Journal: Frontiers in Toxicology

Article Title: Monocyte migration assay using a vascular-on-a-chip model and its utilization for the evaluation of a heated tobacco product

doi: 10.3389/ftox.2025.1658093

Figure Lengend Snippet: Barrier impairment, monocyte adhesion, and migration in VoC with or without proinflammatory cytokines and chemokines. The effect of 10 ng/mL IL-1β or 10 ng/mL TNF-α with or without MCP-1 on (A) the TEER of HCAECs after exposure for 24 h (N = 2, n = 2–4), (B) monocyte adhesion to HCAECs 2 h after the addition of monocytes (N = 2, n = 2–4), and (C) monocyte migration through HCAECs 48 h after the addition of monocytes (N = 2, n = 2–4). Data represent the mean ±95%CI, and open circles in each group represent individual chips. Statistical analysis was performed using the Kruskal–Wallis test followed by Dunn’s test for all pairs of samples for joint rank testing with Bonferroni correction. Asterisks indicate a statistically significant difference, and their colors indicate which groups were compared pairwise (* p < 0.05, ** p < 0.01). VoC, Vascular-on-a-Chip; IL, interleukin; TNF, tumor necrosis factor; MCP, monocyte chemotactic protein; TEER, trans-endothelial electrical resistance; HCAEC, human coronary artery endothelial cell; CI, confidence interval.

Article Snippet: Primary human coronary artery endothelial cells (HCAECs) were purchased from Promocell GmbH (Heidelberg, Germany) and cultured in a collagen-coated flask with Endothelial Cell Growth Medium MV2 (Promocell) containing 1% penicillin-streptomycin (FUJIFILM Wako Pure Chemical Corporation).

Techniques: Migration

Journal: Frontiers in Toxicology

Article Title: Monocyte migration assay using a vascular-on-a-chip model and its utilization for the evaluation of a heated tobacco product

doi: 10.3389/ftox.2025.1658093

Figure Lengend Snippet: Representative images of adhered monocytes and migrated monocytes in VoC treated with proinflammatory cytokines Monocyte adhesion and migration were analyzed by fluorescent imaging. After allowing fluorescently labeled monocytes to flow for 2 h (monocyte adhesion) or 48 h (monocyte migration), images of (A) monocyte adhesion and (B) monocyte migration were obtained. Scale bar = 100 μm. White dashed line indicates the boundary between the tubule structure of HCAECs and the ECM region. VoC, Vascular-on-a-Chip; HCAEC, human coronary artery endothelial cell; ECM, extracellular matrix.

Article Snippet: Primary human coronary artery endothelial cells (HCAECs) were purchased from Promocell GmbH (Heidelberg, Germany) and cultured in a collagen-coated flask with Endothelial Cell Growth Medium MV2 (Promocell) containing 1% penicillin-streptomycin (FUJIFILM Wako Pure Chemical Corporation).

Techniques: Migration, Imaging, Labeling

Journal: Frontiers in Toxicology

Article Title: Monocyte migration assay using a vascular-on-a-chip model and its utilization for the evaluation of a heated tobacco product

doi: 10.3389/ftox.2025.1658093

Figure Lengend Snippet: Effects of LPS-conditioned medium on the barrier integrity of HCAECs, and adhesion and migration of monocytes. (A) The TEER of HCAECs after 24 h of exposure to LPS-conditioned medium (N = 4, n = 5 or 6). (B) Monocyte adhesion to HCAECs 2 h after the addition of monocytes (N = 4, n = 5 or 6), and (C) monocyte migration through HCAECs 48 h after the addition of monocytes (N = 4, n = 5 or 6). Data represent the mean ±95%CI, and open circles in each group represent the value of each biological replicate. Statistical analysis was performed using the Mann–Whitney U -test. Asterisks indicate a statistically significant difference, and their colors indicate which groups were compared pairwise (*** p < 0.001). LPS, lipopolysaccharide; HCAEC, human coronary artery endothelial cell; TEER, trans-endothelial electrical resistance; DMSO, dimethyl sulfoxide; CI, confidence interval.

