Covalently conjugated VEGF-fibrin matrices for endothelialization
Covalently conjugated VEGF-fibrin matrices for endothelialization
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DOI：
10.1016/s0168-3659(01)00266-8
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发表时间：
2001-05-14
影响因子：
10.8
通讯作者：
Hubbell, JA
中科院分区：
文献类型：
--
作者：
Zisch, AH;Schenk, U;Hubbell, JA
血管内皮生长因子（VEGF）是内皮细胞生物学和血管形成的关键因素，也是刺激血管生成依赖性组织再生的候选治疗剂。本研究的目的是将 VEGF(121) 赋予生物材料纤维蛋白的血管生成活性，纤维蛋白是内皮细胞生长的天然底物，临床上被称为“纤维蛋白胶”。为了实现这一目标，我们设计了基于纤维蛋白的水凝胶，并用 VEGF(121) 进行了共价修饰。我们的实验室最近开发了新的方法，允许在凝血过程中通过因子 XIIIa 的转谷氨酰胺酶活性将外源生物活性肽共价掺入纤维蛋白中。在这里，因子 XIIIa 交联纤维蛋白内其他蛋白质的这种能力被用来共价掺入 VEGF(121)。通过重组DNA方法，突变型VEGF(121)变体α(2)-PI1-8-VEGF(121)在氨基末端含有额外的因子XIIIa底物序列NQEQVSPL，在大肠杆菌中表达。在可溶形式下，突变蛋白完全保留了其对内皮细胞的有丝分裂活性。使用 I-125 标记的 α (2)-PI1-8-VEGF(121)，证明并表征了其共价掺入以及掺入纤维蛋白的效率。对于在二维 VEGF(121) 修饰的纤维蛋白表面上生长的人内皮细胞来说，固定的纤维蛋白缀合的 VEGF(121) 蛋白仍然是一种活性且非常有效的有丝分裂原，并且掺入越来越多的 α(2)-PI1-8-VEGF(121) 导致内皮细胞生长的剂量依赖性增强。 VEGF 修饰的纤维蛋白基质可以在体内生理条件下通过一步反应形成可注射凝胶。当用作向内生长基质时，此类掺入VEGF的材料可用于需要对缺血区域或植物体内产生血管生成反应的各种临床情况。 (C) 2001 Elsevier Science B.V. 保留所有权利。
Vascular endothelial growth factor (VEGF) is a key factor in endothelial cell biology and blood vessel formation and a candidate therapeutic for the stimulation of angiogenesis-dependent tissue regeneration. The objective of this study was to confer the angiogenic activity of VEGF(121) upon the biomaterial fibrin, a natural substrate for endothelial cell growth and clinically accepted as 'fibrin glue'. To achieve this, we engineered fibrin-based hydrogels that were covalently modified with VEGF(121). Our laboratory has recently developed novel methodology that allows the covalent incorporation of exogenous bioactive peptides by the transglutominase activity of factor XIIIa into fibrin during cogulation. Here, this ability of factor XIIIa to crosslink additional proteins within fibrin was employed to covalently incorporate VEGF(121). By recombinant DNA methodology, a mutant VEGF(121) variant, alpha (2)-PI1-8-VEGF(121), which contains an additional factor XIIIa substrate sequence NQEQVSPL at the aminoterminus, was expressed in E. coli. In soluble form, the mutant protein fully retained its mitogenic activity for endothelial cells. Using I-125-labeled alpha (2)-PI1-8-VEGF(121), its covalent incorporation and the efficiency of incorporation into fibrin was demonstrated and characterized. The immobilized, fibrin-conjugated VEGF(121) protein remained an active and very efficient mitogen for human endothelial cells grown on two-dimensional VEGF(121)-modified fibrin surfaces, and the incorporation of increasing amounts of alpha (2)-PI1-8-VEGF(121) resulted in dose-dependent enhancement of endothelial cell growth. The VEGF-modified fibrin matrices can be formed as injectable gels in a single-step reaction under physiological conditions in vivo. When used as a ingrowth matrix, such VEGF incorporating materials could be useful in a variety of clinical situations that require an angiogenic response into an ischemic region or inplant. (C) 2001 Elsevier Science B.V. All rights reserved.