Response of the rat lung to respirable fractions of composite fiber-epoxy dusts.

It is unknown whether respirable dusts derived from the machining of composite fiber-epoxy materials pose a health risk. To evaluate the potential pulmonary toxicity of these materials, we studied the effects of six samples of dusts previously well characterized physically, chemically, and morphologically in the lungs of specific pathogen-free rats. A single intratracheal bolus of 5 mg of each sample was injected into separate groups of five rats each. For comparison, free crystalline silica (quartz) and aluminum oxide were used as positive and negative controls in additional animals. One month later, the lungs were fixed and sectioned for light microscopy. Histopathology scores for the six composite-epoxy samples showed a continuum of lung injury that was between the negative and positive controls. None of the composite dusts had effects that paralleled those of quartz; however, four of the composite dusts produced reactions that were more severe than that seen with aluminum oxide. Therefore, respirable fractions of some types of composite materials can induce pathological changes in lungs of experimental animals. The features responsible for the variation in host response between samples are, as yet, unclear.

Carcinogenicity studies on natural and man-made fibres with the intraperitoneal test in rats.

Female Wistar rats were injected intraperitoneally (i.p.) with a suspension of 11 fibrous and 3 granular dusts. A dose of 0.25 mg actinolite or UICC chrysotile induced tumours of the peritoneum in more than 50% of the animals. Even 0.05 and 0.01 mg proved to be carcinogenic, although no adhesions of the abdominal organs could be observed. The findings are in conflict with the hypothesis that a scar is always the morphological precondition for the development of an asbestos-induced tumour. Actinolite injected i.p. in a solution of polyvinylpyridine-N-oxide gave a lower tumour incidence than when suspended only in saline, possibly due to inactivation of the fibre surface. Persistent glass fibres were less effective than actinolite having a similar fibre size distribution. On the other hand, relatively thick basalt fibres and ceramic fibres gave higher tumour incidences than expected. Wollastonite fibres were not carcinogenic, probably because of their low durability. Large amounts of polyvinylchloride, alpha-ferric oxide hydrate and wood dust also led only to adhesions of the abdominal organs and fibrosis; a definite carcinogenic effect was not detected.

Formation of DNA adduct 8-hydroxy-2'-deoxyguanosine induced by man-made mineral fibres.

Two man-made mineral fibres, rockwool and glasswool, were found to mediate hydroxylation of deoxyguanosine and calf thymus DNA to form the DNA adduct 8-hydroxy-2'-deoxyguanosine. The modification of the nucleoside is probably mediated by hydroxyl radicals and may play a role in fibre-induced carcinogenesis.

Effects of in vitro asbestos exposure on natural killer and antibody-dependent cellular cytotoxicity.

Human peripheral blood lymphocytes (PBL) were exposed in vitro to asbestos fibers. Antibody-dependent cellular cytotoxicity (ADCC) activity and natural killer (NK) activity were examined by a chromium-51 release assay. There was a statistically significant enhancement of ADCC and NK activity by chrysotile and crocidolite fibers when cultured together with PBL for a period of 42 hr in medium containing a concentration of at least 2.5% fetal calf serum (P less than 0.05). Isolation of large granular lymphocytes to measure NK activity, however, showed the opposite effect when exposed to asbestos fibers (P less than 0.05). Our results indicate that asbestos fibers can directly affect lymphoid cytotoxic responses in vitro and may provide clues to immunopathogenic mechanisms for the occurrence of neoplasms in vivo.

Role of phagocytosis in Syrian hamster cell transformation and cytogenetic effects induced by asbestos and short and long glass fibers.

We have shown previously that asbestos and other mineral dusts, including glass fibers, induce cell transformation and chromosomal mutations in Syrian hamster embryo cells in culture. In the present study, we observed that both asbestos and glass fibers were phagocytized by these cells and accumulated in the perinuclear region of the cytoplasm. In order to understand the mechanism of fiber length-dependent cellular effects, we examined the phagocytosis and intracellular distribution of glass fibers of differing lengths in cells at various times after treatment. Glass fiber length was decreased by milling with a mortar and pestle. Cells treated with an equal dose of milled glass fibers (on a weight per surface area basis) were exposed to 7-fold more fibers since milling of glass fibers resulted in a 7-fold decrease in length with little change in diameter. However, cells exposed to milled glass fibers phagocytized a similar number of fibers as cells exposed to an equal mass of unmilled glass fibers, indicating that milled fibers were less readily phagocytized. In cells treated with either unmilled or milled glass fibers, the length of the intracellular fibers was more than 2-fold greater than the length of the fibers on the surface, suggesting that cells selectively internalized longer fibers. Fiber length, however, did not appear to affect the migration of intracellular fibers to the perinuclear region of the cytoplasm. Even though cells treated with milled glass fibers contained a similar number of fibers as those treated with unmilled glass fibers, the resulting cytotoxicity, transformation frequency, and frequency of micronuclei were greatly reduced in the cultures treated with milled glass fibers. Thus, fiber length appears to affect the phagocytosis of fibers as well as the ability of intracellular fibers to induce cytogenetic damage and the resultant transformation.

