Peri-implantitis Dynamic Study On Continuous Culture Of The Beneficial Bacteria And Suspicious Pathogens

[Objective]Study on growth curves of peri-implantitis focus beneficial bacteria and suspicious pathogens, interaction between bacteria, through the improvement of existing Chemostat, designed to build a plant of peri-implantitis Subgingival bacteria biofilm in vitro model of the new type of Chemostat,measure and observe biomass on the biofilm, provide a theoretical basis for the microecology treatment of peri-implant inflammation.[Method]Experimental choose S.s as useful bacteria, P.g, and P.i as suspicious pathogens, respectively measurement its growth curve, determines bacteria of logarithmic growing phase, by the anti bacteria experiments to observe the interaction between bacteria, titanium tablets with micro-arc oxidation processing, three species of bacteria are divided into four groups:single bacteria P.g group, and single bacteria P.i group, and double bacteria P.g+S.s group, and double bacteria P.i+S.s group, installed Chemostat device, continuous cultured for72h, respectively at24, and48, and72h, by counting live bacteria on the biofilm to determine the biomass of titanium tablets, through scanning electron microscopy, confocal laser scanning microscopy, fluorescence microscopy to observe the structure of the biofilm on titanium films.[Results]1. Growth curve shows the following growth rate of bacterial:single bacteria P.g> double bacteria P.g+S.s> single bacterium P.i> double bacteria P.i+S.s. The logarithmic growth phase of bacteria:bacteria P.g2-8h; single bacteria P.i4-18h; double bacteria P.g+S.s4-12h; double bacteria P.i+S.s4-8h. Maximum absorption peak of bacteria:bacteria P.g28h; single bacteria P.i12h; double bacteria P.g+S.s22h; double bacteria P.i+S.s12h.2. Antitheses on blank zone shows no bacterial bacteriostatic zone cycle; and diameter of bacterial bacteriostatic zone cycle increased with the increase of the concentration of Ss, reaches its maximum value when the concentration of S.s1.0x108CFU/mL; bacteriostatic zone diameter of the same concentration of Ss, Pg group are larger than the Pi group.3. At the same time point, bacterial colony number are10-2>10-3>10-4. At the same concentration, the number of colonies are72h>48h>24h. After joining the S.s, P.g at the same time with the same concentration, the number of colonies are less than a single bacteria group; After joining the S.s, P.i at the same time with the same concentration, the number of colonies are less than a single bacteria group.4. The number of plaque in each group are increased with the increasing of time. Group one and group two of bacteria plaque biofilm morphology observed no significant differences, Group three and group four of bacteria plaque biofilm morphology observed no significant differences, but compared with the first one and the second one in each different group, observed there is significant difference in morphology, in same time points the generation volume of bacteria plaque in group three less than group one, in same time points the generation volume of bacteria plaque in group four less than group two.5. Biofilm thickness is between5.92-30.52μm. The thickness of biofilm in each group will be increased with time, under the fluorescence microscope, bacteria shows blue-green fluorescence, by using CLSM bacteria shows blue fluorescence, cocci were arranged in chains and intertwined with the group sheet shape bacteria, surrounded by scattered single cocci.[Conclusion]1. Bacterial growth curve shows that after8hours incubation, single and double bacteria groups are entering logarithmic growth phase, which provide data support for subsequent experiments.2. Bacteriostatic experiment, CFU colony count after continuous culture shows that the S.s has the growth inhibition function on P.g, P.i.3. Scanning electron microscopy to detect the biofilm surface morphology shows that mixed culture of Ss has an effect on biofilm morphology of Pg and Pi. 4. Laser confocal microscope measures the thickness of the biofilm increased with time and continued thickening, the thickness of biofilm formed on the surface of the titanium plate in each group at the same point of time are distinctly different from each other. The effect of mixed culture of S.s to the biofilm thickness of P.g, P.i is thinning first then thickening.