Introducing chemical functionality in Fmoc-peptide gels for cell culture
Introducing chemical functionality in Fmoc-peptide gels for cell culture
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DOI：
10.1016/j.actbio.2009.01.006
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发表时间：
2009-03-01
影响因子：
9.7
通讯作者：
Ulijn, Rein V.
中科院分区：
文献类型：
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作者：
Jayawarna, Vineetha;Richardson, Stephen M.;Ulijn, Rein V.
芳族短肽衍生物，即用芳族基团如9-芴基甲氧羰基（Fmoc）修饰的肽，可以自组装成自支撑水凝胶。这些水凝胶由于其高水合、相对刚度和纳米纤维结构而与细胞外基质具有一些相似性。我们以前证明，Fmoc-二苯基丙氨酸（Fmoc-F-2）提供了一个合适的基质二维（2D）或三维（3D）培养的原代牛软骨细胞。在本文中，我们调查是否引入化学功能，如NH 2，COOH或OH，增强与不同细胞类型的兼容性。一系列水凝胶组合物由Fmoc-F-2和n-保护的Fmoc氨基酸、赖氨酸（K，侧链R =（CH 2）（4）NH 2）、谷氨酸（D，侧链R = CH 2 COOH）和丝氨酸（S.具有侧链R = CH_2 OH）的化合物。所有组合物产生纤维直径在32-65 nm范围内的纤维支架，如通过低温扫描电子显微镜和原子力显微镜评估的。傅立叶变换红外光谱分析表明，肽段主要采用反平行β-折叠构象。振荡流变学结果表明，所有四种水凝胶具有软粘弹性材料的机械特性，其弹性模量取决于化学组成，范围从502 Pa（Fmoc-F-2/D）到21.2 KPa（Fmoc-F-2）。所有凝胶均支持牛软骨细胞的活力，如通过活-死染色测定所评估的。此外，Fmoc-F-2/S和Fmoc-F-2/D水凝胶支持人真皮成纤维细胞（HDF）的活力，而Fmoc-F-2/S水凝胶是支持所有三种测试细胞类型活力的唯一凝胶类型。因此，通过在2D培养中研究细胞增殖、细胞骨架组织和组织学分析来进一步研究Fmoc-F-2/S。此外，显示Fmoc-F-2/S凝胶支持牛软骨细胞3D培养中细胞形态的保留。这些结果表明，将化学官能团引入Fmoc-肽支架中可以为体外细胞培养提供具有可调化学和机械性质的凝胶。(c)2009 Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
Aromatic short peptide derivatives, i.e. peptides modified with aromatic groups such as 9-fluorenylmethoxycarbonyl (Fmoc), can self-assemble into self-supporting hydrogels. These hydrogels have some similarities to extracellular matrices due to their high hydration, relative stiffness and nanofibrous architecture. We previously demonstrated that Fmoc-diphenylalanine (Fmoc-F-2) provides a suitable matrix for two-dimensional (2D) or three-dimensional (3D) culture of primary bovine chondrocytes. In this paper we investigate whether the introduction of chemical functionality, such as NH2, COOH or OH, enhances compatibility with different cell types. A series of hydrogel compositions consisting of combinations of Fmoc-F-2 and n-protected Fmoc amino acids, lysine (K, with side chain R = (CH2)(4)NH2), glutamic acid (D, with side chain R = CH2COOH), and serine (S. with side chain R = CH2OH) were studied. All compositions produced fibrous scaffolds with fibre diameters in the range of 32-65 nm as assessed by cryo-scanning electron microscopy and atomic force microscopy. Fourier transform infrared spectroscopy analysis suggested that peptide segments adopt a predominantly anti-parallel beta-sheet conformation. Oscillatory rheology results show that all four hydrogels have mechanical profiles of soft viscoelastic materials with elastic moduli dependent on the chemical composition, ranging from 502 Pa (Fmoc-F-2/D) to 21.2 KPa (Fmoc-F-2). All gels supported the viability of bovine chondrocytes as assessed by a live-dead staining assay. Fmoc-F-2/S and Fmoc-F-2/D hydrogels in addition supported viability for human dermal fibroblasts (HDF) while Fmoc-F-2/S hydrogel was the only gel type that supported viability for all three cell types tested. Fmoc-F-2/S was therefore investigated further by studying cell proliferation, cytoskeletal organization and histological analysis in 2D culture. In addition, the Fmoc-F-2/S gel was shown to support retention of cell morphology in 3D culture of bovine chondrocytes. These results demonstrate that introduction of chemical functionality into Fmoc-peptide scaffolds may provide gels with tunable chemical and mechanical properties for in vitro cell culture. (c) 2009 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.