Osteogenic Evaluation of 3D-printed PLA scaffolds Integrated with Khellin-loaded Chitosan-Alginate Sponges for Bone Tissue Engineering

Bone tissue engineering (BTE) offers promising strategies for bone regeneration, yet the effective delivery of bioactive molecules remains a challenge. Khellin (KH), a plant-derived furanochromone, possesses various biological properties, though its potential in promoting osteogenesis has not been thoroughly investigated. In this study, 3D-printed polylactic acid (PLA) scaffolds were integrated with KH-loaded chitosan-alginate sponges (PLA/ALG/CS-KH) to facilitate controlled and sustained delivery of KH. Using sol-gel and freeze-drying techniques, KH was incorporated at varying concentrations (60, 70, and 80 mu M) to enhance its bioavailability. Comprehensive physicochemical analyses demonstrated that KH incorporation did not alter the scaffolds' porosity, swelling capacity, protein adsorption, degradation rates, or biomineralization potential. In vitro studies revealed that the PLA/ALG/CS-KH scaffolds were biocompatible with mesenchymal stem cells and effectively promoted osteogenic differentiation, particularly at a concentration of 70 mu M KH. These results suggest that PLA/ALG/CS-KH scaffolds have significant potential as osteoinductive platforms for BTE applications, providing a novel approach for enhancing bone regeneration through the sustained release of KH.