Preparation Of Egg York Antibody Aganist Prevotella Intermedia And Its Preventive Effect On Expeimental Pregnancy Gingivitis

Objective To prepare and purify anti Prevotella intermedia yolk antibody(anti P.intermedia-Ig Y),to observe its preventive effect on rat experimental pregnancy gingivitis.Methods P.intermedia was cultured anaerobically,inactivated with 2% formaldehyde,with final concentration of 1 x 109 CFU/m L bacteria,1m L(0.5 m L liquid +0.5 m L adjuvant)/ times,4 times the immune layer.Removed the egg white with yolk separator,sterile syringe punctured the vitelline membrane,collecting the yolk,two step ammonium sulfate precipitation and purification of anti P.intermedia-Ig Y.SDS-PAGE gel electrophoresis,Western blot,ELISA on the molecular weight and specificity of the identification,and detection of changes in the immune titer.30 female SD rats(8-10weeks),15 male SD rats(8-10 weeks),female: male =2:1 caged for 12 hours.24 pregnant mice were randomly divided into 4 groups: 0.12% chlorhexidine incubation group(A),anti P.intermedia-Ig Y incubation group(B),physiological saline incubation group(C),control group(D).During modeling,P.intermedia was incubated with different treatment solutions and then inoculated in gingival sulcus of rats,different groups were vaccinated with different P.intermedia,1 time / day for 7 days.By detecting Gingival index(GI),P.intermedia content in plaque and histological observation of gums between different groups to observe the preventive effects of anti P.intermedia-IgY on experimental pregnancy gingivitis.Results After 4 immunization,the highest titer of antibody was 1:512,000,which was maintained at 45 d.1 g yolk by two step ammonium sulfate precipitation was Ig Y about18 mg.SDS-PAGE gel electrophoresis showed that the purity of purified Ig Y was64.5%.Western blot showed anti P.intermedia-Ig Y recognition and specificity combined with P.intermedia bacterial cleavage fragment.Observation of 0.12% chlorhexidine incubation group and anti P.intermedia-Ig Y incubation group gingival index(gingival index,GI)were significantly lower than that of the saline incubation group(P < 0.01),but there was no difference between the two GI(P > 0.05).Gingival tissue pathology was observed to show that the inflammatory state of the anti P.intermedia-IgY group was less than that of the saline group.Conclusion After 4 times of immunization,sustaining high titer anti P.intermedia-IgY can be gained,and anti P.intermedia-Ig Y can reduce the experimental pregnancy gingivitis inflammatory state of rats,which has a certain preventive effect on experimental pregnancy gingivitis of rats.