The threshold at which substrate nanogroove dimensions may influence fibroblast alignment and adhesion
The threshold at which substrate nanogroove dimensions may influence fibroblast alignment and adhesion
复制标题
DOI：
10.1016/j.biomaterials.2007.05.030
复制
发表时间：
2007-09-01
期刊：
影响因子：
14
通讯作者：
Jansen, J. A.
中科院分区：
文献类型：
--
作者：
Loesberg, W. A.;te Riet, J.;Jansen, J. A.
体外评估了在光滑和纳米凹槽基质（凹槽深度：5-350 nm，宽度：20-1000 nm）上培养的成纤维细胞之间形态行为的差异。该研究的目的是阐明细胞引导在越来越小的地形上发生的程度。使用电子束光刻技术制作图案模板，随后使用溶剂浇铸在聚苯乙烯细胞培养材料中复制。使用原子力显微镜（AFM）对重复进行研究。接种成纤维细胞后，使用扫描电子显微镜（SEM）和光学显微镜研究形态特征，以获得细胞排列的定性和定量信息。 AFM 显示纳米凹槽/脊的宽度被完美复制，尽管在更深的水平上凹槽变得更加凹。除了 1 nm 的粗糙度幅度外，光滑基质没有任何可区分的图案。有趣的是，显微镜和图像分析表明，4 小时后，成纤维细胞已根据纳米形貌特征将其形状调整至 100 nm 宽度和 75 nm 深度的截止值。培养24小时后，成纤维细胞甚至可以在浅至35 nm的凹槽上排列。看来深度是节距比为 1:1 的凹槽图案上细胞排列的最重要参数。在光滑的基质上，细胞总是以随机方式展开。方差分析 (ANOVA) 表明，主要参数（地形和培养时间）都很显着。我们得出结论，在纳米形貌上培养的成纤维细胞经历 35 nm 的阈值特征尺寸，低于该值接触引导不再存在。 (C) 2007 Elsevier Ltd. 保留所有权利。
The differences in morphological behaviour between fibroblasts cultured on smooth and nanogrooved substrata (groove depth: 5-350 nm, width: 20-1000 nm) have been evaluated in vitro. The aim of the study was to clarify to what extent cell guidance occurs on increasingly smaller topographies. Pattern templates were made using electron beam lithography, and were subsequently replicated in polystyrene cell culture material using solvent casting. The replicates were investigated with atomic force microscopy (AFM). After seeding with fibroblasts, morphological characteristics were investigated using scanning electron microscopy (SEM) and light microscopy, in order to obtain qualitative and quantitative information on cell alignment. AFM revealed that the nanogroove/ridge widths were replicated perfectly, although at deeper levels the grooves became more concave. The smooth substrata had no distinguishable pattern other than a roughness amplitude of 1 nm. Interestingly, microscopy and image analysis showed that fibroblast after 4h had adjusted their shape according to nanotopographical features down to cut-off values of 100 nm width and 75 nm depth. After 24h culturing time, fibroblasts would even align themselves on groove depths as shallow as 35 nm. It appears depth is the most essential parameter in cellular alignment on groove patterns with a pitch ratio of 1: 1. On the smooth substrata, cells always spread out in a random fashion. Analysis of variance (ANOVA) demonstrated that both main parameters, topography and culturing time, were significant. We conclude that fibroblast cells cultured on nanotopography experience a threshold feature size of 35 nm, below this value contact guidance does no longer exist. (C) 2007 Elsevier Ltd. All rights reserved.