Promoting Angiogenesis/Osteogenesis by a New Copper/Magnesium Hydroxide Hybrid Nanoparticle: In Vitro and In Vivo Investigation

Abstract

In this study, a new hybrid nanoparticle composed of magnesium hydroxide and copper oxide (Mg(OH)₂/CuO) with an optimized ratio of magnesium (Mg) to copper (Cu) was designed and incorporated into a 3D-printed scaffold made of polycaprolactone (PCL) and gelatin. These hybrid nanostructures (MCNs) were prepared using a green, solvent-free method. Their topography, surface morphology, and structural properties were characterized using scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). The fabricated 3D-printed PCL/Gelatin/MCN scaffolds were investigated in vitro and in vivo. Cell viability tests on murine calvarial preosteoblasts (MC3T3-E1) and human umbilical vein endothelial cells (HUVECs) demonstrated that the scaffolds could induce proper cell proliferation. Additionally, the angiogenic and osteogenic properties of the constructs were evaluated using alkaline phosphatase (ALP) activity, osteogenesis-related, and angiogenesis-related gene expression tests. The in vivo study was conducted using a rat calvarial defect model, which confirmed the superior angiogenic and osteogenic properties of the PCL/gelatin/MCN scaffolds compared to PCL/Gelatin and PCL/Gelatin/Mg(OH)₂ scaffolds. Overall, the PCL/Gelatin/MCN scaffolds showed promising potential for bone regeneration, particularly for critical-sized defects where proper angiogenesis is essential for tissue reconstruction.