Functional Modification of Fibrous PCL Scaffolds with Fusion Protein VEGF-HGFI Enhanced Cellularization and Vascularization
Functional Modification of Fibrous PCL Scaffolds with Fusion Protein VEGF-HGFI Enhanced Cellularization and Vascularization
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用融合蛋白 VEGF-HGFI 增强细胞化和血管化对纤维 PCL 支架进行功能修饰
DOI：
10.1002/adhm.201600226
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发表时间：
2016
影响因子：
10
通讯作者：
Kong Deling
中科院分区：
文献类型：
--
作者：
Zhao Liqiang;Ma Shaoyang;Pan Yiwa;Zhang Qiuying;Wang Kai;Song Dongmin;Wang Xiangxiang;Feng Guowei;Liu Ruming;Xu Haijin;Zhang Jun;Qiao Mingqiang;Kong Deling
常用的聚己内酯（PCL）支架缺乏有效的血管化，阻碍了其在组织工程中的应用。疏水蛋白HGFI是一种两亲性蛋白质，可以在PCL支架表面形成自组装层并改变其润湿性。本研究利用毕赤酵母表达系统制备了HGFI与血管内皮生长因子（VEGF）的融合蛋白。十二烷基磺酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质印迹证实成功分离和纯化了VEGF-HGFI。透射电子显微镜和水接触角测量表明，VEGF-HGFI可以在静电纺丝PCL纤维上形成约25 nm厚的自组装层，并增加其亲水性。VEGF-HGFI修饰能有效增强人脐静脉内皮细胞的粘附、迁移和增殖。近红外荧光成像显示，VEGF-HGFI修饰的PCL支架在体外可存在至少21 d，在体内可存在至少14 d。生物发光成像显示，VEGF-HGFI可有效激活血管内皮生长因子受体2受体。小鼠和大鼠皮下植入表明，VEGF-HGFI修饰的PCL支架的细胞化和血管化显着改善。这些结果表明，VEGF-HGFI是一种有用的分子，可用于支架的功能修饰，以增强组织工程中的细胞化和血管化。
The lack of efficient vascularization within frequently used poly(ε‐caprolactone) (PCL) scaffolds has hindered their application in tissue engineering. Hydrophobin HGFI, an amphiphilic protein, can form a self‐assembly layer on the surface of PCL scaffolds and convert their wettability. In this study, a fusion protein consisting of HGFI and vascular endothelial growth factor (VEGF) is prepared byPichia pastorisexpression system. Sodium dodecyl sulface‐polyacrylamide gel electrophoresis (SDS‐PAGE) and western blotting confirm that the VEGF‐HGFI is successfully isolated and purified. Transmission electron microscope and water contact angle measurement demonstrate that VEGF‐HGFI can form a self‐assembly layer with about 25 nm in thickness on electrospun PCL fibers and increase their hydrophilicity. VEGF‐HGFI modification can effectively enhance the adhesion, migration, and proliferation of human umbilical vein endothelial cells. Near‐infrared fluorescence imaging shows that the VEGF‐HGFI modification on PCL scaffolds can exist at least 21 d in vitro and at least 14 d in vivo. Bioluminescence imaging shows that VEGF‐HGFI can effectively activate vascular endothelial growth factor receptor 2 receptors. Subcutaneous implantation in mice and rats reveal that cellularization and vascularization are significantly improved in VEGF‐HGFI modified PCL scaffolds. These results suggest that VEGF‐HGFI is a useful molecule for functional modification of scaffolds to enhance cellularization and vascularization in tissue engineering.