MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, China; Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cell and Regenerative Medicine, Zhejiang University, Hangzhou 310058, China.
MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, China.
Acta Biomater. 2019 Sep 15;96:137-148. doi: 10.1016/j.actbio.2019.07.004. Epub 2019 Jul 5.
Capture of endothelial progenitor cells (EPCs) in situ has been considered as a promising strategy for the rapid endothelialization and long-term patency of artificial blood vessels and implant devices. In this study, a CD133 EPC capture surface was fabricated by grafting CD133 antibody (a more specific EPC surface marker than CD34) and Arg-Glu-Asp-Val (REDV) peptideon the methacrylate-grafted hyaluronic acid (MA-HA) and heparin-hybridized (MA-HA&Heparin) resisting layer. Vascular endothelial growth factor (VEGF) was further conjugated to the immobilized heparin. This engineered surface showed good hemocompatibility and significantly higher ability of capturing CD133 EPCs from human peripheral blood mononuclear cells (PBMCs) and obviously upregulated the expression of endothelial cell (EC) marker genes of EPCs such as VEGF receptor 2 (VEGFR2), CD31, VE-cadherin, and von Willebrand factor (vWF), facilitating the differentiation of EPCs into ECs. The dramatically enhanced EPC proliferation on this surface was dependent on the integrin-VEGFR synergistic signaling, as ERK1/2 phosphorylation was only significantly enhanced on the REDV and VEGF co-immobilized surface. This study highlights a new surface coating strategy for blood-contact materials based on the specific EPC capturing and rapid endothelialization. STATEMENT OF SIGNIFICANCE: Capture of endothelial progenitor cells (EPCs) in situ is a promising strategy for the rapid endothelialization and long-term patency of artificial blood vessels and scaffolds. More specific capture of EPCs by targeting CD133 rather than CD34 can better reduce the risk of inflammation and restenosis. On the other hand, an appropriate microenvironment for EPC proliferation is equally important for endothelialization, which is rarely considered by the existing EPC capture strategies. In this study, the capture ratio of EPCs was significantly increased by simultaneously grafting CD133 antibody and VEGF on a MA-HA and heparin-hybridized antifouling layer. Further, proliferation of EPCs after capture was significantly promoted by grafting VEGF and REDV peptide through the integrin-VEGFR synergistic signaling. This study highlights a new strategy for the surface coating of blood-contact materials based on specific EPC capture and rapid endothelialization.
捕获原位内皮祖细胞 (EPCs) 已被认为是实现人工血管和植入装置快速内皮化和长期通畅的有前途的策略。在这项研究中，通过将 CD133 抗体（比 CD34 更特异的 EPC 表面标志物）和 Arg-Glu-Asp-Val (REDV) 肽接枝到甲基丙烯酸酯接枝透明质酸 (MA-HA) 和肝素杂交（MA-HA&Heparin）抗污层上，制备了 CD133 EPC 捕获表面。进一步将血管内皮生长因子 (VEGF) 偶联到固定化肝素上。该工程表面表现出良好的血液相容性，能够显著提高从人外周血单核细胞 (PBMCs) 中捕获 CD133 EPC 的能力，并且明显上调 EPC 的内皮细胞 (EC) 标志物基因的表达，如 VEGF 受体 2 (VEGFR2)、CD31、VE-钙粘蛋白和血管性血友病因子 (vWF)，促进 EPC 向 EC 分化。EPC 在该表面上的显著增殖依赖于整合素-VEGFR 协同信号，因为 ERK1/2 磷酸化仅在 REDV 和 VEGF 共固定表面上显著增强。这项研究强调了一种基于特定 EPC 捕获和快速内皮化的新型血液接触材料表面涂覆策略。
原位捕获内皮祖细胞 (EPCs) 是实现人工血管和支架快速内皮化和长期通畅的有前途的策略。通过靶向 CD133 而不是 CD34 来更特异的捕获 EPCs 可以更好地降低炎症和再狭窄的风险。另一方面，为 EPC 增殖提供适当的微环境对于内皮化同样重要，但现有的 EPC 捕获策略很少考虑这一点。在这项研究中，通过同时将 CD133 抗体和 VEGF 接枝到 MA-HA 和肝素杂交的抗污层上，显著提高了 EPC 的捕获比例。此外，通过整合素-VEGFR 协同信号，进一步将 VEGF 和 REDV 肽接枝到表面上，显著促进了捕获后 EPC 的增殖。这项研究强调了一种基于特定 EPC 捕获和快速内皮化的新型血液接触材料表面涂覆策略。
