Cytotoxicity and Spectroscopic Investigations of Organic Free Radicals in Fresh and Stale Coal Dust

"INTRODUCTIONThe mechanistic details of the biological events leading to coal workers pneumoconiosis (CWP) are not yet fully understood, despite several decades of extensive epidemiologic1.2 and laboratory studies.2-6 Epidemiologic studies1.2 have shown, for example, that the incidence and severity of CWP differ markedly in the different regions and mines at comparable exposures, but laboratory investigations2-6 have demonstrated only partial correlation of the epidemiologic data with the differences in the mineral composition and the rank of coal mined. In particular, while epidemiologic data indicate1.2 direct correlation of the· prevalence and severity of CWP with the rank (i.e., % carbon content) of coal, this correlation has not been established by laboratory4'6 and animal exposure3 studies. In order to explain these results, in 1980 Artemov and Reznik' suggested that perhaps they arise from the fundamental differences in the surface properties of the coal. dusts inhaled by the coal miners and those used in the laboratory studies: while the miners inhale freshly fractured coal particles, henceforth called the 'fresh' coal dust, the laboratory studies generally utilize 'aged' coal dusts (i.e., dusts that have been stored for many days or even longer). Reznik and Artemov' used electron spin resonance (ESR) spectroscopy to show that the mechanical crushing of some Soviet Union coals generated organic free radicals and that the concentration and the decay times (hence the reactivity) of these radicals were higher for the coals of higher ranks. Since some of the radicals decayed within a few minutes in air, the authors surmised that while these radical species could lead to certain specific pathogenic reactions at the sites of mining operations, this might not be the case in the laboratory studies, due to the conventional use of 'stored' dusts which would be expected to contain significantly smaller free radical concentrations. This is consistent with the recent findings that, standardized, aged coal dusts exhibit minimal cytotoxicity. 8While the above mentioned work of Artemov and Reznik7 did suggest a possible new clue to the pathogenesis of CWP (i.e., the role of the coal based free radical species), no direct biological/cytotoxicity data were provided to show that 'fresh' coal dusts were, indeed, more pathogenic than the 'stale' ones. Because of the rather significant implications of the Artcmov-Reznik hypothesis to the understanding of 1hc biochemical mechanism and, hence, the strategies for the eventual containment of CWP, we have initiated a comparative study of the free radical formation and cytotoxicity properties of freshly crushed coal ·particles. As done by Artemov and Reznik, 7 we have used the ESR technique as the direct method of measuring the concentration and decay kinetics of the coal based free radicals. Our preliminary ESR studies on two Pennsylvania coals, a bituminous (carbon content 72 % ) and an anthracite (95 % carbon) coal, have confirmed the Artemov-Reznik finding that the crushing-induced free radical sites are higher on the coals with higher carbon content (i.e., higher rank).9-11 In the present work we describe our more recent results of a parallel study of the time dependence of the decrease in the free radical content of a freshly made anthracite dust (as measured via ESR) and that of the dust's cytotoxicity potential as measured by the extent of hemolysis of (sheep) erythrocytes. We have also investigated the effects of free radical scavengers on the cytotoxicity potential of the dust and deduce that indeed the free radicals could play a significant role in the initial events in the mechanism of the cytotoxic effects due to the inhalation of coal dust inhalation."