Vascularization of LBL structured nanofibrous matrices with endothelial cells for tissue regeneration
Vascularization of LBL structured nanofibrous matrices with endothelial cells for tissue regeneration
复制标题
用于组织再生的内皮细胞 LBL 结构纳米纤维基质的血管化
DOI：
10.1039/c6ra26931a
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发表时间：
2017-01-01
期刊：
影响因子：
3.9
通讯作者：
Li, Xueyong
中科院分区：
文献类型：
--
作者：
Cui, Lei;Li, Jing;Li, Xueyong
为了在组织工程中设计用于重建的功能性血管结构，我们在这项研究中评估了通过静电纺丝和LBL技术逐层（LBL）各向同性和各向异性结构的聚（ε-己内酯）（PCL）/纤维素基纳米纤维的可行性。使用场发射扫描电子显微镜（FE-SEM）和原子力显微镜（AFM）分析两种纤维的形态。对齐的纳米纤维支架表面是nonthrombogenic使用血小板粘附试验进行评估，和抗血栓形成的改性纳米纤维垫大大增加涂层双层。将人脐静脉内皮细胞（HUVECs）接种于LBL构建的血管网上，通过场发射扫描电镜（FE-SEM）、四甲基偶氮唑蓝（MTT）比色法、细胞追踪法和细胞迁移实验检测HUVECs的粘附、增殖和迁移能力。此外，通过免疫荧光染色研究了HUVECs在具有各向同性或各向异性纤维组织的LBL结构纳米纤维基质上的表型表达。我们的数据发现，对齐的纳米纤维可以引导形态发生和调节细胞骨架组织的HUVECs，并通过促进表型相关蛋白的表达和毛细血管样管的形成相比，随机取向的纳米纤维，进一步促进体外预血管化。此外，体内植入对齐的复合支架与血管内皮细胞接种证明，促进宿主血管渗透到支架深处，并与体外预制的血管结构与增加涂层双层。总之，这些发现支持了我们的观点，即对齐的纳米纤维支架和预血管化的组合因此可以通过促进快速血管化来作为开发可植入功能性血管移植物的有前途的策略。
To engineer functional vascular structures for reconstruction in tissue engineering, we evaluated the feasibility of layer-by-layer (LBL) isotropic and anisotropic structured poly(e-caprolactone) (PCL)/cellulose based nanofibers via electrospinning and LBL techniques in this study. The morphology of both fibers was analyzed using field emission scanning electron microscopy (FE-SEM) and atomic force microscopy (AFM). The aligned nanofibrous scaffold surface was nonthrombogenic as assessed using a platelet adhesion test, and the antithrombogenicity of modified nanofibrous mats was increased greatly with increased coating bilayers. Besides, human umbilical vein endothelial cells (HUVECs) were then seeded onto the LBL structured nanofiber meshes and analyzed for cell adhesion, proliferation and migration by FE-SEM, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), cell tracking and cell migration assay. Moreover, the phenotypic expressions of HUVECs on LBL structured nanofibrous matrices with either isotropic or anisotropic fiber organizations were studied by immunofluorescent staining. Our data found that aligned nanofibers could guide morphogenesis and regulate cytoskeleton organization of HUVECs, and further promote in vitro prevascularization by facilitating phenotype-related protein expression and capillary-like tube formation as compared to randomly oriented nanofibers. Furthermore, the implantation in vivo of aligned composite scaffolds seeded with VECs demonstrated that the promoted host vessel infiltrated deep into the scaffolds and integrated with in vitro prefabricated vascular structures with increasing coating bilayers. Together, these findings supported our notion that the combination of aligned nanofibrous scaffolds and prevascularization could therefore serve as a promising strategy for the development of implantable functional vascular grafts by promoting rapid vascularization.