The production of reactive oxygen species by irradiated camphorquinone-related photosensitizers and their effect on cytotoxicity.

Camphorquinone (CQ) is widely used as an initiator in modern light-cured resin systems but there are few reports about its effects on living cells. To clarify the mechanism of photosensitizer-induced cytotoxicity, the production of initiator radicals and subsequent reactive oxygen species (ROS) by CQ, benzil (BZ), benzophenone (BP), 9-fluorenone (9-F) in the presence of the reducing agent (2-dimethylaminoethyl methacrylate or N,N-dimethyl-p-toluidine, DMT) with visible-light irradiation was examined in a cell or cell-free system. Initiator radical production was estimated by the reduction rate of 1,1-diphenyl-2-picrylhydrazyl and by the conversion of poly-triethyleneglycol dimethacrylate; the results indicated that CQ/DMT had the highest activity among them. The cytotoxic effects of the photosensitizers on both human submandibular gland (HSG) adenocarcinoma cell line and primary human gingival fibroblast (HGF) showed that the 50% toxic concentration (TC(50)) declined in the order: CQ>BP>9-F>BZ. ROS produced in HSG or HGF cells by elicited, irradiated photosensitizers were evaluated in two different assays, one using adherent cell analysis and sorting cytometry against adherent cells and the other, flow cytometry against floating cells, with fluorescent probes. ROS production was dose- and time- dependent, and declined in the order: BZ>9-F>BP>CQ. Cytotoxic activity was correlated with the amount of ROS. Cytotoxicity and ROS generation in HGF cells was significantly lower than in HSG cells. ROS induced by aliphatic ketones (CQ) were efficiently scavenged by hydroquinone and vitamin E, whereas those by aromatic ketones (9-F) were diminished by mannitol and catalase, suggesting that OH radicals were involved in ROS derived from 9-F. A possible link between the cytotoxic activity and ROS is suggested.