| Part 1 Blood Compatibility of a Valved Bovine Pericardium Patch on Animal Experimental StudyObjective By comparing a valved bovine pericardium patch(VBP)after modification complex cross-linked treatment with a valved autologous pericardium patch(VAP)in the results of in vivo and in vitro blood compatibility,to show whether VBP has good blood compatibility;Through the observation of existent state and serious complication of animal models,to show the feasibility of surgical operation by using VBP.Methods(1)To establish the animal models,15 male sheep were randomly divided into experimental group(group VBP,n=9)and control group(group VAP,n=6).To cut off a pulmonary valve ring with part of pulmonary artery and myocardium of right ventricular outflow tract,then reconstructed with the VBP(experimental group)and VAP(control group)in general anesthesia and cardiopulmonary bypass;(2)The in vivo blood compatibility testing,to detect and compare the results of f Hb and routine blood(including RBC,HGB,PLT,WBC,NEUT,NEUT% index)of experimental group and control group;(3)The in vitro blood compatibility testing,the venous blood in 15 experimental animal will contact with the VBP(experimental group,n=15)and VAP(control group,n=15)in vitro,to detect and compare the results of hemolysis test,complement activation test(C3a,C5 a,TCC,Bb,C4d),plasma soluble P-selectin test and in vitro dynamic cruor time test.Results(1)All the experimental animals were survived and without serious complications;(2)In vivo blood compatibility testing,the results of f Hb were no significant difference in pre-operation,post-operation 1day,3day,7day between VBP group and VAP group(p > 0.05),and the content of f Hb in post-operation 1day was obviously increased compared with(p < 0.05);The results of blood routine test(including RBC,HGB,PLT,WBC,NEUT,NEUT%)were no significant difference in the corresponding time points between VBP group and VAP group(p > 0.05),and all the results of RBC and HGB in post-operation 3day was lower than pre-operation(p < 0.001),all the results of WBC,NEUT and NEUT% in post-operation 3day was higher than pre-operation(p < 0.05);(3)In vitro blood compatibility testing,there were no significant difference between VBP group(1.28±0.14%)and VAP group(1.26±0.06)(p > 0.05)in the results of hemolysis test,and all the results were less than 5%;there were no significant difference between VBP group and VAP group(p > 0.05)in the results of complement activation test(C3a,C5 a,TCC,Bb,C4d);there were no significant difference between VBP group and VAP group(p > 0.05)in the results of plasma soluble P-selectin test;the OD value were no significant difference between VBP group and VAP group(p > 0.05)in each time point of in vitro dynamic cruor time test.Conclusions The VBP has good blood compatibility;The operation has the feasibility by using VBP.Part 2 Tissue Compatibility and Anti-calcification of a Valved Bovine Pericardium Patch on Animal Experimental StudyObjective To confirm whether VBP has the operation feasibility through comparing a valved bovine pericardium patch(VBP)after modification complex cross-linked treatment with a valved autologous pericardium patch(VAP)in the operation of surgery process;To confirm the valve function through measuring the valvular regurgitation area and pulmonary arterial pressure difference by UCG;To show the tissue and organs compatibility by pathological observation;To show the anti-calcification by determination of calcium content.Methods(1)To establish the animal models,15 male sheep were randomly divided into experimental group(group VBP,n=9)and control group(group VAP,n=6).To cut off a pulmonary valve ring with part of pulmonary artery and myocardium of right ventricular outflow tract,then reconstructed with the VBP(experimental group)and VAP(control group)in general anesthesia and cardiopulmonary bypass,and recording the time of operation,cardiopulmonary bypass,aortic occlusion;(2)To contrast the pulmonary valve function between VBP group and VAP group by measuring the pulmonary valvular regurgitation area and pulmonary arterial pressure difference by UCG;(3)The VBP,VAP and related organs(including heart,liver,spleen,lung,kidney and brain)were removed to do pathological observation(through naked eyes,light microscope,scanning electron microscope)after 6 months;(4)To contrast the anti-calcification between VBP group and VAP group by measuring the calcium content of specimens.Results(1)All the experimental animals were survived and without serious complications,the time of operation in VBP group(126±13.2min)was less than the VAP group(151±15.6min)(p<0.05),the time of cardiopulmonary bypass in VBP group(73±8.6min)was less than the VAP group(98±11.5min)(p<0.05);the time of aortic occlusion in VBP group(33±6.8min)was less than the VAP group(58±7.4min)(p < 0.05);(2)The results of pulmonary valvular regurgitation area were no significant difference between VBP group and VAP group in the operation and in post-operation 1month time points(p > 0.05),the pulmonary valvular regurgitation area of VBP group were smaller than VAP group in post-operation 3 month and 6month time points(p < 0.05),The valvular regurgitation area of VBP group were no significant difference between in the operation(1.76±0.53cm2)and in post-operation 6 month(1.87±0.32cm2)time points(P > 0.05),the valvular regurgitation area of VAP group in post-operation 6 month(2.54±0.35cm2)were bigger than in the operation(1.81±0.48cm2)time points(P < 0.05);(3)The pulmonary arterial pressure by UCG were no significant difference between VBP group and VAP group in the operation,post-operation 1 month,3 month and 6 month time points(p > 0.05);(4)In pathological observation through naked eyes,the patch valve of VBP and VAP were soft,the valves function were normal and had no thrombus.The color of patch inwall was similar with the pulmonary artery inwall,the anastomotic was smooth,and has no hyperplasia and necrosis.Other organs were normal.Through light microscope,there were dense fibrous connective tissue,fibroblast proliferation,focal lymphocyte,plasma cell infiltration,surgical suture,no thrombus,endothelial cells in VBP and VAP.Other organs were normal.Through scanning electron microscope,the fiber structure was compact and compact and less adherent cells in pre-operation VBP;The fiber structure was also compact and compact,more endothelial cells on the fiber in post-operation VBP;(5)The results of calcium content were no significant difference between VBP group(1834±53?g/g)and VAP group(1795±61?g/g)(p > 0.05)in post-operation 6 month.Conclusions The VBP has the feasibility of Surgical operation because it had advantages of simple operation and less time-consuming than VAP;The VBP has good function of anti-reflux;The VBP had good Tissue Compatibility and Anti-calcification. |
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