Hemocompatibility improvement of perfusion-decellularized clinical-scale liver scaffold through heparin immobilization.
Hemocompatibility improvement of perfusion-decellularized clinical-scale liver scaffold through heparin immobilization.
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DOI：
10.1038/srep10756
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发表时间：
2015-06-01
影响因子：
4.6
通讯作者：
Bu H
中科院分区：
文献类型：
--
作者：
Bao J;Wu Q;Sun J;Zhou Y;Wang Y;Jiang X;Li L;Shi Y;Bu H
全肝灌注脱细胞化是一种制备临床可移植肝组织的有吸引力的支架制备技术。然而，支架的血液相容性差构成了主要障碍。本研究旨在通过肝素的固定化改善灌注脱细胞猪肝支架的血液相容性。肝素通过使用逐层自组装技术的静电结合（h-LBL支架）、经由多点连接的共价结合（h-MPA支架）或端点连接（h-EPA支架）固定在脱细胞肝脏支架（DLS）上。研究肝素化对细胞抗凝血能力和细胞相容性的影响。肝素含量和释放试验结果表明，EPA法固定肝素的效率高于其他两种方法。然后，凝血酶原时间（PT）、凝血酶时间（TT）、活化部分凝血活酶时间（APTT）、血小板粘附和人血小板因子4（PF 4，指示血小板活化）的系统体外研究证实肝素化支架，尤其是h-EPA对应物，表现出超低的血液组分活化和优异的血液相容性。此外，肝素治疗成功地防止了体内植入后血液灌注的DLS中的血栓形成。同时，肝素处理后，原代肝细胞和内皮细胞的活力也保持良好，这表明肝素处理具有改善的生物相容性，可以扩展到各种血液灌流全器官支架的制备。
Whole-liver perfusion-decellularization is an attractive scaffold–preparation technique for producing clinical transplantable liver tissue. However, the scaffold’s poor hemocompatibility poses a major obstacle. This study was intended to improve the hemocompatibility of perfusion-decellularized porcine liver scaffold via immobilization of heparin. Heparin was immobilized on decellularized liver scaffolds (DLSs) by electrostatic binding using a layer-by-layer self-assembly technique (/h-LBL scaffold), covalent binding via multi-point attachment (/h-MPA scaffold), or end-point attachment (/h-EPA scaffold). The effect of heparinization on anticoagulant ability and cytocompatibility were investigated. The result of heparin content and release tests revealed EPA technique performed higher efficiency of heparin immobilization than other two methods. Then, systematic in vitro investigation of prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT), platelet adhesion and human platelet factor 4 (PF4, indicates platelet activation) confirmed the heparinized scaffolds, especially the /h-EPA counterparts, exhibited ultralow blood component activations and excellent hemocompatibility. Furthermore, heparin treatments prevented thrombosis successfully in DLSs with blood perfusion after implanted in vivo. Meanwhile, after heparin processes, both primary hepatocyte and endothelial cell viability were also well-maintained, which indicated that heparin treatments with improved biocompatibility might extend to various hemoperfusable whole-organ scaffolds’ preparation.