Proteomic identification of serum proteins to induce osteoconductivity of hydroxyapatite

We performed proteomic analysis of rat serum proteins adsorbed on hydroxyapatite (HAp) and α-alumina (α-Al₂O₃) in order to identify proteins that specifically adsorb onto HAp and control cellular responses. Proteins with either or both molecular weight of 22-32 kDa and computed isoelectric point of 5.0-5.5 were preferentially adsorbed on HAp. In total, 182 proteins were adsorbed on both HAp and α-Al₂O₃, of which 14 were highly enriched on HAp, whereas 68 were adsorbed only on HAp. Therefore, 82 (14+68) proteins were further evaluated by bioinformatics and literature-based analyses. We predicted that hepatocyte growth factor and angiopoietin-like protein 3 (ANGPTL3) are candidate proteins responsible for the osteoconductivity of HAp. Although ANGPTL3 promoted the attachment and spreading of MC3T3-E1 cells, it did not promote their proliferation and differentiation. Our results suggest that specific adsorption of ANGPTL3 on HAp induced osteoconductivity by enhancing the attachment and spreading of osteoblasts.