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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- Microbial tale: using chaotic bioprinting to create a structured multi-strain probiotic(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2025-06-18) Flores Loera, Francisco Javier; Alvarez, Mario Moises; emipsanchez; Luna Aguirre, Claudia Maribel; Rocha Pizaña, María del Refugio; School of Engineering and Sciences; Campus Monterrey; Trujillo de Santiago, GrisselProbiotic therapies offer great potential for addressing gut dysbiosis, but current approaches are limited by low strain diversity, high production costs, and the challenges of culturing strict anaerobes. To overcome these limitations, this work introduces a novel strategy combining based on continuous chaotic bioprinting to create structured (micro-architected) co-cultures of probiotic bacteria. Using a Kenics static mixer–based printhead, we fabricated alginate hydrogel filaments with an internal multilayered microarchitecture containing four probiotic strains: Bifidobacterium bifidum, Bacteroides fragilis, Lactobacillus rhamnosus, and Streptococcus thermophilus. The spatial arrangement of the multilayered architecture was designed to promote cooperative interactions, particularly by embedding strict anaerobes between facultative anaerobes to create self-sustaining hypoxic niches. The printed constructs were characterized over 72 hours using fluorescence microscopy, colony-forming unit counts, LIVE/DEAD assays, and qPCR. Results showed that structured co-cultures exhibited higher viability, enhanced growth, and more balanced population dynamics than the monocultures of each bacterial strain and unstructured (scrambled) co-cultures. This study demonstrates that chaotic bioprinting enables precise spatial control over microbial ecosystems, allowing the rational design of microbial communities with tailored interactions. The approach presents a powerful and scalable platform for next-generation probiotic production and opens new opportunities for engineered microbiomes, synthetic biology, and living material design.
- Design for manufacturing and assembly of a multi material bioprinting system towards tissue engineering applications(2025-06-13) Lera Julián, Miguel Ángel; Martínez López, José Israel; emipsanchez; Vázquez Lepe, Elisa Virginia; López Botello, Omar; Chuck Hernández, Cristina; School of Engineering and Sciences; Campus MonterreyLight-based techniques have great potential in bioprinting for tissue engineering, given their inherent advantages in high spatial resolution (10–100 μm) and improved cell viability (>85%) compared to traditional extrusion-based systems. However, current apparatuses found in the state of the art are limited in usability and functionality due to legacy single-material design constraints and the early development stage of photopolymerization-based bioprinters. As tissue constructs become increasingly complex, there is a need to establish a new framework for light-based equipment tailored to specific tissue engineering applications. This work presents the development of multi-material bioprinting equipment that integrates 4K digital light projection with an automated rotating four-vat system, enabling sequential use of bioinks with distinct mechanical and biochemical properties. For this endeavor, the scalability and manufacturability of the apparatus were addressed using Function Tree analysis, Quality Function Deployment (QFD), and Design for Manufacturing and Assembly (DFM&A) principles. These tools guided the definition of a feature set for meniscal tissue regeneration, including layered constructs with stiffness gradients and bioactive cues. The system was designed in Fusion and fabricated using a combination of rapid prototyping techniques. This included the 3D printing of custom resin vats, CNC machining of structural elements, and the development of bespoke electronic components for control and actuation. Initial validation was carried out using a single-vat configuration and Anycubic clear photopolymer resin. Printing trials demonstrated the resolution capacity of the optical system and successful layer-by-layer polymerization using 405 nm light exposure. These results confirm the operational feasibility of the system and establish a baseline for future multi-material implementation using photocurable bioinks
