Bioinspired Hydroxyapatite-Incorporated Polyacrylonitrile Nanofiber and Amoxicillin-Loaded Cellulose Acetate Bilayer Membrane with Tunable Osteoconductivity for Bone Regeneration

Abstract

Nanofiber-based materials show significant potential in bone tissue engineering, offering an ideal environment for cell adhesion, migration, and osteoinduction. Herein, hydroxyapatite (HA) is synthesized from Sepia officinalis, which is a naturally rich source of calcium and magnesium, and incorporated into polyacrylonitrile (PAN) nanofibers through electrospinning. The dual-layer membrane consists of an electrospun HA/PAN nanofiber top layer and an amoxicillin-loaded cellulose acetate (aCA) bottom layer, designed for controlled antibiotic release to provide antimicrobial protection essential for osteogenesis. The uniform distribution of HA nanoparticles within the PAN nanofibers enhances porosity and reduces hydrophobicity. The physicochemical properties and morphology of the membrane are characterized using scanning electron microscopy, X-ray photoelectron spectroscopy, tensile strength analysis, and water contact angle measurements. In vitro studies confirm that the HA/PAN@aCA membrane supports the adhesion, proliferation, and differentiation of L929 fibroblasts and MG-63 osteosarcoma-derived cells, promoting mineralized nodule formation. Additionally, the scaffold demonstrates significant antimicrobial activity with controlled amoxicillin release, effectively preventing microbial contamination and facilitating bone regeneration. These findings highlight the potential of the HA/PAN@aCA dual-layered membrane as a promising biomaterial for bone tissue engineering, offering both structural integrity and bifunctionality for enhanced osteogenesis and infection control.