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Structural characterization of porous tissue scaffolds is challenging due to their complexity. Most investigators report the porosity of the material together with an estimate of the mean pore size and the pore size distribution. The usefulness of these measures is limited, especially in predicting the time-dependent permeation characteristics of a biodegradable, cell-seeded scaffold. A potential solution to this problem is to measure the permeability of the matrix and determine the Darcy permeability coefficient. Darcy permeability coefficients of 3.1 x 10(-14) and 6.3 x 10(-14) m(2) were measured for air and water, respectively, in microporous polycaprolactone scaffolds by monitoring fluid flow in response to a range of pressure differentials. Permeability coefficients for phosphate-buffered saline (5.3 x 10(-14) m(2)), glucose (5.7 x 10(-14) m(2)), and bovine serum albumin (1.8 x 10(-14) m(2)) were obtained by monitoring the change in concentration of molecular probes. This approach revealed the efficiency of transport of glucose molecules through the porous material and the existence of protein-scaffold interactions that resulted in protein retention and a reduction in fluid permeation rate. Darcy permeability measurements can provide valuable insights concerning the transport properties of nutrients, metabolites, and polypeptide growth factors in porous tissue engineering scaffolds and a method of quality assurance in scaffold processing.

Poly(epsilon-caprolactone) (PCL) fibers produced by wet spinning from solutions in acetone under lowshear (gravity-flow) conditions resulted in fiber strength of 8 MPa and stiffness of 0.08 Gpa. Cold drawing to an extension of 500% resulted in an increase in fiber strength to 43 MPa and stiffness to 0.3 GPa. The growth rate of human umbilical vein endothelial cells (HUVECs) (seeded at a density of 5 x 10(4) cells/mL) on as-spun fibers was consistently lower than that measured on tissue culture plastic (TCP) beyond day 2. Cell proliferation was similar on gelatin-coated fibers and TCP over 7 days and higher by a factor of 1.9 on 500% cold-drawn PCL fibers relative to TCP up to 4 days. Cell growth on PCL fibers exceeded that on Dacron monofilament by at least a factor of 3.7 at 9 days. Scanning electron microscopy revealed formation of a cell layer on samples of cold-drawn and gelatin-coated fibers after 24 hours in culture. Similar levels of ICAM-1 expression by HUVECs attached to PCL fibers and TCP were measured using RT-PCR and flow cytometry, indicative of low levels of immune activation. Retention of a specific function of HUVECs attached to PCL fibers was demonstrated by measuring their immune response to lipopolysaccharide. Levels of ICAM-1 expression increased by approximately 11% in cells attached to PCL fibers and TCP. The high fiber compliance, favorable endothelial cell proliferation rates, and retention of an important immune response of attached HUVECS support the use of gravity spun PCL fibers for three-dimensional scaffold production in vascular tissue engineering.