- Biofabrication of anisotropic constructs using extrusive chaotic printing(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2023-12-01) Hernandez Medina, David Hyram; Trujillo de Santiago, Grissel; mtyahinojosa, emipsanchez; Bolívar Monsalve, Edna Johana; Castillo, Jimmy; Esquivel Alfaro, Marianelly; Escuela de Ingeniería y Ciencias; Campus Monterrey; Álvarez, Mario MoisésAligned tissue constitutes a considerable percentage of the body mass, and it is this anisotropic characteristic which confers certain mechanical and functional properties to the tissue. For creating tissue-like structures that resembles the body, one relevant challenge lies on using biomaterials and shaping them to create aligned structures. These constructs serve as scaffolding materials that promote cell proliferation and differentiation that can eventually become a working tissue. In this study, continuous chaotic printing was used to fabricate highly oriented printed scaffolds and bioprinted cell-laden constructs. First, we assessed the effectiveness of a chaotic extrusion printhead, containing a Kenics Static Mixer (KSM), as a tool to align fibrillar inks. In short, soft fibrillar materials (i.e., alginate-cellulose and collagen) were chaotically printed into 1 mm thick filaments and scaffold anisotropy was assessed in terms of printed microstructures orientation. Filaments showed orientation up to 68% in a -15° to 15° region where the main axis (i.e., aligned fiber) correspond to 0°. Moreover, we assessed the capability of chaotic bioprinting to produce aligned and pre-vascularized skeletal-muscle-like tissues. Briefly, fibers were bioprinted using three inks: a hydrogel loaded with myoblasts (C2C12 cells), a non-crosslinkable hydrogel to create inner vessels inside the fiber, and a high viscosity hydrogel loaded with mesoporous bioactive glass (MBG) to provide mechanical robustness to the fiber. A comparison was made between homogeneous (pre mixed) fibers and pre-vascularized fibers with a layered inner structure. The constructs were cultured up to 21 days and demonstrated high viability (>85%) and a significant relation in the orientation trend of the F-actin filaments with the stratification. Overall, we demonstrated that chaotic printing is a practical tool for fabricating anisotropic constructs with both, fibrillar inks and cell-laden constructs.
- Dual bio-printing system for cell deposition of hydrogels using a piston-based controlled nozzle(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2022) Castillo Madrigal, Victor; Chairez Oria, Jorge Isaac; mtyahinojosa, emipsanchez; García González, Alejandro; Escuela de Ingeniería y Ciencias; Campus Guadalajara; Perfecto, YocanxóchitlIn recent years, cell culture has increasingly utilized various 3D scaffolds and hydrogels to promote advanced additive manufacturing within cell culture. Additionally, various types of cell lines have been employed, with mesenchymal stem cells (MSCs) and fibroblasts being among the most commonly used. The aim of this project is to design an automated dual bioprinting system for cell culture. This system will deposit both hydrogels and cells sequentially, creating a foundation for biological tissue.Therefore, reducing the probability of cell culture contamination and decreasing human interaction. To achieve the goal, the project was first designed following specific criteria specifications. This was facilitated by previous cell culture training, which helped in better understanding the necessities of the project.After that, the next step involved simulating the device using CAD software (Solidworks) to create a 3D representation of the system's prototype. This prototype consists of three linear actuators (X-Y-Z axis) and two extruders for each deposited material.Then, the fully virtual device is exported to Matlab Simulink in order to simulate a control process with a PD controller in each actuator and extruder using a sine signal as a reference. Finally the crafting of the prototype was achieved operating tools from the metal crafting laboratory, and experimental processes were started. The study evaluated the feasibility of developing a 3D bioprinting machine. The experiment proved that it is possible to replicate the behavior from the simulated space into a real experiment, using a dual bioprinting system for depositing cells with controlled processes. The error from the control process was below 1% in the virtual enviroment meanwhile in reality the error mantain around 3%. As future work is still to validate the system performing biotic tests with a hydrogel made of alginate and as crosslinker calcium chloride. This system can be further applied in automatized cell culture system, bioprinting on no linear surfaces or even as part of a bioreactor system.
