Action of microparticles of heparin and alginate crosslinked gel when used as injectable artificial matrices to stabilize basic fibroblast growth factor and induce angiogenesis by controlling its release

Abstract

Alginate is an acidic polysaccharide like the glycosaminoglycans and is a candidate for use as an artificial matrix. We developed a novel alginate gel sheet that is crosslinked with heparin (H/A gel sheet) and discovered its properties of releasing biologically active basic fibroblast growth factor (bFGF), a representative member of the heparin-binding growth factors (HBGFs), for about 1 month in vitro and of inducing angiogenesis in vivo. In the present study, the H/A gel sheet was mechanically broken up to produce easily injectable 50- to 200 microm microparticles of the gel (H/A gel particles), the properties of which were analyzed. The H/A gel particles cumulatively released 2.8 times as much bFGF as the H/A gel sheet, despite both having the same amount of bFGF adsorbed onto their gels. In addition, the bFGF-adsorbed H/A gel particles released a significant amount of bFGF, which stimulated cellular growth in a culture of human umbilical venous endothelial cells for up to 5 weeks. The subcutaneous injection of the bFGF-adsorbed H/A gel particles induced the formation of numerous microvessels in the tissue surrounding the gel. These results indicate that the H/A gel particles not only stabilize bFGF by preventing the occurrence of proteolysis or denaturation but also modulate its release from the gel. Because the H/A gel particles can be easily injected into the target tissues, this artificial matrix may be useful for the local delivery of HBGFs in the treatment of ischemic arterial diseases, as well as for regenerating or constructing tissues.