Eco-friendly synthesis of eggshell-derived nano-hydroxyapatite: physicochemical characterization, hemocompatibility, and bone regeneration potential
Huda Jabbar Abdulhussein, Mayyadah H. Mohsin, Majid S. Jabir, Ghassan M. Sulaiman, Hamdoon A. Mohammed, Raid A. Ismail, Asmiet Ramizy, M. H. Eisa, Nasir A. Ibrahim, Nosiba S. Badher
Abstract
Hydroxyapatite (HA) nanoparticles synthesized from bio-waste sources like eggshells offer a sustainable and biocompatible alternative for bone regeneration. This study successfully synthesized nano-hydroxyapatite from chicken eggshell-derived calcium precursors and comprehensively characterized it. X-ray diffraction (XRD) confirmed the formation of a highly crystalline, phase-pure HA structure, while Fourier-transform infrared spectroscopy (FTIR) revealed characteristic phosphate, hydroxyl, and carbonate bands, resembling biological apatite. Scanning electron microscopy (SEM) showed spherical nanoparticles (~ 11.2 nm) with uniform distribution, enhancing bioactivity. Hemocompatibility assays demonstrated concentration-dependent hemolysis, with minimal RBC disruption (< 7%) at 12.5 mg/ml, mitigated further by protein corona formation. In vitro studies revealed excellent osteoblast (MC3T3-E1) adhesion and spreading on HA surfaces, indicating osteoconductivity. Additionally, HA nanoparticles exhibited dose-dependent free radical scavenging activity (up to 84.7% at 200 µg/ml) and significantly suppressed pro-inflammatory cytokines (IL-1β and IL-18) in LPS (Lipopolysaccharide)/ATP (Adenosine triphosphate)-stimulated macrophages, highlighting anti-inflammatory potential. These findings collectively underscore the suitability of eggshell-derived nano-HA for bone tissue engineering, combining eco-friendly synthesis, structural biomimicry, and multifunctional bioactivity to promote osteogenesis and mitigate inflammation.