Promotion of angiogenesis by sustained release of rhGM-CSF from heparinized collagen/chitosan scaffolds

A novel dermal substitute of combining recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) with a porous heparinized collagen/chitosan scaffolds was developed, considering the inadequate angiogenesis during repair of full-thickness skin defects. The physicochemical properties of heparinized collagen/chitosan scaffolds were examined and in vitro release pattern of rhGM-CSF from scaffolds was measured by ELISA. Four groups of composite scaffolds (heparinized or unheparinized scaffolds loaded with or without rhGM-CSF) were fabricated for subcutaneous implantation in young adult male Sprague-Dawley (SD) rats. Tissue specimens were harvested at different time points after implantation for histopathological, immunohistochemical observation, and Western blotting analysis. The heparinized scaffolds (H(1)E) showed slower biodegradation and sustained release of rhGM-CSF in vitro, although no significantly different release pattern was observed between the H(1)E and unheparinized scaffolds (H(0)E). In vivo investigation revealed that the heparinized scaffolds loaded with rhGM-CSF (H(1)E/rhGM-CSF) had the best cellular adhesion and migration, new vessel formation, and highest expression of VEGF and TGF-β1, indicating promoted angiogenesis. This study demonstrated that composite dermal substitute of combining rhGM-CSF with a porous heparinized collagen/chitosan scaffolds could be a potential therapeutic agent for full-thickness skin defects because of its sustained delivery of rhGM-CSF.