Diffusion of rhodamine B and bovine serum albumin in fibrin gels seeded with primary endothelial cells

Scaffolds with adequate mass transport properties are needed in many tissue engineering applications. Fibrin is considered a good biological material to fabricate such scaffolds. However, very little is known about mass transport in fibrin. Therefore, a method based on the analysis of fluorescence intensity for measuring the apparent diffusion coefficient of rhodamine B and fluorescein-labelled bovine serum albumin (FITC-BSA) is described. The experiments are performed in fibrin gels with and without human umbilical vein endothelial cells (HUVEC). The apparent diffusion coefficients of rhodamine B and FITC-BSA in fibrin (fibrinogen concentration of 4 mg/mL) with different cell densities are reported. A LIVE/DEAD® assay is performed to confirm the viability of HUVEC seeded at high densities. Diffusion coefficients for rhodamine B remain more or less constant up to 5 × 105 cells/mL and correlate well with literature values measured by other methods in water systems. This indicates that the presence of HUVEC in the fibrin gels (up to 5 × 105 cells/mL) has almost no effect on the diffusion coefficients. Higher cell densities (>5 × 105 cells/mL) result in a decrease of the diffusion coefficients. Diffusion coefficients of rhodamine B and FITC-BSA obtained by this method agree with diffusion coefficients in water predicted by the Stokes–Einstein equation. The experimental design used in this study can be applied to measure diffusion coefficients in different types of gels seeded or not with living cells.

Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► A technique for measuring diffusion of fluorescent dyes and proteins is described. ► Diffusion coefficients of rhodamine B and BSA in fibrin are estimated. ► Endothelial cell seeding densities can affect rhodamine B diffusion coefficient. ► Experimental results are compared to theoretical and literature values.