Desarrollo y análisis de un protocolo de descelularización para la obtención de válvulas cardiacas acelulares

Résumé

Valvular heart disease is one of the most dynamic areas into cardiovascular medicine. Tissue engineering has been one of the most important research fields applied to this pathology. In the present doctoral research, the main goal was to develop a new decellularization technique to obtain acellular heart valves. We analyzed the combination of two potential factors in a decellularization process: the pH and the osmolarity. Those experimental conditions were the key in the selection of two protocols that were able to eliminate cellular components and to preserve the tridimensional tissue structure. An acellular matrix characterization was carried out through microstructural studies, tissue viability, biomechanical and biochemical assays, citotoxicity and biocompatible tests. We selected the optimal decellularization protocol, verifying that it was efficient and reproducible in order to eliminate nucleic acid and to preserve extracellular matrix proteins. Results showed that this new combination of pH and osmolarity shocks were useful in a high cellular density tissue as that of ovine heart valves. Also, it could be a tool to generate no toxic acellular tridimensional structures, free of genetic material and biocompatible.