Journal: Nature Communications

Article Title: Periodic asymmetric field enhances electrofusion of nanoscale lipid systems

doi: 10.1038/s41467-025-65961-z

Figure Lengend Snippet: a Schematic of inhalable delivery and lung epithelial cells (LECs) targeting of TGF-β1 -siRNA-encapsulated MSC-cLNPs to the PF model mice. The schematic figure was created in BioRender with proof permission. b Fluorescence signal of DiD-labeled MSC-cLNPs prepared by different methods in lungs of PF mice at 4, 8, 12 and 24 h after inhalation. The MSC-cLNPs was labeled by DiD at 40 μM·kg −1 . The inhalation was operated at 24 h after BLM challenging. c LECs staining (green, labeled by FITC-E-cad) and MSC-cLNPs (red, labeled by DiD) prepared by different methods in lungs 24 h post-inhalation delivery. The white arrow indicated the co-localization of LECs and cLNPs around the bronchus. Scale bar, 200 μm. d Quantified analysis of fluorescence signals of MSC-cLNPs prepared by different methods in lungs of PF mice at 4, 8, 12 and 24 h after inhalation ( n = 3, biological independent replicates). e Treatment regime and group assignment of PF mice. TGF-β1 -siRNA-encapsulated MSC-cLNPs prepared by different methods were inhaled delivery into PF mice on day 1 and 7 post PF induction ( n = 6, biological independent replicates). f TGF-β1 expression in lungs of PF model mice and received treatments by TGF-β1 -siRNA-encapsulated MSC-cLNPs prepared by different methods at 21-day post-modeling ( n = 5, biological independent replicates, data are presented as mean values ± SD, exact p -values < 0.00001 are presented in comparison order, 3 × 10 −7 ). g Matrix metalloproteinase 9 (MMP9) expression in lungs of PF model mice and received treatments by TGF-β1- siRNA-encapsulated MSC-cLNPs prepared by different methods at 21-day post-modeling ( n = 5, biological independent replicates, data are presented as mean values ± SD, exact p -values < 0.00001 are presented in comparison order, 7 × 10 −6 , 3 × 10 −6 ). h Hydroxyproline (HYP) expression in lungs of PF model mice and received treatments by TGF-β1- siRNA-encapsulated MSC-cLNPs prepared by different methods at 21-day post-modeling ( n = 5, biological independent replicates, data are presented as mean values ± SD). i Hematoxylin and eosin (H&E) and Masson staining of lungs of PF model mice and received treatments by TGF-β1- siRNA-encapsulated MSC-cLNPs prepared by different methods at 21-day post-modeling. j Quantified analysis of fibrotic ration in lungs of PF model mice and received treatments by TGF-β1- siRNA-encapsulated cLNPs from different methods ( n = 5, biological independent replicates, data are presented as mean values ± SD). TGF-β1 -siRNA-encapsulated LNPs were prepared by D-Lin-MC3-DMA:DOTAP:DSPC:cholesterol:DOPE at a molar ratio of 30:20:30:10:10. The P -value in the figure was calculated by an unpaired Student’s t test.

Article Snippet: The concentrations of TGF-β1 and MMP9 in the supernatant were quantified using specific ELISA kits: Mouse TGF-β1 ELISA Kit (EK0515, Boster, China) and Mouse MMP9 ELISA Kit (EK0466, Boster, China), following the manufacturer’s protocols.

Techniques: Fluorescence, Labeling, Staining, Expressing, Comparison