A chimeric epidermal growth factor with fibrin affinity promotes repair of injured keratinocyte sheets
A chimeric epidermal growth factor with fibrin affinity promotes repair of injured keratinocyte sheets
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DOI：
10.1016/j.actbio.2009.03.022
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发表时间：
2009-09-01
影响因子：
9.7
通讯作者：
Ito, Yoshihiro
中科院分区：
文献类型：
--
作者：
Kitajima, Takashi;Sakuragi, Makoto;Ito, Yoshihiro
本研究的目的是创建一种具有纤维蛋白亲和力的新型表皮生长因子（EGF）嵌合蛋白，并证明其通过固定纤维蛋白来修复受损组织的潜力。嵌合蛋白 (FBD-EGF) 是通过纤连蛋白纤维蛋白结合结构域 (FBD) 与 EGF 融合而产生的。它与纤维蛋白的结合呈剂量依赖性，并且其结合稳定至少 7 天，而天然 EGF 几乎没有亲和力。 FBD-EGF 在纤维蛋白结合状态以及可溶状态下促进成纤维细胞和角质形成细胞的生长。在角质形成细胞培养系统中进一步研究了其活性，其中纤维蛋白在细胞片层损伤时暴露。与天然 EGF 相比，纤维蛋白结合的 FBD-EGF 以显着更快的速度（大约八倍）促进受伤区域上的片层生长（p < 0.01）。 2 分钟宽的伤口在 7-9 天内闭合。该修复过程被抗-EGF 抑制。角质形成细胞在与 FBD-EGF 接触的纤维蛋白片的前缘处更广泛地增殖，大约是邻近区域的 1.7 倍。这些结果表明嵌合EGF与纤维蛋白的稳定结合对于修复受损的角质形成细胞片是有效的，这表明其在组织工程中的潜在用途。 (C) 2009 Acta Materialia Inc. 由 Elsevier Ltd 出版。保留所有权利。
The aim of the present study is to create a novel chimeric protein of epidermal growth factor (EGF) with fibrin affinity and demonstrate its potential for repairing injured tissues by immobilization to fibrin. The chimeric protein (FBD-EGF) was produced by the fusion of the fibronectin fibrin-binding domain (FBD) to EGF. It showed dose-dependent binding to fibrin and its binding was stable for at least 7 days, while native EGF showed little affinity. FBD-EGF promoted the growth of fibroblasts and keratinocytes in the fibrin-bound state as well as in the soluble state. Its activity was further studied in a keratinocyte culture system in which fibrin was exposed upon injury of cell sheets. Fibrin-bound FBD-EGF promoted growth of the sheets over the injured area at a significantly faster rate (approximately eightfold) than native EGF (p < 0.01). Wounds 2 min wide were closed in 7-9 days. This repair process was inhibited by anti-EGF. Keratinocytes proliferated more extensively in the leading edges of sheets contacting fibrin with FBD-EGF, approximately 1.7-fold more than in the adjacent regions. These results imply that the stable binding of chimeric EGF to fibrin is effective for the repair of injured keratinocyte sheets, suggesting a potential use in tissue engineering. (C) 2009 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.