Ciencias Exactas y Ciencias de la Salud
Permanent URI for this collectionhttps://hdl.handle.net/11285/551039
Pertenecen a esta colección Tesis y Trabajos de grado de las Maestrías correspondientes a las Escuelas de Ingeniería y Ciencias así como a Medicina y Ciencias de la Salud.
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- Nanovesículas similares a exosomas (NVEs) vegetales para mejorar la entrega dirigida de carotenoides(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2022-06-15) Cisneros Pardo, Anahí Guadalupe; GONZALEZ VALDEZ, JOSE GUILLERMO; 234501; González Valdez, José Guillermo; tolmquevedo, emipsanchez; Marilena, Antunes Ricardo; Luis Alberto, Mejía Manzano; Escuela de Ingeniería y Ciencias; Campus Monterrey; Ramos Parra, Perla AzucenaLos carotenoides juegan un papel importante en la salud ocular al tener la capacidad de proteger a la retina de lesiones fotoquímicas. Sin embargo, existen diversos obstáculos para proveer un efecto terapéutico específico en el tejido, por lo que existe la necesidad de buscar nuevas alternativas que permitan incrementar su biodisponibilidad, como los son las nanovesículas extracelulares similares a exosomas (NVEs) de origen vegetal, las cuales desempeñan un papel importante en la comunicación celular. Con lo anterior, se generó la hipótesis de que la carga mediante electroporación de las NVEs derivadas de jugo y callo de zanahoria con carotenoides podría incrementar la eficiencia de suministro además de la biodisponibilidad de estos compuestos. La metodología desarrollada constó de tres fases principales: La primera, referente al establecimiento del material inicial para la obtención de las NVEs, seleccionándose jugo y callos de zanahoria. La segunda fase consistió en el aislamiento, purificación, caracterización y carga con carotenoides de las NVEs. Finalmente, la tercera fase tuvo como objetivo el ensayo de bioactividad de carotenoides en la línea celular ARPE-19 del epitelio pigmentario de la retina humana utilizando estas nanovesículas. Dentro de los principales resultados se obtuvieron NVEs aislados mediante ultracentrifugación de un tamaño entre 231 y 1631 nm, y una carga del potencial zeta negativo entre 1.26 y 11.13 mV. Se obtuvo el perfil de los carotenoides nativos en los NVEs de jugo de zanahoria en donde se encontraron hasta 12 carotenoides, siendo identificadas 3 xantófilas y 5 carotenos. Se evaluaron 4 métodos de carga distintos y se obtuvo como resultado que la electroporación (200 mV, 50 µF) tuvo una mayor eficiencia de carga (80%) para zeaxantina. En el ensayo de viabilidad celular para analizar la biodisponibilidad de los carotenoides, se obtuvo que las NVEs derivadas de callo cargadas con xantófilas presentan una mayor protección (< 95%) contra agentes oxidantes en comparación con las NVEs derivadas de jugo cargadas. Los resultados obtenidos en el presente trabajo son considerados de alto impacto debido al creciente interés en encontrar nuevas formas de administración de biocompuestos que tengan propiedades terapéuticas, que puedan potencialmente producirse a gran escala y que sean provenientes de una fuente segura para el consumo humano, de modo que los productos a base de zanahoria, como los NVEs, podrían ser considerados como una atractiva posibilidad para dichas necesidades.
- Optimization in the isolation and purification procedure of pristimerin, an anticancerigen, by Aqueous Two-phase Systems(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-06-11) González Félix, Martha Alicia; González Valdez, José Guillermo; puemcuervo; Ibarra Herrera, Celeste Concepción; González Torres, Maykel; School of Engineering and Sciences; Campus Monterrey; Mejía Manzano, Luis AlbertoThe natural triterpene pristimerin possesses interesting potential against several types of cancer. However, its production at industrial levels requires novel environmentally friendly and efficient extraction procedures. Here we analyze an ethanol-phosphate Aqueous Two-Phase Systems (ATPS) and its design parameters such as Tie-Line Length (TLL) and volume ratio (VR) on the direct extraction of pristimerin, concluding that high TLL 70 % (w/w) and a low VR (0.33) values provide the best extraction yield and purity. Partition experiments provided a compound KP value of 3.12 ± 1.21, reaffirming its preference for the ethanolic-rich phase. Besides, green assessment positioned the extraction in the classification A with 90.93 of 100 points. Furthermore, analysis at different laboratory scales (1X, 10X and 100X) demonstrated the scalability and robustness. Finally, a complete bioprocess based on the obtained results is suggested as a tool to address the pharmaceutical potential of this molecule in a sustainable and efficient manner.
