Use of Chemiluminescence Assays to Monitor the Surface Characteristics and Biological Reactivity of Freshly Fractured VS Aged Silica

"IntroductionInhalation of crystalline silica is often associated with the development of pulmonary disease (Ziskind et al., 1976). Recently, it has been proposed that freshly sheared silica .may have surface properties that make it more reactive with lung tissue than aged silica, and that this unique reactivity ·of freshly crushed silica enhances its pathogenicity. A recent study in our laboratory strongly supports this theory (Vallyathan, et al., 1988a; 1988b). We found that grinding of silica cleaves bonds in a silica tetrahedral lattice generating reactive free radicals sites on the surface of the silica particles. The presence of surface radicals on freshly ground silica was monitored by electron spin resonance (ESR) spectroscopy. The ESR signal generated from fresh silica in air is characteristic of Si-0 and Si radicals. Evidence, i.e., the ESR signal after the addition of a spin trap (DMPO) and its sensitivity to ethanol, indicates that these silicon-based surface radicals can react with water to form hydroxyl radicals (OH). Results also indicate that these surface radicals make fresh silica more cytotoxic than aged silica; i.e., fresh silica causes more membrane damage as evidenced by hemolysis and lipid peroxidation. Therefore, the presence of reactive sites on the surface of freshly cleaved silica may play a significant role in its pathogenicity.If surface· free radicals are related to cytotoxicity, then screening dusts for the existence of such radicals may predict their relative pathogenicity. Since an ESR spectrometer is not available to many laboratories, the objective of this study was to develop a quick and simple technique to monitor the existence of reactive surface sites on dust. The technique chosen was the measurement of chemiluminescence using a liquid scintillation counter; i.e., an instrument which·would be accessible to most investigators."