Zisch Andreas H, Zeisberger Steffen M, Ehrbar Martin, Djonov Valentin, Weber Cornelia C, Ziemiecki Andrew, Pasquale Elena B, Hubbell Jeffrey A
Department of Materials Science, Institute for Biomedical Engineering, ETH Zurich, University of Zurich, Moussonstr. 18, 8044 Zurich, Switzerland.
Biomaterials. 2004 Jul;25(16):3245-57. doi: 10.1016/j.biomaterials.2003.10.015.
With the rapid increase in approaches to pro- or anti-angiogenic therapy, new and effective methodologies for administration of cell-bound growth factors will be required. We sought to develop the natural hydrogel matrix fibrin as platform for extensive interactions and continuous signaling by the vascular morphogen ephrin-B2 that normally resides in the plasma membrane and requires multivalent presentation for ligation and activation of Eph receptors on apposing endothelial cell surfaces. Using fibrin and protein engineering technology to induce multivalent ligand presentation, a recombinant mutant ephrin-B2 receptor binding domain was covalently coupled to fibrin networks at variably high densities. The ability of fibrin-bound ephrin-B2 to act as ligand for endothelial cells was preserved, as demonstrated by a concomitant, dose-dependent increase of endothelial cell binding to engineered ephrin-B2-fibrin substrates in vitro. The therapeutic relevance of ephrin-B2-fibrin implant matrices was demonstrated by a local angiogenic response in the chick embryo chorioallontoic membrane evoked by the local and prolonged presentation of matrix-bound ephrin-B2 to tissue adjacing the implant. This new knowledge on biomimetic fibrin vehicles for precise local delivery of membrane-bound growth factor signals may help to elucidate specific biological growth factor function, and serve as starting point for development of new treatment strategies.
随着促血管生成或抗血管生成治疗方法的迅速增加，将需要新的有效方法来施用细胞结合生长因子。我们试图开发天然水凝胶基质纤维蛋白，作为血管形态发生素ephrin-B2进行广泛相互作用和持续信号传导的平台，该因子通常存在于质膜中，需要多价呈递以连接和激活相邻内皮细胞表面上的Eph受体。利用纤维蛋白和蛋白质工程技术诱导多价配体呈递，将重组突变型ephrin-B2受体结合域以可变的高密度共价偶联到纤维蛋白网络上。纤维蛋白结合的ephrin-B2作为内皮细胞配体的能力得以保留，体外实验表明，内皮细胞与工程化的ephrin-B2-纤维蛋白底物的结合呈剂量依赖性增加。通过将基质结合的ephrin-B2局部且长时间呈递给植入物附近的组织，在鸡胚绒毛尿囊膜中引发局部血管生成反应，证明了ephrin-B2-纤维蛋白植入基质的治疗相关性。这种关于用于精确局部递送膜结合生长因子信号的仿生纤维蛋白载体的新知识，可能有助于阐明特定生物生长因子的功能，并作为开发新治疗策略的起点。
