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.

Perfusion flow bioreactor for 3D in situ imaging: investigating cell/biomaterials interactions.

The capability to image real time cell/material interactions in a three-dimensional (3D) culture environment will aid in the advancement of tissue engineering. This paper describes a perfusion flow bioreactor designed to hold tissue engineering scaffolds and allow for in situ imaging using an upright microscope. The bioreactor can hold a scaffold of desirable thickness for implantation (>2 mm). Coupling 3D culture and perfusion flow leads to the creation of a more biomimetic environment. We examined the ability of the bioreactor to maintain cell viability outside of an incubator environment (temperature and pH stability), investigated the flow features of the system (flow induced shear stress), and determined the image quality in order to perform time-lapsed imaging of two-dimensional (2D) and 3D cell culture. In situ imaging was performed on 2D and 3D, culture samples and cell viability was measured under perfusion flow (2.5 mL/min, 0.016 Pa). The visualization of cell response to their environment, in real time, will help to further elucidate the influences of biomaterial surface features, scaffold architectures, and the influence of flow induced shear on cell response and growth of new tissue.

Interfacial shear strengths of dental resin-glass fibers by the microbond test.

OBJECTIVES: The aim of this study was to investigate the feasibility of using the microbond test (MBT) to probe the durability of the bond between a polymerized dental resin with differently silanized E-glass fibers. METHODS: The E-glass fibers were silanized with equivalent amounts of two types of acrylic-silane coupling agents: 3-methacryloxypropyltrimethoxysilane (MPTMS) and 10-methacryloxydecyltrimethoxysilane (MDTMS), a more hydrophobic silane coupling agent than MPTMS. Unsilanized E-glass fibers were used as the control. Microdroplets of a photo-activated dental resin were applied on the fiber and photocured with visible light irradiation (470 nm). Subsequently, the specimens were tested in shear after 24h storage in air at 23 degrees C or water at 60 degrees C. RESULTS: The mean interfacial shear strength (tau) and the standard deviation in MPa for the three systems in 23 degrees C in air (n>7) were: 33.8(10.1), 33.7(8.9) and 15.3(4.2) for the MPTMS silanized, MDTMS silanized, and unsilanized fibers, respectively. When the three types of fibers were first exposed to 60 degrees C water for 24h prior to having the microdroplets of the resin bonded to them, the strength values of the MDTMS silanized fibers and the control fibers remained essentially unchanged at (n> or =7) 31.8(7.7) and 17.5(4.9)MPa respectively; the MPTMS specimens showed a significant decrease to 15.8(4.8)MPa. Similar trends were observed when the fibers had microdroplets of the resin bonded to them prior to aqueous exposure. SIGNIFICANCE: These results indicate that the microbond test has the sensitivity to measure changes at the interface between polymerized dental resins and variously silanized E-glass fibers. It appears that surface modification of the fibers with the more hydrophobic silane coupling agent MDTMS promotes enhanced resistance to degradation from exposure to water. The microbond test has the potential for studying dental adhesion involving small bonded areas under a variety of conditions with different adhesive systems and substrates.