The interaction of cells with solid surfaces is important in many settings, including the response of tissue to implanted materials. Protein adsorption to the surface plays a critical role in controlling cell interactions with surfaces. However, few comprehensive studies of both cell behavior and protein adsorption in complex protein mixtures (e.g., serum) have been done so the connection between these events is not well understood. In particular, methods to systematically perturb both protein adsorption and cell behavior in order to understand their relationship have been lacking. To induce changes in cell and protein behavior, the effects of serum dilution and substrate surface chemistry were studied. Surface chemistry was varied by using a series of polymers and copolymers of hydroxyethyl methacrylate (HEMA) and ethylmethacrylate (EMA) varying in their hydrophobic/hydrophilic balance. Large changes in cell spreading and fibronectin adsorption were observed when either serum concentration or polymer type was varied. The spreading of 3T3 cells in serum was found to be well correlated with the amount of fibronectin adsorption to the substrates. Attachment was not correlated with fibronectin adsorption, especially on glass preadsorbed with diluted serum. For 3T3 cells and perhaps other cells that have a receptor for a protein which is present in the medium, the amount of adsorption of this protein to the substrate appears to be a critical factor controlling cell interactions with the substrate.
A trypan blue inclusion assay was used to measure cell death on poly(dimethyl siloxane) (PDMS), polyethylene (PE), poly(methyl methacrylate) (PMMA), polyurethanes, glass, and glow-discharge-treated polystyrene or poly(ethylene terephthalate). Cell lines used were bovine aortic endothelial, 3T3, mouse peritoneal macrophage, and BHK cells. In the absence of proteins in the media, PDMS, PE, PMMA, and some polyurethanes were consistently found to induce cell death. This toxic effect disappeared if the cells were seeded in serum-containing medium or if concentrated solutions of proteins (albumin, IgG, or fibronectin) were preadsorbed on the materials. The substrate toxicity appeared to be due to the physical properties of the substrate and not to the release of toxic leachables.
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