摘要
目的:采用肝素铁复合物纳米修饰去细胞异种血管以提高其血液相容性。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,构建一种新型的多层肝素铁复合物(HICMs)抗凝表面,并通过扫描电镜(SEM)检测其表面微结构、拉力器检测其生物力学稳定性、抗凝血活性实验和血小板黏附实验检测其血液相容性。结果:每组装一次,约有(808±86)μg/cm2肝素固定在BJV表面;SEM图片显示HICMs是均匀地包裹在胶原纤维表面并形成纳米膜;组织切片甲苯胺蓝染色提示肝素主要结合在DC-BJV的表层;拉力试验提示实验组的生物力学稳定性有显著提高;经过1 d及1,2,3,4,6,8周的洗脱,肝素的释放量分别为(281±43),(422±60),(729±81),(1053±116),(1317±157),(1618±187),(1945±228)μg/cm2;抗凝血活性检测显示实验组凝血酶原时间(PT)和活化部分凝血酶原时间(APTT)高于正常值范围;血小板黏附试验显示每10000μm2HICMs层层自组装修饰的牛颈静脉(LBL-BJV)和DC-BJV的血小板计数分别为8±4和48±16。结论:HICMs牢固地结合在DC-BJV表层,形成纳米厚度的抗凝表面,并长时间地缓慢释放肝素。HICMs纳米修饰能够提高去细胞异种血管的血液相容性。
Objective： To improve the hemocompatibility of decellular vascular matrix via heparin-iron complex multilayers （HICMs） nanomodification. Methods： A novel thrombo-resistant surface for decellular xenograft was developed by alternating linkage of dihydroxy-iron and heparin to decellular bovine jugular vein （DC-BJV）, and its surface characterization, biomechanical stability and hemocompatibility were detected by scanning electron microscop}5 tensile test and hemocompatibility evaluation, respectively. Results： A toluidine blue colorimetric method indicated the amount of linked heparin was about （808±86）μg/cmz per assembly-cycle. Scanning electron microscopic （SEM） images proved that HICMs were uniformly linked to and formed nanoscale films around the fibrils of DC-BJV. Toluidine blue staining histologic images showed that HICMs were linked mainly to DC-BJV surfaces. Washing test showed that the release of heparin was （281±43）, （422 ± 60）, （729±81）, （1053±116）, （1317＋157）, （1618＋187） and （1945 ＋ 268 ） μg/cm2 at 1 day 1, 2, 3, 4, 6 and 8 week washing, respectively. Tensile tests showed an increased biomechanical stability. Hemocompatibility evaluations showed that PT and APTT of all the trial groups were above the normal reference ranges and that mean platelet count per 10000μm2 area was 8±4 for HICMs layer-by-layer modified BJV （LBL-BJV） vs 48＋16 for DC-BJV. Conclusion： HICMs are firmly linked to DC-BJV, and can form nanoscale thrombo-resistant films, which yield a sustained release of heparin. HICMs nanomodification improves the hemocompatibility of decellular xenograft.
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2012年第3期260-266,共7页
Journal of Central South University ：Medical Science
基金
国家高新技术研究发展计划项目(2007AA071900)
国家自然科学基金(81071275)~~
关键词
肝素
自组装
纳米修饰
血液相容性
牛颈静脉
heparin
self-assembly
nanomodification
hemocompatibility
bovine jugular vein
