血小板影响内皮细胞在心血管植入物表面生长增殖的实验研究

目的探索血小板对内皮细胞在心血管植入物表面生长增殖的影响。方法利用生物材料钛（Ti）和低温各向同性热解碳（LTIC）分别制成试片作为A、B组,钛片表面涂层肝素（Hep）-血管内皮细胞生长因子（VEGF）-内皮祖细胞抗体（CD34）-胶原（Col）作为C组。采用乳酸脱氢酶（LDH）试验法测定试片表面黏附血小板的量,GMP-140试验测定试片表面血小板被激活的量;试片表面黏附的血小板作扫描电镜（SEM）观察,记录其形貌。取3只动物犬左右股动脉分别置入A组及C组试片,另3只犬左右股动脉内分别置入B组及C组试片,3个月后取出试片,SEM观察其表面内皮细胞生长情况。结果 C组黏附血小板的量和激活血小板的量均少于A组与B组（均P〈0.05）,A、B组黏附的血小板形貌有明显变形并有伪足。3个月后动物体内取出的试片SEM观察,A、B组表面未见到内皮细胞生长,C组表面有排列整齐成熟的内皮细胞。结论内皮细胞必须在无血凝的条件下生长,减少血小板的黏附与激活以及构建具有抗凝作用的细胞外基质是内皮细胞在心血管植入物表面生长增殖的关键。

Experimental research about platelet influencing the endothelial cell which was growing and proliferating on the surface of cardiovascular implants

Objective To study the influence of platelet（PL） on endothelial cell（EC） growing and proliferating on the surface of cardiovascular implants.Methods Biological material Ti film as the vascular stent and low temperature isotropic pyrolytic carbon（LTIC） film as the artificial heart value material in clinic were prepared as group A and B,group C was Ti film coated with collagen-heparin-vascular endothelial growth factor（VEGF）-endothelial progenitor cell membrane protein anti-cluster of differentiation 34（CD34）.The amount of PL adhesion on all films surface of each group was counted by using LDH test.The amount of PL activation was tested by euzymelinked immunosorbent assay（ELISA）.Adhesion PL was observed by scanning electron microscope（SEM）.The Ti films were implanted into the right femoral artery and the films of group C（coated） were implanted into the left femoral artery in 3 dogs.The LTIC films and the films of third group（coated） were implanted into the other 3 dogs.Then the samples were taken out 3 months later and observed by SEM.Result The amount of PL adhension and PL activation on all films surface were less in group C than that in group A and B（P0.05）.There are ECs growth and differentiation on the surface of coated films in group C,and no ECs growth on the surface of Ti,LTIC films.Conclusion The growth of EC needs a non coagulating environment.Reducing adhesion and activation of PL and constructing extracellular matrix which has ability of anticoagulation are necessary for the growth and proliferation of EC.