Lithography Technique for Topographical Micropatterning of Collagen-Glycosaminoglycan Membranes for Tissue Engineering Applications

[+] Author and Article Information
Vijayakumar Janakiraman, Brian L. Kienitz

Department of Chemical Engineering, Case Western Reserve University, Cleveland, OH 44106

Harihara Baskaran1

Department of Chemical Engineering and Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH 44106hari@case.edu


Corresponding author.

J. Med. Devices 1(3), 233-237 (Jul 10, 2007) (5 pages) doi:10.1115/1.2775937 History: Received January 25, 2007; Revised July 10, 2007

An adaptable technique for micropatterning biomaterial scaffolds has enormous implications in controlling cell function and in the development of tissue-engineered (TE) microvasculature. In this paper, we report a technique to embed microscale patterns onto a collagen-glycosaminoglycan (CG) membrane as a first step toward the creation of TE constructs with built-in microvasculature. The CG membranes were fabricated by homogenizing a solution of type-I bovine collagen and chondroitin-6-sulfate in acetic acid and vacuum filtering the solution subsequently. The micropatterning technique consisted of three steps: surface dissolution of base matrix using acetic acid solution, feature resolution by application of uniform pressure, and feature stability by glutaraldehyde cross-linking. Application of the new technique yielded patterns in CG membranes with a spatial resolution on the order of 23μm. We show that such a patterned matrix is conducive to the attachment of bovine aortic endothelial cells. The patterned membranes can be used for the development of complex three-dimensional TE products with built-in flow channels, as templates for topographically directed cell growth or as a model system to study various microvascular disorders where feature scales are important. The new technique is versatile; topographical patterns can be custom made for any predetermined design with high spatial resolution, and the technique itself can be adapted for use with other scaffold materials.

Copyright © 2007 by American Society of Mechanical Engineers
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Figure 2

(A) SEM image of flow networks cast onto a CG membrane using micropatterning. (B) SEM image of the smallest “island” on the patterned CG membrane. (C) corresponding AUTOCAD design of the flow networks used for micropatterning. (D) SEM image of the individual collagen fibers on the surface of a patterned CG membrane (with inset at higher magnification).

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Figure 3

(A) Fluorescent image of the cross section of a patterned CG membrane perpendicular to the channel direction, showing the morphological characteristics of the patterned features. The insets show the wide and narrow width channels at higher magnification. (B) SEM image of bovine aortic endothelial cells (BAEC’s) seeded onto a sample patterned CG flow channel network (the pattern is marked by an arrow). (C) SEM image of an individual cell with extension of processes and establishment of cell-cell junctions.

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Figure 1

Schematic of the patterning technique. (1) Surface dissolution: Acetic acid is applied to the CG membrane surface. (2) Feature resolution: After the top layer of the CG membrane has been dissolved, the silicon wafer with photoresist is positioned on the membrane surface and pressure is applied. (3) Feature stabilization: The system is immersed in glutaraldehyde to cross-link the membrane and to stabilize the patterned features. (4) Separation: The silicon wafer and CG membrane are separated resulting in a patterned CG membrane.




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