Dosage Effectiveness of the Combination Calcium Hydroxide-Propolis as an Inhibitor for Aggregatibacter actinomycetemcomitans Biofilm

Abstract

Objective: Calcium hydroxide is widely used as pulp-capping material, but it is resistant to Aggregatibacter
actinomycetemcomitans bacteria. Because of this deficiency, many researchers are considering using
alternative natural ingredients, such as propolis. Propolis has been widely used in dentistry due to its
properties and because it has active ingredients that can act as antibacterial agents. It is hoped that when
calcium hydroxide and propolis are combined, the combination can overcome the disadvantages. The aim of
this study was to determine the dosage effectiveness of the combination calcium hydroxide-propolis as an
inhibitor for A. actinomycetemcomitans biofilm. Material and methods: The sample was divided into five
groups. The positive control group is a combination of calcium hydroxide-sterile aquadest, and the negative
control group is A. actinomycetemcomitans bacteria. Group I is a combination of calcium hydroxidepropolis with a ratio of 1:1, group II has a ratio of 1:1.5, and group III has a ratio of 1:2. Each group consists
of five replications. Combination of calcium hydroxide-propolis and A. actinomycetemcomitans bacterial
suspension was incubated for 2x24 hours then washed three times using PBS. Furthermore, 2% of crystal
violet was used for staining and covered with aluminum foil and then incubated for 15 minutes. Isopropanol
200µL was added, followed by a measurement of OD with an ELISA reader, and the OD values were
calculated using an inhibition biofilm formula. Results: Group II has the largest mean inhibition with 76%
compared to groups one and three. Conclusion: A combination of calcium hydroxide and propolis with a
ratio of 1: 1.5 is more effective than 1:1 or 1:2 to inhibit A. actinomycetemcomitans biofilm.