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Cell Adhesion on Micro-Structured Fibronectin Gradients Fabricated by Multiphoton Excited Photochemistry

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Abstract

Concentration gradients of ECM proteins play active roles in many areas of cell biology including wound healing and metastasis. They may also form the basis of tissue engineering scaffolds, as these can direct cell adhesion and migration and promote new matrix synthesis. To better understand cell–matrix interactions on attractive gradients, we have used multiphoton excited (MPE) photochemistry to fabricate covalently linked micro-structured gradients from fibronectin (FN). The gradient design is comprised of a parallel series of individual linear gradients with overall dimensions of approximately 800 × 800 µm, where a linear dynamic range of nearly 10-fold in concentration was achieved. The adhesion dynamics of 3T3 fibroblasts were investigated, where the cell morphology and actin cytoskeleton became increasingly elongated and aligned with the direction of the gradient at increasing protein concentration. Moreover, the cell morphologies are distinct when adhered to regions of differing FN concentration but with similar topography. These results show that the fabrication approach allows investigating the roles of contact guidance and ECM cues on the cell–matrix interactions. We suggest this design overcomes some of the limitations with other fabrication methods, especially in terms of 3D patterning capabilities, and will serve as a new tool to study cell–matrix interactions.

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Acknowledgments

We gratefully acknowledge support under NIH 1U54-RR022232 and NSF CBET--1057766. Y.-D. Su and S.-J. Chen acknowledge support from National Cheng Kung University. We also thank Prof. Vladimir Rodionov for the use of the imaging system for live cell studies.

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Authors and Affiliations

  1. Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, 53717, USA

    Xiyi Chen, Visar Ajeti & Paul J. Campagnola

  2. Department of Engineering Science, National Cheng Kung University, Tainan, Taiwan

    Yuan-Deng Su & Shean-Jen Chen

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  1. Xiyi Chen
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Correspondence to Paul J. Campagnola.

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Associate Editor Joyce Wong oversaw the review of this article.

Xiyi Chen and Yuan-Deng Su contributed equally to this work.

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Chen, X., Su, YD., Ajeti, V. et al. Cell Adhesion on Micro-Structured Fibronectin Gradients Fabricated by Multiphoton Excited Photochemistry. Cel. Mol. Bioeng. 5, 307–319 (2012). https://doi.org/10.1007/s12195-012-0237-8

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