Ciencias Exactas y Ciencias de la Salud
Permanent URI for this collectionhttps://hdl.handle.net/11285/551014
Pertenecen a esta colección Tesis y Trabajos de grado de los Doctorados correspondientes a las Escuelas de Ingeniería y Ciencias así como a Medicina 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, GrisselPatients 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.
- 3D printed tumor on chips for the culture and maturation of heterotypic cancer spheroids as a platform for drug testing(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2022-12-05) Gallegos Martínez, Salvador; GALLEGOS MARTINEZ, SALVADOR; 814051; puemcuervo, emipsanchez; Shrike Zhang, Yu; Gonzáles Meljem, José Mario; Luna Aguirre, Claudia Maribel; Olvera Posadas, Daniel; School of Engineering and Sciences; Campus Monterrey; Trujillo de Santiago, GrisselThe recapitulation of cancer environment in tumor-on-chip systems will greatly contribute to accelerate cancer studies in the fronts on fundamental research, pharmacological testing of new therapeutic compounds, and personalized medicine. Here we describe the development of two microfluidic platforms aimed to contribute to the advance of tumor-on-chip research. First, we describe a simple and robust method for the fabrication, maturation, and extended culture of large heterotypic cancer (MCF7 and MCF7/fibroblasts) spheroids (~900 µm in diameter) in a 3D-printed mini continuous stirred tank reactor (mini-CSTR). In brief, MCF7 and MCF7/BJ cell suspensions (5×104 cells) were incubated in batch culture to form discoid cell aggregates (600 µm in diameter). These microtissues were then transferred into the mini-CSTR and continuously fed with culture media for an extended time (~30 days). The spheroids progressively increased in size during the first 20 days of perfusion culture to reach a steady diameter. We characterized the spheroid morphology, architecture and the evolution of expression of relevant tumor-related genes (i.e., ER, VEGF, Ki67, Bcl2, LDHA, and HIF-1α) in spheroids cultured for 30 days. This mini-CSTR culture strategy enables the simple and reproducible fabrication of relatively large spheroids and offers great potential for studying the effects of diverse effectors on tumor progression. In addition, we introduce a 3D-printed microfluidic system that can be generically used to culture tumor-tissues under well-controlled environments. The system is composed by three compartments. The left and right compartments have two inlets and two outlets which provide means to continuously feed liquid streams to the system. The central compartment is designed to host a hydrogel where a microtissue can be confined and cultured. A transparent lid can be adapted to enable visual inspection under a microscope. We conducted fluorescent and FITC dextran tracer experiments to characterize the hydraulic performance of the system. In addition, we cultured MCF7 and MCF7/BJ spheroids embedded in a GelMA hydrogel constructs (placed in the central chamber), to illustrate the use of this system to sustain long term micro-tissue culture experiments. We also present experimental results that illustrate the flexibility and robustness of this 3D-printed device for tumor-on-chip experiments including pharmacological testing of anticancer compounds. These “open-source” organ-on-chip systems are intended to be a general-purpose resource to facilitate and democratize the development of tumor-on-chip applications. We also explored the use of these cell aggregates and some of the characterization techniques to develop educational activities in the context of tissue engineering. Students fabricated a DIY (do it yourself) incubator and cultured spheroids for 7 days on average. They evaluated glucose consumption, size progression and change in color of the culture media. In this proposed activity students were exposed to concepts and basic experimental duties commonly use in a tissue engineering lab.
- Development of a novel zein-derived peptide: in vitro assays, and 2D- and 3D-cell models(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2022-12-02) Trinidad Calderón, Plinio Alejandro; TRINIDAD CALDERON, PLINIO ALEJANDRO; 627107; García Lara, Silverio; puemcuervo, emipsanchez; Trujillo De Santiago, Grissel; López Castillo, Laura Margarita; Puente Garza, César Armando; Acosta Cruz, Erika Yanneth; School of Engineering and Sciences; Campus Monterrey; Álvarez, Mario MoisésMaize is the most consumed staple crop worldwide. Its composition, abundant in prolamins corresponding to α-zein, represents an opportunity for the search of bioactive candidates combating diseases such as cancer. In particular, zein hydrolysates have showed activity against this disease in vitro. Nevertheless, the need for specific peptides with known peptide sequence remains. In response, this thesis work attempted to develop a new-to-nature peptide derived from zein and test its effects in vitro. First, acetone precipitation was introduced to the purification train of zein hydrolysates of blue maize. , After 48 h, the resulting hydrolysates (BmapZDH) diminished cancer cell viability to 4.3% and reduced fibroblasts’ viability to 77.3% at 125 and 93.75 ng/mL, respectively. Hypothesizing an attributing specific peptide in silico, a new-to-nature peptide from the 22-kDa α-zein was developed. The resultant peptide, nurP28, had the sequence Ac-LALLALLRLRRRATTAFIIP-NH2 after the iterations. When tested alone, nurP28 was not cytotoxic to human fibroblasts or cancer cells. Notably, higher cytotoxic effects were observed in MCF7 spheroids when combining nurP28 at 3 ng/mL and 1 µM docetaxel than 10 µM alone. Additional testing highlighted that nurP28 inhibits the migration of MDA-MB-231 breast cancer cells by ~50 %, is not-hemolytic for human red blood cells, does not alter the ACE activity when administered at 300 ng/mL, and is innocuous for murine myoblasts cells. In toto, this thesis presents the bases of the potential and capability of nurP28 peptide as a safe molecule to be tested in vivo promptly, aiming to boost breast cancer treatment.