肝素／二羟基铁纳米修饰对异种血管生物相容性的影响  

Effect of heparin/dihydroxy-iron complex muitilayeres nanomodification on biocompatibilities of decellular xenograft

作　　者：陶运明[1] 胡铁辉[1] 吴忠仕[1] 唐浩[1] 胡野荣[1] 谭琦[1] 

机构地区：[1]中南大学湘雅二医院心外科,长沙410011

出　　处：《中华胸心血管外科杂志》2012年第2期94-98,共5页Chinese Journal of Thoracic and Cardiovascular Surgery

基　　金：基金项目：国家自然科学基金（81071275）、国家高技术研究发展基金（2007AA071900）

摘　　要：目的探讨肝素／二羟基铁多层复合物（HDcms）表面纳米修饰对去细胞异种血管生物相容性的影响。方法采用层层自组装技术将二羟基铁和肝素交替固定在去细胞牛颈静脉（DC-BJV）表面，构建一种新型的抗凝表面，并检测其表面微结构和生物相容性。结果甲苯胺蓝比色法显示，每组装一次约有（808±86）μg／cm^2肝素固定在DC—BJV表面。扫描电镜（SEM）和洗脱试验显示HDcms均匀稳定的包裹在胶原纤维表面，形成纳米膜并持续缓慢地释放肝素。抗凝血活性检测显示，实验组凝血酶原时间（PT）和部分活化凝血酶原时间（APTT）明显高于正常值。血小板黏附实验显示实验组和对照组每10000μm^2血小板计数分别为（8±4）个和（48±16）个。皮下包埋4周和8周时实验组钙离子含量分别为（8．5±1．9）μg/mg和（21．5±6．8）μg／mg，对照组钙离子含量分别为（26．6±3．7）μg/mg和（112．6±16．9）μg/mg。结论HDcms能够牢固地结合在去细胞异种血管表面形成纳米厚度的抗凝表面，并能够提高其生物相容性。Objective Xenografts have poor biocompatibilities, the aim of this study was to improve the biocompatibilities of decellular xenografts via heparin/dihydroxy-iron complex muhilayeres （HDCMs） nanomodification. Methods A novel thrombo-resistant surface for decellular xenograft had been developed by alternating linkage of dihydroxy-iron and heparin to de- cellular bovine jugular vein （DC-BJV）, and surface characterization and biocompatibility of HDCMs nanomodified BJV （HDCMs-BJV） were detected. Results Toluidine blue colorimetric method showed the amount of linked heparin was about （808 ± 86） μg/cm2 per assembly-cycle. SEM images proved HDCMs were uniformly linked to and formed nanoscale films around the fibrils of DC-BJV. Washing test proved HDCMs were firmly hnked to BJV and sustainedly released heparin for a long time. Tensile test showed that biomechanical stability was increased. Antithrombogenicity test showed that the activated partial thrombin time （APTT） and prothrombin time （PT） of all trial groups were above the normal reference ranges. Platelet adhesion test evaluated mean platelet count per 10 000 μm^2 area was 8 ±4 for HDCMs-BJV vs. 48 - 16 for DC-BJV. Endothelial ceils （ECs） proliferation test showed the number and activity of ECs on luminal surface of HDCMs-BJV were very similar to DC-BJV at 7-day incubation. Calcium content assay evaluated mean calcium content was （ 8.5 ± 1.9 ） μg/mg dry weight for HDCMs-BJV vs. （26.6 ± 3.7） μg/mg dry weight for DC-BJV at 4 weeks and （21.5 ± 6.8） μg/mg dry weight for HDCMs- BJV vs. （ 112.6 ± 16.9 ） μg/mg dry weight for DC-BJVs at 8 weeks, respectively. Conclusion These results demonstrate HDCMs were firmly linked to BJV and formed nanoscale thrombo-resistant films, and HDCMs nanomodification improves biocompatibilities of decellular xenograft.

关 键 词：人工血管 肝素 移植 异种 纳米医学 组织相溶性 牛颈静 

分 类 号：R318.08[医药卫生—生物医学工程]