Engineering fibrin-binding TGF-β1 for sustained signaling and contractile function of MSC based vascular constructs.
Engineering fibrin-binding TGF-β1 for sustained signaling and contractile function of MSC based vascular constructs.
复制标题
DOI：
10.1016/j.biomaterials.2011.07.079
复制
发表时间：
2011-11
期刊：
影响因子：
14
通讯作者：
Andreadis, Stelios T.
中科院分区：
文献类型：
--
作者：
Liang, Mao-Shih;Andreadis, Stelios T.
关键词：
我们提出了一种将TGF-β1结合到纤维蛋白水凝胶中的策略，以模拟生长因子在3D环境中的体内呈递。为此，我们设计了TGF-β1和由因子XIII结构域和纤溶酶切割位点组成的双功能肽之间的融合蛋白。在另一个版本中，蛋白酶切割位点被省略，以检查不能被细胞从支架释放的生长因子是否对组织构建体具有不同的影响。在潜伏相关肽和成熟TGF-β1结构域之间发现了产生正确加工的功能蛋白的最佳插入位点。在溶液中，融合蛋白表现出与天然TGF-β1相似的生物活性，如通过细胞增殖抑制和启动子活性测定所证明的。免疫沉淀实验表明，融合TGF-β1蛋白以因子XIII依赖的方式与纤维蛋白原结合，并可通过纤溶酶的作用从肽中释放。与推注递送相反，固定化TGF-β1在纤维蛋白包埋的细胞中诱导持续信号传导数天，如通过Smad 2磷酸化所证明的。延长的通路激活与由毛囊间充质干细胞或骨髓来源的平滑肌细胞制备的血管构建体的增强的收缩功能相关。我们的研究结果表明，纤维蛋白固定的TGF-β1可用于增强局部微环境，并改善体外工程组织的功能，并可能在体内植入后，生长因子递送面临压倒性的挑战。
We present a strategy to conjugate TGF-β1 into fibrin hydrogels to mimic the in vivo presentation of the growth factor in a 3D context. To this end, we engineered fusion proteins between TGF-β1 and a bi-functional peptide composed of a Factor XIII domain and a plasmin cleavage site. In another version the protease cleavage site was omitted to examine whether the growth factor that could not be released from the scaffold by cells had different effects on tissue constructs. The optimal insertion site which yielded correctly processed, functional protein was found between the latency associated peptide and mature TGF-β1 domains. In solution the fusion proteins exhibited similar biological activity as native TGF-β1 as evidenced by inhibition of cell proliferation and promoter activity assays. Immunoprecipitation experiments demonstrated that the fusion TGF-β1 protein bound to fibrinogen in a Factor XIII dependent manner and could be released from the peptide by the action of plasmin. In contrast to bolus delivery, immobilized TGF-β1 induced sustained signaling in fibrin-embedded cells for several days as evidenced by Smad2 phosphorylation. Prolonged pathway activation correlated with enhanced contractile function of vascular constructs prepared from hair follicle mesenchymal stem cells or bone marrow derived smooth muscle cells. Our results suggest that fibrin-immobilized TGF-β1 may be used to enhance the local microenvironment and improve the function of engineered tissues in vitro and potentially also after implantation in vivo where growth factor delivery faces overwhelming challenges.
登录
查看更多内容
影响因子：
2.9
作者：
ABE, M;HARPEL, JG;RIFKIN, DB
通讯作者：
RIFKIN, DB
影响因子：
10.8
作者：
Drinnan, Charles T.;Zhang, Ge;Suggs, Laura J.
通讯作者：
Suggs, Laura J.
影响因子：
4.8
作者：
Gong, Zhaodi;Niklason, Laura E.
通讯作者：
Niklason, Laura E.
影响因子：
4.6
作者：
Hu, Xiaohong;Ma, Lie;Gao, Changyou
通讯作者：
Gao, Changyou
影响因子：
2.9
作者：
IKEDA, T;LIOUBIN, MN;MARQUARDT, H
通讯作者：
MARQUARDT, H