The Role Of Enterococcus Faecalis EfaA In Rat Periodontitis And Related Endocarditis

Objective:To establish the model of infective endocarditis caused by experimental periodontitis in rats,to explore the role of Enterococcus faecalis endocarditis antigen(Efa A)in rat periodontitis and related infective endocarditis,On the prevention and treatment of chronic periodontitis and infective endocarditis lay the foundation.Method:1.Establishment and evaluation of periodontitis-infective endocarditis model:(1)Ginging with gingival-Wire ligation-Periodontitis in patients with bacterial culture Rinse-Normal diet method for rat rat periodontitis model.Left maxillary second molar to do experimental periodontitis model,the right to do their own control.24 hours after ligation of the wire,the oral culture of severe periodontitis patients and 0.5 ml of Enterococcus faecalis were washed with the oral cavity of the experimental rats.Four weeks later,rats were sacrificed by intraperitoneal anesthesia,and the periodontitis was measured by maxillary second molar;(2)The PE-50 catheter was inserted from the right common carotid artery of the rat,subcutaneous fixation,suture and incision.After disinfection,the PE-50 catheter was placed in the cage and the rats were sacrificed;(3)Observation of the formation of the left maxillary second molar periodontitis: The formation of the periodontal pocket and the distance between the enamel-bone mass(CEJ)and the alveolar ridge;(4)To observe the infection of endocarditis in rats: Anatomical rat stripping the heart,removing the heart valve neoplasm,weighing the humidium of the supernatant,the homogenate bacterial count;Comprehensive evaluation of periodontitis-infective endocarditis model was established successfully.2.Comparison of Periodontitis of Enterococcus faecalis efaA-wild and efa A + transgenic ratsRats were divided into groups(6 rats in each group),24 hours after ligation of the wire,the culture of severe periodontitis was carried out overnight.Bacterial cultures and enterococcus faecalis efa A-wild plants,Enterococcus faecalis efa A+ the rats in the experimental rats were rinsed.The rats in the maxillary second molar were sacrificed at 4 weeks after abdominal anesthesia.The rats were divided into two groups: HE staining,periodontal tissue C reaction protein(CRP)immunohisto-chemistry.3.Comparison of Infective Endocarditis Related to Periodontitis in Escherichia coli Efa A-Wild and Efa A + TransformantsCatheters were inserted 24 hours after successful,Animals were incubated with 0.5 ml of Enterococcus faecalis efa A— wild strain,Enterococcus faecalis efa A+ transformant strain,overnight,and the rats were sacrificed after one week of abdominal anesthesia,Heart blood,4000 rpm centrifugation 10 minutes,serum collection was completed by enzyme-linked immunosorbent assay to detect C-reactive protein(CRP)levels;Anatomical rats were sacrificed from the heart and the heart valve neoplasms were removed.The bacterial counts were compared between the two groups.Results:1.Periodontitis-Establishment of Infective Endocarditis Model:Successful establishment of periodontitis-endocarditis rat model: left periodontal tissue in the gingival cut 2 days after the emergence of0.5mm periodontal pocket depth,The depth of the periodontal pocket reached 3.0 mm at the end of the fourth week(cardiac intubation),Experimental alveolar bone showed pits-like absorption,and even deep root tip;The rats were sacrificed and dissected,and the heart valves were seen to form vegetation.The average weight of the vegetation was19.27 mg and the average colony count was 2.01 × 1010 CFU / mg.2.Comparison of Periodontitis of Enterococcus faecalis efa A-wild andefa A+ transgenic ratsPeriodontal tissue in periodontal tissues of periodontitis rats.Efa A-wild strain: the epithelium to the root movement,epithelial lamina propria inflammatory cell infiltration,alveolar bone resorption;Efa A+ transfor-mants:inflammatory response is more significant,showing a large number of binding epithelium to the root movement,spinous hyperplasia,epithelial nail protrusion elongation is more obvious,alveolar bone destruction significantly.Immunohistochemical staining of CRP in rat model of periodontitis,Efa A-wild strain,CRP is not expressed in osteoblasts,only weakly positive in partial fibroblasts;Efa A+ transformants,CRP in fibroblasts and osteoblasts were expressed.3.Comparison of Infective Endocarditis Related to Periodontitis in Escherichia coli Efa A-Wild and Efa A+ TransformantsEfa A+ transformants cause periodontitis-associated infective endocarditis,The average wet weight of the vegetation was 19.27 mg,the bacterial count was 2.01 × 1010 CFU / mg,the serum CRP level was 4.69 ng / ml;Efa A-wild-induced periodontitis-associated infective endocarditis,The average wet weight of neoplasm was 8.03 mg,the bacterial count was0.48 × 1010 CFU / mg,the serum CRP was 2.21 ng / ml.Compared with the wild strain,the wet weight and the bacterial count of the supernatant were higher(P<0.01),Serum CRP is more(P<0.05).Conclusions:1.Successful establishment of chronic periodontitis associated with infective endocarditis rat model.2.Enterococcus endocarditis antigen Efa A is beneficial to the occurrence and development of rat periodontitis and related infective endocarditis.