Division of Respiratory Disease, Teikyo University Chiba Medical Center, Japan Abstract: Chronic Pulmonary Toxicity Study of 3 out of 15 of the ITO 6 mg/kg-treated hamsters. Indium-Tin Oxide and Indium Oxide Following Conclusions: The present results clearly demonstrate Intratracheal Instillations into the Lungs of that ITO and In2O3 particles caused chronic pulmonary Hamsters: Akiyo TANAKA, et al. Department of toxicity when repeated intratracheal instillations were Environmental Medicine, Graduate School of given to hamsters. Medical Sciences, Kyushu University—Objectives: (J Occup Health 2010; 52: 14–22) The aim of this study was to clarify the chronic toxicological effects of indium-tin oxide (ITO) and indium Key words: Chronic pulmonary toxicity, Hamsters, oxide (In2O3) on laboratory animals. Methods: Male Indium oxide, Indium-tin oxide Syrian golden hamsters were intratracheally administered 3 mg/kg or 6 mg/kg of ITO particles, or Indium-tin oxide (ITO), which is a sintered alloy 2.7 mg/kg or 5.4 mg/kg of In2O3 particles, containing containing 90% indium oxide and 10% tin oxide, is used 2.2 mg/kg or 4.5 mg/kg of indium, twice a week, for 8 for liquid-crystal displays (LCDs) or plasma displays and wk. Control hamsters were given vehicle of distilled mobile phone displays. In 2007, the indium demand for water only. The hamsters were euthanized serially up ITO is said to have accounted for 90% of the total indium to 78 wk after the final instillation and the toxicological effects were determined. Results: Body weight gain demand in Japan and ITO production is expected to rise was significantly suppressed in the ITO 6 mg/kg-treated significantly due to its increasing use in electronic hamsters compared with the control group, but not in devices, such as LCDs1). Due to the increasing production of ITO, the potential health hazards of occupational or the ITO 3 mg/kg-treated or In2O3-treated hamsters. Relative lung weights among all the indium-treated environmental exposure to this material have began to groups were significantly increased compared to that cause great concern. Since the mid 1990’s, data have in the control group throughout the observation period. become available indicating that indium compounds can The serum indium concentration among all the indium- be toxic to animals2–12), although there is little information treated groups gradually increased up to the end of regarding the adverse health effects for workers exposed the observation period. Histopathologically, foci of slight to indium compounds. In 2003, the first case of interstitial to severe pulmonary inflammatory response with diffuse pneumonia caused by occupational exposure to ITO was alveolar or bronchiolar cell hyperplasia, expansion of 13) the alveolar spaces and interstitial fibrotic proliferation reported . Interstitial pneumonia and numerous fine ITO were present in all the indium-treated hamsters and particles were recognized in the lung of a worker who the severity of these lesions worsened with the passage was engaged in the metal-processing industry as an of time. Lung benign adenomas were only manifest in operator of a wet surface grinder dealing with ITO sputtering targets. An extremely high indium concentration (290 µg/l) was detected in the worker’s Received Jul 1, 2009; Accepted Oct 6, 2009 serum. However, the question as to whether or not Published online in J-STAGE Nov 25, 2009 exposure to indium, including ITO, is a potential Correspondence to: A. Tanaka, Department of Environmental occupational health hazard for employees handling these Medicine, Graduate School of Medical Sciences, Kyushu materials has not been fully resolved. University, Venture Business Laboratory 2F, 6–10–1 Hakozaki, In our previous study, which has been the only study Higashi-ku, Fukuoka 812-8581, Japan dealing with the subchronic toxicity of ITO, we (e-mail: [email protected]) Akiyo TANAKA, et al.: Toxicity of Indium-Tin Oxide and Indium Oxide in Hamsters 15 demonstrated the pulmonary toxicity of 6 mg/kg of ITO at the Laboratory of Animal Experiments, Graduate (4.5 mg In/kg) and 6 mg/kg of indium phosphide (InP, School of Medical Sciences, Kyushu University. Five 4.8 mg In/kg) per one instillation, when given to hamsters hamsters were housed in one stainless steel cage and in repeated intratracheal instillations over a 4 mo period11). provided with a commercial diet (CE-2 pellets, Clea Japan Significant depression of body-weight gain was observed Inc., Tokyo, Japan) and tap water ad libitum. The in the InP-treated hamsters compared with the controls, intratracheal