Development of Extrusion-Based 3D-Printed Chitosan/Magnesium-Doped Nanohydroxyapatite Scaffolds Functionalized With Icariin, Lithium Chloride and Naringin for Enhanced Bone Regeneration in Vitro and in Vivo Publisher Pubmed



Hefzollesan S ; Musayeva H ; Aghazadeh H ; Mammadov A ; Ajdary S ; Baylarov R ; Hefzollesan S ; Mammadov R
Source: International Journal of Biological Macromolecules Published:2026

Abstract

The topological characteristics of bioscaffold materials can be manipulated using 3D printing technology. This has allowed the scaffolds to have a more complex structure that is similar to that of real bone, therefore, allowing more development and differentiation of bone cells. This was aimed at producing a novel 3D-printed nanocomposite scaffold composed of chitosan, magnesium-doped hydroxyapatite (Mg-HAp), and a bioactive blend (icariin, lithium chloride, and naringin) to enhance bone regeneration. The work entailed the production of a tissue-engineered porous composite bone scaffold using chitosan reinforced with icariin, lithium chloride, naringin, and magnesium-doped hydroxyapatite (Mg-HAp) as raw materials, to be 3D printed with high precision. In order to determine the properties of the scaffold, FTIR spectroscopy, porosity and pore size, scaffold swelling, scaffold bioactivity, and mechanical stress testing were done. The viability of the cells on the scaffold was determined using the Kit-8 (CCK-8) assay. The Minimum Inhibitory Concentration (MIC) method was used to evaluate antimicrobial activity against Staphylococcus aureus. The Real-Time PCR method was used to determine the levels of Wnt/β-catenin genes. Finally, 16 male Wistar rats were placed under histological evaluation of the lesion area to promote bone growth. All the research data were analyzed using SPSS version 24. The optimally engineered printed scaffolds were found to be compatible with cells and capable of undergoing osteogenic differentiation in vitro. This is favorable for bone regeneration and helps prevent graft rejection during the regenerative process. © 2026 Published by Elsevier B.V.