Poly(vinylmethylsiloxane) Elastomer Networks as Functional Materials for Cell Adhesion and Migration Studies
Poly(vinylmethylsiloxane) Elastomer Networks as Functional Materials for Cell Adhesion and Migration Studies
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DOI：
10.1021/bm101549y
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发表时间：
2011-04-01
影响因子：
6.2
通讯作者：
Haugh, Jason M.
中科院分区：
文献类型：
--
作者：
Ahmed, Shoeb;Yang, Hyun-kwan;Haugh, Jason M.
细胞迁移对于损伤和感染的生理反应以及生物材料植入物的设计至关重要。调节粘附材料的性质并将这些性质以定量的方式与细胞内过程的动力学相关的能力仍然是细胞迁移操作中的一个明确的挑战。在这里，我们建议使用聚（乙烯基甲基硅氧烷）（PVMS）网络作为新的基质细胞粘附和迁移。这些材料提供了独立调节化学官能度和弹性模量的能力。重要的是，PVMS网络与全内反射荧光（TIRF）显微镜兼容，这是询问细胞-基质界面的理想选择;后者的特性比聚丙烯酰胺凝胶和其他遇水膨胀的材料具有明显的优势。为了证明这些能力，使用羧基封端的硫醇作为接头，将含有乙酰基-甘氨酰-天冬氨酸（RGD）三肽基序的粘附肽成功地接枝到PVMS网络的表面。肽特异性粘附，扩散，和随机迁移的NIH 3 T3小鼠成纤维细胞的特点。这些实验表明，除了RGD之外还含有纤连蛋白（PHSRN）的协同序列的肽促进更有生产力的细胞迁移，而不显著增强细胞粘附强度。使用TIRF显微镜，通过磷脂酰肌醇3-激酶途径的信号转导的动力学进行监测，因为它们在肽接枝PVMS表面迁移的细胞。这种方法为在细胞内过程水平上研究定向迁移和机械转导提供了一个有前途的途径。
Cell migration is central to physiological responses to injury and infection and in the design of biomaterial implants. The ability to tune the properties of adhesive materials and relate those properties in a quantitative way to the dynamics of intracellular processes remains a definite challenge in the manipulation of cell migration. Here, we propose the use of poly(vinylmethylsiloxane) (PVMS) networks as novel substrata for cell adhesion and migration. These materials offer the ability to tune independently chemical functionality and elastic modulus. Importantly, PVMS networks are compatible with total internal reflection fluorescence (TIRF) microscopy, which is ideal for interrogating the cell-substratum interface; this latter characteristic presents a distinct advantage over polyacrylamide gels and other materials that swell with water. To demonstrate these capabilities, adhesive peptides containing the arginyl-glycyl-aspartic acid (RGD) tripeptide motif were successfully grafted to the surface of PVMS network using a carboxyl-terminated thiol as a linker. Peptide-specific adhesion, spreading, and random migration of NIH 3T3 mouse fibroblasts were characterized. These experiments show that a peptide containing the synergy sequence of fibronectin (PHSRN) in addition to RGD promotes more productive cell migration without markedly enhancing cell adhesion strength. Using TIRF microscopy, the dynamics of signal transduction through the phosphoinositide 3-kinase pathway were monitored in cells as they migrated on peptide-grafted PVMS surfaces. This approach offers a promising avenue for studies of directed migration and mechanotransduction at the level of intracellular processes.