Physiological pulsatile flow culture conditions to generate functional endothelium on a sulfated silk fibroin nanofibrous scaffold
Physiological pulsatile flow culture conditions to generate functional endothelium on a sulfated silk fibroin nanofibrous scaffold
复制标题
生理脉动流培养条件在硫酸化丝素蛋白纳米纤维支架上产生功能性内皮
DOI：
10.1016/j.biomaterials.2014.02.050
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发表时间：
2014-06-01
期刊：
影响因子：
14
通讯作者：
Fan, Yubo
中科院分区：
文献类型：
--
作者：
Gong, Xianghui;Liu, Haifeng;Fan, Yubo
许多研究表明，内皮细胞种植的小直径血管移植物的体外剪切应力调节可以改善细胞保留和功能。然而，在血管组织工程和血液动力学研究中常用的层流和脉动流条件与实际的生理脉动流有很大的不同，实际的生理脉动流本质上是脉动的，具有典型的压力和流量波形。实际的生理脉动流导致的壁面切应力的时间和空间的变化可能导致不同的表型和功能的EC。因此，本研究的目的是找出最佳的体外动态培养条件，以产生功能性内皮细胞的硫酸化丝素蛋白纳米纤维支架的小直径血管组织工程。将大鼠主动脉内皮细胞（RAEC）接种在硫酸化丝素蛋白纳米纤维支架上，并在三种不同的流动调节模式下培养，稳定层流（SLF）、正弦流（SF）或生理脉动流（PPF），代表体内长达24小时的典型股骨远端脉搏波。细胞形态，细胞骨架排列，纤连蛋白组装，细胞凋亡，并保留在支架上进行了研究，并比较了三种不同模式的流动调节。结果表明，内皮细胞对不同的暴露时间和不同的流动模式有不同的反应。实际的PPF调理表现出优异的EC保留硫酸化丝素蛋白支架相比，SLF和SF，除了对齐的细胞在流体流动的方向，形成更密集和规则的F-肌动蛋白微丝束在同一方向，组装更厚和高度交联的纤连蛋白，并显着抑制细胞凋亡。因此，实际的PPF调节可能对在硫酸化丝素蛋白纳米纤维支架上产生功能性内皮细胞做出重要贡献，从而产生抗血栓的管腔表面。(C)2014爱思唯尔有限公司版权所有。
Many studies have demonstrated that in vitro shear stress conditioning of endothelial cell-seeded small-diameter vascular grafts can improve cell retention and function. However, the laminar flow and pulsatile flow conditions which are commonly used in vascular tissue engineering and hemodynamic studies are quite different from the actual physiological pulsatile flow which is pulsatile in nature with typical pressure and flow waveforms. The actual physiological pulsatile flow leading to temporal and spatial variations of the wall shear stress may result in different phenotypes and functions of ECs. Thus, the aim of this study is to find out the best in vitro dynamic culture conditions to generate functional endothelium on sulfated silk fibroin nanofibrous scaffolds for small-diameter vascular tissue engineering. Rat aortic endothelial cells (RAECs) were seeded on sulfated silk fibroin nanofibrous scaffolds and cultured under three different patterns of flow conditioning, e.g., steady laminar flow (SLF), sinusoidal flow (SF), or physiological pulsatile flow (PPF) representative of a typical femoral distal pulse wave in vivo for up to 24 h. Cell morphology, cytoskeleton alignment, fibronectin assembly, apoptosis, and retention on the scaffolds were investigated and were compared between three different patterns of flow conditioning. The results showed that ECs responded differentially to different exposure time and different flow patterns. The actual PPF conditioning demonstrated excellent EC retention on sulfated silk fibroin scaffolds in comparison with SLF and SF, in addition to the alignment of cells in the direction of fluid flow, the formation of denser and regular F-actin microfilament bundles in the same direction, the assembly of thicker and highly crosslinked fibronectin, and the significant inhibition of cell apoptosis. Therefore, the actual PPF conditioning might contribute importantly to the generation of functional endothelium on a sulfated silk fibroin nanofibrous scaffold and thereby yield a thromboresistant luminal surface. (C) 2014 Elsevier Ltd. All rights reserved.