title: Microfluidic Genipin Deposition Technique for Extended Culture of Micropatterned Vascular Muscular Thin Films.
abstract: The chronic nature of vascular disease progression requires the development of experimental techniques that simulate physiologic and pathologic vascular behaviors on disease-relevant time scales. Previously, microcontact printing has been used to fabricate two-dimensional functional arterial mimics through patterning of extracellular matrix protein as guidance cues for tissue organization. Vascular muscular thin films utilized these mimics to assess functional contractility. However, the microcontact printing fabrication technique used typically incorporates hydrophobic PDMS substrates. As the tissue turns over the underlying extracellular matrix, new proteins must undergo a conformational change or denaturing in order to expose hydrophobic amino acid residues to the hydrophobic PDMS surfaces for attachment, resulting in altered matrix protein bioactivity, delamination, and death of the tissues. Here, we present a microfluidic deposition technique for patterning of the crosslinker compound genipin. Genipin serves as an intermediary between patterned tissues and PDMS substrates, allowing cells to deposit newly-synthesized extracellular matrix protein onto a more hydrophilic surface and remain attached to the PDMS substrates. We also show that extracellular matrix proteins can be patterned directly onto deposited genipin, allowing dictation of engineered tissue structure. Tissues fabricated with this technique show high fidelity in both structural alignment and contractile function of vascular smooth muscle tissue in a vascular muscular thin film model. This technique can be extended using other cell types and provides the framework for future study of chronic tissue- and organ-level functionality.
authors.affiliation: Department of Biomedical Engineering, University of Minnesota.
authors.forename: Eric S
authors.initials: ES
authors.lastname: Hald
authors.affiliation: Department of Biomedical Engineering, University of Minnesota.
authors.forename: Kerianne E
authors.initials: KE
authors.lastname: Steucke
authors.affiliation: Department of Biomedical Engineering, University of Minnesota.
authors.forename: Jack A
authors.initials: JA
authors.lastname: Reeves
authors.affiliation: Department of Biomedical Engineering, University of Minnesota.
authors.forename: Zaw
authors.initials: Z
authors.lastname: Win
authors.affiliation: Department of Biomedical Engineering, University of Minnesota; pwalford@umn.edu.
authors.forename: Patrick W
authors.initials: PW
authors.lastname: Alford
mesh_terms: D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D005353:Fibronectins / Q000737:chemistry; D006801:Humans; D039823:Iridoids / Q000737:chemistry*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D009131:Muscle, Smooth, Vascular / Q000166:cytology*
chemical_list: D004129:Dimethylpolysiloxanes; D005353:Fibronectins; D039823:Iridoids; C013830:baysilon; C007834:genipin