Multi-omics guided self-assembled nanoparticles co-delivering dalbavancin and shTnf-α for synergistic anti-infection and immunomodulation in osteomyelitis

The treatment of Osteomyelitis (OM) faces challenges due to the harmful cycle of bacterial persistence and pathogen-triggered cytokine storms. Conventional antibiotic therapy centers on eliminating bacteria, yet frequently neglects the immunopathological harm following bacterial breakdown. Therefore, there exists an immediate requirement for dual-action treatments capable of simultaneously alleviating multidrug-resistant infections and hyper-inflammatory reactions. In this study, single-cell sequencing highlights a TNF/NF-κB-centered transcriptional network which serves as a crucial molecular switch, propelling M1 polarization in macrophages related to OM. Combination of network toxicology and transcriptomic investigations uncovers the key regulatory function of TNF in LTA-triggered cytokine storms in OM. Metabolic sequencing reveals that LTA triggers reprogramming of pyrimidine metabolism in macrophages and synergistically boosts Tnf-α expression. As a result, we created DB-shTnf-α@Gel, an injectable hydrogel that delivers Dalbavancin and shTnf-α self-assembled nanoparticles. This system demonstrated strong anti- MRSA efficacy and interfered with bacterial resistance genes in vitro. In vivo experiments revealed that DB-shTnf-α@Gel has the capacity to oppose macrophage M1 polarization and notably relieve bone damage. By accurately zeroing in on TNF-induced excessive inflammation and eliminating multidrug-resistant infections, this nanoplatform presents a synergistic, phased therapeutic approach to surmount the intricate pathology of OM.