Lack of marked cyto- and genotoxicity of cristobalite in devitrified (heated) alkaline earth silicate wools in short-term assays with cultured primary rat alveolar macrophages.

Alkaline earth silicate (AES) wools are low-biopersistence high-temperature insulation wools. Following prolonged periods at high temperatures they may devitrify, producing crystalline silica (CS) polymorphs, including cristobalite, classified as carcinogenic to humans. Here we investigated the cytotoxic and genotoxic significance of cristobalite present in heated AES wools. Primary rat alveolar macrophages were incubated in vitro for 2 h with 200 µg/cm² unheated/heated calcium magnesium silicate wools (CMS1, CMS2, CMS3; heat-treated for 1 week at, or 4 weeks 150 °C below, their respective classification temperatures) or magnesium silicate wool (MS; heated for 24 h at 1260 °C). Types and quantities of CS formed, and fiber size distribution and shape were determined by X-ray diffraction and electron microscopy. Lactate dehydrogenase release and alkaline and hOGG1-modified comet assays were used, ± aluminum lactate (known to quench CS effects), for cytotoxicity/genotoxicity screening. Cristobalite content of wools increased with heating temperature and duration, paralleled by decreases in fiber length and changes in fiber shape. No marked cytotoxicity, and nearly no (CMS) or only slight (MS) DNA-strand break induction was observed, compared to the CS-negative control Al2O3, whereas DQ12 as CS-positive control was highly active. Some samples induced slight oxidative DNA damage, but no biological endpoint significantly correlated with free CS, quartz, or cristobalite. In conclusion, heating of AES wools mediates changes in CS content and fiber length/shape. While changes in fiber morphology can impact biological activity, cristobalite content appears minor or of no relevance to the intrinsic toxicity of heated AES wools in short-term assays with rat alveolar macrophages.

Horizontal diffusion elutriation: a new size-separation technique for preparation of rodent-respirable fibers for animal testing.

Short-and long-term animal experiments are used to examine the toxicology and biopersistence of various types of fibers. In order to ensure an adequate exposure dose for testing, modern experimental protocols specify that the exposure aerosol (in an inhalation test) or the fibers (in an intratracheal instillation [IT] test) must contain at least a minimum concentration of long (> 20 mum) rodent-respirable fibers. As produced and handled, most fibers contain a distribution of diameters and lengths, only some of which are both long and rodent-respirable. Therefore, it is necessary to size-separate the fibers to enrich the proportion of long, rodent-respirable fibers in the material to be tested. This article presents a new and relatively simple method for size separation that avoids some of the difficulties associated with other methods. The method, termed horizontal diffusion elutriation (HDE), is illustrated by size-separating refractory ceramic fiber (RCF) and four polycrystalline alumina (PCA) fibers.