Multifunctional Triple-Layered Composite Scaffolds Combining Platelet-Rich Fibrin Promote Bone Regeneration

There has been substantial progress made in the development of bone regeneration materials, driven by the deficiencies that exist in current clinical products, such as finite sources, donor site complications, and potential for disease transmission. To overcome these shortcomings, multifunctional scaffolds should be developed to integrate the relationship among osteoinduction, osteoconduction, and osseointegration. In this study, we fabricated polycaprolactone/gelatin (PG) nanofiber films by electrospinning, to act as barriers against connective tissue migration into bone defect sites; chitosan/poly (gamma-glutamic acid)/hydroxyapatite (CPH) hydrogels were formed by electrostatic interaction and lyophilization, to exert osteoconduction; and platelet-rich fibrin (PRF) was extracted from rat abdominal aorta and combined with composite scaffolds, to promote bone induction through the release of growth factors. Hydrogels were immersed in simulated body fluid (SBF) for 1 month to investigate mineralization in vitro. Cytocompatibility, cell barrier effect, and osteogenic differentiation were also explored in vitro. The ability to effectively regenerate bone was analyzed by implantation of triple-layered composite scaffolds into rat calvarial defects in vivo. Sizematched hydrogel filled the defect site, and then, fresh PRF was applied to the hydrogel surface. Finally, P2G3 nanofiber films were applied and attached to the surrounding soft tissue. In short, we fabricated multifunctional triple-layered scaffolds by combining the advantages of osteoinduction, osteoconduction, and osseointegration, which could give full play to the role of PRF in bone regeneration and provide new and pragmatic concepts for bone tissue regeneration in clinical applications.