asbestos--crocidolite and titanium-dioxide

The interim results from this 90-day multi-dose, inhalation toxicology study with life-time post-exposure observation has shown an important fundamental difference in persistence and pathological response in the lung between brake dust derived from brake-pads manufactured with chrysotile, TiO

Topics: Animals; Asbestos, Amosite; Asbestos, Crocidolite; Asbestos, Serpentine; Bronchoalveolar Lavage Fluid; Collagen; Dose-Response Relationship, Drug; Dust; Fibrosis; Inhalation Exposure; Lung; Macrophages, Alveolar; Male; Microscopy, Confocal; Pleura; Rats; Titanium; Toxicity Tests, Subchronic; Traffic-Related Pollution

Exposure to mineral fibers is of substantial relevance to human health. A key event in exposure is the interaction with inflammatory cells and the subsequent generation of pro-inflammatory factors. Mast cells (MCs) have been shown to interact with titanium oxide (TiO₂) and asbestos fibers. In this study, we compared the response of rat peritoneal MCs challenged with the asbestos crocidolite and nanowires of TiO₂ to that induced by wollastonite employed as a control fiber.. Rat peritoneal MCs (RPMCs), isolated from peritoneal lavage, were incubated in the presence of mineral fibers. The quantities of secreted enzymes were evaluated together with the activity of fiber-associated enzymes. The ultrastructural morphology of fiber-interacting RPMCs was analyzed with electron microscopy.. Asbestos and TiO₂ stimulate MC secretion. Secreted enzymes bind to fibers and exhibit higher activity. TiO₂ and wollastonite bind and improve enzyme activity, but to a lesser degree than crocidolite.. (1) Mineral fibers are able to stimulate the mast cell secretory process by both active (during membrane interaction) and/or passive (during membrane penetration) interaction; (2) fibers can be found to be associated with secreted enzymes-this process appears to create long-lasting pro-inflammatory environments and may represent the active contribution of MCs in maintaining the inflammatory process; (3) MCs and their enzymes should be considered as a therapeutic target in the pathogenesis of asbestos-induced lung inflammation; and (4) MCs can contribute to the inflammatory effect associated with selected engineered nanomaterials, such as TiO₂ nanoparticles.

Topics: Animals; Asbestos; Asbestos, Crocidolite; Calcium Compounds; Cell Count; Female; Humans; Male; Mast Cells; Microscopy, Electron, Scanning; Microscopy, Electron, Transmission; Mineral Fibers; Nanowires; Peritoneum; Rats; Rats, Wistar; Silicates; Titanium

Identifying and understanding the early molecular events that underscore mineral pathogenicity using in vitro screening tests is imperative, especially given the large number of synthetic and natural fibers and particles being introduced into the environment. The purpose of the work described here was to examine the ability of gene profiling (Affymetrix microarrays) to predict the pathogenicity of various materials in a human mesothelial cell line (LP9/TERT-1) exposed to equal surface area concentrations (15 x 10(6) or 75 x 10(6) microm(2)/cm(2)) of crocidolite asbestos, nonfibrous talc, fine titanium dioxide (TiO(2)), or glass beads for 8 or 24 h. Since crocidolite asbestos caused the greatest number of alterations in gene expression, multiplex analysis (Bio-Plex) of proteins released from LP9/TERT-1 cells exposed to crocidolite asbestos was also assessed to reveal if this approach might also be explored in future assays comparing various mineral types. To verify that LP9/TERT-1 cells were more sensitive than other cell types to asbestos, human ovarian epithelial cells (IOSE) were also utilized in microarray studies. Upon assessing changes in gene expression via microarrays, principal component analysis (PCA) of these data was used to identify patterns of differential gene expression. PCA of microarray data confirmed that LP9/TERT-1 cells were more responsive than IOSE cells to crocidolite asbestos or nonfibrous talc, and that crocidolite asbestos elicited greater responses in both cell types when compared to nonfibrous talc, TiO(2), or glass beads. Bio-Plex analysis demonstrated that asbestos caused an increase in interleukin-13 (IL-13), basic fibroblast growth factor (bFGF), granulocyte colony-stimulating factor (G-CSF), and vascular endothelial growth factor (VEGF). These responses were generally dose-dependent (bFGF and G-CSF only) and tumor necrosis factor (TNF)-alpha independent (except for G-CSF). Thus, microarray and Bio-Plex analyses are valuable in determining early molecular responses to fibers/particles and may directly contribute to understanding the etiology of diseases caused by them. The number and magnitude of changes in gene expression or "profiles" of secreted proteins may serve as valuable metrics for determining the potential pathogenicity of various mineral types. Hence, alterations in gene expression and cytokine/chemokine changes induced by crocidolite asbestos in LP9/TERT-1 cells may be indicative of its increased potential to ca

