The Role of Free Radicals in the Mechanisms of Hemolysis and Lipid Peroxidation by Silica: Comparative ESR and Cytotoxicity Studies

"Electron spin resonance (ESR) and cytotoxicity measurements were made on newly fractured silica to examine specifically the role of the fractured-induced, siliconbased radicals (Si· and SiO·), and silica-generated hydroxyl (·OH) radicals in the mechanism of the cell membrane damage by silica. The concentration of the Si· and SiO· radicals was controlled through decay processes, thermal annealing, and boiling, while that of the ·OH radicals was varied by using catalase, superoxide dismutase, KMnO4, Ha2seO3, ascorbic acid, and metal ions, and monitored via ESR spectroscopy. The dust's cytotozicity potential was evaluated by measuring the silica-induced hemolysis and also by determining lipid peroxidation (using linoleic acid). Prom the comparison of hemolysis and ESR results it is deduced that the radicals play little· or no role in the silica-induced hemolysis. However, the lipid peroxidation data indicate that the radicals might be involved in the initiation of an oxidative chain reaction leading to the macrophage membrane damage through lipid peroxidation. INTRODUCTIONThe biochemical mechanism of Quartz-related pneumoconiosis is still not fully understood. It is believed, nevertheless, that one event of primary importance is the perturbation of cell membranes by the dust particles (Allison et al., 1966; Allison and Harrow, 1971; Bateman et al., 1982; Parazzi, et al., 1968; Reiser and Last, 1979; Shi et al., 1989; Summerton et al., 1977). The ensuing cell-membrane damage unleashes certain lytic enzymes resulting in additional cell injury and eventual fibrosis (Nolan et al., 1981; Summerton et al., 1977). Thus considerable effort is currently being made towards the understanding of the processes by which quartz particles disrupt the membrane of a red blood cell (RBC) or a pulmonary alveolar macrophage (PAM). A simple but important method of evaluating the membrane damage potential of a dust is the hemolysis test (Harington et al., 1971). Even though this test is used routinely for screening dusts for their cytotoxicity potential, the biochemical mechanism underlying the cell lysis by many dusts, including quartz, is still controversial (Kilroe-Smith, 1974; Singh and Rahman, 1987), and this provided us the impetus for the present undertaking."