Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Physical characterization of Apolipoprotein A1 nanoparticles. (A) A1NPs were produced by microfluidic using a chip with 2 inlets for Apolipoprotein A1 (ApoA1) injection at 0.8 mL/min and 1 inlet for phospholipids (POPC) injection at 0.1 mL/min. (B) Three independent productions of A1NPs were characterized by dynamic light scattering to determine their size (7-12 nm). (C) Electron transmission microscopy was used to determine the shape of A1NPs. White triangles indicate stacked disc-like structures, also known as “rolls”.

Article Snippet: Primary anti-human ApoA1 antibody (Calbiochem) was diluted 1:500 in PBS 0.05% triton 0.2% BSA and incubated with the cells overnight at 4°C.

Techniques: Produced, Injection, Transmission Assay, Microscopy

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Apolipoprotein A1 nanoparticle cytotoxicity and uptake assay. (A,B) MTT assay on endothelial (HMEC-1, A) and alveolar epithelial cell lines (A549, B), 24 hours after A1NP incubation. (n=3-5 independent experiments; ANOVA; Tukey’s multiple comparison test; * p<0.05, ** p<0.01, **** p<0,0001). (C) A1NPs [0.05mg/ml] are taken up by both cell lines after 6 and 24 hours of incubation. A rabbit anti human ApoA1 antibody was used for labelling A1NPs (green) and cell nuclei were stained with DAPI (blue). (D,E) Uptake of A1NPs by ABCA1-positive A549 cells. (D) A dot plot illustration for double staining with DilC 18 -A1NPs and ABCA1 and (E) the mean fluorescence intensity of DilC 18 in ABCA1 negative and ABCA1 positive cells. (n=3 independent experiments; Paired t-test; * p < 0.05).

Article Snippet: Primary anti-human ApoA1 antibody (Calbiochem) was diluted 1:500 in PBS 0.05% triton 0.2% BSA and incubated with the cells overnight at 4°C.

Techniques: MTT Assay, Incubation, Comparison, Staining, Double Staining, Fluorescence

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Biodistribution of A1NPs after aerosolization. (A) Experimental design for studying the biodistribution of A1NPs administered intratracheally by aerosolization. (B) Determination of human ApoA1 concentration in mouse plasma at different time points after aerosolization: 0h, 3h, 6h, 12h and 24h (n=6 for PBS, n=10 for A1NPs). (C) A1NPs labeled with DilC18 dye (red) reach the left and right lungs 6 hours after aerosolization and persist in both lungs for up to 24 hours. Representative illustration of 6 mice treated with PBS and 10 mice with A1NPs-DilC18.

Article Snippet: Primary anti-human ApoA1 antibody (Calbiochem) was diluted 1:500 in PBS 0.05% triton 0.2% BSA and incubated with the cells overnight at 4°C.

Techniques: Concentration Assay, Clinical Proteomics, Labeling

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Cellular localization of A1NPs after aerosolization. Immunofluorescence of lung sections from mice 6 hours after A1NP administration. ApoA1 appears in green and cell nuclei are stained with DAPI (blue). (A) AGER (red), specific to type I pneumocytes. (B) SFTPC (orange), marker for type II pneumocytes. (C) EMCN (red), characteristic of pulmonary endothelial cells. Representative illustration of 10 mice with A1NPs-DilC18.

Article Snippet: Primary anti-human ApoA1 antibody (Calbiochem) was diluted 1:500 in PBS 0.05% triton 0.2% BSA and incubated with the cells overnight at 4°C.

Techniques: Immunofluorescence, Staining, Marker

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Passage of A1NPs through A549 epithelial cells grown in air-liquid interface. (A) Schema showing the experimental setup used to study A1NPs migration through lung epithelial cells cultured in air/liquid interface (ALI). (B) Quantification of A1NPs transcytosis under ALI conditions, measured by ELISA specific for human ApoA1, in the basolateral medium. The cumulative concentration of A1NPs in the basolateral medium is determined at different incubation times: 30 min, 1h, 2h, 4h and 6h. (C) Immunofluorescence analysis of ALI membrane after 6h of transcytosis with 0.5 mg/mL A1NPs. Phalloidin (green) labels the actin cytoskeleton, cell nuclei are stained blue, and ApoA1 appear in red. (D) Three-dimensional visualization of the ALI membrane under different experimental conditions.

Article Snippet: Primary anti-human ApoA1 antibody (Calbiochem) was diluted 1:500 in PBS 0.05% triton 0.2% BSA and incubated with the cells overnight at 4°C.

