Effect Of Caesalpinia Sappan, Paris Polyphylla Var.yunnanensis And Syzygium Aromaticum On Plaque Biofilm In Vitro

[Objective] Based on the results of our previous study which focused on the effects of Caesalpinia Sappan (CS), Paris Polyphylla var.yunnanensis (PS) and Syzygium Aromaticum (SA) on the growth and metabolism of cariogenic bacteria in planktonic state, the aim of this study was to evaluate the effects of CS, PS and SA on plaque biofilm in vitro and to explore their potential value for caries prevention.[Methods] The plaque biofilm was established in an artificial mouth model in vitro and was treated by CS, PS and SA respectively.(1) Real-time Quantitative PCR was used to observe the growth of bacteria in plaque biofilm;(2) Scanning Electron Microscopy was used to observe the formation procedure and the surface morphology of plaque biofilm;(3) Laser Scanning Confocal Microscopy and Fluorescence Staining technique were used to observe the thickness and activity of plaque biofilm.[Results](1) Effects of CS, PS and SA on the growth of bacteria in plaque biofilm:①At6h, compaired with the control group, CS could effectively inhibit the growth of S.mutans, S.sanguist, L.acidophilus and A.viscosus (P<0.05); PS could effectively inhibit the growth of S. mutans and L.acidophilus (P<0.05). The results of multiple comparison showed that the inhibition of CS on the growth of L.acidophilus was most obviously.②At24h and48h, compaired with the control group, CS could effectively inhibit the growth of S.mutans, S.sanguist, L.acidophilus and A.viscosus (P<0.05); PS could effectively inhibit the growth of S. mutans, S.sanguist and L.acidophilus (P<0.05); SA could effectively inhibit the growth of S. mutans and L.acidophilus (P<0.05). The results of multiple comparison showed that the inhibition of CS on the growth of S. mutans、A.viscosus were obviously.③At72h, compaired with the control group, CS,PS could effectively inhibit the growth of S.mutans, S. sanguis, L.acidophilus and A.viscosus (P<0.05); SA could effectively inhibit the growth of S. mutans, S. sanguis and L.acidophilus (P<0.05); The inhibition of CS on growth of S. mutans and A.viscosus was obviously. The results of multiple comparison showed that the inhibition of CS on the growth of S. mutans and A.viscosus were obviously.(2) Effects of CS, PS and SA on the surface morphology of plaque biofilm:①At6h:In control group, L.actobacillus was dorminated on the surface of HA and small gaps between bacteria groups were observed. In CS group, only few L.actobacillus with large gaps were discovered. In PS group, the result was nearly identical with control group, but the gaps between L.actobacillus groups were larger. In SA group, bacteria distributed widely.②At24h:More bacteria were observed in four tested groups. In control group, irregular biofilm structure with hole-shape channel was observed. In CS group, no biofilm was formed on the HA surface. In PS group, gaps between bacteria were smaller and no biofilm was formed on the HA surface. In SA group, bacteria were observed to distribute widely or aggregate together.③At48h:In control group, many regular and crn-like structures were observed. In CS group, a few irregular and corn-like structures were observed. In PS group, fewer corn-like structures were observed. In SA group, the result was similar with that of control group, but the biofilm was irregular.④At72h:In control group, three-dimensional reticular structures were observed. In CS group, slice-shape biofilm with hole-shape channel was discovered. Compared with CS group, fewer hole-shape channels were discovered in PS group. In SA group, the density of the biofilm was bigger than that of CS group.(3) Effects of CS,PS and SA on the thickness and activity of plaque biofilm:The thickness of plaque biofilm ranged from5.76μm to19.26μm. At6h,24h,48h and72h, CS,PS and SA could effectively reduce the thickness of plaque biofilm, the effect of CS was much remarkable than PS and SA(P<0.05); At6h,24h, 48h and72h, CS,PS,SA could reduce the activity of plaque biofilm, the effect of CS was much remarkable than PS and SA (P<0.05).[Conclusion] CS could effectively inhibit the growth of S. mutans, L.acdophilus, S. sanguis and A.viscosus. PS could effectively inhibit the growth of S. mutans, L.acidophilus and A.viscosus. SA could effectively inhibit the growth of S. mutans and L.acidophilus. Among them, the inhibition of CS was the strongest. CS, PS and SA could effectively inhibit the formation of normal surface morphology. Among them, the inhibition of PS was the most remarkable. CS, PS and SA could effectively inhibit the thickness and activity of plaque biofilm. Among them, the inhibition of CS was the strongest. Therefore, CS and PS are worth further study on their value of prevention and treatment of dental caries.