The Potency of Non Sucrose Cajuputs Candy to Inhibit the Activity of Streptococcus mutans in Biofilm Formation

Abstract

Cajuputs candy is a confectionery considered as functional product effective for throat lozenges. Flavor compounds of cajuputs candy has been reported as inhibitor of biofilm formation by S. mutans serotype C. Non sucrose cajuputs candy is a product of cajuputs candy developed by replacing sucrose as the main component by isomalt. The aim of this research is to observe the potency of non sucrose cajuputs candy to inhibit the activity of S. mutans in biofilm formation by identifying the volatil components of candy and monitoring the formation of biofilm. This study has three experiment models. Control model (K) is non sucrose candy without flavor compounds, CC1 model is cajuput candy non sucrose with cajuput extract as flavor compound, and CC2 model is cajuputs candy non sucrose with cajuput and peppermint extract as flavor compounds. The volatil components was identified using Gas Chromatography Mass Spectra (GC-MS) with capillary column DB-5 MS in flavor laboratory of Indonesia Rice Research Center, Sukamandi-Subang, West Java. The volatil components of cajuput and peppermint extract is used as former information that could verify volatil components in CC1 and CC2. The inhibition of S. mutans activity in biofilm formation was monitored by analysis of biofilm mass viability and quantification of gtfB-gtfC mRNA with 4 and 6 hours incubation time. The viability of biofilm mass analysis and mRNA quantification was conducted in biology oral laboratory of Dentistry Faculty-University of Indonesia. The viability of biofilm mass obtained by monitoring the absorbance of biofilm suspension using microplate Enzyme Linked Immunosorbent Assay (ELISA) Reader. Expression level of gtfB-gtfC mRNA obtained by monitoring the CT The identification result of volatil components based by LRI value showed that cajuput extract has 22 volatil components inside meanwhile peppermint extract has 17 volatil compponents inside. The main volatil components of cajuput extract, consecutively are 1,8-cineole (47,61%), α-terpineol (10,76%), α-caryophyillene (7,91%), γ-terpinene (5,78%), terpinyl acetate (4,09%), α-pinene (3,90%), p-cymene (3,75%), and terpinolene (3,52%). The main volatil components of peppermint extract, consecutively are menthol (32,53%), menthon (15,60%), isomenthon (11,87%), methyl salicylate (10,86%), D-limonene (10,07%), β-pinene (3,77%), dan α-pinene (3,09%). The volatil components of cajuput and peppermint extract becoming components target of CC1 and CC2. The study showed that CC1 has 14 volatil components with main volatil components consecutively are 1,8-cineole (39,26%), α-terpineol (19,63%), dan α-caryophyillene (17,90%). Meanwhile CC2 has 18 volatil components with main volatil components consecutively are menthol (23,47%), α-terpineol (17,57%), value of gtfB and gtfC mRNA during amplification process using Real Polymerase Chain Reaction (RT-PCR). 1,8-cineole (14,94%), α-caryophyllene (13,46%), menthone (7,80%), dan isomenthone (7,65%). This research showed that the optimum inhibition of biofilm mass formation occured in 4 hours incubation with inhibition potency of K, CC1, and CC2 consecutively are 63,98%, 48,34%, and 65,64%. The inhibition potency of all models are less than 50% in 6 hours incubation, only CC2 has inhibition potency more than 40%. The expression level of gtfB mRNA in K, CC1, and CC2 model is higher than blank during 4 hours incubation. The experiment models K, CC1, and CC2 could inhibit the expression of gtfB mRNA later (6 hours incubation). Meanwhile, the existence of K, CC1, and CC2 could inhibit gtfC mRNA expression since 4 hours incubation. The experiment model CC2 containing mentol, α-terpineol, and 1,8-cineole as main volatil components has the best effect in inhibiting gtfB and gtfC mRNA