Extended interaction of beta1 integrin subunit-deficient cells (GD25) with surfaces modified with fibronectin-derived peptides: Culture optimization, adhesion and cytokine panel studies.
The modification of biomaterials with extracellular matrix-mimicking factors is a common technique used to influence the cellular response through integrin-mediated signaling. The inherent limitations of antibody-inhibition studies necessitate the use of complementary methods to block integrin function to confirm cell-surface interaction. In this study, we employed a beta1 integrin-deficient cell line, GD25, to investigate the role of beta1 subunit in cell adhesion and subsequent cytokine (granulocyte macrophage colony stimulating factor; interleukin (IL)-1alpha; IL-1beta; IL-6; monocyte chemoattractant protein-1; regulated upon activation, normal T-cell expressed, and secreted; tumor necrosis factor-alpha) release kinetics in the presence of tissue culture polystyrene (TCPS) and semi-interpenetrating polymer networks (sIPN) modified with fibronectin (FN)-mimic peptides (RGD, PHSRN). Culture conditions (i.e. seeding density, medium, serum supplementation) were optimized for long-term observation. Differences in cell adhesion, cell viability and cytokine release behavior were dependent on the presence of the beta1 integrin subunit, FN, sIPN cast method and peptide identity. By comparing two complementary techniques for assaying integrin function, we observed both similarities (i.e. decreased adhesion to FN-absorbed TCPS and increased IL-1beta release at 96h) and differences (i.e. no difference in adhesion or IL-1beta release in the presence of different sIPN surfaces) when the function of the beta1 subunit was blocked in cell adhesion and signaling in the presence of biomaterials.
Department of Biomedical Engineering, College of Engineering, University of Wisconsin, Madison, WI 53705, USA.
Heather
H
Waldeck
Weiyuan John
WJ
Kao
D000818:Animals; D040761:Biomimetic Materials / Q000737:chemistry*; D002448:Cell Adhesion / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D020099:Coated Materials, Biocompatible / Q000737:chemistry / Q000494:pharmacology*; D016207:Cytokines / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D005353:Fibronectins / Q000737:chemistry / Q000494:pharmacology*; D019012:Integrin beta1 / Q000378:metabolism*; D008422:Materials Testing; D051379:Mice; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D013499:Surface Properties; D023822:Tissue Engineering / Q000379:methods
D020099:Coated Materials, Biocompatible; D016207:Cytokines; D005353:Fibronectins; D019012:Integrin beta1; D010455:Peptides