Desirable bone engineering materials should have a conducive three-dimensional (3D) structure and bioactive mediators for guided bone regeneration. In the present study, hydroxyapatite (HA)/collagen (Col) scaffolds were prepared by an optimized freeze-drying process. The porosity, moisture content, and mechanical properties of the composite have been investigated. The micro-morphology and structure were analyzed with scanning electron microscopy (SEM) and transmission electron microscopy (TEM), confirmed that self-cross-linked HA/Col was evenly distributed and formed a 3D porous scaffold. The physicochemical/mechanical characterization was carried out by Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD). Morphological observation and CCK-8 assay of co-culture cells indicated that HA/Col scaffolds were biocompatible. Then HA/Col scaffolds coupled with recombinant human bone morphogenetic proteins 2 (rhBMP-2) were implanted in the mandibular critical size defect in rats, and histological staining was used to evaluate the bone reconstruction. The result showed that HA/Col coupled with rhBMP-2 could significantly improve the formation of new bone and angiogenesis within the scaffolds as well as the proliferation and differentiation of osteoblasts. Thanks to the encouraging osteogenesis effects, the well-defined 3D scaffolds (HA/Col) cooperating with bioactive agents (rhBMP-2) are expected to be a promising candidate for bone tissue engineering applied to regenerative medicine.
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