title: Cell shape and the presentation of adhesion ligands guide smooth muscle myogenesis.
abstract: The reliable generation of smooth muscle cells is important for a number of tissue engineering applications. Human mesenchymal stem cells (MSCs) are a promising progenitor of smooth muscle, with high expression of smooth muscle markers observed in a fraction of isolated cells, which can be increased by introduction of soluble supplements that direct differentiation. Here we demonstrate a new micropatterning technique, where peptides of different ligand affinity can be microcontact printed onto an inert background, to explore MSC differentiation to smooth muscle through controlled biochemical and biophysical cues alone. Using copper-catalyzed alkyne-azide cycloaddition (CuAAC), we patterned our surfaces with RGD peptide ligands-both a linear peptide with low integrin affinity and a cyclic version with high integrin affinity-for the culture of MSCs in shapes with various aspect ratios. At low aspect ratio, ligand affinity is a prime determinant for smooth muscle differentiation, while at high aspect ratio, ligand affinity has less of an effect. Pathway analysis reveals a role for focal adhesion turnover, Rac1, RhoA/ROCK, and calpain during smooth muscle differentiation of MSCs in response to cell shape and the affinity of the cell adhesion interface. Controlling integrin-ligand affinity at the biomaterials interface is important for mediating adhesion but may also prove useful for directing smooth muscle myogenesis. Peptide patterning enables the systematic investigation of single to multiple peptides derived from any protein, at different densities across a biomaterials surface, which has the potential to direct multiple MSC differentiation outcomes without the need for soluble supplements.
authors.affiliation: Department of Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois.
authors.forename: Douglas
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Sun
authors.affiliation: Department of Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois.
authors.forename: Junmin
authors.initials: J
authors.lastname: Lee
authors.affiliation: Department of Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois.
authors.forename: Amr A
authors.initials: AA
authors.lastname: Abdeen
authors.affiliation: Department of Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois.
authors.forename: Kristopher A
authors.initials: KA
authors.lastname: Kilian
mesh_terms: D001672:Biocompatible Materials / Q000737:chemistry* / Q000378:metabolism; D002448:Cell Adhesion; D002454:Cell Differentiation; D002460:Cell Line; D048430:Cell Shape; D022001:Focal Adhesions / Q000378:metabolism; D006801:Humans; D016023:Integrins / Q000378:metabolism; D008024:Ligands; D040542:Mechanotransduction, Cellular; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D024510:Muscle Development; D009130:Muscle, Smooth / Q000166:cytology* / Q000254:growth & development*; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D010456:Peptides, Cyclic / Q000737:chemistry / Q000378:metabolism; D013499:Surface Properties; D023822:Tissue Engineering
chemical_list: D001672:Biocompatible Materials; D016023:Integrins; D008024:Ligands; D009842:Oligopeptides; D010455:Peptides; D010456:Peptides, Cyclic; C507169:cyclic arginine-glycine-aspartic acid peptide; C047981:arginyl-glycyl-aspartic acid
keywords: mesenchymal stem cells; microcontact printing; peptides; self-assembled monolayers; smooth muscle