title: A fluorophore-tagged RGD peptide to control endothelial cell adhesion to micropatterned surfaces.
abstract: The long-term patency rates of vascular grafts and stents are limited by the lack of surface endothelialisation of the implanted materials. We have previously reported that GRGDS and WQPPRARI peptide micropatterns increase the endothelialisation of prosthetic materials in vitro. To investigate the mechanisms by which the peptide micropatterns affect endothelial cell adhesion and proliferation, a TAMRA fluorophore-tagged RGD peptide was designed. Live cell imaging revealed that the micropatterned surfaces led to directional cell spreading dependent on the location of the RGD-TAMRA spots. Focal adhesions formed within 3 h on the micropatterned surfaces near RGD-TAMRA spot edges, as expected for cell regions experiencing high tension. Similar levels of focal adhesion kinase phosphorylation were observed after 3 h on the micropatterned surfaces and on surfaces treated with RGD-TAMRA alone, suggesting that partial RGD surface coverage is sufficient to elicit integrin signaling. Lastly, endothelial cell expansion was achieved in serum-free conditions on gelatin-coated, RGD-TAMRA treated or micropatterned surfaces. These results show that these peptide micropatterns mainly impacted cell adhesion kinetics rather than cell proliferation. This insight will be useful for the optimization of micropatterning strategies to improve vascular biomaterials.
authors.affiliation: Centre de Recherche sur les Matériaux Avancés, Département de génie des mines, de la métallurgie et des matériaux, Université Laval, Québec G1V 0A6, Canada; Centre de recherche du CHU de Québec, Hôpital Saint-François d'Assise, Québec G1L 3L5, Canada; PROTEO Research Center and Département de génie chimique, Université Laval, Québec G1V 0A6, Canada. Electronic address: corinne.hoesli@gmail.com.
authors.forename: Corinne A
authors.initials: CA
authors.lastname: Hoesli
authors.forename: Alain
authors.initials: A
authors.lastname: Garnier
authors.forename: Pierre-Marc
authors.initials: PM
authors.lastname: Juneau
authors.forename: Pascale
authors.initials: P
authors.lastname: Chevallier
authors.forename: Carl
authors.initials: C
authors.lastname: Duchesne
authors.forename: Gaétan
authors.initials: G
authors.lastname: Laroche
mesh_terms: D001807:Blood Vessel Prosthesis; D002448:Cell Adhesion; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D042783:Endothelial Cells / Q000166:cytology*; D005353:Fibronectins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D009842:Oligopeptides / Q000737:chemistry*; D012235:Rhodamines / Q000737:chemistry*
chemical_list: C114557:5-carboxytetramethylrhodamine succinimidyl ester; D020099:Coated Materials, Biocompatible; D005353:Fibronectins; D005456:Fluorescent Dyes; D009842:Oligopeptides; D012235:Rhodamines; C417707:fibronectin peptide V; C047981:arginyl-glycyl-aspartic acid; C049554:glycyl-arginyl-glycyl-aspartyl-serine
keywords: Biomimetic material; Cell spreading; Endothelialisation; Micropatterning; Surface grafting; Vascular grafts