Time-dependent migratory behaviors in the long-term studies of fibroblast durotaxis on a hydrogel substrate fabricated with a soft band.
Time-dependent migratory behaviors in the long-term studies of fibroblast durotaxis on a hydrogel substrate fabricated with a soft band.
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DOI：
10.1021/la501058j
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发表时间：
2014-06-03
期刊：
影响因子：
--
通讯作者：
Kidoaki S
中科院分区：
文献类型：
--
作者：
Kuboki T;Chen W;Kidoaki S
趋硬性是细胞在培养基质上沿刚度梯度的偏向运动，是操纵细胞在工程生物材料表面迁移的有用趋性行为之一。在这项研究中，研究了凝胶状基底上的长期硬旋转，该基底含有使用弹性图案制作的20、50和150 μm宽的软带;实现了梯度强度为300 kPa/50 μm的尖锐弹性边界。在3天的时间段内观察3 T3成纤维细胞的时间依赖性迁移行为。在第一天，大多数细胞强烈排斥的软带的带宽无关，表现出典型的硬旋转行为。然而，软带的排斥性减弱，更多的细胞越过软带或在观察过程中表现出其他混合迁移行为。结果发现，最宽软带的基底上的硬旋转强度减弱，并且随着时间的推移，最宽软带上的粘附亲和诱导包封变得明显。检查了可能导致延长培养期间硬脊膜扩张明显减少的因素，如表面形貌、弹性和/或化学变化。免疫荧光分析表明，优先胶原蛋白沉积到软带，这是由成纤维细胞分泌，导致随着时间的推移，对软带的haptotaxis的贡献增加。沉积的胶原蛋白不影响表面形貌或表面弹性，但确实改变了表面化学，特别是在软带上。所观察到的时间依赖性硬旋转行为是混合机械和化学线索的结果。在受控刺激下细胞迁移行为的研究和应用中，重要的是彻底检查其他（隐藏的）复合刺激，以便能够准确地解释数据并设计合适的生物材料来操纵细胞迁移。
Durotaxis, biased cell movement up a stiffness gradient on culture substrates, is one of the useful taxis behaviors for manipulating cell migration on engineered biomaterial surfaces. In this study, long-term durotaxis was investigated on gelatinous substrates containing a soft band of 20, 50, and 150 μm in width fabricated using photolithographic elasticity patterning; sharp elasticity boundaries with a gradient strength of 300 kPa/50 μm were achieved. Time-dependent migratory behaviors of 3T3 fibroblast cells were observed during a time period of 3 days. During the first day, most of the cells were strongly repelled by the soft band independent of bandwidth, exhibiting the typical durotaxis behavior. However, the repellency by the soft band diminished, and more cells crossed the soft band or exhibited other mixed migratory behaviors during the course of the observation. It was found that durotaxis strength is weakened on the substrate with the narrowest soft band and that adherent affinity-induced entrapment becomes apparent on the widest soft band with time. Factors, such as changes in surface topography, elasticity, and/or chemistry, likely contributing to the apparent diminishing durotaxis during the extended culture were examined. Immunofluorescence analysis indicated preferential collagen deposition onto the soft band, which is derived from secretion by fibroblast cells, resulting in the increasing contribution of haptotaxis toward the soft band over time. The deposited collagen did not affect surface topography or surface elasticity but did change surface chemistry, especially on the soft band. The observed time-dependent durotaxis behaviors are the result of the mixed mechanical and chemical cues. In the studies and applications of cell migratory behavior under a controlled stimulus, it is important to thoroughly examine other (hidden) compounding stimuli in order to be able to accurately interpret data and to design suitable biomaterials to manipulate cell migration.
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影响因子：
3.4
作者：
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