Integrated experimental and computational insights into the anti-inflammatory potential of flower-derived exosome-like nanoparticles targeting the NF-κB pathway.

Dysregulated inflammation underlies numerous chronic pathologies, with the NF-κB p65-p50 heterodimer acting as a pivotal transcriptional regulator that mediates different inflammatory responses. Consequently, inhibiting NF-κB nuclear translocation has emerged as a promising strategy in anti-inflammatory drug development. While floral extracts have been widely used, recent advances have highlighted the therapeutic potential of flower-derived exosome like nanoparticles as promising cell-free therapeutics owing to their enhanced biocompatibility and stability. Exosome like nanoparticles were isolated from three ethnomedicinal flowers and systematically characterized. Antioxidant potential of ELNs was evaluated through DPPH assay and their anti-inflammatory potential was assessed. Further, to elucidate the molecular mechanisms underlying NF-κB modulation, key ELN-associated metabolites were computationally screened against the crystallized NF-κB p65-p50 heterodimer using molecular docking, followed by molecular dynamics simulations to evaluate binding stability and interaction dynamics. Isolated ELNs demonstrated a strong antioxidant potential and in vitro analysis revealed significant regulation in mRNA expression of inflammatory cytokines and NF-κB transcriptional activity. Molecular docking identified several metabolites with higher binding affinity against NF-κB p65-p50 heterodimer supported by simulation studies confirming stable ligand-protein interaction. Both docking scores and simulation trajectories strongly supported stable, high-affinity interactions consistent with NF-κB pathway inhibition. Overall, the combined experimental and computational findings in the study represent the first comprehensive data of floral ELN bioactives, offering the significant translational potential of floral nanovesicles as a new class of biocompatible, cell-free nanotherapeutics for anti-inflammatory drug discovery.