Compare The Efficiency Of Different Sterilizing Solutions On Multispieces Biofilm

Periapical inflammation is a common disease in clinical, currently the most effectivetreatment is root canal therapy, the key principle is disinfect the infection within the rootcanal system to help healing the periapical lesions. Since the complex of apcial anatomyand some specific types of microbial etc that would cause root canal treatment failure.These disease can be retreated by root canal therapy or apical surgery solved. Which, insome infection, microbial appeared at outside of the apical, form apical biofilm. Then itmust be treated by resection of the apical part. In order to remove the apical biofilmcompletely, it requires removal about3mm of the tooth apical. But this bound to affectthe structure of the teeth. Whether can use physical or chemical means (non-mechanicalremoval) to clean apical biofilm, to exterminate the infection and maximize protect toothstructure?Domestic and international scholars refer this after root canal treatment even hard tocontrol by retreatment periapical disease as Refractory Periapical disease. There areresearch shown Enterococcus faecalis, Streptococcus sanguis, Actinomyces israe-lii aregram-positive facultative anaerobic bacteria, these bacteria can be found in the biofilm that located on the extracular part of apical,actinomyces can cause chronic apicalactinomycosis,Streptococcus sanguis is the common bacteria exist in the periodontal andheart disease. Because of his strong tolerance ability,rEnterococcus faecalis is often foundin root canal disease, The pathogen aspects of these three kinds of pathogenic bacteriaformed biofilm remains to be seen, but obviously there will have a different pathologicalreaction when mixed infection.1.Apical multi-cultured bacterial biofilmsFirst,put three kinds of bacteria Enterococcus faecalis, Streptococcus sanguis andActinomyces israe-lii,co-cultured in BHI and1/2BHI agar to observe are there have theformation of mutual colonies inhibition among the three bacterial colonies.Then inoculatethese three bacteria into96-well plates,Use Crystal violet staining to observe the growthof biofilm.Take a96well plate, the plate was placed in an anaerobic incubator for37℃after48h culture, crystal violet staining was observed with a single bacteria forming biofilm,the culture were terminate at24,48,54,56,58,70,76,82,94,106hour,Washed with distilledwater three times,250μl ethanol solution was added to each well, fixing30min; liquiddiscarded after a fix end, respectively, use1g/L crystal violet staining5min and330ml/Lacetic acid to wash the dye, OD value was measured at a wavelength of570nm using aplate reader, and biofilm growth curve was observed.Three kinds bacteria co-cultured on the apical cementum sheet for5days, Use25ml/L glutaraldehyde solution fix those sheets overnight at4℃, Gradient dehydrationperformed use500ml/L,600ml/L,750ml/L850ml/L,950ml/L,1000ml/Lacetonitrile; coated with gold, take photograph of the result by scanning electronmicroscopy. The above experiments suggest that the three different bacteria can growtogether.2.The the efficiency of different sterilizing solutions on multispieces BiofilmOn the basis of the first experiment, Inoculated cementum sheets and cell climbing slides were washied with strongly acidic electrolyte water (SAEW),differentconcentrations of sodium hypochlorite (NaClO),recombinant human beta defensin－3(rHBD-3),chlorhexidin (CHX) solution for15seconds,1minute and5minutes.52.5g/LNaClO solution and saline respectively as the positive and negative controls.After eachtreatment,fix and dehydrate the sample,Spray with platinum then observe the result withscanning electron microscopyand with confocal laser scanning microscopy(CLSM) toobserve the treatment group which stained with immunofluorescence, the observationresults were analyzed with ImageJ software. Each experimental group and positivecontrol group were significantly lower viable bacteria than the negative control group ateach treatment time point (P <0.05). SAEW/15s groups,5g/L/NaClO15s、1min、5mingroups,50、100、800mg/L rHBD-3solution,15s、1min、5min treatment groups, higherviable bacteria than the positive control group at each treatment time point (P<0.05).SEAW1min、5min groups,10g/L sodium hypochlorite,20g/L chlorhexidine1min、5min treatment group compare with52.5g/L Sodium hypochlorite solution theproportion of viable bactria in treatment groups were not statistically different(P>0.05).Conclusion：SAEW、high concentrate NaCLO and can effectively kill the bacteria withinthe apical biofilm, rHBD-3also have the bactericide ability,consider both NaCLO andCHX irritating and can causeAllergy,SAEW would have a bright future on clinical use.