Journal: Advanced Science

Article Title: 3D Bioprinted Blood–Brain–Tumor Barrier Spheroid‐Based In Situ Evaluation of Anticancer Compounds Against Glioblastoma: ROS‐Producing Ruthenium as a Potential Therapeutic Candidate

doi: 10.1002/advs.78150

Figure Lengend Snippet: Cytotoxicity profiling and Transwell permeability assessment of test compounds across hCMEC/D3 and U‐87 MG cell models. (A1–A4) Cell viability of hCMEC/D3 human cerebral microvascular endothelial cells following 24 h treatment with Rh123 (A1), Ru (A2), Osi (A3), or TMZ (A4), assessed by CellTiter‐Glo luminescent cell viability assay. (B1–B3) Cell viability of U‐87 MG glioblastoma cells following 48 h treatment with Ru (B1), Osi (B2), or TMZ (B3), assessed by MTT assay. (C1–C4) Cumulative apical‐to‐basolateral transport profiles of Rh123 (C1), Ru (C2), Osi (C3), and TMZ (C4) across hCMEC/D3 monolayers in the presence (Ver+) or absence (Ver−) of the P‐glycoprotein inhibitor verapamil (Ver, 100 µM), measured at 15‐min intervals over 60 min. (D1–D4) Apparent permeability coefficients (P app ) of Rh123 (D1), Ru (D2), Osi (D3), and TMZ (D4) in the presence (Ver+) or absence (Ver−) of verapamil. ** p < 0.01, *** p < 0.001, **** p < 0.0001; n.s., not significant. Rh123, Rhodamine‐123; Ru, ruthenium compound; Osi, osimertinib; TMZ, temozolomide; Ver, verapamil. Data in (A1–A4) and (B1–B3) are presented as mean ± SD of n = 3 replicate wells per concentration seeded in parallel; data in (C1–C4) and (D1–D4) are presented as mean ± SD of n = 3 Transwell inserts seeded in parallel per condition. Statistical significance was assessed by ordinary one‐way ANOVA followed by Tukey's multiple‐comparisons test, using a two‐sided alpha of 0.05. Analyses were performed using GraphPad Prism 11.0.2.

Article Snippet: Human cerebral microvascular endothelial cells (hCMECs/D3; Cedarlane, Canada; RRID: CVCL_0M40) were cultured in Endothelial Growth Medium 2 (EGM2; Lonza Bioscience, Switzerland), containing human epidermal growth factor, hydrocortisone, GA 1000, FBS, vascular endothelial growth factor (VEGF), human fibroblastic growth factor (hFGF‐B), R 3 IGF‐I, ascorbic acid, and heparin (Lonza).

Techniques: Profiling, Permeability, Cell Viability Assay, MTT Assay, Ruthenium, Concentration Assay

Journal: bioRxiv

Article Title: Microfluidic analysis reveals ROCK2 regulation of endothelial cilia is essential for blood vessel lumen formation and vascular integrity

doi: 10.64898/2026.05.27.728336

Figure Lengend Snippet: (A) Schematic of a primary cilium, depicting its core compartments (basal body, transition zone, axoneme, ciliary tip) and the localization of key ciliary proteins ARL13B and RPGRIP1L. Intraflagellar transport (IFT) complexes (IFT-A and IFT-B) facilitate cargo trafficking along the axoneme. Adapted from Smith et al. (2020). (B, C) Immunofluorescence images confirming primary cilia (ARL13B, red) in human cerebral microvascular endothelial cells (hCMEC/D3) (B) and mouse bEnd.3 cells (C) . Nuclei are counterstained with DAPI (blue). Scale bars, 50 µm. (D) Primary cilia (ARL13B, red) in a monolayer of human umbilical vein endothelial cells (HUVECs). Nuclei are stained with DAPI (blue). Scale bar, 50 µm. (E) Primary cilia (ARL13B, red) on endothelial tubules (white arrowheads) and fibroblasts (yellow arrowheads) in a HUVEC–human dermal fibroblast (HDF) organotypic coculture. L, lumen. Nuclei are stained with DAPI (blue). Scale bar: 15 µm. (F, G) Endothelial primary cilia in E13.5 mouse embryonic brain vessels. Cilia (arrows, ARL13B, red) are visible on endothelial cells lining the lumen of a superficial vessel (F) and on intraparenchymal vessels (G) . Endomucin (green (F), brown (G)) marks endothelial cells; nuclei are stained with DAPI (blue). Scale bars, 50 µm (F) and 10 µm (G) .

Article Snippet: The following cell lines were cultured in a humidified atmosphere at 37°C with 5% CO 2 . hCMEC/D3 (human cerebral microvascular endothelial cells) (ATCC® CRL-3245TM) were cultured in EndoGRO-MV Complete Media (Merck, SCME004) supplemented with 1 ng/ml basic Fibroblast Growth Factor (bFGF; PeproTech, 100-18B) and used between passages 30-35 for ciliogenesis assays. bEnd.3 (mouse brain endothelial cells) (ATCC® CRL-2299TM) were cultured in DMEM (Gibco, 11995065) supplemented with 10% (v/v) heat-inactivated foetal bovine serum (FBS) (Merck, F9665), 100 U/ml penicillin, 100 μg/ml streptomycin, and 2 mM L-glutamine (all from Gibco). hTERT RPE-1 (human retinal pigment epithelial cells) (ATCC® CRL-4000TM) were cultured in DMEM/F12 (Gibco, 11330032) supplemented with 10% FBS, penicillin/streptomycin, and L-glutamine.

Techniques: Immunofluorescence, Staining

Journal: Virulence

Article Title: Integrated transcriptomic and proteomic analysis reveals inflammatory activation and blood-brain barrier disruption during meningitis-associated extraintestinal pathogenic Escherichia coli infection

doi: 10.1080/21505594.2026.2670939

Figure Lengend Snippet: RS218 infection induces cell death and tight junction disruption in vitro . (A) Relative LDH release in RS218-infected hCMEC/D3 cells pretreated with calcium chelator BAPTA-AM (10 μM), necroptosis inhibitor Necrostatin-1 (10 μM), caspase-1 inhibitor VX-765 (10 μM), or caspase-3 inhibitor Z-DEVD-FMK (20 μM). (B) LDH release in BV2 cells and murine peritoneal macrophages (PMs) infected with RS218 at MOI = 0.1, 1, 10, and 100 for 1 h and 2 h. (C) Immunofluorescence staining of tight junction protein ZO-1 in hCMEC/D3 cells following RS218 infection. Representative images show DAPI (blue), ZO-1 (green), and merged channels in control and infected cells. Scale bars, 50 μm. Bar graphs show ZO-1 positive area and fluorescence intensity in infected cells relative to controls ( n = 3).

Article Snippet: For inhibitor experiments, hCMEC/D3 cells were pretreated for 1 h with BAPTA-AM (10 μM), Necrostatin-1 (10 μM), VX-765 (10 μM), or Z-DEVD-FMK (20 μM) (all from MedChemExpress, USA) prior to RS218 infection.

Techniques: Infection, Disruption, In Vitro, Immunofluorescence, Staining, Control, Fluorescence