In previous studies, we have described the use of demineralized bone matrix (DBM) as a carrier for the localized delivery system of growth factors in vitro and in vivo. The aim of the present work was to develop a direct chemical approach to immobilize the platelet-derived growth factor-BB (PDGF-BB) on DBM with cross-linking reagents. The amount of PDGF-BB covalently immobilized on DBM was significantly increased. The increased proliferation of fibroblasts demonstrated that the biological activity of PDGF-BB was not significantly reduced by cross-linking. Compared with control groups, there was a statistically significant increase in blood vessel density in the PDGF-C-DBM group after having been subcutaneously implanted into the dorsal side of the rats. The surface bioactivity of scaffolds on stimulation cell and new blood vessel invasion was improved. Therefore, the direct chemical cross-linking approach could be used to retain growth factors on collagen scaffolds effectively to develop functional biomaterials.