Metabolic consequences of inflammatory disruption of the blood-brain barrier in an organ-on-chip model of the human neurovascular unitReport as inadecuate

Metabolic consequences of inflammatory disruption of the blood-brain barrier in an organ-on-chip model of the human neurovascular unit - Download this document for free, or read online. Document in PDF available to download.

Journal of Neuroinflammation

, 13:306

First Online: 12 December 2016Received: 09 September 2016Accepted: 07 November 2016


BackgroundUnderstanding blood-brain barrier responses to inflammatory stimulation such as lipopolysaccharide mimicking a systemic infection or a cytokine cocktail that could be the result of local or systemic inflammation is essential to understanding the effect of inflammatory stimulation on the brain. It is through the filter of the blood-brain barrier that the brain responds to outside influences, and the blood-brain barrier is a critical point of failure in neuroinflammation. It is important to note that this interaction is not a static response, but one that evolves over time. While current models have provided invaluable information regarding the interaction between cytokine stimulation, the blood-brain barrier, and the brain, these approaches—whether in vivo or in vitro—have often been only snapshots of this complex web of interactions.

MethodsWe utilize new advances in microfluidics, organs-on-chips, and metabolomics to examine the complex relationship of inflammation and its effects on blood-brain barrier function ex vivo and the metabolic consequences of these responses and repair mechanisms. In this study, we pair a novel dual-chamber, organ-on-chip microfluidic device, the NeuroVascular Unit, with small-volume cytokine detection and mass spectrometry analysis to investigate how the blood-brain barrier responds to two different but overlapping drivers of neuroinflammation, lipopolysaccharide and a cytokine cocktail of IL-1β, TNF-α, and MCP1,2.

ResultsIn this study, we show that 1 during initial exposure to lipopolysaccharide, the blood-brain barrier is compromised as expected, with increased diffusion and reduced presence of tight junctions, but that over time, the barrier is capable of at least partial recovery; 2 a cytokine cocktail also contributes to a loss of barrier function; 3 from this time-dependent cytokine activation, metabolic signature profiles can be obtained for both the brain and vascular sides of the blood-brain barrier model; and 4 collectively, we can use metabolite analysis to identify critical pathways in inflammatory response.

ConclusionsTaken together, these findings present new data that allow us to study the initial effects of inflammatory stimulation on blood-brain barrier disruption, cytokine activation, and metabolic pathway changes that drive the response and recovery of the barrier during continued inflammatory exposure.

KeywordsLipopolysaccharide Cytokine Tight junctions IL-1β TNF-α MCP1,2 Brain-on-chip Micro-organ Mass spectrometry Metabolomics AbbreviationsANOVAAnalysis of variance

BBBBlood-brain barrier

CNSCentral nervous system

ELISAEnzyme-linked immunosorbent assay

FITC-dextranFluorescein isothiocyanate-dextran

GM-CSFGranulocyte-macrophage colony-stimulating factor

hBMVECHuman brain-derived microvascular endothelial cells

HDMSHigh-definition data-independent mass spectrometry acquisition with simultaneous analysis of molecular fragmentation

hiPSCHuman induced pluripotent stem cells

IM-MSIon mobility-mass spectrometry

LCLiquid chromatography


MSMass spectrometry

MSData-independent MS acquisition with simultaneous analysis of molecular fragmentation

NVUNeuroVascular Unit

PCAPrincipal component analysis


ROCKRho-associated coiled-coil kinase

TEERTransendothelial electrical resistance

UPLCUltraperformance liquid chromatography

VIIBREVanderbilt Institute for Integrative Biosystems Research and Education

Electronic supplementary materialThe online version of this article doi:10.1186-s12974-016-0760-y contains supplementary material, which is available to authorized users.

Download fulltext PDF

Author: Jacquelyn A. Brown - Simona G. Codreanu - Mingjian Shi - Stacy D. Sherrod - Dmitry A. Markov - M. Diana Neely - Clayton


Related documents