Growth‐Factor‐Free Bone Regeneration: A Chitosan/Mg‐Doped Hydroxyapatite Nanocomposite Scaffold Synergistically Activated by Icariin, Lithium Chloride, and Naringin

Abstract

This study aimed to develop a cost‐effective bone tissue engineering scaffold that mimics the native bone environment without relying on exogenous recombinant growth factors. A chitosan/magnesium‐doped hydroxyapatite nanocomposite scaffold incorporating icariin, lithium chloride, and naringin was fabricated via freeze‐drying and comprehensively characterized. Physicochemical analyses (FTIR, XRD, SEM) confirmed an interconnected porous architecture with porosity exceeding 85%, appropriate mechanical strength, and controlled biodegradability. In vitro assays using SAOS‐2 cells demonstrated favorable cytocompatibility, enhanced osteoblast proliferation, and notable antibacterial activity. In vivo, a rat calvarial defect model over 12 weeks revealed significantly improved bone defect closure and trabecular bone formation in treated groups compared with blank controls. Histological findings were supported by molecular analyses indicating activation of the Wnt/β‐catenin signaling pathway as a key mechanism underlying the osteogenic response to the incorporated bioactive agents. Collectively, these results indicate that chitosan/Mg‐HAp scaffolds enriched with icariin, lithium chloride, and naringin exhibit an advantageous combination of structural, mechanical, biological, and antimicrobial properties, and effectively promote bone regeneration without the need for expensive recombinant growth factors, highlighting their potential as advanced biomimetic scaffolds for clinical bone repair applications.