Techniques: Migration, Cell Culture, Enzyme-linked Immunosorbent Assay, Concentration Assay, Incubation, Immunofluorescence, Membrane, Staining

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Physical characterization of Apolipoprotein A1 nanoparticles. (A) A1NPs were produced by microfluidic using a chip with 2 inlets for Apolipoprotein A1 (ApoA1) injection at 0.8 mL/min and 1 inlet for phospholipids (POPC) injection at 0.1 mL/min. (B) Three independent productions of A1NPs were characterized by dynamic light scattering to determine their size (7-12 nm). (C) Electron transmission microscopy was used to determine the shape of A1NPs. White triangles indicate stacked disc-like structures, also known as “rolls”.

Article Snippet: Basolateral media were collected after 30 minutes, 1 hour, 2 hours, 4 hours, and 6 hours and analyzed with a human ApoA1 ELISA assay (3710-1HP-2, Mabtech).

Techniques: Produced, Injection, Transmission Assay, Microscopy

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Apolipoprotein A1 nanoparticle cytotoxicity and uptake assay. (A,B) MTT assay on endothelial (HMEC-1, A) and alveolar epithelial cell lines (A549, B), 24 hours after A1NP incubation. (n=3-5 independent experiments; ANOVA; Tukey’s multiple comparison test; * p<0.05, ** p<0.01, **** p<0,0001). (C) A1NPs [0.05mg/ml] are taken up by both cell lines after 6 and 24 hours of incubation. A rabbit anti human ApoA1 antibody was used for labelling A1NPs (green) and cell nuclei were stained with DAPI (blue). (D,E) Uptake of A1NPs by ABCA1-positive A549 cells. (D) A dot plot illustration for double staining with DilC 18 -A1NPs and ABCA1 and (E) the mean fluorescence intensity of DilC 18 in ABCA1 negative and ABCA1 positive cells. (n=3 independent experiments; Paired t-test; * p < 0.05).

Article Snippet: Basolateral media were collected after 30 minutes, 1 hour, 2 hours, 4 hours, and 6 hours and analyzed with a human ApoA1 ELISA assay (3710-1HP-2, Mabtech).

Techniques: MTT Assay, Incubation, Comparison, Staining, Double Staining, Fluorescence

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Biodistribution of A1NPs after aerosolization. (A) Experimental design for studying the biodistribution of A1NPs administered intratracheally by aerosolization. (B) Determination of human ApoA1 concentration in mouse plasma at different time points after aerosolization: 0h, 3h, 6h, 12h and 24h (n=6 for PBS, n=10 for A1NPs). (C) A1NPs labeled with DilC18 dye (red) reach the left and right lungs 6 hours after aerosolization and persist in both lungs for up to 24 hours. Representative illustration of 6 mice treated with PBS and 10 mice with A1NPs-DilC18.

Article Snippet: Basolateral media were collected after 30 minutes, 1 hour, 2 hours, 4 hours, and 6 hours and analyzed with a human ApoA1 ELISA assay (3710-1HP-2, Mabtech).

Techniques: Concentration Assay, Clinical Proteomics, Labeling

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Cellular localization of A1NPs after aerosolization. Immunofluorescence of lung sections from mice 6 hours after A1NP administration. ApoA1 appears in green and cell nuclei are stained with DAPI (blue). (A) AGER (red), specific to type I pneumocytes. (B) SFTPC (orange), marker for type II pneumocytes. (C) EMCN (red), characteristic of pulmonary endothelial cells. Representative illustration of 10 mice with A1NPs-DilC18.

Article Snippet: Basolateral media were collected after 30 minutes, 1 hour, 2 hours, 4 hours, and 6 hours and analyzed with a human ApoA1 ELISA assay (3710-1HP-2, Mabtech).

Techniques: Immunofluorescence, Staining, Marker

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Passage of A1NPs through A549 epithelial cells grown in air-liquid interface. (A) Schema showing the experimental setup used to study A1NPs migration through lung epithelial cells cultured in air/liquid interface (ALI). (B) Quantification of A1NPs transcytosis under ALI conditions, measured by ELISA specific for human ApoA1, in the basolateral medium. The cumulative concentration of A1NPs in the basolateral medium is determined at different incubation times: 30 min, 1h, 2h, 4h and 6h. (C) Immunofluorescence analysis of ALI membrane after 6h of transcytosis with 0.5 mg/mL A1NPs. Phalloidin (green) labels the actin cytoskeleton, cell nuclei are stained blue, and ApoA1 appear in red. (D) Three-dimensional visualization of the ALI membrane under different experimental conditions.

Article Snippet: Basolateral media were collected after 30 minutes, 1 hour, 2 hours, 4 hours, and 6 hours and analyzed with a human ApoA1 ELISA assay (3710-1HP-2, Mabtech).

