Dynamic Substrates for Cell Adhesion & Motility Studies

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DESCRIPTION (provided by applicant): Synthetic materials design for applications such as tissue engineering has benefited from increased understanding of the cell-substrate adhesive interactions that occur during clinical events such as wound healing, tissue morphogenesis, and biomedical device assimilation. Interactions that occur in the context of such events are dynamic, and the receptors and ligands involved in receptor-mediated adhesion possess a high degree of spatial and temporal flexibility. The work proposed seeks to develop a modified polymer hydrogel surface with dynamic adhesion properties utilizing a protein dimer, DF2t1, designed de novo. One protein molecule of the dimer will be functionalized with the common cell-binding domain, RGD; the other molecule will be conjugated to the surface of polyacrylamide gels using a multicomponent heterobifunctional linking scheme. Adhesivity of the surface to cultured bovine aortic endothelial cells (BAEC) will be precipitated by dimerization of the RGD containing component of the dimer to its complement on the surface. Uniform surfaces and patterned surfaces will be produced to provide the opportunity to observe cells as they encounter regions of differing adhesivity, and as the adhesive character of the surface is switched "on" and "off" chemically. The cellular response to surface properties under dynamic adhesion conditions will be monitored using quantitative microscopy methods for measuring single cell motility, force traction generation, and cell population migration. The results will be interpreted in the context of endothelial cell migration during clinically relevant processes such as vasculogenesis and angiogenesis.