Characterizing 3t3 Fibroblast Migration across Gradient Peptide Surfaces

This study focuses on the simple preparation of adhesive peptide gradients on glass substrates and observation of their effect on 3T3 fibroblast migration and dispersion. The trends observed using this method will eventually be applied to the design and construction of hydrogel implants for tissue engineering, specifically wound repair. However, gradient peptide hydrogels are more difficult to construct, and cell dispersion would be harder to quantify in a polymer network. Thus, glass slides are used for each trial. The adhesive peptide, RGDS, is attached to an acrylated PEG polymer and covalently bound to the surface via a thiol-acrylate Michael addition reaction with a deposited thiol self-assembled monolayer. The gradient is prepared and then quantified using water contact angles. Cells are seeded onto the gradient surface in 80, 1 mm diameter wells created by a silicone rubber isolator mold. After detaching the mold, cell groups are photographed at 0, 12 and 24 hours to monitor their migration. Ellipses are drawn around each group with Microsoft PowerPoint and each area calculated. The results of this experiment are inconclusive, and the method requires major sources of error to be eliminated before it can be considered viable.