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Inflammation-directed targeted delivery in vitro. (A) Representative confocal images of the uptake of M1-EXOs and iM1-EXOs-PAN by GB cells. Scale bar, 25 μm. (B) Quantification of the PKH67‐positive cell ratio based on the confocal microscopy images. (C) Schematic illustration of the in vitro BBB model using a trans-well system to evaluate the penetration capability of iM1-EXOs-PAN across the endothelial monolayer. The transwell co-culture system containing HUVEC cells (simulating the vascular endothelial cells of BBB) in the upper chamber and GB cells in the bottom chamber. And, iM1-EXOs-PAN was added to the upper chamber, <t>and</t> <t>FBS</t> free medium with or without <t>fMLP</t> (100 μM, MCE, <t>HY-P0224)</t> was added to the lower chamber. (D) Representative images of HUVECs and LN229 cells up-taking the nanoformulation treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 12 h. Scale bar, 20 μm. (E) The percentage of PKH67 positive cells treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 4, 8, and 12 h. Statistical significance was calculated using two-way ANOVA. n. s = not significant, and ∗∗∗ P < 0.001.
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Inflammation-directed targeted delivery in vitro. (A) Representative confocal images of the uptake of M1-EXOs and iM1-EXOs-PAN by GB cells. Scale bar, 25 μm. (B) Quantification of the PKH67‐positive cell ratio based on the confocal microscopy images. (C) Schematic illustration of the in vitro BBB model using a trans-well system to evaluate the penetration capability of iM1-EXOs-PAN across the endothelial monolayer. The transwell co-culture system containing HUVEC cells (simulating the vascular endothelial cells of BBB) in the upper chamber and GB cells in the bottom chamber. And, iM1-EXOs-PAN was added to the upper chamber, <t>and</t> <t>FBS</t> free medium with or without <t>fMLP</t> (100 μM, MCE, <t>HY-P0224)</t> was added to the lower chamber. (D) Representative images of HUVECs and LN229 cells up-taking the nanoformulation treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 12 h. Scale bar, 20 μm. (E) The percentage of PKH67 positive cells treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 4, 8, and 12 h. Statistical significance was calculated using two-way ANOVA. n. s = not significant, and ∗∗∗ P < 0.001.
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Inflammation-directed targeted delivery in vitro. (A) Representative confocal images of the uptake of M1-EXOs and iM1-EXOs-PAN by GB cells. Scale bar, 25 μm. (B) Quantification of the PKH67‐positive cell ratio based on the confocal microscopy images. (C) Schematic illustration of the in vitro BBB model using a trans-well system to evaluate the penetration capability of iM1-EXOs-PAN across the endothelial monolayer. The transwell co-culture system containing HUVEC cells (simulating the vascular endothelial cells of BBB) in the upper chamber and GB cells in the bottom chamber. And, iM1-EXOs-PAN was added to the upper chamber, <t>and</t> <t>FBS</t> free medium with or without <t>fMLP</t> (100 μM, MCE, <t>HY-P0224)</t> was added to the lower chamber. (D) Representative images of HUVECs and LN229 cells up-taking the nanoformulation treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 12 h. Scale bar, 20 μm. (E) The percentage of PKH67 positive cells treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 4, 8, and 12 h. Statistical significance was calculated using two-way ANOVA. n. s = not significant, and ∗∗∗ P < 0.001.
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Inflammation-directed targeted delivery in vitro. (A) Representative confocal images of the uptake of M1-EXOs and iM1-EXOs-PAN by GB cells. Scale bar, 25 μm. (B) Quantification of the PKH67‐positive cell ratio based on the confocal microscopy images. (C) Schematic illustration of the in vitro BBB model using a trans-well system to evaluate the penetration capability of iM1-EXOs-PAN across the endothelial monolayer. The transwell co-culture system containing HUVEC cells (simulating the vascular endothelial cells of BBB) in the upper chamber and GB cells in the bottom chamber. And, iM1-EXOs-PAN was added to the upper chamber, <t>and</t> <t>FBS</t> free medium with or without <t>fMLP</t> (100 μM, MCE, <t>HY-P0224)</t> was added to the lower chamber. (D) Representative images of HUVECs and LN229 cells up-taking the nanoformulation treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 12 h. Scale bar, 20 μm. (E) The percentage of PKH67 positive cells treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 4, 8, and 12 h. Statistical significance was calculated using two-way ANOVA. n. s = not significant, and ∗∗∗ P < 0.001.
