Alveolar macrophages are considered to play a major role in the pathophysiology of lung diseases caused by exposure to various kinds of pathogens and particles. In this study, the cytotoxic effect of different shapes of titanium dioxide (TiO(2)) was evaluated on macrophages using a unique magnetometry method and was compared with conventional methods of lactate dehydrogenase (LDH) release, apoptosis measurement, and morphological observations. Alveolar macrophages obtained from Fischer rats (F344) by bronchoalveolar lavage were incubated in vitro for 18 h with Fe(3)O(4) as a magnetometric indicator and fibrous and particulate forms of TiO(2) as test materials. In the control and particulate exposed group, rapid attenuation of the residual magnetic field, so-called "relaxation," was observed immediately after cessation of the external magnetic field. In comparison, a delay of relaxation was observed in alveolar macrophages exposed to fibrous TiO(2). LDH released into serum-free medium induced by exposure to TiO(2) increased significantly in a concentration-dependent manner in macrophages exposed to fibrous TiO(2), while negligible LDH release was observed in macrophages exposed to particulate TiO(2). The DNA ladder detection method and morphological examination detected no apoptosis in macrophages exposed to 60 micro g/ml of fibrous or particulate TiO(2). Electron microscopic examination revealed vacuolar changes and cell surface damage in macrophages exposed to fibrous TiO(2), but no significant changes in macrophages exposed to particulate TiO(2). The results of magnetometry, LDH release, and electron microscopy suggest that cytotoxicity of TiO(2) depends on the shape of the material.

Gallium arsenide (GaAs), a chemical compound of gallium and arsenic, causes various toxic effects including pulmonary diseases in animals. Since the toxicity is not completely investigated, GaAs has been used in workplaces as the material of various semiconductor products. The present study was conducted to clarify the toxicity of GaAs particles in the alveolar macrophages of hamsters using magnetometry, enzyme release assays and morphological examinations. Alveolar macrophages obtained from hamsters by tracheobronchial lavage and adhered to the disks in the bottom of wells were exposed to ferrosoferric oxide and GaAs particles. Ferrosoferric oxide particles were magnetized externally and the remanent magnetic field was measured. Relaxation, a fast decline of the remanent magnetic fields radiated from the alveolar macrophages, was delayed and decay constants were decreased dose-dependently due to exposure to GaAs. Because the relaxation is thought to be associated with cytoskeleton, the exposure of GaAs may have impaired the motor function of them. Enzyme release assay and morphological findings indicated the damage to the macrophages. Thus the cytotoxicity causes cytostructural changes and cell death. According to DNA electrophoresis and the TUNEL method, necrotic changes occur more frequently than apoptotic changes.

Photocopier toner has been implicated in the etiology of some pulmonary diseases. We examined here the in vitro toxicity of toner particles to alveolar macrophages. Cell magnetometry revealed that relaxation was not delayed in macrophages exposed to toner, which represents a rapid decrease in the remaining magnetism emitted by phagocytosed magnetite. However, relaxation was delayed in macrophages exposed to silica (positive controls). The release of intracellular LDH enzyme activity to the extracellular space was negligible in cells exposed to toner compared with negative and positive controls. Morphological examinations by light and electron microscopy revealed no abnormal findings in the exposed cells. A histochemical study using TUNEL staining and the electrophoretic profile of DNA obtained from cells exposed to toner and to silica were negative for apoptosis. The results of the present and other investigations into animal exposure indicate that photocopier toner is toxicologically inert. However, although the present study examined only effects in vitro, exposure to toner should be minimized because lung overloading in animals has been reported.

Rock wool (RW), a type of man-made mineral fiber (MMMF), is a building material used as an asbestos substitute for heat insulation, fire resistance, and reinforcement. RW is included in group 3 of the IARC classification. In the present study, the cytotoxicity of RW was investigated by cell magnetometry, enzyme assay, DNA ladder detection, and electron microscopic morphological evaluation in comparison with chrysotile fibers (CF). Specimens were prepared by 18-h incubation of Fischer rat alveolar macrophages in the presence of RW fibers as the study material, CF as positive control, and phosphate-buffered saline (PBS) as negative control, together with a relaxation indicator, Fe3O4, except for morphological evaluation, followed by additional procedures of external magnetization and subsequent 20-min remanent magnetic field measurement for magnetometric evaluation, and macrophage DNA extraction for evaluating possible apoptosis by DNA ladder detection. In magnetometry, relaxation, a marker of cytotoxicity, was rapid in both the RW- and PBS-treated groups, while it was delayed in both the long and short CF-treated groups. Differences in percent lactate dehydrogenase (LDH) release between the RW-treated group and PBS-treated group were not significant, but those between the RW-treated group and short CF-treated group were statistically significant. A DNA ladder was not detected in any of the study groups. Electron micrographs showed that RW did not cause any change, but CF caused changes in macrophages. Thus, magnetometric measurements suggested no cytotoxicity of RW. We plan, in the future, to evaluate the safety of RW by magnetometric measurement and morphological observation of the lungs in in vivo inhalation experiments.

No method has yet been established to evaluate the exposure to tobacco smoke in passive smoking (PS). We therefore conducted a study on the possibility that the levels of urinary trans, trans-muconic acid (MA) and the exhaled carbon monoxide (CO) could be indices of the passive exposure to tobacco smoke. The moderate correlation was observed between urinary MA levels and the number of consumed cigarettes per day in smokers. The mean urinary MA level of the PS (+) group was significantly higher than that with the PS (-) group. Among the PS (+) group, the mean MA level in the urine obtained in the afternoon was higher than that obtained in the morning . A high correlation was observed between the exhaled CO levels and the number of consumed cigarettes per day in smokers. Like the urinary MA level, the mean exhaled CO level in the PS (+) group, too, gave a significantly higher level than in the PS (-) group. Because the biological half life of MA (7.5 ± 0.85 h) was longer than that of CO (3.0 ± 0.36 h), the measurement of urinary MA level is recommended for evaluating the exposure of passive smoking. The measurement of exhaled CO levels is useful only for chain smokers and nonsmokers with PS just before measurement .