- Development of a Low-Cost and High-Resolution Stereolithography Bioprinting System(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-08) Pérez Cortez, Juan Enrique; Martínez López, José Israel; emimmayorquin; Rodríguez González, Ciro Ángel; Chuck Hernández, Cristina; Escuela de Ingeniería y Ciencias; Campus Monterrey; Vázquez Lepe, Elisa VirginiaResearchers that require access to 3D biological constructs for tissue engineering face significant cost barriers when it comes to using light-based bioprinters. The bioprinters that are typically used are either commercial-grade expensive technology or modified DIY extrusion apparatuses. Despite the potential benefits of low-cost equipment, there are few examples of such equipment being used in the context of light-based bioprinters. Additionally, the high cost of bioinks, large volume consumption, and a lack of information on parameter selection for light-based bioprinting restrains the adoption of this technology. In this work, retrofitting a commercial light-based 3D printer is showcased to demonstrate that it is possible to adapt this technology to build low-cost and high-resolution 3D bioprinters. To prove the capacities of this equipment, several manufacturability and biological tests are performed with a worldwide used bioink, Gelatin Methacryloyl (GelMa). A set of experiments performed in two different equipment (Anycubic Photon Mono 2K and 4K) is documented. Moreover, in this work it is showcased the use of C2C12 and Alpha TC1 Clone 6 cells to create muscular fibers and a mini pancreas model, respectively. Actin-Dapi and Live-Dead fluorescence assays at different days of culture show positive results for biocompatibility of the process. This work demonstrate that the presented low-cost retrofitting has great potential to be used as a 3D bioprinting system
- Development of a GelMA-based bioink enhanced with minimally-processed tissue for the fabrication of skeletal muscle(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-06-12) Tavares Negrete, Jorge Alfonso; TAVARES NEGRETE, JORGE ALFONSO; 744164; Trujillo de Santiago, Grissel; ilquio/puemscuervo; Pérez Carrillo, Esther; Mertgen, Anne-Sophie; Santiago Miramontes, Ma. de los Ángeles de; Escuela de Ingeniería y Ciencias; Campus Monterrey; Álvarez, Mario MoisésBioprinting, an emerging technology that uses living cells and biomaterials for fabricating engineered tissues with complex three-dimensional (3D) architecture, has become popular as an alternative to conventional 3D-culture techniques. In typical extrusion bioprinting, a bioink (i.e., generally a suspension of cells in a hydrogel) is extruded through a printing head to build an artificial 3D tissue in a layer-by-layer-fashion. Bioinks are key components of a bioprinting process. Bioinks have to meet appropriate biological and rheological characteristics to be printable and to provide a proper microenvironment to cells. Here, we present a simple strategy to develop cost-effective bioinks based on gelatin methacryloyl (GelMA) enhanced with fetal minimally processed muscle tissue (MPT). This strategy is intended to supplement bioinks with growth factors, glycosaminoglycans and proteins from fetal tissue, due to its biochemical composition rich in growth factors and peptides. As a first demonstrative model, we supplemented GelMA hydrogels with 0.5, 1, 2% dried and powdered MPT derived from a goat or calf fetus. The biochemical characterization of MPT showed that our minimally processed technique preserves more than 65% of GAG content compared to traditional methods (i.e. decellularization). The rheological profile of our hydrogels was analyzed to determine a suitable working-window of printing parameters, all inks have shear thinning behavior. Cell-culture experiments showed that the incorporation of MPT in the hydrogels influences the metabolic activity of the myoblast cells (C2C12) and confers structural stability to the hydrogels for cultures of up to 28 days in comparison to pristine GelMA hydrogels. The cell orientation of the GelMA-MPT bioprinted constructs (measured with image analysis) was up to 60% resembling the tissue coherence and architecture of native tissue. Our results demonstrate that our materials can be used as cost-effective bioinks for the bioprinting of skeletal muscle models.
- Volumetric bioprinting for medical applications(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-06-05) López Franco, Arturo; VARGAS ROSALES, CESAR; 33901; Vargas Rosales, César; RR; Zhang, Yu Shrike; School of Engineering and Sciences; Campus Monterrey; Galaviz Aguilar, José AlejandroAdditive manufacturing (3D printing) has been a widely used tool in a lot of different industries. Among these industries can be found tissue engineering and regenerative medicine, since bioprinting is one of the main techniques applied. The implementations of new technologies for additive manufacturing, have been adapted into the bioprinting area for medical purposes. Additive manufacturing technologies have been evolving from printing point-to-point, layer-by-layer, and more recently volumetric printing, which represents printing a whole volume simultaneously. In this thesis is presented a new technique for bioprinting, the Computed Axial Lithography (CAL) printing, which is a recently additive manufacturing technology based on reconstruct- ing a volume simultaneously, has demonstrated to have advantages against other additive manufacturing techniques, improving the printing speed, the resolution, minimum material waste, and more, and its application in the medicine industry has not been explorer looking very promising for this research field with limitless applications. The bioink used in the experiments presented is a GelMA-based hydrogel, and the 3D structures achieved should be capable of present biocompatibility with living organisms. For this thesis, the reproduction of the CAL printing for biomaterials, in this particular case GelMA, is proved, opening the doors for applying the same concept to different biomaterials, which could have limitless applications in many distinct research areas.
- 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 MoisésMultiple 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.