Study On Inhibitory Of Rubrus Suarissimus S.Lee Dental Care Buccal Tablets On S.Mutans

To assess the effects of RSL dental care buccal tablets on Glucosyltransferase(GTF) and Insoluble exopolysaccharides(IS) of S.mutans to provide the experimental basis for the marketing of RSL dental care buccal tablets on prevention of dental caries in the future.METHODS1. Preparation of S.mutans bacterial suspensionThe freeze-dried S.mutans standard strain powder was activated in TSB for 3h, then inoculated in MSB for 48h cultivation under anaerobic condition(37℃). Typical colonies were picked to make of bacterial suspension in normal saline. At last adapted the optical density (OD) at 540nm wavelength to l,then used bacteria count board to calculate the concentration of bacteria.2. Preparation of the RSL dental care buccal tablets solution Lapped into powder under aseptic condition, the RSL dental care buccal tablets were diluted into TTY liquid medium by two-fold dilution, with concentration ranging from 20g/50ml�'0.165g/50ml.3. Effect of RSL dental care buccal tablets on the growth of S.mutansUsing tube dilution method for determination of minimal inhibitory concentration(MIC)of the growth of S.mutans, each tube of bacteria was collected by centrifugation and placed in 2ml saline, then mixed to measure OD540nm.4. Extract Glucosyltransferase(GTF) of S.mutans and measure its activitySelected five different concentration RSL dental care buccal tablets 3 ml which were sub-MIC�'0.3ml S.Mutans were added into experimental groups and the control group�'anaerobic cultured for 48h�'centrifugate to collect supernatant�'added by solid ammonium sulfate�'centrifugated and collected the precipitation which was dissolved into sodium phosphate buffer, and dialyzed for obtaining crude GTF. Respectively Bradford method and Neson-Somogyi method were used to measure the content of total protein of GTF and glucan polysaccharide of the enzyme-substrate reaction solution. Dynamic unit of GTF was defined that under standard assay conditions the amount of enzyme required to release 1μmol glucan polysaccharide from glucose per minute, and specific activity was computed.5. Measure insoluble exopolysaccharides (IS)The part of bacteria was added with 0.5M NaOH. Centrifugation was followed. The supernatant was collected, and the ethanol which was 3 times volume of the supernatant was added. Followed by centrifugation, precipitation was the reaction of IS. Anthron method was used to measure the contents of IS.RESULT1. In test of tube dilution method, S.mutans didn’t appear in TTY which concentration of RSL dental care buccal tablets was over 20g/50ml. S.mutans grew differently in TTY which concentration was sub 10g/50ml. The lower concentration, the more S.mutans. MIC of RSL on S.mutans was 20g/50ml by experimental results. When the concentration greater than 0.313g/50ml, the OD of RSL dental care buccal tablets were lower than the control group(P<0.05).2. In the tests of S.mutans impacting on GTF dynamic, the total protein content was increased with the increasing concentration of RSL dental care buccal tablets. The total protein content of different concentrations of RSL dental care buccal tablets were higher than the ones of negative control group(P<0.05), Each experimental group was different from the other(P<0.05).The activity of GTF was decreased with the increasing of concentrations of RSL dental care buccal tablets. The content of glucan poly saccharide、the activity of GTF and the specific activity were different among each concentration of RSL dental care buccal tablets, of which were all lower than the negative control group.3. In the tests of S.mutans impacting on IS, the IS of S.mutans decreased gradually with the increasing concentration of RSL dental care buccal tablets. The content of IS in experimental groups were different from those of control group. There was highly significant difference among each group of RSL dental care buccal tablets. CONCLUSION1. RSL dental care buccal tablets can inhibit the growth of S.mutans.2. RSL dental care buccal tablets can significantly inhibit the GTF activity of S.mutans.3. RSL dental care buccal tablets can inhibit the synthesis of IS of S.mutans.4. In the aspect of caries prevention, RSL dental care buccal tablets is expected to be a new, inexpensive dental-care products, with a certain economic developmental value.