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Inflammation-directed targeted delivery in vitro. (A) Representative confocal images of the uptake of M1-EXOs and iM1-EXOs-PAN by GB cells. Scale bar, 25 μm. (B) Quantification of the PKH67‐positive cell ratio based on the confocal microscopy images. (C) Schematic illustration of the in vitro BBB model using a trans-well system to evaluate the penetration capability of iM1-EXOs-PAN across the endothelial monolayer. The transwell co-culture system containing HUVEC cells (simulating the vascular endothelial cells of BBB) in the upper chamber and GB cells in the bottom chamber. And, iM1-EXOs-PAN was added to the upper chamber, <t>and</t> <t>FBS</t> free medium with or without <t>fMLP</t> (100 μM, MCE, <t>HY-P0224)</t> was added to the lower chamber. (D) Representative images of HUVECs and LN229 cells up-taking the nanoformulation treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 12 h. Scale bar, 20 μm. (E) The percentage of PKH67 positive cells treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 4, 8, and 12 h. Statistical significance was calculated using two-way ANOVA. n. s = not significant, and ∗∗∗ P < 0.001.
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Inflammation-directed targeted delivery in vitro. (A) Representative confocal images of the uptake of M1-EXOs and iM1-EXOs-PAN by GB cells. Scale bar, 25 μm. (B) Quantification of the PKH67‐positive cell ratio based on the confocal microscopy images. (C) Schematic illustration of the in vitro BBB model using a trans-well system to evaluate the penetration capability of iM1-EXOs-PAN across the endothelial monolayer. The transwell co-culture system containing HUVEC cells (simulating the vascular endothelial cells of BBB) in the upper chamber and GB cells in the bottom chamber. And, iM1-EXOs-PAN was added to the upper chamber, <t>and</t> <t>FBS</t> free medium with or without <t>fMLP</t> (100 μM, MCE, <t>HY-P0224)</t> was added to the lower chamber. (D) Representative images of HUVECs and LN229 cells up-taking the nanoformulation treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 12 h. Scale bar, 20 μm. (E) The percentage of PKH67 positive cells treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 4, 8, and 12 h. Statistical significance was calculated using two-way ANOVA. n. s = not significant, and ∗∗∗ P < 0.001.
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Inflammation-directed targeted delivery in vitro. (A) Representative confocal images of the uptake of M1-EXOs and iM1-EXOs-PAN by GB cells. Scale bar, 25 μm. (B) Quantification of the PKH67‐positive cell ratio based on the confocal microscopy images. (C) Schematic illustration of the in vitro BBB model using a trans-well system to evaluate the penetration capability of iM1-EXOs-PAN across the endothelial monolayer. The transwell co-culture system containing HUVEC cells (simulating the vascular endothelial cells of BBB) in the upper chamber and GB cells in the bottom chamber. And, iM1-EXOs-PAN was added to the upper chamber, <t>and</t> <t>FBS</t> free medium with or without <t>fMLP</t> (100 μM, MCE, <t>HY-P0224)</t> was added to the lower chamber. (D) Representative images of HUVECs and LN229 cells up-taking the nanoformulation treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 12 h. Scale bar, 20 μm. (E) The percentage of PKH67 positive cells treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 4, 8, and 12 h. Statistical significance was calculated using two-way ANOVA. n. s = not significant, and ∗∗∗ P < 0.001.
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Inflammation-directed targeted delivery in vitro. (A) Representative confocal images of the uptake of M1-EXOs and iM1-EXOs-PAN by GB cells. Scale bar, 25 μm. (B) Quantification of the PKH67‐positive cell ratio based on the confocal microscopy images. (C) Schematic illustration of the in vitro BBB model using a trans-well system to evaluate the penetration capability of iM1-EXOs-PAN across the endothelial monolayer. The transwell co-culture system containing HUVEC cells (simulating the vascular endothelial cells of BBB) in the upper chamber and GB cells in the bottom chamber. And, iM1-EXOs-PAN was added to the upper chamber, <t>and</t> <t>FBS</t> free medium with or without <t>fMLP</t> (100 μM, MCE, <t>HY-P0224)</t> was added to the lower chamber. (D) Representative images of HUVECs and LN229 cells up-taking the nanoformulation treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 12 h. Scale bar, 20 μm. (E) The percentage of PKH67 positive cells treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 4, 8, and 12 h. Statistical significance was calculated using two-way ANOVA. n. s = not significant, and ∗∗∗ P < 0.001.
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Inflammation-directed targeted delivery in vitro. (A) Representative confocal images of the uptake of M1-EXOs and iM1-EXOs-PAN by GB cells. Scale bar, 25 μm. (B) Quantification of the PKH67‐positive cell ratio based on the confocal microscopy images. (C) Schematic illustration of the in vitro BBB model using a trans-well system to evaluate the penetration capability of iM1-EXOs-PAN across the endothelial monolayer. The transwell co-culture system containing HUVEC cells (simulating the vascular endothelial cells of BBB) in the upper chamber and GB cells in the bottom chamber. And, iM1-EXOs-PAN was added to the upper chamber, and FBS free medium with or without fMLP (100 μM, MCE, HY-P0224) was added to the lower chamber. (D) Representative images of HUVECs and LN229 cells up-taking the nanoformulation treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 12 h. Scale bar, 20 μm. (E) The percentage of PKH67 positive cells treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 4, 8, and 12 h. Statistical significance was calculated using two-way ANOVA. n. s = not significant, and ∗∗∗ P < 0.001.

