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首页> 外文学位 >Well-Defined Topographically and Peptide-Functionalized Hydrogel Arrays for Investigating Corneal Epithelial Cell Behavior.
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Well-Defined Topographically and Peptide-Functionalized Hydrogel Arrays for Investigating Corneal Epithelial Cell Behavior.

机译：定义良好的地形图和肽功能化水凝胶阵列，用于研究角膜上皮细胞的行为。

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摘要

Epithelial cells reside on specialized extracellular matrices that provide instructive cues to regulate and support cell function. We have previously demonstrated that substrate topography with dimensions similar to the native extracellular matrix (submicrometer and nano-scale features) significantly impacts corneal epithelial proliferation and migration. In this work, we included additional instructive cues by incorporating specific peptide ligands into topographically nano-patterned synthetic hydrogels. Peptides RGD and AG73 were conjugated to monodisperse poly(ethylene glycol) spacers that separated the peptide from the monomeric functionality which reacts during hydrogel polymerization. These PEG-peptide conjugates were then copolymerized with PEG diacrylate to form an inert hydrogel network decorated with peptide ligands for cell interactions. The efficient, systematic study of multiple instructive cues (peptide, peptide concentration, topographic dimensions), however, was also contingent on the development of enhanced throughput platforms. Towards this goal, we developed a hydrogel array platform to systematically and rapidly evaluate combinations of the two different peptide motifs and a range of nano-scale topographic dimensions. Elastomeric stencils with arrays of millimeter-scale regions were used to spatially confine hydrogel precursor solutions on elastomeric stamps with nano-scale patterns generated by soft lithography. The resulting topographically and peptide-functionalized hydrogel arrays were used to characterize single cell behavior, including morphology, proliferation, and migration. We showed that epithelial cell migration speed and persistence was governed by both the biochemical and topographical cues of the underlying substrate. We also developed a wound healing assay for these hydrogel arrays to study collective epithelial cell migration. Similar to single-cell studies, collective cell migration and wound healing rates were faster on topographic surfaces with the addition of AG73. The strategy and synthetic techniques developed here for well-defined hydrogel arrays functionalized with both peptides and topographic features provides unparalleled control for investigating synergistic interactions of both physical and chemical cues.
机译：上皮细胞驻留在专门的细胞外基质上，这些基质提供了指导性信号以调节和支持细胞功能。我们先前已经证明，尺寸类似于天然细胞外基质（亚微米和纳米级特征）的基质形貌会显着影响角膜上皮的增殖和迁移。在这项工作中，我们通过将特定的肽配体掺入地形纳米图案的合成水凝胶中，从而获得了其他指导性提示。肽RGD和AG73与单分散的聚（乙二醇）间隔基偶联，该间隔基将肽与在水凝胶聚合过程中发生反应的单体官能团分开。然后将这些PEG-肽共轭物与PEG二丙烯酸酯共聚，形成惰性水凝胶网络，该网络装饰有用于细胞相互作用的肽配体。但是，对多种指导性线索（肽，肽浓度，形貌尺寸）的有效，系统的研究还取决于增强的通量平台的开发。为了实现这一目标，我们开发了一种水凝胶阵列平台，可系统快速评估两种不同肽基序和一系列纳米尺度地形尺寸的组合。具有毫米级区域阵列的弹性模版用于将水凝胶前体溶液在空间上限制在具有通过软光刻产生的纳米级图案的弹性体印模上。所得的地形图和肽官能化水凝胶阵列用于表征单细胞行为，包括形态，增殖和迁移。我们表明，上皮细胞的迁移速度和持久性受基础底物的生化和地形线索的支配。我们还针对这些水凝胶阵列开发了伤口愈合测定法，以研究集体上皮细胞迁移。与单细胞研究相似，加入AG73后，在地形表面上集体细胞迁移和伤口愈合速度更快。此处开发的策略和合成技术，用于利用肽和地形特征功能化的定义明确的水凝胶阵列，为研究物理和化学线索的协同相互作用提供了无与伦比的控制。

著录项

  • 作者

    Wilson, Michelle Juliette.;

  • 作者单位

    The University of Wisconsin - Madison.;

  • 授予单位 The University of Wisconsin - Madison.;
  • 学科 Engineering Chemical.;Nanotechnology.;Engineering Biomedical.
  • 学位 Ph.D.
  • 年度 2012
  • 页码 148 p.
  • 总页数 148
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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