Teixeira Ana I, Abrams George A, Bertics Paul J, Murphy Christopher J, Nealey Paul F
Department of Chemical Engineering, School of Veterinary Medicine, University of Wisconsin, Madison, WI 53706, USA.
J Cell Sci. 2003 May 15;116(Pt 10):1881-92. doi: 10.1242/jcs.00383.
The human corneal basement membrane has a rich felt-like surface topography with feature dimensions between 20 nm and 200 nm. On the basis of these findings, we designed lithographically defined substrates to investigate whether nanotopography is a relevant stimulus for human corneal epithelial cells. We found that cells elongated and aligned along patterns of grooves and ridges with feature dimensions as small as 70 nm, whereas on smooth substrates, cells were mostly round. The percentage of aligned cells was constant on substrate tomographies with lateral dimensions ranging from the nano- to the micronscale, and increased with groove depth. The presence of serum in the culture medium resulted in a larger percentage of cells aligning along the topographic patterns than when no serum was added to the basal medium. When present, actin microfilaments and focal adhesions were aligned along the substrate topographies. The width of the focal adhesions was determined by the width of the ridges in the underlying substrate. This work documents that biologic length-scale topographic features that model features encountered in the native basement membrane can profoundly affect epithelial cell behavior.
人角膜基底膜具有丰富的毡状表面形貌，其特征尺寸在20纳米至200纳米之间。基于这些发现，我们设计了光刻定义的基底，以研究纳米形貌是否是人类角膜上皮细胞的相关刺激因素。我们发现，细胞沿特征尺寸小至70纳米的沟槽和脊图案延长并排列，而在光滑基底上，细胞大多呈圆形。在横向尺寸从纳米级到微米级的基底断层扫描上，排列细胞的百分比是恒定的，并且随着沟槽深度增加。与基础培养基中不添加血清时相比，培养基中血清的存在导致更大比例的细胞沿形貌图案排列。当存在时，肌动蛋白微丝和粘着斑沿基底形貌排列。粘着斑的宽度由下层基底中脊的宽度决定。这项工作证明，模拟天然基底膜中遇到的特征的生物长度尺度形貌特征可深刻影响上皮细胞行为。
