8/3/2023 0 Comments Chamber 10 portal reloadedWe constructed our 3D BBB model in the form of a tube inside a single layered microfluidic platform. We demonstrate the potential relevance of this model in studying neuroinflammation and cerebral ischemia, by measuring the effect of antioxidant scavenger, edaravone 22, 23, 24 and Rho Kinase (ROCK) inhibitor, Y-27632 25, 26, on BBB permeability and protection against hypoxia-inducing oxidative damage. We validate the tightness of the constructed model by visualizing the delayed dye leakage across our BBB, the expression of endothelial junction proteins, Zonula Occluden-1 (ZO-1) and VE-Cadherin along the cellular boundaries on top/bottom membranes and by observing the blockage of neutrophil migration across the side membrane in the presence of standard chemoattractant. Here, we construct an in vitro BBB model by arranging endothelial monolayers in tube-like structures on a single-layered microfluidic platform 21. Recent models, relying on microscale technologies, could manipulate vertically stacked and planar BBB models, with limited capability for visualizing cellular interaction across the BBB 5, 19, 20. They rely on 2 dimensional cellular contructs and are valuable only for end-point assays and the study of simple stimuli 17, 18. However, their relevance is often limited due to over-simplification. In vitro BBB models provide microenvironment conditions that are usually easier to control and quantify. The complexity of brain microenvironment reduces the ability to isolate the specific roles of the endothelial cells modulating the BBB tightness during neurovascular disorders from various factors in the blood stream or CNS. Despite their importance for health and disease, studies that disrupt BBB in animal models pose significant challenges. These processes could have relevance to various neurological diseases including Alzheimer’s disease (AD) 9, 10, Parkinson’s disease (PD) 11, 12, 13, amyotrophic lateral sclerosis (ALS), brain edema, dementia and multiple sclerosis 14, 15, 16. Recent studies reported that various neuropathology including neuroinflammation 5 and cerebral ischemia 6, 7, 8 can induce the loss of the tightness or the destruction of BBB, allowing the leakage of serum proteins and the entrance of blood cells into the brain tissue. The tight junctions between endothelial cells in the blood-brain barrier (BBB) allow only small nutrient molecules and gases to diffuse across the BBB and limit the entrance of larger, potentially neurotoxic macromolecules, bacteria and leukocytes from the blood 3, 4. Overall, our 3D BBB model provides a robust platform, adequate for detailed functional studies of BBB and for the screening of BBB-targeting drugs in neurological diseases.Įndothelial cells in the brain form an impermeable barrier, which delineates a unique chemical, functional and immunologic environment in a central nervous system (CNS) 1, 2. To validate the functionality of the BBB model, we probed its disruption by neuro-inflammation mediators and ischemic conditions and measured the protective function of antioxidant and ROCK-inhibitor treatments. Moreover, we verified the localization at endothelial cell boundaries of ZO-1 and VE-Cadherin, two components of tight and adherens junctions. We verified the tightness of the BBB by showing its ability to reduce the leakage of dyes and to block the transmigration of immune cells towards chemoattractants. To address these limitations, we developed a 3-dimensional (3D) model of BBB on a microfluidic platform. In vitro models of BBB are simpler, however they have limited functionality and relevance to disease processes. In vivo animal models are highly relevant, however they are hampered by complex, multi-cellular interactions that are difficult to decouple. However, the study of BBB pathology is difficult in the absence of models that are simple and relevant. Blood–brain barrier (BBB) pathology leads to neurovascular disorders and is an important target for therapies.
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