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Characterization of Atherosclerotic Plaque Coating for Thrombosis Microfluidics Assays
INTRODUCTION: Studying arterial thrombus formation by in vitro flow assays is a widely used approach. Incorporating human atherosclerotic plaque material as a thrombogenic surface in these assays represents a method to model the pathophysiological environment of thrombus formation upon plaque disrup...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer International Publishing
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8761191/ https://www.ncbi.nlm.nih.gov/pubmed/35087606 http://dx.doi.org/10.1007/s12195-021-00713-9 |
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author | Karel, M. F. A. Lemmens, T. P. Tullemans, B. M. E. Wielders, S. J. H. Gubbins, E. van Beurden, D. van Rijt, S. Cosemans, J. M. E. M. |
author_facet | Karel, M. F. A. Lemmens, T. P. Tullemans, B. M. E. Wielders, S. J. H. Gubbins, E. van Beurden, D. van Rijt, S. Cosemans, J. M. E. M. |
author_sort | Karel, M. F. A. |
collection | PubMed |
description | INTRODUCTION: Studying arterial thrombus formation by in vitro flow assays is a widely used approach. Incorporating human atherosclerotic plaque material as a thrombogenic surface in these assays represents a method to model the pathophysiological environment of thrombus formation upon plaque disruption. Up until now, achieving a homogeneous coating of plaque material and subsequent reproducible platelet adhesion has been challenging. Here, we characterized a novel method for coating of plaque material on glass coverslips for use in thrombosis microfluidic assays. METHODS: A homogenate of human atherosclerotic plaques was coated on glass coverslips by conventional manual droplet coating or by spin coating. Prior to coating, a subset of coverslips was plasma treated. Water contact angle measurements were performed as an indicator for the hydrophilicity of the coverslips. Homogeneity of plaque coatings was determined using profilometric analysis and scanning electron microscopy. Thrombogenicity of the plaque material was assessed in real time by microscopic imaging while perfusing whole blood at a shear rate of 1500 s(−1) over the plaque material. RESULTS: Plasma treatment of glass coverslips, prior to spin coating with plaque material, increased the hydrophilicity of the coverslip compared to no plasma treatment. The most homogeneous plaque coating and highest platelet adhesion was obtained upon plasma treatment followed by spin coating of the plaque material. Manual plaque coating on non-plasma treated coverslips yielded lowest coating homogeneity and platelet adhesion and activation. CONCLUSION: Spin coating of atherosclerotic plaque material on plasma treated coverslips leads to a more homogenous coating and improved platelet adhesion to the plaque when compared to conventional droplet coating on non-plasma treated coverslips. These properties are beneficial in ensuring the quality and reproducibility of flow experiments. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12195-021-00713-9. |
format | Online Article Text |
id | pubmed-8761191 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Springer International Publishing |
record_format | MEDLINE/PubMed |
spelling | pubmed-87611912022-01-26 Characterization of Atherosclerotic Plaque Coating for Thrombosis Microfluidics Assays Karel, M. F. A. Lemmens, T. P. Tullemans, B. M. E. Wielders, S. J. H. Gubbins, E. van Beurden, D. van Rijt, S. Cosemans, J. M. E. M. Cell Mol Bioeng Methods Paper INTRODUCTION: Studying arterial thrombus formation by in vitro flow assays is a widely used approach. Incorporating human atherosclerotic plaque material as a thrombogenic surface in these assays represents a method to model the pathophysiological environment of thrombus formation upon plaque disruption. Up until now, achieving a homogeneous coating of plaque material and subsequent reproducible platelet adhesion has been challenging. Here, we characterized a novel method for coating of plaque material on glass coverslips for use in thrombosis microfluidic assays. METHODS: A homogenate of human atherosclerotic plaques was coated on glass coverslips by conventional manual droplet coating or by spin coating. Prior to coating, a subset of coverslips was plasma treated. Water contact angle measurements were performed as an indicator for the hydrophilicity of the coverslips. Homogeneity of plaque coatings was determined using profilometric analysis and scanning electron microscopy. Thrombogenicity of the plaque material was assessed in real time by microscopic imaging while perfusing whole blood at a shear rate of 1500 s(−1) over the plaque material. RESULTS: Plasma treatment of glass coverslips, prior to spin coating with plaque material, increased the hydrophilicity of the coverslip compared to no plasma treatment. The most homogeneous plaque coating and highest platelet adhesion was obtained upon plasma treatment followed by spin coating of the plaque material. Manual plaque coating on non-plasma treated coverslips yielded lowest coating homogeneity and platelet adhesion and activation. CONCLUSION: Spin coating of atherosclerotic plaque material on plasma treated coverslips leads to a more homogenous coating and improved platelet adhesion to the plaque when compared to conventional droplet coating on non-plasma treated coverslips. These properties are beneficial in ensuring the quality and reproducibility of flow experiments. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12195-021-00713-9. Springer International Publishing 2021-10-27 /pmc/articles/PMC8761191/ /pubmed/35087606 http://dx.doi.org/10.1007/s12195-021-00713-9 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Methods Paper Karel, M. F. A. Lemmens, T. P. Tullemans, B. M. E. Wielders, S. J. H. Gubbins, E. van Beurden, D. van Rijt, S. Cosemans, J. M. E. M. Characterization of Atherosclerotic Plaque Coating for Thrombosis Microfluidics Assays |
title | Characterization of Atherosclerotic Plaque Coating for Thrombosis Microfluidics Assays |
title_full | Characterization of Atherosclerotic Plaque Coating for Thrombosis Microfluidics Assays |
title_fullStr | Characterization of Atherosclerotic Plaque Coating for Thrombosis Microfluidics Assays |
title_full_unstemmed | Characterization of Atherosclerotic Plaque Coating for Thrombosis Microfluidics Assays |
title_short | Characterization of Atherosclerotic Plaque Coating for Thrombosis Microfluidics Assays |
title_sort | characterization of atherosclerotic plaque coating for thrombosis microfluidics assays |
topic | Methods Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8761191/ https://www.ncbi.nlm.nih.gov/pubmed/35087606 http://dx.doi.org/10.1007/s12195-021-00713-9 |
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