Abstract
TNFα signaling in the vascular endothelium elicits multiple inflammatory responses that drive vascular destabilization and leakage. Bioactive lipids are main drivers of these processes. In vitro mechanistic studies of bioactive lipids have been largely based on two-dimensional endothelial cell cultures that, due to lack of laminar flow and the growth of the cells on non-compliant stiff substrates, often display a pro-inflammatory phenotype. This complicates the assessment of inflammatory processes. Three-dimensional microvessels-on-a-chip models provide a unique opportunity to generate endothelial microvessels in a more physiological environment. Using an optimized targeted liquid chromatography-tandem mass spectrometry measurements of a panel of pro- and anti-inflammatory bioactive lipids, we measure the profile changes upon administration of TNFα. We demonstrate that bioactive lipid profiles can be readily detected from three-dimensional microvessels-on-a-chip and display a more dynamic, less inflammatory response to TNFα, that resembles more the human situation, compared to classical two-dimensional endothelial cell cultures.
Highlights
Tumor necrosis factor-a (TNFa) is a central mediator of the inflammatory response (Sedger and McDermott, 2014)
We demonstrate bioactive lipid profiles can be readily detected from single microvessels and display a more dynamic, less inflammatory response to TNFa, that resembles more the human situation, compared to classical 2D endothelial cell cultures
Conditioned medium perfused through TNFa treated and control microvessels was sampled, pooled and measured with a UPLC-MS/ MS metabolomics method developed recently by us to study inflammation and oxidative stress (Figure 1; Schoeman et al, 2018)
Summary
Tumor necrosis factor-a (TNFa) is a central mediator of the inflammatory response (Sedger and McDermott, 2014). Chronic exposure to adverse metabolic and hemostatic risk factors (Masi et al, 2018), obesity (Engin, 2017), or disease states such as kidney disease (Rabelink et al, 2010) or rheumatoid arthritis (van Zonneveld et al, 2010) are all associated with a systemic inflammatory condition and elevated circulating levels of TNFa. As a consequence, TNFa signaling induces the generation of high levels of free radicals in the vascular endothelium that, when excessive, can deplete the cellular anti-oxidant defense systems and lead to a state of oxidative stress and vascular dysfunction (Pisoschi and Pop, 2015). TNFa signaling in endothelial cells involves the activation of NFkB and results in the increased synthesis of reactive oxygen species (ROS) from a number of sources such as mitochondria, NADPH oxidase, uncoupled eNOS, xanthine oxidase, and peroxidases (Cai and Harrison, 2000; Blaser et al, 2016).
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