Ten years of epidemiologic investigations on man-made mineral fibers and health.

This paper introduces the reports that follow it in a supplement dealing with the extended follow-up of mortality and cancer incidence among workers in the man-made mineral fiber (MMMF) production industry in Europe, the largest international study of its kind thus far initiated. In addition it reviews the prior epidemiologic evidence on this subject, both in terms of the earlier results of the same study and the results of other studies on the long-term effects of occupational exposure to MMMF. The conclusion drawn from this prior evidence was that the possibility that some excess of lung cancer may be causally related to exposure to MMMF could not be ruled out of the range of reasonable interpretations. When the findings of the extended follow-up, which this paper introduces, are added to the existing evidence, the overall results were considered to endorse the indication that no adverse long-term health effects have been detectable in terms of mortality throughout almost all segments of the MMMF production industry and to support the inference that MMMF--as present in the environmental conditions of early slag-wool/rock-wool production--may have played a role in the causation of lung cancer.

Glass fibers and vapor phase components of cigarette smoke as cofactors in experimental respiratory tract carcinogenesis.

Syrian golden hamsters were given intratracheal instillations of glass fibers with or without BP suspended in saline, once a fortnight for 52 weeks; the experiment was terminated at week 85. No tumors of the respiratory tract were observed in hamsters treated with glass fibers alone. There was no indication that glass fibers enhanced the development of respiratory tract tumors induced by BP. In another study Syrian golden hamsters were exposed to fresh air or to a mixture of 4 major vapor phase components of cigarette smoke, viz. isoprene (800----700 ppm), methyl chloride (1000----900 ppm), methyl nitrite (200----190 ppm) and acetaldehyde (1400----1200 ppm) for a period of at most 23 months. Some of the animals were also given repeated intratracheal instillations of BP or norharman in saline. Laryngeal tumors were found in 7/31 male and 6/32 female hamsters exposed only to the vapor mixture, whereas no laryngeal tumors occurred in controls. The tumor response of the larynx most probably has to be ascribed entirely to the action of acetaldehyde. Simultaneous treatment with norharman or BP did not affect the tumor response of the larynx. Acetaldehyde may occur in the vapor phase of cigarette smoke at levels up to 2000 ppm. Chronic inhalation exposure of rats to acetaldehyde at levels of 0 (controls), 750, 1500 or 3000----1000 ppm resulted in a high incidence of nasal carcinomas, both squamous cell carcinomas of the respiratory epithelium and adenocarcinomas of the olfactory epithelium. It was discussed that acetaldehyde may significantly contribute to the induction of bronchogenic cancer by cigarette smoke in man. No evidence was obtained for a role of isoprene, methyl chloride or methyl nitrite in the induction of lung cancer by cigarette smoke.

Macrophage functions after exposure to mineral fibers.

UICC, other well-defined asbestos samples and different man-made mineral fibers (MMM) such as glass fiber and synthetic amphibole asbestos were studied in vitro by using rat and guinea pig lung macrophages. These samples had relatively narrow length and diameter spectra. Most of the fiber samples were added to the cultures on a gravimetric basis, although some were added on a numerical basis. Electrocorundum and DQ12 (Dorentruper Quartz) were used as controls at comparable gravimetrical concentrations. The assays used were the release of lactate dehydrogenase (to demonstrate plasma membrane permeability) and the release of beta-glucuronidase (to indicate lysosomal permeability). Carbohydrate metabolism was monitored by the measurement of lactic acid production and, as one of the tests for macrophage function, the production of lysozyme was determined. The phagocytic ability of the cells was measured, after the addition of opsonized zymosan, by bioluminescence following luminol enhancement. Only some results could be evaluated, however, due to technical difficulties. A length- and dose-dependent cytotoxicity of the fibers was found in this system which was similar to that previously described with permanent cell lines. No great differences were found between fibers having different physicochemical compositions if their geometric dimensions were similar. Long, very thin fibers of glass, chrysotile, crocidolite and synthetic fluoroamphiboles were all toxic in the test system.

Mesotheliomata in rats after inoculation with asbestos and other materials.

Four experiments in which SPF Wistar rats were inoculated intrapleurally with asbestos or other materials are described. Mesotheliomata were observed in a considerable proportion of animals with all the samples of asbestos used and with a sample of brucite. A few were produced with synthetic aluminium silicate fibres and single ones with barium sulphate, glass powder and aluminium oxide. The risk of developing a mesothelioma at a given time after injection was approximately proportional to the dose. Of the UICC standard reference samples, crocidolite was the most carcinogenic and removal of the oils by benzene extraction did not alter the carcinogenicity of these samples. Chemical properties also seem unlikely to be the main factor producing mesotheliomata but the results support the hypothesis that the finer fibres are the more carcinogenic, and this is additional to the known aerodynamic advantage which the finer fibres have in penetrating to the periphery of the lung.