Effect of surface modification on protein retention and cell proliferation under strain.

When culturing cells on flexible surfaces, it is important to consider extracellular matrix treatments that will remain on the surface under mechanical strain. Here we investigate differences in laminin deposited on oxidized polydimethylsiloxane (PDMS) with plasma treatment (plasma-only) vs. plasma and aminopropyltrimethoxysilane treatment (silane-linked). We use specular X-ray reflectivity (SXR), transmission electron microscopy (TEM), and immunofluorescence to probe the quantity and uniformity of laminin. The surface coverage of laminin is approximately 45% for the plasma-only and 50% for the silane-linked treatment as determined by SXR. TEM and immunofluorescence reveal additional islands of laminin aggregates on the plasma-only PDMS compared with the relatively smooth and uniform silane-linked laminin surface. We also examine laminin retention under strain and vascular smooth muscle cell viability and proliferation under static and strain conditions. Equibiaxial stretching of the PDMS surfaces shows greatly improved retention of the silane-linked laminin over plasma-only. There are significantly more cells on the silane-linked surface after 4 days of equibiaxial strain.

Evaluation of polydimethylsiloxane modification methods for cell response.

Many methods exist in the literature to modify surfaces with extracellular matrix (ECM) proteins prior to cell seeding. However, there are few studies that systematically characterize and compare surface properties and cell response results among modification methods that use different bonding mechanisms. In this work, we compare cell response and physical characterization results from fibronectin or laminin attached to polydimethylsiloxane (PDMS) elastomer surfaces by physical adsorption, chemisorption, and covalent attachment to determine the best method to modify a deformable surface. We evaluate modification methods based on completeness and uniformity of coverage, surface roughness, and hydrophilicity of attached ECM protein. Smooth muscle cell adhesion, proliferation, morphology, and phenotype were also evaluated. We found that chemisorption methods resulted in higher amounts of protein attachment than physical adsorption and covalent bonding of the ECM proteins. Cell response to protein-modified surfaces was similar with respect to cell adhesion, area, aspect ratio, and phenotype. When all the data are considered, the chemisorption methods, most notably silane_70, provide the best surface properties and highest cell proliferation.