Water-soluble C-60 and macrophages: Morphologic features of FC4S-treated peritoneal macrophages in vitro and in vivo - A preliminary report

Morphologic investigation of water-soluble hexasulfonated C-60 (FC4S) effec ts on rat peritoneal macrophages in vitro and in vivo was conducted by tran smission electron microscopy. In the case of in vitro studies, two groups o f spontaneous peritoneal macrophages were separately cocultured with FC4S i n a concentration of either 0, or 100 ng/ml for 24 h. The other two groups were cocultured with either FC4S (100 ng/ml) for 12 h, or Pseudomonas aerug inosa exotoxin A (ETA) 100 ng/ml for 24 h. The former 12 h FC4S-treated gro up was subsequently cultured with ETA; for an another 12 h. All these 4 gro ups 24 h of cocultured monolayer cells were fixed in glutaraldehyde 2.5%, p ost fixed in 1% OsO4 and processed for morphologic evaluation. The results revealed that the viability of FC4S-treated cells in a healthy state, conta ining numerous cytoplasmic electron-dense (ED) bodies. The origin and natur e of these ED bodies were not known. Comparable observation was obtained wi th the FC4S-pretreated and ETA-treated the cells as viable and healthy, Thi s suggested the existence of plausible antioxidative bio-reactions involvin g phagocytosed hexa(sulfobutyl)[60] fullerenes in cocultured macrophages. I n the case of in vivo studies, the cells removed from rats that were ip inj ected with 60 mg/kg of FC4S were found to be typical active macrophages tha t bore numerous lysosomes and phagolysosomes in a variable degree of phagoc ytosing state. The presence of phagolysosomes were attributable to the FC4S exposure. The affected cells were diagnosed as "phagolysosomal macrophagop athy".