Techniques: Migration, Cell Culture, Enzyme-linked Immunosorbent Assay, Concentration Assay, Incubation, Immunofluorescence, Membrane, Staining

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Physical characterization of Apolipoprotein A1 nanoparticles. (A) A1NPs were produced by microfluidic using a chip with 2 inlets for Apolipoprotein A1 (ApoA1) injection at 0.8 mL/min and 1 inlet for phospholipids (POPC) injection at 0.1 mL/min. (B) Three independent productions of A1NPs were characterized by dynamic light scattering to determine their size (7-12 nm). (C) Electron transmission microscopy was used to determine the shape of A1NPs. White triangles indicate stacked disc-like structures, also known as “rolls”.

Article Snippet: Primary anti-human ApoA1 antibody (178422; Calbiochem) was diluted 1:500 in PBS triton 0.5% BSA 1% and incubated for 45 minutes at RT.

Techniques: Produced, Injection, Transmission Assay, Microscopy

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Apolipoprotein A1 nanoparticle cytotoxicity and uptake assay. (A,B) MTT assay on endothelial (HMEC-1, A) and alveolar epithelial cell lines (A549, B), 24 hours after A1NP incubation. (n=3-5 independent experiments; ANOVA; Tukey’s multiple comparison test; * p<0.05, ** p<0.01, **** p<0,0001). (C) A1NPs [0.05mg/ml] are taken up by both cell lines after 6 and 24 hours of incubation. A rabbit anti human ApoA1 antibody was used for labelling A1NPs (green) and cell nuclei were stained with DAPI (blue). (D,E) Uptake of A1NPs by ABCA1-positive A549 cells. (D) A dot plot illustration for double staining with DilC 18 -A1NPs and ABCA1 and (E) the mean fluorescence intensity of DilC 18 in ABCA1 negative and ABCA1 positive cells. (n=3 independent experiments; Paired t-test; * p < 0.05).

Article Snippet: Primary anti-human ApoA1 antibody (178422; Calbiochem) was diluted 1:500 in PBS triton 0.5% BSA 1% and incubated for 45 minutes at RT.

Techniques: MTT Assay, Incubation, Comparison, Staining, Double Staining, Fluorescence

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Biodistribution of A1NPs after aerosolization. (A) Experimental design for studying the biodistribution of A1NPs administered intratracheally by aerosolization. (B) Determination of human ApoA1 concentration in mouse plasma at different time points after aerosolization: 0h, 3h, 6h, 12h and 24h (n=6 for PBS, n=10 for A1NPs). (C) A1NPs labeled with DilC18 dye (red) reach the left and right lungs 6 hours after aerosolization and persist in both lungs for up to 24 hours. Representative illustration of 6 mice treated with PBS and 10 mice with A1NPs-DilC18.

Article Snippet: Primary anti-human ApoA1 antibody (178422; Calbiochem) was diluted 1:500 in PBS triton 0.5% BSA 1% and incubated for 45 minutes at RT.

Techniques: Concentration Assay, Clinical Proteomics, Labeling

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Cellular localization of A1NPs after aerosolization. Immunofluorescence of lung sections from mice 6 hours after A1NP administration. ApoA1 appears in green and cell nuclei are stained with DAPI (blue). (A) AGER (red), specific to type I pneumocytes. (B) SFTPC (orange), marker for type II pneumocytes. (C) EMCN (red), characteristic of pulmonary endothelial cells. Representative illustration of 10 mice with A1NPs-DilC18.

Article Snippet: Primary anti-human ApoA1 antibody (178422; Calbiochem) was diluted 1:500 in PBS triton 0.5% BSA 1% and incubated for 45 minutes at RT.

Techniques: Immunofluorescence, Staining, Marker

Journal: bioRxiv

Article Title: Aerosolized ApoA1 Nanoparticles Synthesized by Microfluidics Cross the Lung Barrier and Modulate Inflammation

doi: 10.1101/2025.07.09.663869

Figure Lengend Snippet: Passage of A1NPs through A549 epithelial cells grown in air-liquid interface. (A) Schema showing the experimental setup used to study A1NPs migration through lung epithelial cells cultured in air/liquid interface (ALI). (B) Quantification of A1NPs transcytosis under ALI conditions, measured by ELISA specific for human ApoA1, in the basolateral medium. The cumulative concentration of A1NPs in the basolateral medium is determined at different incubation times: 30 min, 1h, 2h, 4h and 6h. (C) Immunofluorescence analysis of ALI membrane after 6h of transcytosis with 0.5 mg/mL A1NPs. Phalloidin (green) labels the actin cytoskeleton, cell nuclei are stained blue, and ApoA1 appear in red. (D) Three-dimensional visualization of the ALI membrane under different experimental conditions.

Article Snippet: Primary anti-human ApoA1 antibody (178422; Calbiochem) was diluted 1:500 in PBS triton 0.5% BSA 1% and incubated for 45 minutes at RT.

Techniques: Migration, Cell Culture, Enzyme-linked Immunosorbent Assay, Concentration Assay, Incubation, Immunofluorescence, Membrane, Staining