Angiogenin-loaded fibrin/bone powder composite scaffold for vascularized bone regeneration

Angiogenin (ANG) is a potent stimulator of angiogenesis. The aim of this study was to fabricate an ANG-loaded scaffold and to evaluate its angiogenic and osteogenic effects. In this study, we fabricated an ANG-loaded scaffold using bovine bone powder and fibrin glue. We then evaluated the structural, morphological, and mechanical properties of the scaffold and the in vitro release profile of ANG. Cell proliferation, viability, and adhesion were evaluated using endothelial cells in vitro, and angiogenesis and new bone formation were evaluated using a rabbit calvarial defect model in vivo.

Micro-computed tomography imaging showed that the bone powder was uniformly distributed in the scaffold, and scanning electron microscopy showed that the bone powder was bridged by polymerized fibrin. The porosity and compressive strength of the scaffolds were ~60 % and ~0.9 MPa, respectively, and were not significantly altered by ANG loading. In vitro, at 7 days, approximately 0.4 μg and 1.3 μg of the ANG were released from the FB/ANG 0.5 and FB/ANG 2.0, respectively and sustained slow release was observed until 25 days. The released ANG stimulated cell proliferation and adherence and was not cytotoxic. Furthermore, in vivo implantation resulted in enhanced angiogenesis, and new bone formation depended on the amount of loaded ANG.

These studies demonstrate that a fibrin and bone powder scaffold loaded with ANG might be useful to promote bone regeneration by enhanced angiogenesis.