A Dynamic <i>in vitro</i> BBB Model for the Study of Immune Cell Trafficking into the Central Nervous System

Journal of Cerebral Blood Flow and Metabolism - Tập 31 Số 2 - Trang 767-777 - 2011
Luca Cucullo1,2, Nicola Marchi1,2, Mohammed Hossain1,2, Damir Janigro1,2,3
1Cerebrovascular Research, Cleveland Clinic Lerner College of Medicine, Cleveland, Ohio, USA
2Department of Cell Biology, Cleveland Clinic Lerner College of Medicine, Cleveland, Ohio, USA
3Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine, Cleveland, Ohio, USA

Tóm tắt

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.

Từ khóa


Tài liệu tham khảo

10.1016/j.nbd.2009.07.031

10.1016/j.brainres.2004.04.034

Cavender DE, 1989, Am J Pathol, 134, 551

10.1161/STROKEAHA.109.551341

Cucullo L, 2005, Curr Opin Drug Discov Devel, 8, 89

10.1038/sj.jcbfm.9600525

10.1111/j.1528-1167.2006.00960.x

10.1016/S0006-8993(02)03167-0

Davson H, Segal MB (1996) Blood-brain barrier. In: Physiology of the CSF and blood–brain barriers (Davson H, Segal MB, eds), Boca Raton, pp 49–91

10.1080/10623320212004

10.1016/j.it.2005.07.004

Ghosh C, Gonzalez-Martinez J, Hossain M, Cucullo L, Fazio V, Janigro D, Marchi N (2010) Pattern of P450 expression at the human blood-brain barrier: Roles of epileptic condition and laminar flow. Epilepsia; e-pub ahead of print

Greenwood J, 1994, Lab Invest, 70, 39

10.1016/j.surneu.2005.12.028

10.1186/1471-2202-11-12

10.1016/S0074-7742(07)79004-9

Kalaria RN, 1992, Cerebrovasc Brain Metab Rev, 4, 226

10.1002/jcp.20429

Lochhead JJ, McCaffrey G, Quigley CE, Finch J, Demarco KM, Nametz N, Davis TP (2010) Oxidative stress increases blood-brain barrier permeability and induces alterations in occludin during hypoxia-reoxygenation. J Cereb Blood Flow Metab; e-pub ahead of print

10.1016/S0006-8993(01)02418-0

10.1191/1352458503ms965oa

10.1016/S0006-8993(03)02980-9

Patsalos PN, 1996, Acta Neurochir Suppl, 67, 59

10.1023/A:1007049806660

10.1517/13543784.10.10.1809

10.1161/01.STR.29.10.2189

10.1023/A:1015187410909

10.1016/j.brainres.2006.06.027

10.1016/S0361-9230(03)00097-2

10.1007/3-211-30714-1_91

Stanness KA, 1996, Neurotoxicology, 17, 481

10.1016/S0006-8993(97)00829-9

10.1371/journal.pone.0003037

10.1097/01.inf.0000131634.57368.45

10.1111/j.1528-1157.1996.tb00586.x

Thông tin
Thông tin xuất bản

Journal of Cerebral Blood Flow and Metabolism

Tập 31 Số 2

767-777

Thông tin tác giả