Anin vitro study on the cytotoxicity of chlorhexidine digluconate to human gingival cells

Chlorhexidine digluconate is the active ingredient in mouthrinses used to prevent dental plaque and gingivitis. Thein vitro cytotoxicity of chlorhexidine was evaluated with the Smulow-Glickman (S-G) gingival epithelial cell line. The potency of chlorhexidine was dependent on the length of exposure and composition of the exposure medium. The midpoint cytotoxicity values for 1-, 24-, and 72-h exposures were 0.106, 0.011, and 0.0045 mmol/L, respectively. S-G cells exposed for 2 h to chlorhexidine and then maintained for 48 h in chlorhexidine-free medium were unable to recover from the initial insult. The adverse effects of chlorhexidine on the plasma membrane were suggested by the leakage of lactic acid dehydrogenase from chlorhexidine-treated S-G cells and by the increased permeability of chlorhexidine-treated liposomes to Ca²⁺. The toxicity of a 24-h exposure to chlorhexidine to the S-G cells was progressively lessened as the content of fetal bovine serum (FBS) in the exposure medium was increased from 2% to 8%. The potency of a 1-h exposure to chlorhexidine was reduced in medium amended with albumin, lecithin, and heat-killedEscherichia coli. These reductions in toxicity were presumably due to the binding of the cat onic chlorhexidine to the negatively charged chemical moieties of the components of FBS and of albumin and lecithin and of sites on the surfaces of bacteria. Combinations of chlorhexidine and carbamide peroxide were additive in their cytotoxicities.Abbreviations ANOVA analysis of variance - [Ca²⁺]i calcium concentration in internal medium of liposomes - DMEM Dulbecco's modified Eagle medium - EDTA ethylenediamine tetraacetic acid - FBS fetal bovine serum - Hepes N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid - HGF-1 human gingival fibroblast cell line - HSD honestly significant differences - KB cell line derived from a human epidermoid carcinoma in the mouth - LDH lactic acid dehydrogenase - NADH nicotinamide adenine dinucleotide, reduced form - NR neutral red - NR₅₀ concentration inhibiting neutral red uptake by 50% - PBS phosphate-buffered saline - SEM standard error of the mean - S-G Smulow-Glickman human gingival epithelial cell line