Preparation and preliminary in vitro evaluation of a bFGF-releasing heparin-conjugated poly(ε-caprolactone) membrane for guided bone regeneration

Abstract

In an effort to improve guided bone regeneration (GBR), we successfully fabricated a novel basic fibroblast growth factor (bFGF)-releasing heparin-conjugated poly(ε-caprolactone) membrane (hep-PCL/bFGF). This material has a porous microstructure with smooth and rough pore walls before and after heparinization, respectively. Our FTIR analyses indicated that chemical bonds were formed between PCL and heparin with a new amide C=O band at 1660 cm−1 and a band at 3400 cm−1 that can be attributed to –OH stretching in cross-linked heparin. We showed that bFGF was released from hep-PCL/bFGF in a continuous pattern, which remained for 3 weeks. We evaluated MG63 cell proliferation and biocompatibility of GBR membrane by a CCK-8 assay and a live/dead assay. The CCK-8 results revealed that the hep-PCL/bFGF group had superiority compared to other groups. Furthermore, cell morphology of hep-PCL membrane exhibited larger projected areas than those of PCL surfaces based on scanning electron microscopy analysis and immunofluorescent staining of cell cytoskeleton and vinculin expression. Our alkaline phosphatase activity assay also confirmed better performance of the hep-PCL/bFGF group. These results suggested that this novel hep-PCL/bFGF membrane is suitable for osteoblast-like cells to attach, proliferate, and differentiate. Therefore, the hep-PCL/bFGF membrane has potential to be a biodegradable membrane for GBR and warrants further investigation.