Ciencias Exactas y Ciencias de la Salud

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Rational design & engineering of cost-efficient Point-of-Care (POC) systems for rapid diagnostics of emergent and chronic degenerative diseases
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2024) Bravo Gonzalez, Sergio; Alvarez, Mario Moises; emipsanchez; Schultz-Cherry, Stacey; González González, Everardo; Hernández Torre, Martín Virgilio; Álvarez, Claudio; Escuela de Ingeniería y Ciencias; Campus Monterrey; Trujillo de Santiago, Grissel
Patients require proper access to diagnostics to benefit from medicine and obtain proper treatment. However, diagnostics availability is one of the biggest challenges concerning global public healthcare. The recent Covid-19 pandemic has highlighted the consequences of the lack of fast and widely available diagnostics. This dissertation aims to propose a novel solution for addressing this complex healthcare challenge. We propose the engineering of cost-efficient Point-of-Care (POC) systems for the rapid diagnostics of emergent and chronic degenerative diseases. We chose SARS-CoV-2 as a model for emergent infectious diseasse due to the recent pandemic in winter 2019. In addition, cancer was selected as the representative of chronic degenerative diseases tdue to its incidence and mortality rate. To assess the relative importance of intervention strategies such as vaccination, social distancing, and rapid diagnostic, first we developed a population level surveillance model based on ordinary differential equations that simulate the effect of vaccine rate against Covid-19 spreading. The model revealed the benefits of rapid interventions such as fast vaccination campaigns and widespread diagnostics, and that in the absence of vaccines, rapid diagnostic followed by the quarantine of infected subjects. In alignment with this finding, we developed cost-effective and portable diagnostic methods to identify SARS-CoV-2 nucleic acids based on isothermal amplification strategies. First, we developed and characterized the performance of a point of-care (POC) do-it-yourself (DiY) device to identify SARS-CoV-2 RNA in less than 45 minutes. We also conducted a pilot study in Monterrey to evaluate the effectiveness of this DiY POC testing strategy based on a colorimetric LAMP & polyethylene-sulfonate membrane. We determined a sensitivity and specificity of 100% and 87%, respectively, highlighting the value of utilizing quick and accurate diagnostic responses. Furthermore, we engineered an Arduino-based detection system for the rapid diagnostics of SARS-CoV-2 in 5 saliva using a nucleic acid amplification strategy. We tested our Arduino-based detection system ability to discriminate between and positive saliva samples spiked with SARS-CoV 2 genetic material. We decided to further challenge the versatility of our system by testing its ability to discriminate between cancer and non-cancerous tissue spheroids. We were able to identify clusters based on the expression of selected genetic biomarkers by implementing our detection strategy. Overall, our solutions present with viable alternatives to alleviate the lack of accessible and cost-effective diagnostic platforms for infectious and chronic diseases. By implementing our systems and models we would increase early detection and offer easy to-implement population surveillance models to increase disease monitoring.
Bioprinting of spatially organized cancer models using chaotic flows
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2024) Ceballos Gonzalez, Carlos Fernando; Alvarez, Mario Moises; emimmayorquin; Weiss, Paul S.; Magaña Aguirre, Johathan Javier; Luna Aguirre, Claudia Maribel; Sánchez Salazar, Mónica Gabriela; Escuela de Ingeniería y Ciencias; Campus Monterrey; Trujillo de Santiago, Grissel
Human tissues exhibit a balance between aggregation and segregation, but achieving such microscale organization is challenging for most extrusion-based bioprinting techniques. Conventional strategies often require smaller nozzles to generate microscopic features. However, smaller nozzles decrease production yields, and increase cellular damage due to a higher shear stress. Recently, our research group introduced the concept of chaotic bioprinting for the high-throughput fabrication of tissue models exhibiting microscale organization. The focus of this PhD thesis was the development of spatially-organized cancer models using chaotic bioprinting. This document is organized into four (4) chapters. The first chapter introduces the concept of chaos-assisted production of architected spheres (CAPAS). This system allows to modulate the sphere diameter (0.6 to 3.5 mm) and production yield (up to ~2,000 droplets per minute) by adjusting simple process parameters: flow rate, printhead outlet diameter, polymer concentration (sodium alginate or GelMA), and crosslinking bath composition. CAPAS was used to create a breast cancer model where malignant cells were surrounded by healthy stromal cells. This multicellular cancer model exhibited a higher susceptibility to paclitaxel doses than monocultures of breast cancer cells. The second chapter presents an expanded portfolio of chaotic printheads by the incorporation of superior and lateral inlets. The number and type of biomaterials extruded through a printhead can be adjusted to meet different requirements. For example, chaotic printheads were used to produce fibers exhibiting axial microgradients, structured emulsions, micro-channeled hydrogel filaments, and a tumor niche containing breast cancer microtumors. Cells from embedded spheroids migrated through the micropatterns created by the lateral inlets of the printhead. The third chapter outlines a strategy to fabricate a pre-vascularized breast cancer model using a multimaterial chaotic printhead. In this model, breast cancer microtumors were surrounded by inner channels that served as corridors for cell migration. The length of the migratory fronts was higher in multichannel filaments (~1,400 μm in length) than in solid scaffolds (~400 μm in length). Genes involved in cell migration (N-cadherin, vimentin) were highly upregulated in multichannel filaments. The fourth chapter introduces a novel set of static mixers designed to mimic the architecture of some human tissues. Each mixer generates a specific inner structure in the printed filament that resembles, for example, the intestinal villi, the human breast tissue, or marbled meat. I fabricated a colon cancer model that exhibited a high post-printing viability (~75% viable cells) over an extended cultivation period (14 days). I also adapted these novel printheads to a cartesian bioprinter to produce three-dimensional scaffolds exhibiting inner microarchitecture. In conclusion, I envision that the work derived from my PhD thesis will help to fabricate disease models that better recapitulate the spatial organization of human tissues. One limitation of the current work is that the effective generation of microstructure by chaotic printing is strongly dependent on the rheology of the biomaterials used. This limitation may be addressed by optimizing biomaterial formulations for chaotic printing.

Ciencias Exactas y Ciencias de la Salud

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