School of Life Science and Technology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8502, Japan.
Biomed Mater. 2017 Aug 9;12(4):045021. doi: 10.1088/1748-605X/aa7616.
Self-assembling peptides are attractive materials for tissue engineering applications because of their functionality including high biocompatibility and biodegradability. Modification of self-assembling peptides with functional motifs, such as the cell-adhesive tripeptide sequence RGD leads to functional artificial extracellular matrices (ECMs). In this study, we developed an artificial self-assembling ECM protein tethered with a growth factor via heterotrimer triple-helix (helix A/B/C) formation. The helix A and helix C peptides, which are capable of forming a heterodimer coiled-coil structure, were fused to both ends of a matrix protein composed of the elastin-derived structural unit (APGVGV) with an RGD motif. The helix B peptide, which constituents the third helix of the triple-helix structure, was fused with basic fibroblast growth factor (bFGF) for tethering to the artificial ECM proteins. Each recombinant protein exhibited cell adhesion and cell proliferation activities similar to the original, while the designed bFGF-tethered ECM protein exhibited superior cell proliferation activity. These results demonstrate that the approach of creating growth factor-tethered self-assembling proteins via triple-helix formation can be applied to develop functional ECMs for tissue engineering applications.
自组装肽因其高生物相容性和可生物降解性等功能而成为组织工程应用的有吸引力的材料。通过将具有功能性基序（例如细胞黏附三肽序列 RGD）的自组装肽进行修饰，可以得到具有功能性的人工细胞外基质（ECM）。在本研究中，我们通过异三聚体三螺旋（A/B/C 螺旋）形成，开发了一种通过生长因子将人工自组装 ECM 蛋白连接起来的方法。能够形成异二聚体螺旋结构的 A 螺旋和 C 螺旋肽被融合到由弹性蛋白衍生的结构单元（APGVGV）和 RGD 基序组成的基质蛋白的两端。构成三螺旋结构的第三个螺旋的 B 螺旋肽与碱性成纤维细胞生长因子（bFGF）融合，用于连接到人工 ECM 蛋白上。每个重组蛋白均表现出与原始蛋白相似的细胞黏附和增殖活性，而设计的 bFGF 连接的 ECM 蛋白则表现出优异的细胞增殖活性。这些结果表明，通过三螺旋形成创建生长因子连接的自组装蛋白的方法可用于开发用于组织工程应用的功能性 ECM。
