A double-blind crossover study on 4 adult volunteers was performed to evaluate the effect of hexetidine on Cu2+ accumulation in dental plaque as well as a possible enhanced effect of copper on inhibition of acid production in the presence of hexetidine. The experimental period was 5 days. No oral hygiene was allowed, and sucrose-containing chewing gum was used to enhance plaque formation during the test period. In order to evaluate the effect on pH, the test persons rinsed with a 15% glucose (w/v) solution on the 5th experimental day. Plaque pH values recorded before and 5 min after the rinse served as control values. One hour later the test persons rinsed with 10 ml of the test solutions for 1 min. Glucose rinses with pH measurements 5 min after the rinse were carried out 0, 3, and 7 h after the test agents were used. The combination of 1.0 mM copper and 2.0 m M hexetidine gave a significant (p < 0.05) inhibition of acid production at all times compared both to the controls and to each of the test agents separately. Plaque samples were collected with a toothpick immediately before a 1-min rinse with 10 ml of the test solutions. Subsequent plaque samples were taken 5 min and 3 and 8 h after a rinse from corresponding tooth surfaces. Dry weight was estimated, the plaque bacteria digested by HNO3, and the amount of Cu2+ determined by atomic absorption. Presence of hexetidine resulted in a significantly (p < 0.05) higher amount of Cu2+ retained by plaque bacteria compared both to the control values and the amount measured after rinsing with 1.0 mM CuSO4 alone.

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