instillations were started on 8-wk-old but not in the ITO-treated hamsters. Concerning lung hamsters after an acclimatization period of 2 wk. lesions, both pneumonia and hyperplastic lesions with squamous cell metaplasia were observed in the InP group. Intratracheal instillation of materials Pneumonia was recognized in the ITO group, but The hamsters, mean ± SD weight of 112.2 ± 6.9 g, hyperplastic lesions were not. Both ITO and InP particles were randomly divided into 5 groups: the control group are toxic to the lung of hamsters, and InP has greater (n=40), those treated by a dose of 3 mg/kg of ITO (ITO 3 systemic or pulmonary toxic effects than ITO. mg group, n=40), 6 mg/kg of ITO (ITO 6 mg group, Accordingly, we considered that indium was the main n=40), 2.7 mg/kg of In2O3 (In2O3 2.7 mg group, n=23), cause of the lung lesions. and 5.4 mg/kg of In2O3 (In2O3 5.4 mg group, n=23). There In this study, we evaluated the chronic pulmonary toxic was no significant difference in body weight among the effect of ITO when administered to the trachea of groups at the start of the experiment. ITO and indium hamsters twice a week for 8 wk according to the method oxide were suspended in sterile distilled water just prior of Yamazaki et al.9), who demonstrated the chronic to each instillation. Each material was instilled into the pulmonary toxicity of indium arsenide (InAs) and InP trachea of hamsters which had been anesthetized with which are semiconductor materials containing indium. ether, twice a week, a total of 16 times, during an 8-wk ITO was administered at a dose of 6 mg/kg or 3 mg/kg, period. Each instillation per animal comprised 3 mg/kg the same and half the dose used in our previous study11). or 6 mg/kg ITO (2.2 mg In/kg or 4.5 mg In/kg) or 2.7 Indium oxide, which is the main material of ITO, was mg/kg or 5.4 mg/kg as In2O3 (2.2 mg In/kg or 4.5 mg In/ also used in order to compare the toxicity of indium kg), these doses being either the same as, or half the compounds, and it was administered in equimolar quantity of the molar indium dose used in our earlier amounts of indium to those of ITO. study11). The control hamsters received 1.0 ml/kg of distilled water only. Six to eight surviving hamsters in Materials and Methods each group were euthanized by carbon dioxide gas 0, 8, Test materials 16, 40 or 78 wk after the final instillation in the two ITO ITO particles were donated by a company. They groups and the control group, and 0, 16 or 40 wk after contained 74.4% (wt %) indium and 7.8% tin, the the final instillation in the two In2O3 groups, and then remainder being oxygen. Indium oxide, over 99.99% autopsied. The blood was taken from the posterior vena pure, was purchased from Katayama Chemicals, Osaka, cava and the serum was separated after centrifugation. Japan. The mean particle diameter for ITO was 0.95 µm Resected lungs were weighed and fixed in 10% neutral [σg (geometric standard deviation): 2.42], as measured buffered formalin and processed in paraffin. Specimens by an image analyzer (Nikon Co. Ltd, Tokyo Japan) using were cut at a thickness of 6 µm and each section was scanning electron microscopy (T-220, JEOL Ltd, Tokyo, stained with hematoxylin-eosin. These sections were Japan). The mean particle diameter for indium oxide examined by light microscopy. Histopathological was 0.14 µm, which was found from the ratio of the findings in the lung were scored as present or absent; if surface area of the particle by the BET adsorption method. they were absent, findings were expressed as negative. ITO particles were analyzed in an energy dispersive X- In the case of pulmonary lesions, the severity of each of ray fluorescence element analyzer (MESA-500, Horiba the lung lesions was graded on a 4-item scale of slight to Ltd., Kyoto, Japan). The ITO particles contained 0.16% severe, indicating the approximate fraction of the lung zirconium and 0.02% silicon. ITO and indium oxide or structure judged to be involved (slight=1–10%, particles were suspended in sterile distilled water just mild=11–24%, moderate=25–50%, and severe=51– prior to each instillation. 100%). Animals Quantifying of serum indium concentration by inductively One hundred sixty-six male Syrian golden hamsters, coupled plasma mass spectrometry (ICP-MS) from the colony of Japan SLC Inc. (Shizuoka, Japan) Serum samples were stored at –80°C until analysis. were purchased at 6 wk of age and housed under For pretreatment, 1 ml of serum was digested with 5 ml temperature conditions of between 22°C and 25°C.
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