Topics: Asbestos, Crocidolite; Cell Line; Epithelium; Fibroblast Growth Factor 2; Gene Expression; Gene Expression Profiling; Granulocyte Colony-Stimulating Factor; Humans; Interleukin-13; Particulate Matter; Proteomics; Talc; Telomerase; Titanium; Toxicity Tests; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Human mesothelial cells (LP9/TERT-1) were exposed to low and high (15 and 75 microm(2)/cm(2) dish) equal surface area concentrations of crocidolite asbestos, nonfibrous talc, fine titanium dioxide (TiO2), or glass beads for 8 or 24 hours. RNA was then isolated for Affymetrix microarrays, GeneSifter analysis and QRT-PCR. Gene changes by asbestos were concentration- and time-dependent. At low nontoxic concentrations, asbestos caused significant changes in mRNA expression of 29 genes at 8 hours and of 205 genes at 24 hours, whereas changes in mRNA levels of 236 genes occurred in cells exposed to high concentrations of asbestos for 8 hours. Human primary pleural mesothelial cells also showed the same patterns of increased gene expression by asbestos. Nonfibrous talc at low concentrations in LP9/TERT-1 mesothelial cells caused increased expression of 1 gene Activating Transcription Factor 3 (ATF3) at 8 hours and no changes at 24 hours, whereas expression levels of 30 genes were elevated at 8 hours at high talc concentrations. Fine TiO2 or glass beads caused no changes in gene expression. In human ovarian epithelial (IOSE) cells, asbestos at high concentrations elevated expression of two genes (NR4A2, MIP2) at 8 hours and 16 genes at 24 hours that were distinct from those elevated in mesothelial cells. Since ATF3 was the most highly expressed gene by asbestos, its functional importance in cytokine production by LP9/TERT-1 cells was assessed using siRNA approaches. Results reveal that ATF3 modulates production of inflammatory cytokines (IL-1 beta, IL-13, G-CSF) and growth factors (VEGF and PDGF-BB) in human mesothelial cells.

Topics: Activating Transcription Factor 3; Asbestos, Crocidolite; Cell Line; Cytokines; Dose-Response Relationship, Drug; Epithelial Cells; Female; Gene Expression Profiling; Gene Expression Regulation; Glass; Humans; Minerals; Oligonucleotide Array Sequence Analysis; Ovary; Particle Size; Pleura; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; RNA, Messenger; RNA, Small Interfering; Silicon Dioxide; Talc; Time Factors; Titanium

Cocultures of human pulmonary epithelial cells (BEAS-2B) and lung fibroblasts (WISTAR-38), representing two cell types of central regulatory potential in (chronic) lung disease, were used as an in vitro model to study the role of interleukin 6 (IL-6) and of granulocyte macrophage-colony stimulating factor (GM-CSF) in early fibrogenesis. For this purpose, epithelial cells were pre-exposed to UICC crocidolite asbestos fibers or titanium dioxide (TiO2) particles for 96 h and subsequently cocultured with fibroblasts for additional 72 h. Gene expression of IL-6 or GM-CSF in both cell types as well as of alpha1 procollagens types I and III in fibroblasts was determined by RT-PCR. Synthesis of IL-6, GM-CSF or collagen I was quantified using IL-6 bioassay or ELISA tests, respectively. Both mediators were directly induced in bronchoepithelial cells by crocidolite but not by TiO2. Likewise, steady-state mRNA levels of procollagens as well as collagen synthesis were upregulated in cocultured fibroblasts. As a result of coculture, cytokine concentrations were synergistically enhanced and further increased by crocidolite in a dose-dependent manner. Suppression of cytokine induction by corresponding neutralizing antibodies consistently abrogated collagen enhancement. Direct stimulation of fibroblast monocultures with recombinant human IL-6 or GM-CSF significantly increased collagen synthesis and transcription in a dose-dependent manner. Thus, our results demonstrate that crocidolite selectively stimulated production of IL-6 and GM-CSF in bronchoepithelial cells. In epithelial-fibroblast interactions, these mediators appear to play a key role in regulating fibroblast activity, indicating a close correlation between these cytokines and the fibrogenic potential of particulates.

Topics: Asbestos, Crocidolite; Biomarkers; Carcinogens; Cell Communication; Cell Line; Coculture Techniques; Epithelial Cells; Fibroblasts; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-6; Lung; Particle Size; Reverse Transcriptase Polymerase Chain Reaction; Titanium

The cytotoxic and oxidative responses of crocidolite and chrysotile asbestos fibers and ultrafine titanium dioxide (UF-TiO2) particles were measured in alveolar macrophages (AM) and peripheral red blood cells (RBC) of rat after 30 days with a single intratracheal exposure (5 mg). The following responses were observed one month after fiber/particle instillation: (1) AM population increased; (2) lactate dehydrogenase and acid phosphatase activities in cell free lung lavage fluid increased; (3) substances that react with hydrogen peroxide or thiobarbituric acid were elevated in both AM and peripheral RBC; (4) glutathione peroxidase, glutathione reductase, and catalase were altered in both AM and peripheral RBC; (5) glutathione and ascorbic acid decreased in both AM and peripheral RBC. A significant difference from negative controls was noted in all responses of the two fiber-exposed groups, and in most responses of the UF-TiO2-exposed group. The level of responses to the three test substances suggested a decreasing order of toxicity, with crocidolite > chrysotile > UF-TiO2.

