Two-week inhalation toxicity of polymeric diphenylmethane-4, 4'-diisocyanate (PMDI) in rats: analysis of biochemical and morphological markers of early pulmonary response.
The pulmonary response of Wistar rats to respirable polymeric diphenylmethane-4,4'-diisocyanate (PMDI) aerosol was examined in a 2-wk repeated nose-only inhalation exposure study. Exposure concentrations were 1.1, 3.3, and 13.7 mg PMDI/m(3) (6 h/day, 15 exposures). The level of 13.7 mg/m(3) was actually a combination of an initial target concentration of 10 mg/m(3) in wk 1, which was raised to 16 mg/m(3) in wk 2, due to a lack of signs suggestive of pulmonary irritation. An acute sensory irritation study on rats served as basis for selection of these concentrations. Shortly after the 2-wk exposure period, rats were subjected to pulmonary function and arterial blood gas measurements. Lungs were examined by light and transmission electron microscopy, and labeling indices in terminal bronchioles were measured. Bronchoalveolar lavage (BAL) was performed to assess various indicators of pulmonary inflammation, including neutrophil and macrophage numbers, protein, lactate dehydrogenase (LDH), gamma-glutamyltranspeptidase (gamma-GT), alkaline phosphatase (APh), acid phosphatase (ACPh), and beta-N-acetylglucosaminidase (beta-NAG). Phosphatidylcholine in BAL fluid and BAL cells was determined as aggregated endpoint suggestive of changes in pulmonary surfactant. Rats exposed to 3.3 and 13.7 mg/m(3) experienced concentration-dependent signs of respiratory tract irritation. Determination of arterial blood gases, lung mechanics, and carbon monoxide diffusing capacity did not demonstrate specific effects. Analysis of BAL fluid and BAL cells revealed changes indicative of marked inflammatory response and/or cytotoxicity in rats exposed to 13.7 mg/m(3), and the changes were characterized by statistically significantly increased activities of LDH, beta-NAG, and protein. Phospholipid concentrations were increased in rats exposed to 1.1 mg/m(3) and above (elevated levels of lipid material in alveolar macrophages demonstrated by polychrome stain) and 3.3 mg/m(3) and above (increased intracellular ACPh activity and intracellular phospholipids). In these groups, gamma-GT was statistically significantly increased. These findings suggest that changes in phospholipid homeostasis appear to occur at lower levels than those eliciting inflammation and cytotoxicity. Light and transmission electron microscopy suggest that exposure to 3.3 and 13. 7 mg/m(3) resulted in focal inflammatory lesions and an accumulation of refractile, yellowish-brownish material in alveolar macrophages with concomitant activation of type II pneumocytes. In the terminal bronchioles a concentration-dependent increase of bromodeoxyuridine (BrdU)-labeled epithelial cells was observed in all PMDI exposure groups. In summary, it appears that respirable PMDI aerosol interacts with pulmonary surfactant, which, in turn, may stimulate type II pneumocytes to increase their production of surfactant and to proliferate.
Pauluhn
J
J
Institute of Toxicology, BAYER AG, Building No. 514, 42096 Wuppertal, Germany. juergen.pauluhn.jp@bayer-ag.de.
Emura
M
M
Mohr
U
U
Popp
A
A
Rosenbruch
M
M
D000485:Allergens / Q000633:toxicity*; D000818:Animals; D001273:Atmosphere Exposure Chambers; D015415:Biomarkers; D001784:Blood Gas Analysis; D001835:Body Weight / Q000187:drug effects; D001988:Bronchiolitis / Q000139:chemically induced / Q000473:pathology; D001992:Bronchoalveolar Lavage Fluid; D004261:DNA Replication / Q000187:drug effects; D005260:Female; D019570:Inhalation Exposure / Q000009:adverse effects*; D017953:Isocyanates / Q000008:administration & dosage / Q000633:toxicity*; D008171:Lung Diseases / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology*; D008297:Male; D009929:Organ Size / Q000187:drug effects; D011140:Polyurethanes / Q000633:toxicity*; D051381:Rats; D017208:Rats, Wistar; D012129:Respiratory Function Tests; D015656:Respiratory Mechanics / Q000187:drug effects; D014692:Ventilation-Perfusion Ratio
D000485:Allergens; D015415:Biomarkers; D017953:Isocyanates; D011140:Polyurethanes; C087867:polymeric 4,4'-diphenylmethane diisocyanate