Tesis

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Colección de Tesis y Trabajos de grado (informe final del proyecto de investigación, tesina, u otro trabajo académico diferente a Tesis, sujeto a la revisión y aceptación de una comisión dictaminadora) presentados por alumnos para obtener un grado académico del Tecnológico de Monterrey.

Para enviar tu trabajo académico al RITEC, puedes consultar este Infográfico con los pasos generales para que tu tesis sea depositada en el RITEC.

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Now showing 1 - 5 of 5
  • Tesis de maestría
    Culture of Euglena gracilis in photoautotrophy for paramylon production: effect of pH and media composition
    (Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-11-15) Martín Roldán, María; ; Pacheco Moscoa, Adriana; puemcuervo/tolmquevedo; Senés Guerrero, Carolina; School of Engineering and Sciences; Campus Monterrey; Valledor González, Luis
    Microalgae have been widely studied in biotechnology for their application in various areas such as bioremediation, production of biofuels, or use in nutrition. More specifically, certain species are recognized to produce compounds of high commercial value. Euglena is a group of microalgae characterized by the generation of the reserve polysaccharide paramylon, with promising prospects for its application in pharmacology, nutrition, to produce bioplastics, or biodiesel. Paramylon is accumulated mainly in the presence of organic carbon in the culture medium; however, this represents a challenge when establishing a large-scale culture due to the risk of biological contamination. In this study, an extensive study of the literature was carried out with respect to the autotrophic culture of Euglena gracilis destined to paramylon production. As a result, we evaluated the effect of culture pH, vitamin supplementation, and nitrogen source in the culture medium in order to reach the maximum biomass productivity. A pH of 7.5 and ammonium as nitrogen source were optimum for the autotrophic culture of E. gracilis, while an improvement in productivity was not observed with vitamin supplementation. Finally, it was possible to scale up the photoautotrophic culture of E. gracilis to a 1-L airlift photobioreactor. In conclusion, there is still much optimization work to achieve the biomass and paramylon productivity reported for the heterotrophic cultures of E. gracilis, but the results of this study reveal its viability.
  • Tesis de doctorado
    Corneal endothelium produced by tissue engineering
    (Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-12-14) Montalvo Parra, María Dolores; Zavala Arcos, Judith; tolmquevedo; Ortega Lara, Wendy; Brunck, Marion; Valdez García, Jorge E.; Merayo Llóves, Jesús; School of Engineering and Sciences; Campus Monterrey; Aguilar Yañez, José Manuel
    Engineered corneal endothelium (ECE) must assure cell morphology and physiology. To do so, a biocompatible, transparent, scaffold is the best option. The purpose of this thesis is to produce an ECE with a collagen scaffold and corneal endothelium cells (CECs) harvested in a two-phase system that resembles healthy corneal endothelium (CE) characteristics. Collagen based scaffolds were produced in two steps: 1) Gelification; ~2 µl/mm2 of collagen type I, HEPES solutions, and fetal bovine serum mixture per well were placed on a 12 well plate, 37°C, 5% CO2 for 2 hrs. 2) Vitrification in a Matryoshka System: sealed desiccation chamber with a saturated solution of K2CO3 was placed in an oven set to 40°C. Collagen gels were left inside for 37 days to decrease relative humidity up to 40%. Scaffolds were characterized with confocal microscopy, SEM and spectrophotometry. CECs were isolated from young New Zealand rabbits and from human donor corneas independently. Descement’s membrane was peeled from cornea, digested and, CECs obtained were cultured until confluence in proliferative media (OptiMEM I, FBS 8%, nerve growth factor 20 ng/ml, endothelial growth factor 5ng/ml, CaCl2 200 µg/ml, ascorbic acid 20 µg/ml, chondroitin sulfate 0.08%, antibiotic 1%). Passages 1-2 were carried out in resting media (OptiMEM I 8%FBS). At passage 3, ~24,000 CECs were planted onto 8 mm Ø CV membranes. The alternate use of Proliferative and Resting media conforms the two phase culture system. SEM and confocal microscopy tests showed CV membranes yielded a ~4 µm thickness and smooth surface upon 20 min hydration. SEM also showed collagen fibers merge to form a mesh-like laminar structure. Spectrophotometric scan from 450-700 nm showed a 94-95.5% transmittance. CECs seeded on CV membranes showed adhesion and proliferation at 24 hours; 72 hours served to reach confluence in a ~5 mm Ø. Culture on scaffolds reached canonical CE shape. We produced 12 rabbit and 5 human ECE with desired morphology and specific molecular marker expression. In conclusion, our collagen membrane synthesis method, along with the two phase CECs culture system, offers an option to produce ECE with healthy endothelium characteristics.
