Bone marrow micronucleus assay in Brown-Norway rats exposed to diphenyl-methane-4,4′-diisocyanate

Abstract.

Four groups of young adult male Brown-Norway rats (strain: BN/RijHsd) were either exposed whole-body (WB) to filtered air (negative control) or to respirable aerosols of monomeric diphenylmethane-4,4′-diisocyanate (MDI) at actual breathing zone concentrations of 9.2±1.5 and 118±8.6 mg/m³. One additional group was exposed to 11,0±14.4 mg/m³ MDI using a nose-only (NO) mode. Exposure was 1 h/day, one exposure per week on 3 consecutive weeks. MDI aerosols were generated using either a condensation (WB) or a dispersion-condensation (NO) principle with resultant MMADs of 2.4–3.1 µm and 1.2 µm (GSD≈1.5), respectively. Humidity ranged from ≈40% (WB) to ≈5% (NO). Positive controls received cyclophosphamide and colcemid. Micronuclei in polychromatic erythrocytes (MN-PCE) were counted in bone marrow smears prepared after the final exposure on post-exposure days 1, 2 and 7 and stained with acridine orange or Wright-Giemsa. Both the WB-exposure regimen and the 7-day sampling time point were based upon a previous study in which a significant increase in MN-PCE was reported to occur. Rats exposed to 118 (WB) and 110 mg/m³ MDI (NO) exhibited signs of respiratory distress, including hypothermia, and increased lung weights when compared to WB-exposed rats. The intensity of changes appeared to be slightly more pronounced in NO-exposed rats. At no time point did this study provide any evidence of an MDI-induced effect on the frequency of MN-PCE. No differences in outcome existed following staining with acridine orange or Wright-Giemsa. There was an absence of any effect on the frequency of mast cells and their frequency was low enough not to interfere with the outcome of study. Positive control groups exhibited significant increases in MN-PCE. In summary, monomeric MDI aerosol did not induce cytogenetic damage in Brown-Norway rats when investigated according to current testing guidelines.