The In Vitro Effect Of Erythritol-milk Mixture In A Biofilm-enamel Model

Purposes：Erythritol is a new type of sugar alcohols sweetener, has low quantityof heat, high resistance, non-toxic side effects, hygroscopicityadvantages, in recent years, the study found that it also has the functionof prevention of caries disease. Milk contains rich nutrition, be called"white blood", it contains calcium, phosphorus inorganic salt ion andcasein and other material can play a role of preventing caries.Thisresearch try to make a combination of erythritol and milk, hope the mixtureboth play erythritol sugar substitutes attributes and milk nutritioncharacteristics, to meet the child to eat sugar quantity and growth ofnutrition requirements,the most important is to prevent caries disease.Make a study about the mixture’s feasibility and anticaries mechanism,toprovide the experimental evidences for it be safety and widely used.Methods：1.Constructing an artificial caries model in vitroS.mutans were choosed to incubate the caries model. The optical densitywere measured at6,12,24,36,48,72hours, following a curve describedbiofilm growth.Measure enamel surface microhardness value and stop theexperiment when statistical significance was found between before,judging whether the model formed early enamel caries according to thechange of hardness value.2.The in vitro effect of Erythritol-milk mixture to S.mutans biofilm Using BHI, fluorine milk, milk,2%erythritol-milk,4%erythritol-milk,6%erythritol-milk deal with established caries model. Stripping S.mutansbiofilm after modeling120hours, viable count method to observe biofilmtotal viable, CLSM to observe the bacterial biofilm structure andcalculate the viable percentage in different parts of the biofilm.3.The in vitro effect of Erythritol-milk mixture to take off mine enamelMeasuring enamel microhardness value in the modeling25day found thestatistical significance was meaningful, terminate the experiment.Remove surface biofilm, measuring enamel block microhardness reactionmineral loss situation. Using ESEM to observe the enamel surfacemicrostructure.Results：1.S.mutans biofilm showed slow nonlinear growth trend,6-12hour curvestraight steep,12-48hour slow,48-72hour curve was very gentle.Comparedwith before, artificial caries group enamel block processing of hardnesssignificantly (P<0.05), whereas the control hardness value no significantchange (P>0.05).2.BHI group viable quantity was significantly higher than other groups(P<0.05);2%erythritol-milk group more than4%erythritol-milk group and6%erythritol-milk group (P<0.05);4%erythritol-milk group,6%erythritol-milk group, and fluorine milk group viable count had nostatistical significance (P>0.05).CLSM results showed that the viable percentage of fluorine milk group anderythritol milk group were higher than the BHI group and milk group ineach layer, but biological membrane structure loose and bacteriadistribution. 3.All group made normal enamel hardness value decrease (P<0.05); BHI group,milk group,2%erythritol-milk group hardness loss most;4%erythritol-milk group,6%erythritol-milk group and fluorine milk groupmicrohardness loss small.ESEM pictures showed that BHI group enamel surface rough and uneven inthe most, serious destruction; fluorine milk group enamel surface leveloff, deposit the crystal particles; Milk group enamel surface a smallamount of particles sedimentation;2%,4%,6%erythritol-milk groupenamel surface were flat, mineral particles cover like membrane,microporous reduced;6%erythritol-milk group sediment most full,microporous almost disappear.Conclusions：1.Application of the experimental method can induce the S.mutans biofilmwhich was necessary in established this in vitro model, at the end ofmodeling the caries enamel owed the basic caries performance. This modelcan be used for subsequent experiments.2.Erythritol-milk mixture can inhibition S.mutans biofilm growth andadhesion at certain extent.3.All the concentration of erythritol-milk mixture can reduce surfaceenamel demineralization, promote again the occurrence of mineralization,and6%erythritol-milk mixture had strongest mineralization ability.