摘要
背景:冠状动脉内置入药物涂层支架显著解决了支架内再狭窄,但是,支架内晚期及超晚期血栓带来新的难题加快支架内皮化,预防血栓形成成为研究的关键。目的:观察内皮细胞在经刻蚀并构建仿生的生物活性表面,整合细胞外基质成分的金属钛表面的生长情况。设计、时间及地点:对比观察,于2007-08/2009-01在西南交通大学先进材料与技术教育部重点实验室完成。材料:成年杂交犬6只,体质量15～20kg,纯钛(99.997%)为西安宝鸡有色金属有限公司产品,切割成5mm×5mm×2mm圆片试样。方法:在已刻蚀的钛表面通过自组装技术构建胶原-肝素-血管内皮生长因子-内皮祖细胞膜蛋白抗体为组分的多层膜作为涂层组;未涂层的钛为未涂层组。成年杂交犬6只,涂层组样品置入右侧股动脉,未涂层组置入左侧股动脉内,获得2h,15d,3个月的样品。主要观察指标:乳酸脱氢酶实验测定黏附血小板的数量;酶联免疫法测定GMP-140以检测黏附血小板激活程度;扫描电镜观察黏附血小板的形貌;采用SysmexCA-600凝血仪测定活化部分凝血活酶时间及凝血酶原时间。扫描电镜观察植入物在不同时间点,内皮细胞在其表面的生长;免疫荧光法观察置入2h的试片表面分化成熟的内皮细胞,其血管内皮生长因子受体2的表达。结果:①乳酸脱氢酶实验显示涂层样片的乳酸脱氢酶值比未涂层显著减少,说明涂层钛可以减少血小板的黏附(P<0.05)。②黏附血小板激活程度的定量分析结果显示在涂层上黏附血小板P-选择素的表达显著小于未涂层上的表达(P<0.05)。③扫描电镜可见涂层钛表面黏附血小板形貌的变形程度小于未涂层钛。④涂层的活化部分凝血活酶时间和凝血酶原时间分别是(134.2±13.3),(34.2±5.7)s;未涂层的活化部分凝血活酶时间和凝血酶原时间分别是(44.9±2.3),(17.3±2.4)s,可见涂层的凝血时间显著延长(P<0.05)。⑤扫描
BACKGROUND： Coronary artery drug-eluting stent implantation significantly resolves the in-stent restenosis. However, the late and very late in-stent thrombosis makes new problems. Accelerating stent endothelialization to prevent thrombosis is the key in related study. OBJECTIVE： To observe the growth and propagation of vascular endothelial cells （ECs） on the surface of intravascular stent material. DESIGN, TIME AND SETTING： Comparative observation was performed at the Key Laboratory of Advancad Materials Technology, Ministry of Education, Southwest Jiaotong University from August 2007 to January 2009. MATERIALS： Atotal of 6 adult hybrid dogs, weighing 15-20 kg, were used. Pure titanium （99.97%） was purchased from Xi＇an Baoji Non-ferrous Metals, and cut into round samples, 5min×5mm×2mm. METHODS： The multilayer of collagen/heparin/vascular growth factor （VEGF）-endothelial progenitor cell membrane protein anti-cluster of differentiation 34（CD34） was constructed by a layer-by-layer self-assembly （LbL）on the surface of＇litanium testing slice as coated group. Uncoated group was the bare Titanium testing slice. In 6 adult hybrid dogs, the coated group samples were inserted into the dght femoral artery, the uncoated group samples were inserted into the left femoral artery. The samples of 2 hours, 15 days and 3 months were harvested. MAIN OUTCOME MEASURES： Lactate dehydrogenase（LDH） experiment determined the number of platelet adhesion. Enzyme-linked immunosorbent assay was used to detect the degree of platelet-activating on adhesion. The morphology of adhesive platelet was tested by scanning electron microscope （SEM）. Activated partial thromboplastin time （aPTT） and Prothrombin time （PT） tests were measured on Sysmex CA-600 Blood Coagulation Analyzer. SEM evaluated the growth on the surface of implant testing titanium slice at different time points. The expression of vascular endothelial growth factor receptor-2 （VEGF-R2） of maturation EC on the surface of testing
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第43期8425-8431,共7页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
“973”项目(国家重点基础研究发展计划《组织诱导性生物医用材料的基础研究》项目下属子项目“诱导抗凝血生物化表面的设计和构建”)(2005CB623904)
国家自然科学基金资助项目(20603027)
博士后基金资助项目(20060401019)~~
