Non-irradiated campherquinone induces DNA damage in human gingival fibroblasts.
OBJECTIVE
Camphorquinone (CQ) is cytotoxic in cell cultures. The mechanism of this toxic action, however, is not yet clearly understood. Aim of this investigation was to analyze the effects of non-irradiated CQ on intracellular formation of reactive oxygen species (ROS), intracellular glutathione (GSH) content, and the integrity of DNA in cultured primary human gingival fibroblasts (HGF).


METHODS
Cells were exposed to CQ at concentrations ranging between 0.05mM and 2.5mM. Intracellular levels of ROS were detected by the fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH-DA) and GSH was determined by the fluorescent probe monobromobimane (MBBr). Genotoxicity was measured quantitatively by the alkaline comet assay. The cytotoxic effects of CQ were investigated by means of the fluorescent dye propidium iodide and the Cytotoxicity Detection Kit.


RESULTS
CQ generated an increase of intracellular ROS, a depletion of intracellular GSH level, decreased cells' viability and total cell number dependent on the applied CQ concentration: 0.5-2.5mM (ROS upward arrow, GSH downward arrow) and 0.125-2.5mM CQ (cytotoxicity upward arrow). Increased DNA damage was observed at all concentrations (0.05-2.5mM, p<0.05). The ROS-scavenger N-acetylcysteine (NAC) reduced CQ-induced ROS formation at CQ concentrations higher than 0.5mM (p<0.05).


CONCLUSIONS
Our data indicate that non-irradiated CQ induces oxidative stress, DNA damage and cytotoxicity as well in primary HGF.
Volk
Joachim
J
Clinic of Conservative/Preventive Dentistry and Periodontology, Hannover Medical School, 30625 Hannover, Germany.
Ziemann
Christina
C
Leyhausen
Gabriele
G
Geurtsen
Werner
W
D000111:Acetylcysteine / Q000494:pharmacology; D001643:Bridged Bicyclo Compounds; D002452:Cell Count; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D020552:Comet Assay; D004249:DNA Damage; D003764:Dental Materials / Q000633:toxicity*; D005347:Fibroblasts / Q000187:drug effects*; D005452:Fluoresceins; D005456:Fluorescent Dyes; D016166:Free Radical Scavengers / Q000494:pharmacology; D005881:Gingiva / Q000166:cytology / Q000187:drug effects*; D005978:Glutathione / Q000187:drug effects; D006801:Humans; D011419:Propidium; D017382:Reactive Oxygen Species / Q000032:analysis; D013439:Sulfhydryl Reagents; D013729:Terpenes / Q000633:toxicity*; D013997:Time Factors
D001643:Bridged Bicyclo Compounds; D003764:Dental Materials; D005452:Fluoresceins; D005456:Fluorescent Dyes; D016166:Free Radical Scavengers; D017382:Reactive Oxygen Species; D013439:Sulfhydryl Reagents; D013729:Terpenes; D011419:Propidium; C048339:camphoroquinone; C037631:2',7'-dichlorofluorescein; D005978:Glutathione; C028381:monobromobimane; D000111:Acetylcysteine