Journal: Materials Today Bio

Article Title: Co-delivery of panobinostat and siSTAT3 using engineered M1 exosomes to establish a one-two punch therapeutic strategy for glioblastoma recurrence

doi: 10.1016/j.mtbio.2025.102680

Figure Lengend Snippet: Inflammation-directed targeted delivery in vitro. (A) Representative confocal images of the uptake of M1-EXOs and iM1-EXOs-PAN by GB cells. Scale bar, 25 μm. (B) Quantification of the PKH67‐positive cell ratio based on the confocal microscopy images. (C) Schematic illustration of the in vitro BBB model using a trans-well system to evaluate the penetration capability of iM1-EXOs-PAN across the endothelial monolayer. The transwell co-culture system containing HUVEC cells (simulating the vascular endothelial cells of BBB) in the upper chamber and GB cells in the bottom chamber. And, iM1-EXOs-PAN was added to the upper chamber, and FBS free medium with or without fMLP (100 μM, MCE, HY-P0224) was added to the lower chamber. (D) Representative images of HUVECs and LN229 cells up-taking the nanoformulation treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 12 h. Scale bar, 20 μm. (E) The percentage of PKH67 positive cells treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 4, 8, and 12 h. Statistical significance was calculated using two-way ANOVA. n. s = not significant, and ∗∗∗ P < 0.001.

Article Snippet: Monolayered HUVEC with TEER values greater than 300 Ω cm 2 were adopted as the BBB model. Then, 0.2 μg/μL of PKH67-labeled iM1-EXOs-PAN was added to the upper chamber, and FBS-free medium with or without fMLP (100 μΜ, HY-P0224, MCE) was added to the lower chamber [ ].

Techniques: In Vitro, Confocal Microscopy, Co-Culture Assay