Serum Change Of ICAM-1, VEGF And E-sectin In Experimental Rat Model Of Periodontitis With Atherosclerosis

Objective: To establish the animal model of periodontitis withatherosclerosis in male rats, and detect the serum change of inflammatoryfactor: ICAM-1, VEGF and E-selectin of periodontitis with atherosclerosis.Explore the interrelationship between periodontitis and atherosclerosis.Methods:40Wistar rats (6-month-old, male and body weight~300g)were randomly diveded into four groups (10rats per group): normal group,periodontitis group (group PD), atherosclerosis group(group AS), andperiodontitis with atherosclerosis group (group PD+AS). Group PD, groupPD+AS were anaesthetized by intraperitoneal (IP) injection of10%chloralhydrate, and ligated bilaterally of their first maxillary molar teeth with0.2mmorthodonticwire (as far as possible placed in the gingival sulcus). Meanwhile,buccal palatal gingival sulcus infusion toxin10μl of (using the cotton ballstrictly separated from the wet to prevent endotoxin spillover), intervalinfusion once a week for a total infusion of three times. Group PD+AS andgroup AS were IP injected Vitamin D37×105IU/kg for two weeks, graduallyreplaced by the fat feed the standard diet, feeding four weeks. Four weeksafter the establishment of the rat model draw-off animal randomly in eachgroup, the maxilla and arteria carotis communis were histological investigatedwith H&E stainto confirm the periodontitis and atheroslerosis modelsuccessfully established. Twelve weeks after the rat model established, all ratswere bled to death by femoral arteries bleeding, and the serum was seperatedform whole blood for the detecion of ICAM-1, VEGF and E-selectin. Themaxillae and common carotid artery were immediately fixed in10%formaldesolution and changed to10%EDTA decalcification solution after24hrs. Thetissue sections were stained with H&E for histological investigation in8weeks. Results:1The model groups showed poor appetite and lost weight for the firstweek and were gradually got normal appetite back in four weeks, but the bodyweight were lower in model groups than normal group. By the end of twelveweeks, the body weight of normal group increased the most among the groups,while the periodontitis with atherosclerosis model group were the leastincreased in body weight.2After four weeks establishing the models, the histological investigationon H&E stained maxilla on random samples showed periodontal inflammationwere obvious in periodontitis group and periodontitis with atherosclerosisgroup. The inflammatory changes included lower alveolar crest, irregulargingival periodontal ligament fibers, epithelial proliferation and infusion to theroot side, the formation of shallow periodontal pocket, absorption lacuna andinflammatory cell infiltration. All the above indicated the successfulestablishment of periodontitis animanl model.3Common carotid artery sections from draw-off group normal, groupatherosclerosis and group periodontitis with atherosclerosis around four weeksshowed the arterial vescular tissue structure was integrated and withoutinflammation in normal group, whereas the other two groups with infiltrationof inflammatory cells, desthructive lining structure, hyperplasis of smoothmuscles cells and atherosclerotic plaques containing necrotic material andcalcified plaque. The results indicated the animal model of atherosclerosis wasestablished successfully.4Clinical examination results revealed that the periodontal tissue of thefirst maxillary molar teeth in all rats was normal without bleeding and otherinflammatory changes at the beginning of experiment. However, the teethcheck of the rats showed foof scraps, swollen gingival, shallow periodontalpocket, exposed fang and recessive gingival in group PD and group PD+ASon12weeks, while no obvious change in group AS and group normal. TheSulcus Bleeding Index(SBI) and Probing Depth(PD) of group PD and groupPD+AS were significantly higher than group normal (P<0.01); group PD and group PD+AS were higher than group AS(P<0.05); group PD+AS was morehigher than group(P<0.05).5Periodontal tissue morphology (H&E stain) on twelve weeks: groupNormal: junctional epithelium was attached to the joint of cementum andenamel, periodontal ligament fibers was arranged regularly and there is noabsorption of alvolar crest. Group PD: inflammatory cell infiltrated to gingivalepithelium, junctional epithelial proliferated or infused to the root side;periodontal pocket formatted and alvelolar crest was absorbed and lower.Group PD+AS: compare to other groups, much more inflammatory cellinfiltration abserved in gingival epithelium and more junctional epithelialproliferation and infusion to the root side deeply; deep periodontal pocketformatted and alveolar crest was much lower and more absorption withosteoclast absorption lacuna. For vascular tissue morphology, group normal:tissue structure of the arterial was integrated without inflammation; group PD:no obvious pathological changes; group AS: destructive lining structure,endometrial thickening and middle membrane thinning, plaque formation;group PD+AS: the pathological change was more severe than group AS,which revealed atherosclerotic plaques containing necrotic material andcalcified plaque with lots of infitration of inflammatory cells.6The level of serum ICAM-1, VEGF and E-selectin by enzyme linkedimmunoabsorbent assay (ELISA): ICAM-1and E-selectin of group modeswas significantlu higher than the group normal (P<0.01); group PD+AS wassignificantly higher than group AS and group PD (P<0.05); group AS wassigificantly higher than group PD (P<0.05). The level of VEGF was higher ingroup PD+AS and group AS than group normal (P<0.01), group AS wassigificantly higher than group PD (P<0.05); group PD+AS was higher thangroup AS, but no obvious (P>0.05); group PD was higher than group normal,but no obvious (P>0.05).Conclusion:1The periodontitis rat model was successfully established by maxillarybilateral ligation combined with endotoxic drip；The atherosclerosis model was successfully established by feeding Vitamin D3with high fat.2The periodontitis with atherosclerosis model was successfullyestablished.3The serum ICAM-1, VEGF and E-selectin increased in periodontitismodel, atherosclerosis model and combined model compare to the normal rat,while the combined model showed a higher increase than single models,which indicated periodontitis and atherosclerosis accelerated the effect oninflammation to each other, indicated the influence lie in between periodontitisand atherosclerosis.