Future bioengineering applications in female reproductive medicine. From basic science to animal models
- Autores: Emilio Frances Herrero
- Directores de la Tesis: Irene Cervelló Alcaraz (dir. tes.), Vicente Serra Serra (codir. tes.)
- Lectura: En la Universitat de València ( España ) en 2024
- Idioma: inglés
- Número de páginas: 317
- Tribunal Calificador de la Tesis: Pedro Baptista (presid.), Manuel Mata Roig (secret.), Christiani Andrade Amorim (voc.)
- Programa de doctorado: Programa de Doctorado en Biomedicina y Biotecnología por la Universitat de València (Estudi General)
- Materias:
- Enlaces
- Tesis en acceso abierto en: TESEO
- Resumen
Extracellular matrix (ECM) hydrogels are promising and remarkable tissue engineering biomaterials. They aim to recreate the 3D native environment that cells need to differentiate and develop specific functions. ECM hydrogels have emerging applications in the field of gynecology. These include the development of next generation in vitro platforms, such as 2D coating or 3D encapsulation systems, as well as in vivo applications involving ECM hydrogels as cell carriers or drug depots.
The five objectives of this thesis were (1) to design oviductal ECM (OviECM) hydrogels from decellularized (DC) rabbit tissue, (2) to study the effect of an OviECM hydrogel coating during in vitro embryonic development, (3) to design ovarian ECM (OvaECM) hydrogels from DC bovine ovarian cortex (OC), (4) to test the potential of the bovine OvaECM hydrogel to promote and sustain murine folliculogenesis in vitro, and (5) to evaluate the feasibility and effectivity of using OvaECM hydrogels as carriers of growth factors (GFs) into the murine ovarian niche to reverse premature ovarian failure (POF).
To achieve these objectives, oviductal and ovarian tissue were isolated from rabbit and bovine species respectively, subjected to different decellularization protocols, and processed to create OviECM and OvaECM hydrogels, whose physicochemical properties were studied. The effect of an OviECM hydrogel coating during in vitro embryonic development was assessed in terms of viability, metabolic behavior, and gene expression, as well as health and phenotype of the offspring. Later, development of murine follicles on OvaECM hydrogels was assessed through analysis of viability, growth, hormone production, and oocyte competence. Finally, the feasibility and effectivity of these hydrogels as carriers of GFs into the murine ovarian niche to reverse POF was evaluated. We generated and characterized in depth oviduct and ovarian acellular hydrogels, preserving a large part of the biochemical richness of the native tissues. Our results showed that culturing in vitro preimplantation rabbit embryos on OviECM coatings improved their metabolic performance and showed different gene expression profiles compared to those cultured under standard conditions. These embryos also exhibited higher implantation and birth rates, and gave rise to offspring with a higher weight. OvaECM hydrogel coatings were able to support murine folliculogenesis in vitro, inducing more physiological growth than 2D culture conditions, and improving rates of oocyte maturation and embryo development. Furthermore, the OvaECM hydrogel proved to be an excellent carrier of GFs, allowing prolonged release of GFs into the ovarian niche of a murine POF model. Improvements in follicular populations, cell proliferation and vasculature were observed, while apoptosis and fibrosis were reduced. In addition, more oocytes and embryos were obtained by ovarian stimulation, and the natural fertility of the mice was increased in the long term.
