title: Subcellular stretch-induced cytoskeletal response of single fibroblasts within 3D designer scaffolds.
abstract: In vivo, cells are exposed to mechanical forces in many different ways. These forces can strongly influence cell functions or may even lead to diseases. Through their sensing machinery, cells are able to perceive the physical information of the extracellular matrix and translate it into biochemical signals resulting in cellular responses. Here, by virtue of two-component polymer scaffolds made via direct laser writing, we precisely control the cell matrix adhesions regarding their spatial arrangement and size. This leads to highly controlled and uniform cell morphologies, thereby allowing for averaging over the results obtained from several different individual cells, enabling quantitative analysis. We transiently deform these elastic structures by a micromanipulator, which exerts controlled stretching forces on primary fibroblasts grown in these scaffolds on a subcellular level. We find stretch-induced remodeling of both actin cytoskeleton and cell matrix adhesions. The responses to static and periodic stretching are significantly different. The amount of paxillin and phosphorylated focal adhesion kinase increases in cell matrix adhesions at the manipulated pillar after static stretching whereas it decreases after periodic stretching.
authors.affiliation: Institute of Applied Physics (APH), Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe, Germany.
authors.forename: Andrea C
authors.initials: AC
authors.lastname: Scheiwe
authors.affiliation: Institute of Functional Interfaces (IFG), Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe, Germany.
authors.forename: Stephanie C
authors.initials: SC
authors.lastname: Frank
authors.affiliation: Institute of Functional Interfaces (IFG), Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe, Germany.
authors.forename: Tatjana J
authors.initials: TJ
authors.lastname: Autenrieth
authors.affiliation: Institute of Functional Interfaces (IFG), Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe, Germany; Zoological Institute, Department of Cell and Neurobiology, Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe, Germany. Electronic address: bastmeyer@kit.edu.
authors.forename: Martin
authors.initials: M
authors.lastname: Bastmeyer
authors.affiliation: Institute of Applied Physics (APH), Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe, Germany; DFG-Center for Functional Nanostructures (CFN), and Institute of Nanotechnology (INT), Karlsruhe Institute of Technology (KIT), 76128 Karlsruhe, Germany. Electronic address: martin.wegener@kit.edu.
authors.forename: Martin
authors.initials: M
authors.lastname: Wegener
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena; D049109:Cell Proliferation; D021982:Cell-Matrix Junctions / Q000378:metabolism; D002642:Chick Embryo; D003599:Cytoskeleton / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology*; D008846:Micromanipulation; D013314:Stress, Mechanical; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors; D054457:Tissue Scaffolds / Q000737:chemistry*
keywords: Cell adhesion; Direct laser writing; Focal contact; Mechanical properties; Scaffolds