A tubular, hierarchically structured hybrid vascular tissue composed of vascular cells and collagen was prepared. First, a cold mixed solution of bovine aortic smooth muscle cells (SMCs) and Type I collagen was poured into a tubular glass mold composed of a mandrel and a sheath (example of dimensions: inner diameter, 1.5 mm; outer diameter, 7 mm; length, 7 cm). Upon incubation at 37°C, an SMC-incorporated collagenous gel was formed. After the sheath was removed, the resulting fragile tissue, when cultured in medium, thinned in a time-dependent manner to form an opaque, dense tissue. Higher SMC seeding density and lower initial collagen concentration induced more rapid and prominent shrinkage of the tissue. Morphologic investigation showed that over time, bipolarly elongated SMCs and collagen fiber bundles became positioned around the mandrel. Both components became circumferentially oriented. When the mandrel was removed, a tubular hybrid medial tissue was formed. A hybrid vascular tissue with a hierarchical structure was constructed by seeding endothelial cells onto the inner surface of the hybrid medial tissue. Prepared tissues tolerated luminal pressures as great as 100 mmHg and mechanical stress applied during an anastomotic procedure. This method allowed us to prepare a tubular hybrid medial tissue of predetermined size (inner diameter, wail thickness, and length) by selecting appropriate mold design, initial collagen concentration, and SMC seeding density. Such hybrid vascular tissues may provide physiological functions when implanted into the venous system.
Functional role of cGMP-dependent VASP phosphorylation in vascular cells
