The use of poly(ethylene glycol) hydrogels to investigate the impact of ECM chemistry and mechanics on smooth muscle cells
The use of poly(ethylene glycol) hydrogels to investigate the impact of ECM chemistry and mechanics on smooth muscle cells
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DOI：
10.1016/j.biomaterials.2006.05.012
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发表时间：
2006-10-01
期刊：
影响因子：
14
通讯作者：
Putnam, Andrew J.
中科院分区：
文献类型：
--
作者：
Peyton, Shelly R.;Raub, Christopher B.;Putnam, Andrew J.
基于聚（乙二醇）（PEG）的水凝胶对于再生医学应用越来越感兴趣，并且由于能够在PEG的惰性骨架上赋予天然细胞外基质（ECM）的关键功能，因此是指导细胞功能的理想材料。鉴于ECM顺应性影响多种细胞功能的最近大量证据，基于PEG的水凝胶也是有吸引力的，因为它们的机械性质可以容易地控制。在这些研究中，我们利用基于PEG的凝胶的化学和机械可调性来研究ECM化学和力学对2-D和3-D模型系统中的平滑肌细胞（SMC）的影响。首先，通过控制交联程度，从而控制基于PEG的水凝胶的机械性能（拉伸模量为13.7至423.9 kPa），我们在这里报告说，F-肌动蛋白应力纤维和粘着斑的组装（指示肌动蛋白收缩性的状态）受到影响用短粘附肽或全长ECM蛋白官能化的2-D PEG凝胶的顺应性。不同的ECM配体密度和身份独立的凝胶顺应性影响的物理性质的局灶性粘连，也影响SMC的扩散在2-D。此外，随着凝胶硬度的增加，SMCs增殖的程度更大。与此相反，SMC分化的程度，这是定性评估的程度平滑肌α-肌动蛋白捆绑和协会的钙调蛋白和钙调蛋白与α-肌动蛋白原纤维，被发现减少与基板刚度在2-D文化。在3-D中，尽管它们的活力和扩散程度大大降低，但当在PEG-RGDS构建体中培养时，SMC确实表达一些指示其分化表型的收缩标记物。结合，这些数据表明，PEG水凝胶的机械和化学性质可以调整，以影响SMC表型在2-D和3-D。(c)2006爱思唯尔有限公司保留所有权利。
Hydrogels based on poly(ethylene glycol) (PEG) are of increasing interest for regenerative medicine applications and are ideal materials to direct cell function due to the ability to confer key functionalities of native extracellular matrix (ECM) on PEG's otherwise inert backbone. Given extensive recent evidence that ECM compliance influences a variety of cell functions, PEG-based hydrogels are also attractive due to the ease with which their mechanical properties can be controlled. In these studies, we exploited the chemical and mechanical tunability of PEG-based gels to study the impact of ECM chemistry and mechanics on smooth muscle cells (SMCs) in both 2-D and 3-D model systems. First, by controlling the extent of crosslinking and therefore the mechanical properties of PEG-based hydrogels (tensile moduli from 13.7 to 423.9 kPa), we report here that the assembly of F-actin stress fibers and focal adhesions, indicative of the state of actin contractility, were influenced by the compliance of 2-D PEG gels functionalized with either short adhesive peptides or full-length ECM proteins. Varying ECM ligand density and identity independent of gel compliance affected the physical properties of the focal adhesions, and also influenced SMC spreading in 2-D. Furthermore, SMCs proliferated to a greater extent as gel stiffness was increased. In contrast, the degree of SMC differentiation, which was qualitatively assessed by the extent of smooth muscle alpha-actin bundling and the association of calponin and caldesmon with the alpha-actin fibrils, was found to decrease with substrate stiffness in 2-D cultures. In 3-D, despite the fact that their viability and degree of spreading were greatly reduced, SMCs did express some contractile markers indicative of their differentiated phenotype when cultured within PEG-RGDS constructs. Combined, these data suggest that the mechanical and chemical properties of PEG hydrogels can be tuned to influence SMC phenotype in both 2-D and 3-D. (c) 2006 Elsevier Ltd. All rights reserved.