title: A dynamic in vitro BBB model for the study of immune cell trafficking into the central nervous system.
abstract: Although there is significant evidence correlating overreacting or perhaps misguided immune cells and the blood-brain barrier (BBB) with the pathogenesis of neuroinflammatory diseases, the mechanisms by which they enter the brain are largely unknown. For this purpose, we revised our humanized dynamic in vitro BBB model (DIV-BBBr) to incorporate modified hollow fibers that now feature transmural microholes (2 to 4 μm Ø) allowing for the transendothelial trafficking of immune cells. As with the original model, this new DIV-BBBr reproduces most of the physiological characteristics of the BBB in vivo. Measurements of transendothelial electrical resistance (TEER), sucrose permeability, and BBB integrity during reversible osmotic disruption with mannitol (1.6 mol/L) showed that the microholes do not hamper the formation of a tight functional barrier. The in vivo rank permeability order of sucrose, phenytoin, and diazepam was successfully reproduced in vitro. Flow cessation followed by reperfusion (Fc/Rp) in the presence of circulating monocytes caused a biphasic BBB opening paralleled by a significant increase of proinflammatory cytokines and activated matrix metalloproteinases. We also observed abluminal extravasation of monocytes but only when the BBB was breached. In conclusion, the DIV-BBBr represents the most realistic in vitro system to study the immune cell trafficking across the BBB.
authors.affiliation: Cerebrovascular Research, Cleveland Clinic Lerner College of Medicine, Cleveland, Ohio 44195, USA.
authors.forename: Luca
authors.initials: L
authors.lastname: Cucullo
authors.forename: Nicola
authors.initials: N
authors.lastname: Marchi
authors.forename: Mohammed
authors.initials: M
authors.lastname: Hossain
authors.forename: Damir
authors.initials: D
authors.lastname: Janigro
mesh_terms: D000328:Adult; D001812:Blood-Brain Barrier / Q000166:cytology* / Q000276:immunology*; D001932:Brain Neoplasms / Q000473:pathology; D002478:Cells, Cultured; D002490:Central Nervous System / Q000166:cytology* / Q000276:immunology*; D002560:Cerebrovascular Circulation / Q000502:physiology; D016207:Cytokines / Q000378:metabolism; D004232:Diuretics / Q000494:pharmacology; D042783:Endothelial Cells / Q000502:physiology; D004730:Endothelium, Vascular / Q000502:physiology; D005434:Flow Cytometry; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007111:Immunity, Cellular / Q000502:physiology*; D019344:Lactic Acid / Q000378:metabolism; D007962:Leukocytes / Q000502:physiology; D008353:Mannitol / Q000494:pharmacology; D020782:Matrix Metalloproteinases / Q000378:metabolism; D008833:Microcirculation / Q000502:physiology; D008959:Models, Neurological; D057705:Transendothelial and Transepithelial Migration / Q000502:physiology*
chemical_list: D016207:Cytokines; D004232:Diuretics; D019344:Lactic Acid; D008353:Mannitol; D020782:Matrix Metalloproteinases; D005947:Glucose