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

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• 作者：Andriy Shkilnyy^a ; Pierre Proulx^b ; Jamie Sharp^a ; Martin Lepage^c ; Patrick Vermette^a ; ^{patrick.vermette@usherbrooke.ca}
• 关键词：Diffusion ; Rhodamine B ; Bovine serum albumin (BSA) ; Fluorescence ; Fibrin gel ; Endothelial cells (HUVEC)
• 刊名：Colloids and Surfaces B ; Biointerfaces
• 出版年：2012
• 期刊代码：14_09277765
• 类别：ms
• 出版时间：1 May, 2012
• 卷：93
• 期：Complete
• 页码：202-207
• 文件大小：586 K

摘要

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^庐 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 脳 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 脳 10⁵ cells/mL) has almost no effect on the diffusion coefficients. Higher cell densities (>5 脳 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.