Extended interaction of β1 integrin subunit-deficient cells (GD25) with surfaces modified with fibronectin-derived peptides: culture optimization adhesion and cytokine panel studies

摘要

The modification of biomaterials with extracellular matrix-mimicking factors to influence the cellular response through mainly integrin-mediated signaling is a common technique. 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 β1 integrin-deficient cell line, GD25, to investigate the role of β1 subunit in cell adhesion and subsequent cytokine (granulocyte macrophage colony stimulating factor, interleukin (IL)-1α, IL-1β, IL-6, granulocyte macrophage colony stimulating factor -1, regulated upon activation, normal T-cell expressed, and secreted, tumor necrosis factor-α) 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 β1 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-1β release at 96 h) and differences (i.e. no difference in adhesion or IL-1β release in the presence of sIPN surfaces) when the function of the β1 subunit was blocked in cell adhesion and signaling in the presence of biomaterials.