Article Snippet: Primary human coronary artery endothelial cells (HCAECs) were purchased from Promocell GmbH (Heidelberg, Germany) and cultured in a collagen-coated flask with Endothelial Cell Growth Medium MV2 (Promocell) containing 1% penicillin-streptomycin (FUJIFILM Wako Pure Chemical Corporation).

Techniques: Migration, MANN-WHITNEY

Journal: Frontiers in Toxicology

Article Title: Monocyte migration assay using a vascular-on-a-chip model and its utilization for the evaluation of a heated tobacco product

doi: 10.3389/ftox.2025.1658093

Figure Lengend Snippet: Effects of 1R6F TPM- and DT3.0a ACM-conditioned medium on the barrier integrity of HCAECs, and adhesion and migration of monocytes. VoC were exposed to 1R6F TPM-conditioned medium or DT3.0a ACM-conditioned medium for 24 h. (A) The TEER of HCAECs after 24 h of exposure to 1R6F TPM-conditioned medium or DT3.0a ACM-conditioned medium (N = 4, n = 5 or 6). After 24 h of exposure and the measurement of TEER, fluorescently labeled monocytes were added. (B) The number of monocytes adhered to HCAECs at 2 h (N = 4, n = 5 or 6) and (C) the number of monocytes migrated through HCAECs 48 h after the perfusion of monocytes (N = 4, n = 5 or 6). Data represent the mean ±95%CI, and open circles in each group represent the value of each biological replicate. Statistical analysis was performed using the Kruskal–Wallis test followed by Dunn’s test for all pairs of samples for joint rank testing with Bonferroni correction. Asterisks indicate a statistically significant difference, and their colors indicate which groups were compared pairwise (** p < 0.01, *** p < 0.001). TPM, total particulate matter; ACM, aerosol collected mass; DT3.0a, Direct Heating Tobacco System Platform 3 Generation 0 version a; HCAEC, human coronary artery endothelial cell; TEER, trans-endothelial electrical resistance; DMSO, dimethyl sulfoxide; CI, confidence interval.

Article Snippet: Primary human coronary artery endothelial cells (HCAECs) were purchased from Promocell GmbH (Heidelberg, Germany) and cultured in a collagen-coated flask with Endothelial Cell Growth Medium MV2 (Promocell) containing 1% penicillin-streptomycin (FUJIFILM Wako Pure Chemical Corporation).

Techniques: Migration, Labeling, Aerosol

Journal: iScience

Article Title: Comparative effects of hemodialysis modalities on microparticle induction and neutrophil activation in a randomized cross-over study

doi: 10.1016/j.isci.2025.113485

Figure Lengend Snippet: Effects of post-hemodialysis microparticles from the mixed dilution online-HDF (OL-HDF) or hemodialysis with median cut-off dialyzer (MCO-HDX) on human coronary artery endothelial cells (HCAECs) The representation of carboxyfluorescein succinimidyl ester (CSFE)-stained MPs (green color) represented by CSFE fluorescent intensity with the representative immunofluorescent pictures (A and B) are demonstrated ( n = 6/group, scale bar = 10 μ m). The red color in A is zona occluden-1 (ZO-1; a tight junction molecule) and the blue color is DNA-stained by 4′,6-diamidino-2-phenylindole (DAPI). The influences of the post-dialysis MPs from OL-HDF (C) or MCO+HDX (D) with 1x10 4 (light color) or 1x10 5 (heavy color) particles/μL at 24 h after inoculation, compared with control (only media but not MPs) in the condition with or without phorbol myristate acetate (PMA, the positive control activator) on HCAECs as indicated by vascular cell adhesion molecule 1 (VCAM-1) and cell apoptosis, are shown. The effect of MPs on tight junction molecules ZO-1 is also demonstrated through the intensity score of the red color with representative immunofluorescent (E, F) ( n = 6/group, scale bar = 10 μ m). Notably, the MPs were not stained to avoid the noise from the green-fluorescent color. ∗ p < 0.05 vs. control within group; # p < 0.05 vs. same condition between groups.

Article Snippet: Primary Human Coronary Endothelial Cells (HCAECs), Female , ATCC , PCS-100-020 Lot No. 70001400.

Techniques: Staining, Control, Positive Control