Topics: Aerosols; Animals; Asbestos, Crocidolite; Asbestos, Serpentine; Bronchoalveolar Lavage Fluid; Carcinogens; Dust; Erythrocytes; Female; Lipid Peroxidation; Macrophage Activation; Macrophages, Alveolar; Mineral Fibers; Particle Size; Rats; Reactive Oxygen Species; Titanium

Refractory ceramic fiber has been developed for industrial processes requiring materials with high thermal and mechanical stability. To evaluate the biological activity of this fiber, rat alveolar macrophages were exposed for < or = 24 h to 0-1,000 micrograms/ml of refractory ceramic fiber, crocidolite asbestos, silica (fibrogenic particles), or titanium dioxide (a nonfibrogenic particle), and eicosanoid, tumor necrosis factor-alpha (TNF), and lactate dehydrogenase release were measured. Particle dimensions were determined by electron microscopy. Radioactivity coeluting with leukotriene B4 (LTB4) and immunoreactive LTB4 and TNF release increased after refractory ceramic fiber and were similar in magnitude after asbestos but less than after silica. For example, the total [3H]eicosanoid release increased 3.9-fold after refractory ceramic fiber, 4.6-fold after asbestos, and 8.7-fold after silica. Refractory ceramic fiber and asbestos also have similar particle dimensions (diameter, length, and surface area). Inasmuch as macrophage-derived LTB4 and TNF are potent mediators in inflammatory events, including migration and activation of neutrophils, these findings suggest that refractory ceramic fiber can activate macrophages in vitro to release mediators relevant to in vivo findings of inflammation and fibrotic lung disease in laboratory animals.

Topics: Animals; Asbestos, Crocidolite; Bronchoalveolar Lavage Fluid; Ceramics; Chemotaxis, Leukocyte; Cytokines; Eicosanoids; In Vitro Techniques; Leukotriene B4; Lipoxygenase; Macrophages, Alveolar; Male; Particle Size; Prostaglandin-Endoperoxide Synthases; Radioimmunoassay; Rats; Rats, Inbred F344; Silicon Dioxide; Titanium; Tumor Necrosis Factor-alpha

The cytogenetic effects of asbestos fibers on rat pleural mesothelial cells were studied in vitro. Crocidolite UICC significantly enhanced aneuploidy and produced few structural chromosome aberrations, whereas anatase, an isomorphic particle, induced no numerical or structural changes. Mitomycin C (300 nM) produced a tenfold increase in abnormal anaphases compared with controls. Asbestos produced anaphase/telophase abnormalities in a concentration-dependent manner. The majority of the abnormalities involved lagging chromosomes. Crocidolite UICC induced abnormalities at a dose of 7.0 micrograms/cm2, whereas Canadian chrysotile did so at 1.0 to 2.0 micrograms/cm2. When the response was assessed by the number of long and thin fibers per cm2 (length > 8 microns; diameter < or = 0.25 microns), crocidolite UICC produced more abnormalities than Canadian chrysotile at all concentrations. On a per-weight basis, these findings differ from those obtained after intrapleural inoculation, as crocidolite induced more mesotheliomas than chrysotile; however, on a per-fiber basis, the in vitro and in vivo effects were similar. These results show that anaphase/telophase analysis is sensitive and complementary to metaphase analysis, and suggest that asbestos might produce cell transformation by inducing chromosome missegregation and aneuploidy.

Topics: Anaphase; Aneuploidy; Animals; Asbestos; Asbestos, Crocidolite; Cell Cycle; Cells, Cultured; Chromosome Aberrations; Epithelial Cells; Epithelium; Metaphase; Mutagens; Pleura; Rats; Telophase; Titanium

