Collagen Matrix Alignment Using Inkjet Printer Technology
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Collagen Matrix Alignment Using Inkjet Printer Technology Sandra Deitch1, Catherine Kunkle2, Xiaofeng Cui1, Thomas Boland1, and Delphine Dean1 1 Bioengineering, Clemson University, Clemson, SC, 29634 2 Presbyterian College, Clinton, SC, 29325 ABSTRACT Collagen fiber orientation plays an important role in many cell properties and actions in vivo. While it is easy to replicate randomly oriented collagen in vitro, it is much more difficult to create aligned collagen matrices for cell culture. In this work, a novel inkjet printer-based collagen alignment technique was established. A collagen type I solution was printed in a line pattern onto glass substrates using a modified inkjet printer. Staining studies indicated that the heat involved in the printing process is not great enough to denature the collagen. The extent of alignment was observed using light microscopy, atomic force microscopy (AFM), and polarized light microscopy. Additionally, cardiomyocytes, which require extracellular matrix alignment to maintain their in vivo phenotype, were cultured on the aligned matrices. The cells grew along the lines of collagen and coordinated beating, indicating the success of the aligned matrix. This collagen alignment technique is cheap, fast, precise, and easy to use in comparison to other current techniques. It may be used to align other extracellular matrix proteins and could even be used to create a three dimensional construct with varying fiber orientations. INTRODUCTION In tissue engineering, it is important to mimic in vivo physiological conditions in the in vitro environment as closely as possible. The extracellular matrix (ECM) has been shown to affect cell growth, adhesion, differentiation, morphology, and signaling and is thus an important part of the cellular environment that should be replicated in cell culture [1, 2]. Collagen type I, a triple helical protein, is a major constituent of many ECM and has long been used in cell culture as an approximation to the ECM. The protein is 1.5 nm in diameter and 300 nm in length. It undergoes self-assembly into fibrils with a diameter of 36 nm, which in turn form larger fibers and fiber bundles [3, 4]. Although fibrils have a persistence length of approximately 130 nm, the fiber bundles are known to have much greater stiffness and appear straight over long distances particularly in tissue [5]. Collagen fibers organize in different orientations in different tissues in the body, affecting the tissue properties. For example, the fibers organize into extensive parallel arrays in tendons and ligaments, regular sheets of varying fiber orientation in the transparent cornea of the eye, and randomly in isotropic connective tissues [6]. Aligned fibers are also believed to play a role in cell signaling and development as well as directing cell proliferation and migration after injury [7, 8]. Therefore, it is important to orient the collagen fibers in cell culture to match the in vivo environment for the particular cell type under investigation. For example, some cell t
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