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Dewez_65311700_2023.pdf
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- Heart valves play a crucial role in blood circulation and any failure results in cardiac complications. Tissue engineering could bring a promising solution to the treatment of valve pathologies, but its success relies on a complete understanding of its complex macro- and microstructure. While classical 2D histology has provided valuable insights in the visualization of leaflet microstructure, it does not allow high-resolution 3D imaging. This thesis focused on the optimization of an imaging protocol designed for mitral valves (MVs). The objective was to develop a method that allows high-resolution 2D and 3D ex-vivo imaging to enhance our knowledge of the microstructure of MVs. Micro-computed tomography (micro-CT) is an imaging technique providing highresolution 3D images and allowing the visualization and analysis of microstructure of samples. Throughout this study, we evaluated the advantages and limitations of different imaging techniques, including contrast-enhanced computed tomography (CECT) and cryogenic-CECT (cryo-CECT), as well as two contrast-enhancing staining agents (CESAs), Lugol’s iodine and 1:2 Hafnium-Wells-Dawson polyoxometalate (Hf-WD 1:2 POM). Our results showed better visualization of adipocytes using Lugol’s iodine, both with CECT and cryo-CECT. Lugol’s iodine diffused rapidly (<1h) through the tissue and did not induce a significant change in sample volume. Vessel and connective tissue visualization was enhanced by the use of Hf-WD 1:2 POM combined with cryo-CECT. This enabled 3D representation of the blood vessels and collagen and elastin fibers present in the samples. These results demonstrate the potential of CECT and cryo-CECT to improve our knowledge on the MV leaflet microstructure and this might offer a promising prospect on new treatments for valve pathologies.