human cerebral microvascular endothelial cells  (Cedarlane)

Journal: Journal of Nanobiotechnology

Article Title: Application of an intercellular adhesion molecule-1-targeted nanoparticles loaded with irisin in postoperative neurocognitive disorder through metabolic axis activation and barrier restoration

doi: 10.1186/s12951-026-04271-y

Figure Lengend Snippet: Establishment and characterization of an inflammatory tricellular BBB in vitro model. ( A ) Changes in TEER of hCMEC/D3 cells before and after LPS treatment, with LPS stimulation significantly reducing TEER levels. ( B ) Permeability assay using FITC-dextran, showing an approximately 2.6-fold increase in permeability following LPS stimulation, indicating barrier function impairment. ( C ) Immunofluorescence reveals MAP2⁺ neuronal processes extending along GFAP⁺ astrocytic end-feet, forming synapse-endfoot contacts. ( D ) Top-down view of cellular co-localization in the tricellular co-culture system, displaying neurons, astrocytes, and endothelial cells co-existing within the same field of view. ( E ) Schematic representation of the spatial distribution within the tricellular in vitro BBB model, illustrating the arrangement of different cells in the Transwell system. ( F ) VE-cadherin immunostaining shows changes in endothelial cell junction integrity before and after LPS treatment, with blurred junctional boundaries and reduced expression post-stimulation. Data are presented as mean ± SEM. Statistical significance in panels A and B was determined using a two-tailed unpaired Student’s t-test, **p < 0.01, ***p < 0.001

Article Snippet: The human cerebral microvascular endothelial cell line hCMEC/D3 (ATCC ® CRL-3245 TM ) was cultured at 37 °C in a humidified atmosphere containing 5% CO 2 using EndoGro TM -MV complete medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin-streptomycin, and 1% L-glutamine.

Techniques: In Vitro, Permeability, Immunofluorescence, Co-Culture Assay, Immunostaining, Expressing, Two Tailed Test

Journal: Journal of Nanobiotechnology

Article Title: Application of an intercellular adhesion molecule-1-targeted nanoparticles loaded with irisin in postoperative neurocognitive disorder through metabolic axis activation and barrier restoration

doi: 10.1186/s12951-026-04271-y

Figure Lengend Snippet: Enhanced targeted uptake of ICAM-1-NP@Irisin in inflammatory brain endothelial cells. ( A ) Confocal laser scanning micrographs: Representative fluorescence images of ICAM-1-NP@Irisin and NP@Irisin uptake in inflammatory hCMEC/D3 cells at 1 h, 3 h, and 6 h (red indicates NP fluorescence, blue indicates DAPI nuclear staining, scale bar 25 μm); the ICAM-1-NP@Irisin group exhibits stronger fluorescence signals with higher distribution density. ( B ) Quantitative analysis of fluorescence intensity: At all time points, intracellular fluorescence signals in the ICAM-1-NP@Irisin group are significantly higher than those in the NP@Irisin group (1 h: 1.4 ± 0.1 fold; 3 h: 1.8 ± 0.2 fold; 6 h: 2.1 ± 0.2 fold; data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test, ****p < 0.0001).

Article Snippet: The human cerebral microvascular endothelial cell line hCMEC/D3 (ATCC ® CRL-3245 TM ) was cultured at 37 °C in a humidified atmosphere containing 5% CO 2 using EndoGro TM -MV complete medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin-streptomycin, and 1% L-glutamine.

Techniques: Fluorescence, Staining

Journal: Journal of Nanobiotechnology

Article Title: Application of an intercellular adhesion molecule-1-targeted nanoparticles loaded with irisin in postoperative neurocognitive disorder through metabolic axis activation and barrier restoration

doi: 10.1186/s12951-026-04271-y

Figure Lengend Snippet: Establishment and characterization of an inflammatory tricellular BBB in vitro model. ( A ) Changes in TEER of hCMEC/D3 cells before and after LPS treatment, with LPS stimulation significantly reducing TEER levels. ( B ) Permeability assay using FITC-dextran, showing an approximately 2.6-fold increase in permeability following LPS stimulation, indicating barrier function impairment. ( C ) Immunofluorescence reveals MAP2⁺ neuronal processes extending along GFAP⁺ astrocytic end-feet, forming synapse-endfoot contacts. ( D ) Top-down view of cellular co-localization in the tricellular co-culture system, displaying neurons, astrocytes, and endothelial cells co-existing within the same field of view. ( E ) Schematic representation of the spatial distribution within the tricellular in vitro BBB model, illustrating the arrangement of different cells in the Transwell system. ( F ) VE-cadherin immunostaining shows changes in endothelial cell junction integrity before and after LPS treatment, with blurred junctional boundaries and reduced expression post-stimulation. Data are presented as mean ± SEM. Statistical significance in panels A and B was determined using a two-tailed unpaired Student’s t-test, **p < 0.01, ***p < 0.001

Article Snippet: In the inflammatory hCMEC/D3 model, cells were pretreated with the AMPK inhibitor Compound C (10 μM, MCE) for 1 h prior to nanoparticle exposure.

Techniques: In Vitro, Permeability, Immunofluorescence, Co-Culture Assay, Immunostaining, Expressing, Two Tailed Test

Journal: Journal of Nanobiotechnology

Article Title: Application of an intercellular adhesion molecule-1-targeted nanoparticles loaded with irisin in postoperative neurocognitive disorder through metabolic axis activation and barrier restoration

doi: 10.1186/s12951-026-04271-y

Figure Lengend Snippet: Enhanced targeted uptake of ICAM-1-NP@Irisin in inflammatory brain endothelial cells. ( A ) Confocal laser scanning micrographs: Representative fluorescence images of ICAM-1-NP@Irisin and NP@Irisin uptake in inflammatory hCMEC/D3 cells at 1 h, 3 h, and 6 h (red indicates NP fluorescence, blue indicates DAPI nuclear staining, scale bar 25 μm); the ICAM-1-NP@Irisin group exhibits stronger fluorescence signals with higher distribution density. ( B ) Quantitative analysis of fluorescence intensity: At all time points, intracellular fluorescence signals in the ICAM-1-NP@Irisin group are significantly higher than those in the NP@Irisin group (1 h: 1.4 ± 0.1 fold; 3 h: 1.8 ± 0.2 fold; 6 h: 2.1 ± 0.2 fold; data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test, ****p < 0.0001).

Article Snippet: In the inflammatory hCMEC/D3 model, cells were pretreated with the AMPK inhibitor Compound C (10 μM, MCE) for 1 h prior to nanoparticle exposure.

Techniques: Fluorescence, Staining