title: In vitro elastogenesis: instructing human vascular smooth muscle cells to generate an elastic fiber-containing extracellular matrix scaffold.
abstract: Elastic fibers are essential for the proper function of organs including cardiovascular tissues such as heart valves and blood vessels. Although (tropo)elastin production in a tissue-engineered construct has previously been described, the assembly to functional elastic fibers in vitro using human cells has been highly challenging. In the present study, we seeded primary isolated human vascular smooth muscle cells (VSMCs) onto 3D electrospun scaffolds and exposed them to defined laminar shear stress using a customized bioreactor system. Increased elastin expression followed by elastin deposition onto the electrospun scaffolds, as well as on newly formed fibers, was observed after six days. Most interestingly, we identified the successful deposition of elastogenesis-associated proteins, including fibrillin-1 and -2, fibulin-4 and -5, fibronectin, elastin microfibril interface located protein 1 (EMILIN-1) and lysyl oxidase (LOX) within our engineered constructs. Ultrastructural analyses revealed a developing extracellular matrix (ECM) similar to native human fetal tissue, which is composed of collagens, microfibrils and elastin. To conclude, the combination of a novel dynamic flow bioreactor and an electrospun hybrid polymer scaffold allowed the production and assembly of an elastic fiber-containing ECM.
authors.affiliation: Department of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), 70569 Stuttgart, Germany. Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University Tübingen, 72076 Tübingen, Germany.
authors.forename: Svenja
authors.initials: S
authors.lastname: Hinderer
authors.forename: Nian
authors.initials: N
authors.lastname: Shena
authors.forename: Léa-Jeanne
authors.initials: LJ
authors.lastname: Ringuette
authors.forename: Jan
authors.initials: J
authors.lastname: Hansmann
authors.forename: Dieter P
authors.initials: DP
authors.lastname: Reinhardt
authors.forename: Sara Y
authors.initials: SY
authors.lastname: Brucker
authors.forename: Elaine C
authors.initials: EC
authors.lastname: Davis
authors.forename: Katja
authors.initials: K
authors.lastname: Schenke-Layland
mesh_terms: D019149:Bioreactors; D002478:Cells, Cultured; D004547:Elastic Tissue / Q000502:physiology*; D004595:Electroplating; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D066330:Printing, Three-Dimensional / Q000295:instrumentation; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods*; D054457:Tissue Scaffolds