Characterizing and Patterning Polyacrylamide Substrates Functionalized with N-Hydroxysuccinimide
Characterizing and Patterning Polyacrylamide Substrates Functionalized with N-Hydroxysuccinimide
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DOI：
10.1007/s12195-013-0288-5
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发表时间：
2013-09-01
影响因子：
2.8
通讯作者：
Johnson, Amy J. Wagoner
中科院分区：
文献类型：
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作者：
Poellmann, Michael J.;Johnson, Amy J. Wagoner
聚丙烯酰胺是细胞力生物学研究中广泛使用的材料。在这里，我们验证并优化了一种激活聚丙烯酰胺底物以进行蛋白质图案化的方法，从而形成一个能够精确且独立控制细胞感知的几何和机械因素的系统。掺入水凝胶中的丙烯酸为 N-羟基琥珀酰亚胺的激活提供了功能位点，进而与以微型图案打印的蛋白质形成共价键。为了验证和优化基材制造，我们证明丙烯酸掺入到聚合物中，达到 0.4 wt% 时对杨氏模量没有影响，并且增加丙烯酸浓度会导致基材上结合的蛋白质数量增加。最后，我们证明细胞附着并扩散到带有电流体动力喷射（e-jet）打印图案的蛋白质的基底上。该方法代表了对最广泛使用的用磺基-SANPAH 化学活化聚丙烯酰胺的方法的改进。通过进一步改进，可以真正独立地控制配体密度和刚度。这些基质是探索基质硬度和配体密度对细胞行为的相互作用影响的强大平台。
Polyacrylamide is a widely used material in cell mechanobiology research. Here, we validate and optimize a method to activate polyacrylamide substrates for protein patterning, resulting in a system with precise and independent control over the geometric and mechanical factors that cells perceive. Acrylic acid incorporated into the hydrogel provides functional sites for activation with N-hydroxysuccinimide, which in turn forms covalent bonds with proteins printed in microscale patterns. To validate and optimize substrate fabrication, we demonstrate that acrylic acid incorporates into the polymer, that is has no effect on Young's modulus at up to 0.4 wt%, and that increasing concentrations of acrylic acid result in substrates with increasing amounts of protein bound to them. Finally, we demonstrate that cells attach and spread to substrates with protein patterned with electrohydrodynamic jet (e-jet) printing. The method represents an improvement over the most-widely used method to chemically activate polyacrylamide with sulfo-SANPAH. With further refinement, truly independent control over ligand density and stiffness is possible. These substrates are powerful platforms for exploring the interacting influence of substrate stiffness and ligand density on cell behavior.