- Recovery and purification of exosomes using polymer-polymer aqueous two-phase systems (ATPS)(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-12-03) Torres Bautista, Abril Lorena; González Valdez, José Guillermo; emipsanchez/tolmquevedo; Torres Acosta, Mario Antonio; Mejía Manzano, Luis Alberto; Ramos de la Peña, Ana Mayela; Escuela de Ingeniería y Ciencias; Campus MonterreyExosomes are a type of extracellular vesicles that present natural and attractive characteristics such as nano size (30-200 nm) and unique structure for their use as drug delivery systems for drug therapy, biomarkers for prognostic, diagnostic and personalized treatments. So far, one of the major challenges for therapeutic applications of exosomes is the development of optimized methods for their isolation. In this context, Aqueous Two-Phase Systems (ATPS) have shown to be an alternative method to isolate biological molecules and particles with promising expectations for exosomes. In this work, fractionation of exosomes obtained from a CaCo2 cell line and the contaminants present in their culture media was performed in 20 different polymer-polymer aqueous two-phase systems (ATPSs) in a discrete way. The effect of system design parameters such as polymer composition, molecular weight, and TLL (15-30%) on PEG-Dextran, Dextran-Ficoll and PEG-Ficoll systems were studied keeping phase volume ratio (VR) and pH constant at 1.0 and 7.0, respectively. After partition analysis, 4 out of the 20 systems presented the best exosome fractionation from the contaminants under initial conditions and were selected for further partition optimization via salt addition (NaCl) to observe the behavior of both samples, individually, and improve the efficiency of collection. The PEG 10,000 gmol-¹ – Dextran 10,000 gmol-¹ system at TLL 25% w/w, with the addition NaCl to reach a final concentration of 25 mM, showed the best isolation efficiency resulting in exosome purification factor of 2 and furtherly demonstrated that ATPS has the potential for the selective recovery these promising nanovesicles.
- Exosome-mediated insulin delivery for the treatment of diabetes mellitus(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-11-16) Rodríguez Morales, Belén; GONZALEZ VALDEZ, JOSE GUILLERMO; 234501; González Valdez, José Guillermo; tolmquevedo, emipsanchez; Hernández Pérez, Jesús; Rodríguez Sánchez, Dariana Graciela; School of Engineering and Sciences; Campus Monterrey; Antunes Ricardo, MarilenaExosomes are membrane-bound extracellular nanovesicles of 30-150 nm released by almost all cell types. Since they were discovered in 1983, numerous studies yielded clear evidences that exosomes serve as essential messengers for hundreds of biological-signaling processes as well as in pathological processes. In consequence of the discovery of exosomal natural functions, a wide range of applications have been developed in several fields of study, especially in drug delivery for therapeutic use. Following this growing stream of study, the aim of this work was to test the efficiency of exosome-mediated human insulin delivery using exosomes extracted from three different cell lines: hepatocellular carcinoma (HepG2); primary dermal fibroblasts (HDFa) and pancreatic β cells (RIN-m); all related to the production and/or the ability to sense insulin and, consequently regulate glucose levels in the extracellular medium. The obtained results revealed that the optimal loading efficiency was achieved by a 200 V electroporation, in comparison to an incubation at room temperature. Moreover, the maximum in vitro exosome uptake was reached after 6 h of incubation with loaded exosomes, and lightly decrease 24 h after adding the exosomes. Furthermore, glucose quantification assays revealed that exosome-mediated incorporation of insulin present significative differences in HDFa and HepG2 cells, enhancing the transport in HDFa, in comparison with free human insulin effect in the regulation of extracellular glucose levels. On the other hand, no significative differences were found between both treatments in RIN-cells. Hence, the results suggest that exosomes could be a potentially valuable tool for a biocompatible and stable insulin delivery as part of a future treatment of diabetes mellitus.