Surface modification with fibrin/hyaluronic acid hydrogel on solid-free form-based scaffolds followed by BMP-2 loading to enhance bone regeneration

Bone tissue engineering often requires a well-defined scaffold that is highly porous. The multi-head deposition system (MHDS), a form of solid freeform fabrication, has raised great interest as a method for fabricating scaffolds, since it yields a highly porous inter-connective structure without the use of cytotoxic solvents, and permits the diffusion of nutrients and oxygen. However, this method is not suitable for introducing proteins, as it includes a heating process. Hydrogels incorporated with protein coating of the scaffold surface could overcome this MHDS limitation. In the present study, the surface of a scaffold fabricated using MHDS was coated with a mixture of fibrin and hyaluronic acid (HA) and used as a vehicle for delivery of both bone morphogenetic protein-2 (BMP-2) and adipose-derived stromal cells (ASCs). Fibrin/HA coating of the scaffold significantly enhanced initial cell attachment. Furthermore, the in vitro release of BMP-2 from fibrin/HA-coated scaffolds was sustained for 3 days and it stimulated the alkaline phosphatase activity of ASCs seeded on the scaffold for 10 days more actively and continuously than did the soluble BMP-2 that was added to the culture media, not the scaffold itself. Importantly, the transplantation of undifferentiated ASCs inoculated on BMP-2-loaded, fibrin/HA-coated scaffolds resulted in more improved bone formation and mineralization than did the transplantation of undifferentiated ASCs seeded on uncoated scaffolds or on fibrin/HA-coated scaffolds without BMP-2, but containing BMP-2 in the cell suspension medium. These results show that BMP-2-loaded, fibrin/HA-coated scaffolds fabricated using MHDS may be useful in stimulating bone regeneration from undifferentiated ASCs in vivo.