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Advanced in-vitro models of the blood-brain barrier

Micol Baronio

Advanced in-vitro models of the blood-brain barrier.

Rel. Gianni Ciofani, Elisa Mele. Politecnico di Torino, Corso di laurea magistrale in Ingegneria Biomedica, 2019


This master thesis project focuses on the realization of an advanced in-vitro model of Blood-Brain-Barrier (BBB). The model innovation is the possibility to test drugs and nanovectors permeability, understanding which therapies could be effective in brain cancer treatment, and in particularly for glioblastoma multiforme. The aim of this model is to overcome all the limitations related to ineffective 2D and animal models, the latter usually related also to ethical concerns. The polymeric porous and biocompatible membranes were fabricated, via vapour induced phase separation (VIPS). These supports were mechanically and surface characterized, and their morphological images were taken by FEGSEM microscope. These chemical and physical procedures were performed at the Materials Engineering Department of Loughborough University (UK). In the laboratories of Smart Bio-Interfaces, at the Italian Institute of Technology (IIT), in Pontedera (PI, Italy), the membranes were assembled on stiff PET supports, in order to obtain the final structure in which seed endothelial cells (hCMEC/D3) and human astrocytes to achieve an in-vitro barrier model, very similar to the biological human BBB. In final models were performed tests of integrity (by Transendothelial electrical resistance), permeability and confocal imaging. The thesis consists of abstract, introduction, materials and methods, results and discussion, conclusions. Introduction In the first chapter are summarised the biological information of the BBB and glioblastoma multiforme and the state of art of the current brain cancer treatments. The second chapter describes previous approaches of BBB in-vitro models realization, the concepts of tissue engineering and organ-on-a-chip and conventional fabrication techniques. There is also a description of the vapour induced phase separation (VIPS) and the definitions of the process parameters. Materials & Methods The third chapter is more technical and gives details about the experimental chemical procedures used to produce the membranes, describing the materials and the methods adopted. The inadequate results are also described, in order not to take them into consideration in further projects. The fourth chapter explains the biological procedures of the project, focusing on the assembly of the membranes on stiffer supports and the seeding of endothelial cells and human astrocytes on the final structure. All the protocols adopted in the laboratory for the work with the cells are described. In the fifth chapter are described, in detail, all the adopted instruments and protocols for the different physical and mechanical characterizations of the membranes and the final assembled BBB models. Results & Discussion The sixth chapter contains all the results and their explanations, obtained through the mechanical, surface and microstructural characterizations, trans endothelial electrical resistance (TEER), permeability and diffusion tests, degradation and vitality analysis performed in-vitro, both in absence and presence of cells. Conclusions The conclusions discuss about all the obtained results and the future perspectives of the fabricated BBB in-vitro model.

Relators: Gianni Ciofani, Elisa Mele
Academic year: 2019/20
Publication type: Electronic
Number of Pages: 110
Additional Information: Tesi secretata. Fulltext non presente
Corso di laurea: Corso di laurea magistrale in Ingegneria Biomedica
Classe di laurea: New organization > Master science > LM-21 - BIOMEDICAL ENGINEERING
Ente in cotutela: Materials Department, Loughborough University (REGNO UNITO)
Aziende collaboratrici: UNSPECIFIED
URI: http://webthesis.biblio.polito.it/id/eprint/12254
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