  • Tesis de doctorado
    Implantable platform with in-situ vascularization and localized immunosuppression for allogeneic cell transplantation.
    (Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-04) Páez Mayorga,Jesús; PAEZ MAYORGA, JESUS; 778572; tolmquevedo; Nichols, Joan E.; Cuevas Díaz Durán, Raquel; Guajardo Flores, Daniel; School of Engineering and Sciences; Campus Monterrey; Grattoni, Alessandro
    Allogeneic islet transplantation for diabetes management faces the challenge of preventing immune rejection while maintaining enough graft oxygenation for proper metabolic function. Islet encapsulation within membranes impermeable to immune cells prevents rejection at the expense of optimal graft oxygenation. Conversely, direct vascularization avoids hypoxia but requires systemic administration of toxic immunosuppressive drugs. To overcome this problem, we developed the subcutaneously implantable NICHE platform, which integrates in situ graft vascularization and local immunosuppressant delivery, for long-term islet engraftment. NICHE, 3D-printed in nylon, is comprised of independent drug and cell reservoirs separated by a nanoporous membrane. The membrane allows steady local diffusion of immunosuppressant from the drug reservoir into the cell reservoir. Enhanced NICHE vascularization was assessed using mesenchymal stem cells (MSC) and platelet rich plasma (PRP) in rats and non-human primates (NHP). Tunability of antibody and peptide release from NICHE was tested in vitro. Transplantation of allogeneic islets into prevascularized NICHE with localized co-delivery of anti-lymphocyte serum (ALS) and CTLA4Ig was evaluated in immunocompetent diabetic rats. NICHE loaded with MSC had dense vascularity within the cell reservoir by 4 weeks of subcutaneous implantation in rats and NHP. In vitro drug release was tuned via modification of membrane exchange area and drug concentration loaded. In diabetic rats, localized co-delivery of ALS and CTLA4Ig protected allogeneic islets, resulting in reversal to euglycemic state for at least 60 days. Transplanted rats with NICHE responded to glucose challenge comparable to healthy controls and had significantly higher c-peptide levels than no-implant diabetic controls, demonstrating full graft function. ALS-CLTA4Ig co-delivery via NICHE localized drug at the transplant site with limited accumulation in plasma and peripheral tissues, avoiding potential toxicity. Moreover, circulating lymphocyte population remained intact, indicating rats were not systemically immunosuppressed. NICHE is an effective platform for islet allotransplantation with promising adaptability for use with other cell types and pathologies.
