Human Fresh Fibrin Membrane with Bone Morphogenetic Protein-2 (BMP-2) Induces Bone Formation in the Subcutaneous Tissues of Nude Mice

Autologous blood-derived fibrin glue with platelets, called the concentrated growth factor (CGF), can be prepared immediately by only the decided centrifuge without the addition of coagulation factors. Collagen materials combined with recombinant human BMP-2 have been commercially available for clinical use. The fresh CGF is auto-clot with wettability and elasticity, while most collagen membranes are derived from the cow or pig. The fresh CGF has wettability and elasticity, while collagen membranes are dry materials without elasticity. The aim of this study was to observe the microstructures of human CGF membrane and evaluate its behavior as a delivery scaffold of rhBMP-2 in the subcutaneous tissues of nude mice. Twenty-four nude mice (5-week-old, male) were used for the assessment of in vivo ectopic bone formation. Mice were received the CGF membrane as the controls and the CGF/rhBMP-2 membrane as the experimental group in the subcutaneous tissues, and harvested at 7, 10, and 14 days after the graft. Harvested samples were evaluated for the histological examination and the histomorphometric measurement was conducted to compare the residue of the CGF, as well as the new bone. Mature fibrin fibers assembled from multiple fibrillary elements and platelets with the rhBMP-2 membrane induced several bony islands and cartilage without residues of CGF at 14 days, while the CGF membrane alone was almost absorbed at 10 days and failed to induce bone formation at 14 days. These results demonstrated that the fresh, human CGF membrane could contribute to a short-term, sticky fibrin matrix for the delivery of rhBMP-2.