Journal: Materials Today Bio

Article Title: Redirecting the route: Monocyte-mediated delivery of oHSV-1 across a human BBB-on-chip model

doi: 10.1016/j.mtbio.2025.102458

Figure Lengend Snippet: Microfluidic BBB-on-chip design and validation. A. Schematic representation of the BBB-on-chip, seeded with endothelial cells on the blood side and pericytes and astrocytes on the brain side, equipped with three microwells for 3D GBM spheroid culture (GBM spheroids area). B. Image of the assembled BBB-on-chip showing the upper channel (yellow) mimicking the vascular compartment and main perfusion conduit, and the lower channels (red) leading to the tumor compartment (scale bar = 2 cm). C. Transendothelial electrical resistance (TEER) measurements recorded daily across the membrane between the upper and lower channels and supporting the BBB. D. Permeability coefficients calculated from the diffusion of 40, 70, and 150 kDa FITC-dextrans through the assembled BBB-on-chip in the following conditions: i. No cells (control, membrane without cells), ii. Blood side (membrane + HUVECs endothelial monolayer), and iii. BBB complete (membrane with the three-culture of HUVECs on the blood side and pericytes and astrocytes on the brain side). E . IF staining of the BBB-on-chip model on day 7. On the blood side compartment , endothelial tight junction protein ZO-1 (green) and the endothelial marker CD31 (green) are expressed, confirming the formation of a continuous endothelial layer. On the brain side , astrocytes are identified by GFAP expression (magenta), indicating appropriate localization. DAPI (blue) highlights cell nuclei in both compartments. Scale bar = 50 μm. Values are expressed as median±SEM from at least 3 independent experiments. ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001 vs control. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The assembled BBB-on-chip devices were bound to glass coverslips via further plasma treatment and sterilized with 70 % ethanol followed by UV exposure for 60 min. BBB-on-chip cells seeding and culturing: Human umbilical vein endothelial cells (HUVECs), brain vascular pericytes (hBVPs), and immortalized astrocytes (hAs) were obtained from Innoprot (Derio, Spain).

Techniques: Biomarker Discovery, Membrane, Permeability, Diffusion-based Assay, Control, Staining, Marker, Expressing