title: Pulmonary inflammation in rats after intratracheal instillation of quartz, amorphous SiO2, carbon black, and coal dust and the influence of poly-2-vinylpyridine-N-oxide (PVNO).
abstract: Effects of poly-2-vinylpyridine-N-oxide (PVNO) were investigated in numerous in vivo and in vitro studies published in the nineteen sixties and seventies. These studies showed that PVNO inhibited development of fibrosis from quartz dust and improved lung clearance of quartz after inhalation exposure. Ameliorating effects of PVNO were observed also for pulmonary damage from colloidal SiO2 and organic substances, and the fibrogenic inflammation caused by carrageenan. Although it is not proven that silicosis is a precondition for quartz-induced lung tumours, we investigated the hypothesis that PVNO could reduce the lung tumour risk from quartz in rats. A carcinogenicity study was therefore started in rats with the main focus on the quantitative relationships among pulmonary inflammation, fibrosis and neoplasia caused by intratracheal instillation of 3 mg quartz DQ 12 with or without additional subcutaneous PVNO treatment. Other study groups were treated with multiple dust instillations, i.e. 30 instillations of 0.5 mg amorphous SiO2 at intervals of 2 weeks, 10 instillations of 0.5 mg of ultrafine carbon black or 1 mg coal at weekly intervals. The analyses of the bronchoalveolar lavage fluid (BALF) 9 months after start of the life-time study showed that the aim of producing similar levels of increased enzyme concentrations in the four groups treated with quartz/PVNO, amorphous SiO2, carbon black and coal was achieved. A 2.5- to 7.7-fold increase for lactate dehydrogenase (LDH), total protein, alkaline phosphatase and gamma-glutamyl transferase (gamma-GT) was found in these groups as compared to the control. In contrast, quartz treatment without PVNO increased the LDH level up to 24-fold and of total protein to 13-fold. However, the cell counts in the BALF were not so much different in all five groups, i.e. quartz without PVNO (leukocytes: 480.000, PMN: 190.000), quartz with PVNO (leukocytes: 300.000, PMN: 100.000), amorphous SiO2 (leukocytes: 570.000, PMN: 315.000), carbon black (leukocytes: 390.000, PMN: 150.000) and coal (leukocytes: 200.000, PMN: 65.000). Histopathological investigations after four weeks and three months revealed that the used PVNO sample was active in the quartz and amorphous SiO2 groups and markedly reduced the incidences or severity of several pulmonary changes such as macrophage accumulation, inflammatory cell infiltration, interstitial fibrosis, bronchiolo-alveolar hyperplasia, alveolar lipoproteinosis and amorphous SiO2 -induced granulomatous alveolitis/interstitial fibrotic granulomas. Also in the lung-associated lymph nodes (LALN), PVNO treatment significantly reduced the incidence and severity of inflammation in both quartz and amorphous SiO2 groups as evidenced by the presence of well-circumscribed aggregates of intact particle-laden macrophages without signs of degeneration and accompanying granulocytic infiltration and fibrosis. Immunological investigations at the 9 months timepoint on the in vitro production of reactive nitrogen (RNI) or oxygen (ROI) intermediates and tumour necrosis factor (TNF-alpha) from BALF-derived cells indicated a diminished responsiveness to LPS in all particle treatment groups. A diminished production of ROI was also found in the quartz, carbon black, and coal dust groups, respectively, as compared to the values seen in the quartz/PVNO- and amorphous SiO2 treated groups. Treatment with quartz plus PVNO restored the capability of the cells to respond to LPS as compared to the treatment with quartz alone. TNF-alpha production was diminished in the groups treated with quartz, carbon black, and coal dust alone whereas in the quartz/PVNO- and amorphous SiO2-treated groups an elevated TNF-alpha production was seen. These results led to the conclusion that only amorphous SiO2 did not affect the "normal" ability of the cells to respond to LPS and that PVNO protected the cells from a toxic effect of the quartz particles.
authors.affiliation: Fraunhofer Institute of Toxicology and Aerosol Research, Hannover, Germany. ernst@ita.fraunhofer.de
authors.forename: Heinrich
authors.initials: H
authors.lastname: Ernst
authors.forename: Susanne
authors.initials: S
authors.lastname: Rittinghausen
authors.forename: Wilfried
authors.initials: W
authors.lastname: Bartsch
authors.forename: Otto
authors.initials: O
authors.lastname: Creutzenberg
authors.forename: Clemens
authors.initials: C
authors.lastname: Dasenbrock
authors.forename: Bernd-Detlef
authors.initials: BD
authors.lastname: Görlitz
authors.forename: Matthias
authors.initials: M
authors.lastname: Hecht
authors.forename: Ulf
authors.initials: U
authors.lastname: Kairies
authors.forename: Hartwig
authors.initials: H
authors.lastname: Muhle
authors.forename: Meike
authors.initials: M
authors.lastname: Müller
authors.forename: Uwe
authors.initials: U
authors.lastname: Heinrich
authors.forename: Friedrich
authors.initials: F
authors.lastname: Pott
mesh_terms: D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology; D002244:Carbon / Q000009:adverse effects*; D003031:Coal; D004391:Dust; D005260:Female; D007249:Inflammation; D019570:Inhalation Exposure; D008168:Lung / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000473:pathology*; D008175:Lung Neoplasms / Q000209:etiology* / Q000517:prevention & control*; D011144:Polyvinylpyridine N-Oxide / Q000494:pharmacology*; D011658:Pulmonary Fibrosis / Q000209:etiology / Q000473:pathology; D011791:Quartz / Q000009:adverse effects*; D051381:Rats; D017208:Rats, Wistar; D012307:Risk Factors; D012822:Silicon Dioxide / Q000009:adverse effects*; D014132:Trachea; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000494:pharmacology
chemical_list: D003031:Coal; D004391:Dust; D014409:Tumor Necrosis Factor-alpha; D011791:Quartz; D002244:Carbon; D012822:Silicon Dioxide; D011144:Polyvinylpyridine N-Oxide