Dual-functional inhaled Ebselen liposomes modulate the TWIST2–GPX3 redox axis to suppress fibroblast activation and attenuate pulmonary fibrosis

Idiopathic pulmonary fibrosis is a fatal interstitial lung disease characterized by persistent fibroblast activation and excessive extracellular matrix deposition. Although redox imbalance plays an important role in regulating fibroblast phenotypes, the upstream TWIST2–GPX3-associated redox mechanism and a locally deliverable strategy to suppress fibroblast activation remain incompletely defined. Dense extracellular matrix barriers and impaired local blood perfusion further limit the effective delivery of antifibrotic drugs. This study aimed to define the TWIST2–GPX3 redox axis in fibroblast activation and develop dual-functional inhaled Ebselen liposomes to attenuate fibrotic remodeling. Through multi-omics analyses, we found that GPX3 transcript expression was markedly reduced in fibrotic fibroblast populations. Mechanistically, the transcription factor TWIST2 was associated with increased GPX3 transcription and promoter histone acetylation, involving EP300-associated H3K27 acetylation. TGF-β1 stimulation reduced TWIST2 and GPX3 expression, accompanied by reactive oxygen species accumulation and increased TGF-β1/SMAD2 pathway activity, supporting a TWIST2–GPX3 redox-axis model during fibroblast activation. To modulate this fibrosis-associated redox imbalance and improve lesion-associated delivery, we engineered dual-functional inhaled Ebselen-loaded liposomes as a GPX-mimetic redox-modulating nanotherapeutic. The optimized formulation integrated L-arginine modification to support NO/MMP-associated collagen-barrier penetration and FAPI-04 conjugation to promote FAP-associated fibroblast uptake. In vitro and in vivo experiments showed that dual-functional inhaled Ebselen liposomes improved pulmonary accumulation and supported fibroblast-associated localization, suppressed fibroblast activation, and reduced fibrotic remodeling in bleomycin- and silica-induced mouse models, while exhibiting limited systemic toxicity under the tested conditions. This study identifies a TWIST2–GPX3 redox axis linked to fibroblast redox imbalance, TGF-β1/SMAD2 signaling, and fibroblast activation during pulmonary fibrosis. Dual-functional inhaled Ebselen liposomes modulated this redox imbalance by restoring GPX-like antioxidant activity and improving pulmonary/fibroblast-associated delivery. The optimized Lip@ELF formulation suppressed fibroblast activation and showed antifibrotic activity in preclinical pulmonary fibrosis models, providing a basis for further development of inhaled redox-modulating nanotherapeutic strategies.