Abstract: Osteomyelitis caused by Staphylococcus aureus (S. aureus) is a severe inflammatory bone disease that is difficult to eradicate and can be life-threatening. Traditional treatments relying on high-dose systemic antibiotics often fail due to biofilm resistance and emerging drug-resistant strains. This study proposes a diflunisal-loaded 3D-printed scaffold as a novel therapeutic strategy. A composite biomaterial ink composed of gelatin (Gel), polycaprolactone (PCL), and nanohydroxyapatite (n-HA) was synthesized and used to fabricate customized porous scaffolds via 3D printing. Diflunisal was loaded onto the scaffolds using a Gel swelling method. In vitro experiments showed sustained diflunisal release under different pH conditions (pH?=?6.0 and 7.4) mimicking infection and protection of bone marrow stromal cells (BMSCs) from S. aureus toxins. In vivo studies revealed significant alleviation of infection and promotion of bone regeneration at the defect site. This diflunisal-loaded Gel/PCL/n-HA scaffold integrates anti-virulence therapy with bone regeneration, offering a promising solution for osteomyelitis treatment.

Key words: Osteomyelitis')">Osteomyelitis,  , Staphylococcus aureus')">Staphylococcus aureus,  ')"> , Diflunisal-loaded scaffold, 3D printing, Bone regeneration, Antivirulence therapy