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Development of a three-dimensional in vitro blood-brain barrier using the chitosan-alginate polyelectrolyte complex as the extracellular matrix

Development of a three-dimensional in vitro blood-brain barrier using the chitosan-alginate... Polyelectrolyte complexes (PECs) consist of a spontaneous assembly of oppositely charged polysaccharides. PECs can be used to obtain a hydrogel tissue scaffold in tissue culture. In this study, it is aimed to use PEC as a blood-brain barrier (BBB) model scaffold. By mixing polycationic chitosan and polyanionic alginate solutions at a certain ratio it was obtained a 3D hydrogel scaffold and mimicked in vivo environment of the tissue. The PEC hydrogel scaffold’s chemical, physical, and mechanical characterizations were performed with FTIR, DSC, DMA, and Micro-CT analyses. In order to develop an in vitro BBB model, the human neuroblastoma cell line (SH-SY5Y) and mouse astrocyte cell line (C8-D1A) were mixed into a hydrogel, which is the abluminal side of the BBB. Human microvascular endothelial cells (HBEC-5i) were seeded on the hydrogel, and it was aimed to mimic the luminal side of the BBB. The characterization of the BBB model was determined by measuring the TEER, observation of the cell morphology with SEM, performing the permeability of Lucifer Yellow, and observation of tight junction proteins with immunofluorescence staining. As a result, HBEC-5i cells expressed tight junction proteins (ZO-1 and Claudin-5), showed TEER of 340 ± 22 Ω.cm2, and the Lucifer Yellow permeability of 7.4 × 10−7 ± 2.7 × 10−7 cm/s, which was suitable for use as an in vitro BBB model. Using a hydrogel PEC composed of chitosan and alginate as an extracellular matrix increased the direct interaction of endothelial cells, astrocytes, and neurons with each other and thus obtained a much less permeable model compared to other standard transwell models.Graphical abstract http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Bioactive and Compatible Polymers: Biomedical Applications SAGE

Development of a three-dimensional in vitro blood-brain barrier using the chitosan-alginate polyelectrolyte complex as the extracellular matrix

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Publisher
SAGE
Copyright
© The Author(s) 2023
ISSN
0883-9115
eISSN
1530-8030
DOI
10.1177/08839115231157096
Publisher site
See Article on Publisher Site

Abstract

Polyelectrolyte complexes (PECs) consist of a spontaneous assembly of oppositely charged polysaccharides. PECs can be used to obtain a hydrogel tissue scaffold in tissue culture. In this study, it is aimed to use PEC as a blood-brain barrier (BBB) model scaffold. By mixing polycationic chitosan and polyanionic alginate solutions at a certain ratio it was obtained a 3D hydrogel scaffold and mimicked in vivo environment of the tissue. The PEC hydrogel scaffold’s chemical, physical, and mechanical characterizations were performed with FTIR, DSC, DMA, and Micro-CT analyses. In order to develop an in vitro BBB model, the human neuroblastoma cell line (SH-SY5Y) and mouse astrocyte cell line (C8-D1A) were mixed into a hydrogel, which is the abluminal side of the BBB. Human microvascular endothelial cells (HBEC-5i) were seeded on the hydrogel, and it was aimed to mimic the luminal side of the BBB. The characterization of the BBB model was determined by measuring the TEER, observation of the cell morphology with SEM, performing the permeability of Lucifer Yellow, and observation of tight junction proteins with immunofluorescence staining. As a result, HBEC-5i cells expressed tight junction proteins (ZO-1 and Claudin-5), showed TEER of 340 ± 22 Ω.cm2, and the Lucifer Yellow permeability of 7.4 × 10−7 ± 2.7 × 10−7 cm/s, which was suitable for use as an in vitro BBB model. Using a hydrogel PEC composed of chitosan and alginate as an extracellular matrix increased the direct interaction of endothelial cells, astrocytes, and neurons with each other and thus obtained a much less permeable model compared to other standard transwell models.Graphical abstract

Journal

Journal of Bioactive and Compatible Polymers: Biomedical ApplicationsSAGE

Published: May 1, 2023

Keywords: Chitosan; alginate; brain microvascular endothelial cell; astrocyte; neuron

References