  • Tesis de maestría
    Fabrication of highly perfusable gelatin-methacryloyl (GelMA) constructs using flow-based strategies
    (Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-06-05) Pedroza González, Sara Cristina; TRUJILLO DE SANTIAGO, GRISSEL; 256730; Trujillo de Santiago, Grissel; RR, emipsanchez; González Gamboa, Ivonne; Mertgen, Anne-Sophie; De Santiago Miramontes, Ma. de los Ángeles; Escuela de Ingeniería y Ciencias; Campus Monterrey; Álvarez, Mario Moisés
    One of the most important challenges when engineering tissues in vitro is the creation of viable thick constructs. The diffusion of gas and nutrients severely limits the size of engineered constructs. Therefore, the incorporation of perfusable lumen structures within thick engineered tissues is needed for enabling gas exchange, perfusion of nutrients, and waste removal down to the depth of the tissue. Current biofabrication techniques used to create perfusable networks in thick 3D constructs are limited in resolution and control, and they require sophisticated or expensive tools. In this work, we propose a simple technique to develop perfusable hydrogel constructs based on the use of a 3D flow-based biofabrication technique, namely the mini Journal Bearing (mJB), and by employing sacrificial inks. Through the action of regular flows induced in a mJB and the flow-advection of two different hydrogels, we created constructs with an internal sacrificial structure. We used gelatin methacryloyl (GelMA) as a permanent hydrogel matrix, and a drop (100 µL) of gelatin as a fugitive ink/bioink. Here we present a thorough characterization of the microarchitecture and porosity of these constructs. Especially, we demonstrated how permeability increased within these constructs. Additionally, aiming to mimic the architectural complexity of natural tissues, we added nanotopographical cues to our constructs by the incorporation of elongated flexuous plant viruses, namely Turnip Mosaic Virus (TuMV). We conducted our in vitro experiments using myoblasts cells as a biological model and characterized their biological response through time. We fabricated three different types of cell-laden-constructs: GelMA with suspended cells, GelMA with a gelatin ink loaded with cells, and GelMA with a gelatin ink loaded with cells and TuMV. Cells were able to grow faster and for longer in GelMA/gelatin constructs than in pristine-GelMA constructs. While an intricate network of cells was developed after 28 days of culture within permeabilized GelMA/gelatin constructs, only surface proliferation was observed in dense constructs made exclusively with GelMA. The use of GelMA/gelatin-TuMV had an evident morphological effect on cell attachment and proliferation. TuMV 3D meshes providing additional scaffolding within the lumina. While myoblast alignment was strongly evident in GelMA/gelatin where cells adhered mainly to the lamellae walls, in GelMA/gelatin-TuMV constructs, cells alignment was attenuated by interaction with the 3D micromesh of TuMV.
  • Tesis de maestría
    Biofabrication of nanoenhanced hydrogel fibers for muscle tissue engineering using surface chaotic flows: Chaotic 2D-printing
    (Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020) Frías Sánchez, Ada Itzel; TRUJILLO DE SANTIAGO, GRISSEL; 256730; FRIAS SANCHEZ, ADA ITZEL; 887018; ALVAREZ, MARIO MOISES; 26048; Trujillo de Santiago, Grissel; RR; Tamayol, Ali; Ponz Ascaso, Fernando; Samandari, Mohamadmahdi; School of Engineering and Sciences; Campus Monterrey; Alvarez, Mario Moisés
    Multiple human tissues exhibit a fibrous nature. Therefore, the fabrication of hydrogel filaments for biomedical engineering applications is a trending topic. Current tissue models are made of materials that often require further enhancement for appropriate cell attachment, proliferation and differentiation. Here we present a simple strategy, based on the use of mathematically modelable surface chaotic flows, to fabricate continuous, long and thin filaments of gelatin methacryloyl (GelMA) added with Turnip mosaic virus (TuMV) for enhanced muscle tissue engineering. The fabrication of these filaments was achieved by chaotic advection in a finely controlled and miniaturized version of the journal bearing (JB) system. A drop of a pre-gel solution of GelMA was injected on a higher-density viscous fluid (glycerin) and a chaotic flow was applied through an iterative process. The hydrogel drop exponentially deformed and elongated to generate a fiber, which was then photocrosslinked under exposure to UV light. Computational fluid dynamics (CFD) simulations were conducted for the design and prediction of our results. GelMA fibers were then used as scaffolds for C2C12 myoblast cells, and the effect of adding plant-based viral nanoparticles (VNP) to the hydrogel fibers as nano-scaffolds for cellular growth was evaluated. Chaotic 2D-printing was proven to be a viable method for the fabrication of hydrogel fibers. CFD simulations accurately predicted the lengths of the printed fibers, and a correlation coefficient of R2=0.9289 was determined from the experimental and simulated data of the first two cycles. The hydrogel fibers were effective scaffolds for muscle cells and show potential to be used as cost-effective models for muscle tissue engineering purposes. TuMV significantly increased the metabolic activity of the cell-seeded fibers (p<0.05), strengthened cell attachment throughout the first 28 days, improved cell alignment to ~50%, and promoted the generation of structures that resemble natural mammal muscle tissue.
En caso de no señalar algo distinto de manera particular, los materiales son compartidos bajo los siguientes términos: Atribución-No comercial-No derivadas CC BY-NC-ND http://creativecommons.org/licenses/by-nc-nd/4.0
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