title: Nano-stenciled RGD-gold patterns that inhibit focal contact maturation induce lamellipodia formation in fibroblasts.
abstract: Cultured fibroblasts adhere to extracellular substrates by means of cell-matrix adhesions that are assembled in a hierarchical way, thereby gaining in protein complexity and size. Here we asked how restricting the size of cell-matrix adhesions affects cell morphology and behavior. Using a nanostencil technique, culture substrates were patterned with gold squares of a width and spacing between 250 nm and 2 µm. The gold was functionalized with RGD peptide as ligand for cellular integrins, and mouse embryo fibroblasts were plated. Limiting the length of cell-matrix adhesions to 500 nm or less disturbed the maturation of vinculin-positive focal complexes into focal contacts and fibrillar adhesions, as indicated by poor recruitment of α5-integrin. We found that on sub-micrometer patterns, fibroblasts spread extensively, but did not polarize. Instead, they formed excessive numbers of lamellipodia and a fine actin meshwork without stress fibers. Moreover, these cells showed aberrant fibronectin fibrillogenesis, and their speed of directed migration was reduced significantly compared to fibroblasts on 2 µm square patterns. Interference with RhoA/ROCK signaling eliminated the pattern-dependent differences in cell morphology. Our results indicate that manipulating the maturation of cell-matrix adhesions by nanopatterned surfaces allows to influence morphology, actin dynamics, migration and ECM assembly of adhering fibroblasts.
authors: Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
authors: Roman
authors: R
authors: Lutz
authors: Kristopher
authors: K
authors: Pataky
authors: Neha
authors: N
authors: Gadhari
authors: Mattia
authors: M
authors: Marelli
authors: Juergen
authors: J
authors: Brugger
authors: Matthias
authors: M
authors: Chiquet
mesh_terms: D000818:Animals; D002465:Cell Movement / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D005353:Fibronectins / Q000737:chemistry; D022001:Focal Adhesions / Q000187:drug effects* / Q000378:metabolism; D006046:Gold / Q000737:chemistry*; D039081:Integrin alpha5beta1 / Q000378:metabolism; D051379:Mice; D049329:Nanostructures / Q000737:chemistry*; D036103:Nanotechnology; D009842:Oligopeptides / Q000737:chemistry* / Q000494:pharmacology*; D010316:Particle Size; D055503:Protein Multimerization / Q000187:drug effects; D011554:Pseudopodia / Q000235:genetics*; D054460:rho-Associated Kinases / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
chemical_list: D005353:Fibronectins; D039081:Integrin alpha5beta1; D009842:Oligopeptides; D006046:Gold; C047981:arginyl-glycyl-aspartic acid; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein