Formation of hydrogel-based three-dimensional static blood-brain barrier model
Project no. 20255 · Status: Kapanmış · 2018
Summary
Polyelectrolyte complexes consist of a spontaneous assembly of oppositely charged polysaccharides. Polyelectrolyte complexes can be used to obtain a hydrogel tissue scaffold in cell culture. In this study, it is aimed to use polyanionic and polycationic polymer solutions as a blood-brain barrier (BBB) model scaffold that has not been used before. By mixing polycationic chitosan solution and polyanionic alginic acid solutions at a certain rate, it was ensured that cells form a suitable three-dimensional scaffold to mimic in vivo environment. This scaffold was used to mimic the BBB basal lamina, which has not been used before in the literature. In order to create an in vitro BBB model, the human neuroblastoma cell line (SH-SY5Y) and mouse astrocyte cell line (C8-D1A) were mixed into a polyanionic alginic acid solution and the hydrogel was obtained by crosslinking with the polycationic chitosan solution. This part is the abluminal side imitating the brain part of the BBB. Human microvascular endothelial cells (HBEC-5i) were seeded on the hydrogel, and it was aimed to create the luminal side mimicking the blood part of the BBB model. Whether the model is realistic or not, was determined by measuring the transendothelial electrical resistance (TEER) and comparing it with in vivo values by performing the permeability analysis of some reference molecules passing more or less than the in vivo BBB. In addition, whether the tight junction proteins that BBB endothelial cells should have were expressed or not was analyzed by the immunofluorescence staining method. HBEC-5i cells on the obtained hydrogel BBB model expressed tight junction proteins, showed TEER of 340±22 ?.cm2 at the end of the 3rd day, and that the caffeine permeability was suitable for use as an in vitro model.