Enhanced Osteoinduction, Rheological and Mechanical Performance of 3D-Printed Methylcellulose-Gelatin-Hydroxyapatite...

Complex patient-specific bone defects remain difficult to reconstruct because the regenerative capacity of bone is limited and prefabricated implants cannot readily match defect geometry. In this work, methylcellulose-gelatin-hydroxyapatite (MC/GEL/HA) inks were formulated with varying methylcellulose content and hydroxyapatite incorporation, crosslinked with EDC/NHS, and 3D-printed into porous scaffolds with defined square-pore architectures. Inks were evaluated by oscillatory and steady-shear rheology, and printed scaffolds were characterized for morphology, chemical composition, mechanical performance, physicochemical stability, wettability, apatite-forming bioactivity, and osteogenic responses of human bone marrow mesenchymal stem cells. Methylcellulose content primarily governed ink rheology and printability, and increased compressive strength (up to ~0.38 MPa for 15MC/10GEL/30HA), whereas hydroxyapatite enhanced surface hydrophilicity, promoted apatite nucleation within 7 days in simulated body fluid, and markedly increased alkaline phosphatase activity (>10-fold over HA-free scaffolds), mineralization (~2-fold by Alizarin Red), and the highest osteocalcin expression among the HA-containing formulations (~2.45-fold at day 14). The 15MC/10GEL/30HA formulation showed the most favorable balance of printability, mechanical performance, and osteogenic performance. These complementary functions reconciled structural stability with osteogenic performance, supporting the MC/GEL/HA system as a tunable bioink platform for non-weight-bearing bone regeneration, while warranting further in vivo validation.