Precoating of biphasic calcium phosphate bone substitute with atelocollagen enhances bone regeneration through stimulation of osteoclast activation and angiogenesis.

Type I collagen (Col) is a naturally polymerizing protein and important extracellular matrix bone component. The aim of this study was to improve bone regeneration capacity by precoating the surface of biphasic calcium phosphate (BCP) granules with AT-Col, and evaluating its biological effects. BCP granules were precoated with AT-Col using adsorption and lyophilization method. Morphology of AT-Col precoated surfaces was observed using scanning electron microscopy (SEM). Biocompatibility and osteogenic activity of AT-Col were determined in vitro with human mesenchymal stem cell (hMSCs). In vivo bone healing efficiency and related biological effects were determined using a rabbit calvarial defect model. SEM results revealed numerous irregularly distributed AT-Col polymer clusters on BCP granule surface. Biocompatibility experiments demonstrated that AT-Col was non-cytotoxic, and that cell proliferation, adhesion, and osteogenic activity were improved by AT-Col precoating. After in vivo surgical implantation into bone defects, new bone formation was improved by AT-Col granule precoating. Specifically, 8 weeks post-surgery, percentage bone volume was significantly higher in AT-Col/BCP animals (35.02 ± 1.89%) compared with BCP-treated animals (8.94 ± 1.47%) (p < 0.05). Furthermore, tartrate-resistant acid phosphatase staining and CD31 immunohistochemical staining revealed that osteoclast activation and new blood vessel formation in vivo were also induced by AT-Col precoating. Collectively, these data indicate that AT-Col/BCP may be potentially used as a bone substitute to enable effective bone regeneration through enhanced new blood vessel formation and osteoclast activation.