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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.

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