Engineered microenvironments for synergistic VEGF - Integrin signalling during vascularization.
Engineered microenvironments for synergistic VEGF - Integrin signalling during vascularization.
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DOI：
10.1016/j.biomaterials.2017.02.024
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发表时间：
2017-05
期刊：
影响因子：
14
通讯作者：
Salmerón-Sánchez M
中科院分区：
文献类型：
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作者：
Moulisová V;Gonzalez-García C;Cantini M;Rodrigo-Navarro A;Weaver J;Costell M;Sabater I Serra R;Dalby MJ;García AJ;Salmerón-Sánchez M
我们设计了基于聚合物的微环境，通过协同整合素-生长因子受体信号在体外和体内促进血管生成。聚丙烯酸乙酯(PEA)触发纤维连接蛋白(FN)自发形成纳米网络，从而提供关键结合域的可用性。重要的是，生长因子结合区(FNIII12-14)和整合素结合区(FNIII9-10)同时存在于组装在PEA上的FN纤维上。这个材料平台促进了整合素/血管内皮生长因子信号的协同作用，这对于低浓度的血管内皮生长因子的体外血管形成事件是非常有效的。与对照聚合物(聚甲基丙烯酸甲酯，PMA)相比，在PEA上，VEGF特异性地结合到FN纤维上，其中FN保持球状构象，整合素/GF结合域不同时可用。与较高剂量的可溶性给药相比，种植在这些协同界面上的人内皮细胞(与PEA上组装的FN结合的血管内皮生长因子)的血管生成反应显著改善。只有当血管内皮生长因子与PEA上的FN纳米网络结合时，早期启动的血管内皮生长因子信号(PLCERK1磷酸化)和整合素和血管内皮生长因子信号(γ1/2磷酸化)才增加，而可溶性血管内皮生长因子不影响早期信号转导。用具有受损整合素结合位点(FN-RGE)的突变FN分子的实验证实了FN的整合素结合位点通过整合素/血管内皮生长因子联合信号在血管生成反应中的作用。在体内实验中，使用涂有FN和VEGF的3D支架植入小鼠脂肪垫，显示了通过促进支架毛孔内新组织的形成来促进血管形成的信号。豌豆驱动的FN的组织促进了血管内皮生长因子的高效表达，从而促进了再生医学应用中的血管生成。血管内皮生长因子与聚合体上自发形成的纤维连接蛋白特异性结合。高效的血管内皮生长因子递呈允许同时传递VEGFR和整合素信号。低水平的血管内皮生长因子可促进血管内皮细胞向原始血管方向重组。纤维连接蛋白和血管内皮生长因子包裹的PEA支架在体内刺激血管生长。
We have engineered polymer-based microenvironments that promote vasculogenesis both in vitro and in vivo through synergistic integrin-growth factor receptor signalling. Poly(ethyl acrylate) (PEA) triggers spontaneous organization of fibronectin (FN) into nanonetworks which provide availability of critical binding domains. Importantly, the growth factor binding (FNIII12-14) and integrin binding (FNIII9-10) regions are simultaneously available on FN fibrils assembled on PEA. This material platform promotes synergistic integrin/VEGF signalling which is highly effective for vascularization events in vitro with low concentrations of VEGF. VEGF specifically binds to FN fibrils on PEA compared to control polymers (poly(methyl acrylate), PMA) where FN remains in a globular conformation and integrin/GF binding domains are not simultaneously available. The vasculogenic response of human endothelial cells seeded on these synergistic interfaces (VEGF bound to FN assembled on PEA) was significantly improved compared to soluble administration of VEGF at higher doses. Early onset of VEGF signalling (PLCγ1 phosphorylation) and both integrin and VEGF signalling (ERK1/2 phosphorylation) were increased only when VEGF was bound to FN nanonetworks on PEA, while soluble VEGF did not influence early signalling. Experiments with mutant FN molecules with impaired integrin binding site (FN-RGE) confirmed the role of the integrin binding site of FN on the vasculogenic response via combined integrin/VEGF signalling. In vivo experiments using 3D scaffolds coated with FN and VEGF implanted in the murine fat pad demonstrated pro-vascularization signalling by enhanced formation of new tissue inside scaffold pores. PEA-driven organization of FN promotes efficient presentation of VEGF to promote vascularization in regenerative medicine applications. VEGF specifically binds to spontaneously formed fibronectin assembled on polymers. Highly efficient VEGF presentation allows simultaneous VEGFR and integrins signalling. Low VEGF amount for endothelial cell reorganization towards primitive vasculature. Fibronectin and VEGF coated PEA scaffold stimulates vessel growth in vivo.