Effect of crosslinking chemistry of albumin/heparin multilayers on FGF-2 adsorption and endothelial cell behavior
Abstract
The biomaterials that efficiently deliver growth factors represent an important challenge in cell-based tissue engineering. Layer-by-layer (LbL) thin films are attractive for incorporating controlled amounts of growth factors and releasing them over time.
Herein, we investigated the effect of a method of cross linking of albumin/heparin layer-by-layer (LbL) assembly ((Alb/Hep)(3)) on the loading and release of basic fibroblast growth factor (FGF-2), and subsequent proliferation of human endothelial cells (HUVECs). The (Alb/Hep)3 assemblies were cross-linked using glutaraldehyde, reductive amination or carbodiimide chemistries, and then biofunctionalized with FGF-2.
The (Alb/Hep)3 assemblies were characterized by the infrared multi-internal reflection spectroscopy, atomic force microscopy, ellipsometry, and surface plasmon resonance (SPR). The FGF-2 loading was quantified by the SPR in situ analysis.
Our results showed that the (Alb/Hep)(3) cross-linking affected the amount of the bound heparin (from 150 to 315 ng/cm(2)), amount of FGF-2 loaded (from 75 to 125 ng/cm(2)), FGF-2 release (from 15 to 53% over 8 days), and consequently the HUVEC cell proliferation (from 50 to 80 x 10(3) cells/cm(2) at day 5). All FGF-2 loaded assemblies stimulated the cell growth more than a soluble FGF-2 added into the cell media.
In particular, the highest HUVECs proliferation was detected on the carbodiimide-cross-linked assembly. Overall, these biocompatible cross-linked assemblies can fine-tune the loading and release of growth factor providing a platform for cell-contacting applications.