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Cell Biology and Toxicology

, Volume 11, Issue 2, pp 79–88 | Cite as

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

  • H. Babich
  • B. J. Wurzburger
  • Y. L. Rubin
  • M. C. Sinensky
  • L. Blau
Article

Abstract

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.

Key words

calcium influx assay chlorhexidine digluconate dental products gingival epithelial cells gingivitis in vitro cytotoxicity liposomes mouthrinse neutral red assay plaque 

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

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Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • H. Babich
    • 1
  • B. J. Wurzburger
    • 2
  • Y. L. Rubin
    • 1
  • M. C. Sinensky
    • 3
  • L. Blau
    • 2
  1. 1.Department of Biology, Stern College for WomenYeshiva UniversityNew YorkUSA
  2. 2.Department of Chemistry, Stern College for WomenYeshiva UniversityNew YorkUSA
  3. 3.New York University College of DentistryNew YorkUSA

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