Substrate stiffness and shear stress collectively regulate the inflammatory phenotype in cultured human brain microvascular endothelial cells

Abstract Brain endothelial cells experience mechanical forces in the form of blood flow-mediated shear stress and underlying matrix stiffness, but intersectional contributions of these factors towards blood–brain barrier (BBB) impairment and neurovascular dysfunction have not been extensively studie...

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Main Authors: Alexis K. Yates, Heather Murray, Andrew Kjar, Daniel Chavarria, Haley Masters, Hyosung Kim, Alexander P. Ligocki, Angela L. Jefferson, Ethan S. Lippmann
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Fluids and Barriers of the CNS
Subjects:
Online Access:https://doi.org/10.1186/s12987-025-00683-4
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author Alexis K. Yates
Heather Murray
Andrew Kjar
Daniel Chavarria
Haley Masters
Hyosung Kim
Alexander P. Ligocki
Angela L. Jefferson
Ethan S. Lippmann
author_facet Alexis K. Yates
Heather Murray
Andrew Kjar
Daniel Chavarria
Haley Masters
Hyosung Kim
Alexander P. Ligocki
Angela L. Jefferson
Ethan S. Lippmann
author_sort Alexis K. Yates
collection DOAJ
description Abstract Brain endothelial cells experience mechanical forces in the form of blood flow-mediated shear stress and underlying matrix stiffness, but intersectional contributions of these factors towards blood–brain barrier (BBB) impairment and neurovascular dysfunction have not been extensively studied. Here, we developed in vitro models to examine the sensitivity of primary human brain microvascular endothelial cells (BMECs) to substrate stiffness, with or without exposure to fluid shear stress. Using a combination of molecular profiling techniques, we show that BMECs exhibit an inflammatory signature at both the mRNA and protein level when cultured on gelatin substrates of intermediate stiffness (~ 30 kPa) versus soft substrates (~ 6 kPa). Exposure to modest fluid shear stress (1.7 dyne/cm2) partially attenuated this signature, including reductions in levels of soluble chemoattractants and surface ICAM-1. Overall, our results indicate that increased substrate stiffness promotes an inflammatory phenotype in BMECs that is dampened in the presence of fluid shear stress.
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series Fluids and Barriers of the CNS
spelling doaj-art-6743328d61c44f6e97c8d1faa9f11fa22025-08-20T03:06:01ZengBMCFluids and Barriers of the CNS2045-81182025-07-0122112110.1186/s12987-025-00683-4Substrate stiffness and shear stress collectively regulate the inflammatory phenotype in cultured human brain microvascular endothelial cellsAlexis K. Yates0Heather Murray1Andrew Kjar2Daniel Chavarria3Haley Masters4Hyosung Kim5Alexander P. Ligocki6Angela L. Jefferson7Ethan S. Lippmann8Interdisciplinary Materials Science Program, Vanderbilt UniversityChemical and Biomolecular Engineering Department, Vanderbilt UniversityBiomedical Engineering Department, Vanderbilt UniversityChemical and Biomolecular Engineering Department, Vanderbilt UniversityChemical and Biomolecular Engineering Department, Vanderbilt UniversityChemical and Biomolecular Engineering Department, Vanderbilt UniversityChemical and Biomolecular Engineering Department, Vanderbilt UniversityVanderbilt Memory and Alzheimer’s Center, Vanderbilt University Medical CenterInterdisciplinary Materials Science Program, Vanderbilt UniversityAbstract Brain endothelial cells experience mechanical forces in the form of blood flow-mediated shear stress and underlying matrix stiffness, but intersectional contributions of these factors towards blood–brain barrier (BBB) impairment and neurovascular dysfunction have not been extensively studied. Here, we developed in vitro models to examine the sensitivity of primary human brain microvascular endothelial cells (BMECs) to substrate stiffness, with or without exposure to fluid shear stress. Using a combination of molecular profiling techniques, we show that BMECs exhibit an inflammatory signature at both the mRNA and protein level when cultured on gelatin substrates of intermediate stiffness (~ 30 kPa) versus soft substrates (~ 6 kPa). Exposure to modest fluid shear stress (1.7 dyne/cm2) partially attenuated this signature, including reductions in levels of soluble chemoattractants and surface ICAM-1. Overall, our results indicate that increased substrate stiffness promotes an inflammatory phenotype in BMECs that is dampened in the presence of fluid shear stress.https://doi.org/10.1186/s12987-025-00683-4Brain microvascular endothelial cellVascular stiffeningFluid shear stressIn vitro modelsInflammation
spellingShingle Alexis K. Yates
Heather Murray
Andrew Kjar
Daniel Chavarria
Haley Masters
Hyosung Kim
Alexander P. Ligocki
Angela L. Jefferson
Ethan S. Lippmann
Substrate stiffness and shear stress collectively regulate the inflammatory phenotype in cultured human brain microvascular endothelial cells
Fluids and Barriers of the CNS
Brain microvascular endothelial cell
Vascular stiffening
Fluid shear stress
In vitro models
Inflammation
title Substrate stiffness and shear stress collectively regulate the inflammatory phenotype in cultured human brain microvascular endothelial cells
title_full Substrate stiffness and shear stress collectively regulate the inflammatory phenotype in cultured human brain microvascular endothelial cells
title_fullStr Substrate stiffness and shear stress collectively regulate the inflammatory phenotype in cultured human brain microvascular endothelial cells
title_full_unstemmed Substrate stiffness and shear stress collectively regulate the inflammatory phenotype in cultured human brain microvascular endothelial cells
title_short Substrate stiffness and shear stress collectively regulate the inflammatory phenotype in cultured human brain microvascular endothelial cells
title_sort substrate stiffness and shear stress collectively regulate the inflammatory phenotype in cultured human brain microvascular endothelial cells
topic Brain microvascular endothelial cell
Vascular stiffening
Fluid shear stress
In vitro models
Inflammation
url https://doi.org/10.1186/s12987-025-00683-4
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