Evaluation of inhibitory effects on sucrosedependent cell adhesion on smooth surfaces by mutans streptococci

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When mutans streptococci are cultured in a medium containing sucrose, they strongly adhere on smooth surfaces. Dental biofilms are formed on teeth surface by interaction between glucan and oral bacteria. Therefore, the measurement of sucrose-dependent cell adhesion is used to evaluate the formation of biofilms on teeth surface. If sucrose-dependent cell adhesion on the smooth surface is inhibited by phytochemicals, the test substance indicates the possibility to reveal anticariogenic effects.

This procedure has been described by Hamada and Torii (Hamada and Torii, 1978). After mutans streptococci (S. sobrinus or S. mutans) are grown in the medium, the collected cells are resuspended in the medium. The cell suspension (0.5 mL), sucrose solution (final concentration, 1%) and 0.3 mL of phytochemical solution are mixed in a new glass test tube, and incubated at angle 20° for 24 h at 37°C. After incubation, the reaction mixture containing nonadherent cell is gently removed by a Pasteur pipette. The glass test tube upon which mutans streptococci adhere to the surface is gently washed with distilled water at angle 20°. In the control, cell and glucan are not peeled by washing because of those are tightly adhere on test tube. Then, the cell and glucan are suspended in 1 N NaOH to measure absorbance at 550 nm.

We have tried to evaluate the inhibitory effect of phytochemicals using this method, described below (Hashiguchi-Ishiguro et al, 2011). The degree of inhibitory effects on cell adhesion correlated roughly with inhibition of glucan production by phytochemicals. Anticariogenic effects of extractives from red wine, apple polyphenols and propolis are evaluated using this method (Furiga et al, 2008; Hayacibara et al, 2005; Yanagida et al, 2000). In this method, the surface of glass test tube is used for smooth surface model of teeth. The glass test tube used in this method must be new one that has very smooth surface with no flaw. Human saliva-coated hydroxyapatite beads are used as teeth surface model in other method (Venkitaraman et al, 1995), because teeth consists mainly of hydroxyapatite. The model of latter method reflects the situation of oral environment. Koo et al have reported the details of the procedure (Koo et al, 2002, 2010).

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