RESUMEN
BACKGROUND: Assisted Reproductive Technologies (ARTs) have been validated in human and animal to solve reproductive problems such as infertility, aging, genetic selection/amplification and diseases. The persistent gap in ART biomedical applications lies in recapitulating the early stage of ovarian folliculogenesis, thus providing protocols to drive the large reserve of immature follicles towards the gonadotropin-dependent phase. Tissue engineering is becoming a concrete solution to potentially recapitulate ovarian structure, mostly relying on the use of autologous early follicles on natural or synthetic scaffolds. Based on these premises, the present study has been designed to validate the use of the ovarian bioinspired patterned electrospun fibrous scaffolds fabricated with poly(ε-caprolactone) (PCL) for multiple preantral (PA) follicle development. METHODS: PA follicles isolated from lamb ovaries were cultured on PCL scaffold adopting a validated single-follicle protocol (Ctrl) or simulating a multiple-follicle condition by reproducing an artificial ovary engrafted with 5 or 10 PA (AO5PA and AO10PA). The incubations were protracted for 14 and 18 days before assessing scaffold-based microenvironment suitability to assist in vitro folliculogenesis (ivF) and oogenesis at morphological and functional level. RESULTS: The ivF outcomes demonstrated that PCL-scaffolds generate an appropriate biomimetic ovarian microenvironment supporting the transition of multiple PA follicles towards early antral (EA) stage by supporting follicle growth and steroidogenic activation. PCL-multiple bioengineering ivF (AO10PA) performed in long term generated, in addition, the greatest percentage of highly specialized gametes by enhancing meiotic competence, large chromatin remodeling and parthenogenetic developmental competence. CONCLUSIONS: The study showcased the proof of concept for a next-generation ART use of PCL-patterned scaffold aimed to generate transplantable artificial ovary engrafted with autologous early-stage follicles or to advance ivF technologies holding a 3D bioinspired matrix promoting a physiological long-term multiple PA follicle protocol.
Asunto(s)
Folículo Ovárico , Poliésteres , Ingeniería de Tejidos , Andamios del Tejido , Femenino , Folículo Ovárico/crecimiento & desarrollo , Folículo Ovárico/citología , Andamios del Tejido/química , Animales , Poliésteres/química , Ingeniería de Tejidos/métodos , Ovinos , Ovario/crecimiento & desarrollo , Ovario/citología , Oogénesis/fisiología , Oogénesis/efectos de los fármacos , Bioingeniería/métodos , Técnicas Reproductivas Asistidas , Fertilización In Vitro/métodosRESUMEN
Mesoporous bioactive glass nanoparticles (MBGNs) are widely recognized for their ability to bond to hard tissue, while the ions released from the BG structure enhance specific cellular pathways. In this study, the SiO2-P2O5-CaO-MgO-ZnO system was used to successfully synthesize MBGNs by a microemulsion-assisted sol-gel method. The MBGNs calcinated at 600 °C/3 h had a typical phosphosilicate structure together with a poorly crystalline hydroxyapatite (HAp). The addition of ZnO not only led to a higher degree of crystallinity of HAp but also induced a higher porosity of the particles. All MBGNs had a mesoporous structure with an interconnected network of slit shape pores. For each type of composition, two families of highly dispersed spherical nanoparticles could be identified. In vitro tests in simulated body fluid (SBF) proved that after only 3 days of immersion all the materials were covered with a layer of brushite whose degree of crystallinity decreases in the presence of Zn2+. The antibacterial assay revealed a strong inhibitory effect for all samples after 40 h of contact. Simultaneously, MBGNs did not increase the intracellular oxidative stress while it stimulated the cell proliferation process.
RESUMEN
Bioglass (BG) is a class of biomaterials increasingly approached in biomedical applications, such as in regeneration of hard tissues, due to the properties of bioactivity, osteoinductivity, osteoconductivity, but also the high rate of biodegradation, both in vitro and in vivo. The present paper addresses the obtaining of bioglasses from the ZnO(MgO)-CaO-SiO2-P2O5 system by the sol-gel method and the use of a surfactant to ensure a specific surface or high open porosity, starting from S53P4 bioglass (53% SiO2, 23% Na2O, 20% CaO, 4% P2O5), also known as BoneAlive®. The precursor powders were analyzed from the phase composition point of view by complex thermal analysis and X-ray diffraction, the vitreous powders were assessed from the compositional point of view by X-ray diffraction, morpho-structural by scanning electron microscopy, specific surface area and the pore size dimension by the Brunauer-Emmett-Teller (BET) analysis, dispersion by laser granulometry, and also cell biology and surface mineralization tests were performed by immersion in SBF (simulated body fluid). The system proposed in this paper ZnO(MgO)-CaO-SiO2-P2O5 was successfully obtained by sol-gel method. The results showed the higher interaction between the samples and the SBF medium for samples containing magnesium (M2) and the lowest degree of mineralization after immersion in SBF was noticed for samples containing zinc (M1). The results also prove that by incorporating different ionic species in bioglass composition-Zn2+ and Mg2+, biocompatibility and antibacterial properties will be significantly enhanced.