Exposure of phagocytic cells to asbestos in vitro results in an augmented production of reactive oxygen metabolites and increased peroxidation of lipids. The aim of this investigation was to assess the extent of lipid peroxidation both in cells and fluid obtained from bronchoalveolar lavage (BAL), and in lungs of rats exposed to crocidolite asbestos or titanium dioxide (TiO2), a nonfibrous particulate control. In comparison to sham and TiO2-exposed rats, the BAL fluid and cells of crocidolite-exposed animals contained significantly elevated levels of malondialdehyde (MDA), a breakdown product of lipid peroxidation detected using high-pressure liquid chromatography (HPLC). In contrast, no significant differences in MDA were detected in lavaged lung tissue from these animals. Inhalation of crocidolite caused an early inflammatory response characterized by elevated numbers of polymorphonuclear leukocytes and lymphocytes, as well as enhanced total protein in BAL. Pulmonary fibrosis and increased lung hydroxyproline also were observed after 20 days of exposure. Exposure to TiO2 did not cause inflammation, pulmonary fibrosis, or elevated amounts of hydroxyproline in the lung. Our results show that exposure to the fibrogenic and inflammatory mineral, crocidolite, results in an enhanced lipid peroxidation in BAL cells and fluid not observed after inhalation of the particulate TiO2. These novel observations suggest that MDA in BAL may be useful as a biomarker of exposure to inhaled asbestos or other oxidants.

Topics: Administration, Inhalation; Animals; Asbestos; Asbestos, Crocidolite; Bronchoalveolar Lavage Fluid; Collagen; Hydroxyproline; Lipid Peroxidation; Lung; Male; Malondialdehyde; Rats; Rats, Inbred F344; Titanium

Experiments were carried out to assess the effect of intraperitoneal injection of the mineral dusts, titanium dioxide, quartz, or asbestos, on splenic lymphocyte antibody forming cells in immunised mice. Titanium dioxide and quartz caused similar, about one third, reductions in plaque forming cells; asbestos caused substantial reduction to about a quarter of the number found in control spleens. The inhibition of antibody forming cells in the spleen found with chrysotile was dose dependent and both chrysotile and crocidolite asbestos were similar in activity. Systemic immunomodulation after local deposition of mineral dust may be important to the development of disease.

Topics: Animals; Antibody-Producing Cells; Asbestos; Asbestos, Crocidolite; Asbestos, Serpentine; Dose-Response Relationship, Immunologic; Mice; Mice, Inbred C57BL; Mice, Inbred CBA; Peritonitis; Quartz; Silicon Dioxide; Spleen; Titanium

Rat mesothelial cell cultures were shown to have considerable plasminogen dependent and independent fibrinolytic activity in vitro using an 125I fibrin degradation assay. At non-toxic doses of the mineral dusts titanium dioxide, quartz and crocidolite asbestos, as assessed by 51Cr release, the fibrinolytic activity of mesothelial cells was inhibited. Quartz had the greatest inhibitory effect and crocidolite asbestos had the least. These results suggests that inhibition of mesothelial cell fibrinolysis does not, on its own, explain pleural fibrosis due to toxic mineral dusts.

Topics: Animals; Asbestos; Asbestos, Crocidolite; Cell Survival; Cells, Cultured; Dust; Female; Fibrinolysis; Plasminogen; Pleura; Quartz; Rats; Rats, Inbred Strains; Titanium

Luminol-dependent chemiluminescence (CL) was used to assess the in vitro production of reactive oxygen species by human neutrophils and monocytes on exposure to six standard respirable mineral dusts. Every dust caused CL production in both phagocytic cell types, although, for each dust, the two cells showed a different pattern of response. Light output was markedly affected by the presence of serum in the system. While the results illustrated the complexity of the interaction between mineral dusts and monocytes and neutrophils, they did not support the hypothesis that pathogenic dusts would induce the production of more reactive oxygen species than non-pathogenic dusts.

Topics: Adult; Aluminum Silicates; Asbestos; Asbestos, Crocidolite; Asbestos, Serpentine; Clay; Dose-Response Relationship, Drug; Dust; Humans; In Vitro Techniques; Luminescent Measurements; Luminol; Male; Minerals; Monocytes; Neutrophils; Opsonin Proteins; Oxygen Consumption; Phagocytosis; Quartz; Time Factors; Titanium

A macrophage-like cell line (P388D1) has been treated with asbestos and the release of arachidonic acid and its metabolites has been studied using two methods. In the first monolayer cultures of the cells were labelled with tritiated arachidonic acid and the release of label into the medium was quantified: secondly the synthesis and release of prostaglandins E2 and F2 alpha were followed using radioimmune assay. Crocidolite asbestos caused the greatest release of tritium while the medium from chrysotile-treated cultures contained more of both prostaglandins. Both of the fibrous dusts were significantly more active in both test systems than were the two 'inert' materials--titanium dioxide and milled sample of crocidolite. It is suggested that these phenomena are due to the effect of mineral dusts on phospholipase activity and that differences in this activity are associated with differences in the pathogenicity of various mineral dusts.

Topics: Arachidonic Acid; Arachidonic Acids; Asbestos; Asbestos, Crocidolite; Asbestos, Serpentine; Cell Line; Humans; Indomethacin; Macrophages; Prostaglandins; Titanium