Yang Dongchuan, Qiu Juhui, Xu Ning, Zhao Yinping, Li Tianhan, Ma Qinfeng, Huang Junli, Wang Guixue
Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400030, P. R. China.
J Mater Chem B. 2018 Jun 28;6(24):4151-4163. doi: 10.1039/c8tb00526e. Epub 2018 Jun 12.
Endothelium is the only known completely non-thrombogenic material. In the present study, a strategy to mimic the adhesive interactions of endothelial cells (ECs) to alter the vascular microenvironment was established and applied to directing the behaviour of cells. To facilitate the regeneration of a functional endothelium in vascular lesions, we designed a recombinant mussel foot protein (Mfp-5) fused with the VE-cadherin extracellular domain EC1-2, termed VE-M. Surface coating analysis showed that recombinant VE-M successfully formed a coating on substrate materials with uniform nanorods, low roughness, and sufficient hydrophilicity. We then evaluated the effects of VE-M on the adhesion of ECs and the capture of endothelial progenitor cells (EPCs). The result demonstrated that VE-M efficiently promoted the adhesion of ECs and EPCs. The number of ECs and EPCs on VE-M was 5.5- and 1.8-fold higher, respectively, than that on bare 316L SS under static conditions, whereas there was no significant difference in the number of captured smooth muscle cells (SMCs) between VE-M and other substrates. In addition, the number of EPCs captured by VE-M was approximately four times higher than that captured by 316L SS under dynamic conditions. In particular, the result of the neutralization test indicated that VE-M specifically triggered ECs' adhesion via the interaction of VE-cadherin EC1-2. Further investigation showed that VE-M significantly increased the levels of endogenous VE-cadherin in HUVECs as well as the endothelial eNOS content, with little or no endothelial inflammation. Our results showed that VE-M could be a promising biomimetic modification for accelerating endothelialization and vascularization in tissue engineering.
内皮是唯一已知的完全抗血栓形成的材料。在本研究中，建立了一种模拟内皮细胞（ECs）黏附相互作用以改变血管微环境的策略，并将其应用于指导细胞行为。为促进血管损伤处功能性内皮的再生，我们设计了一种与VE-钙黏蛋白细胞外结构域EC1-2融合的重组贻贝足蛋白（Mfp-5），称为VE-M。表面涂层分析表明，重组VE-M成功地在基底材料上形成了具有均匀纳米棒、低粗糙度和足够亲水性的涂层。然后，我们评估了VE-M对ECs黏附以及内皮祖细胞（EPCs）捕获的影响。结果表明，VE-M能有效促进ECs和EPCs的黏附。在静态条件下，VE-M上的ECs和EPCs数量分别比裸露的316L不锈钢上的高5.5倍和1.8倍，而VE-M与其他底物捕获的平滑肌细胞（SMCs）数量没有显著差异。此外，在动态条件下，VE-M捕获的EPCs数量比316L不锈钢捕获的高约四倍。特别是，中和试验结果表明，VE-M通过VE-钙黏蛋白EC1-2的相互作用特异性触发ECs的黏附。进一步研究表明，VE-M显著增加了人脐静脉内皮细胞（HUVECs）中内源性VE-钙黏蛋白的水平以及内皮型一氧化氮合酶（eNOS）的含量，且几乎没有内皮炎症。我们的结果表明，VE-M可能是一种有前途的仿生修饰，可加速组织工程中的内皮化和血管化。
