Publicação
Microfluidic hydrogel biofabrication for high-throughput screening, modeling and quantifying 3D biological phenomena
| Resumo: | The functionality of living tissues depends entirely on their intricate 3D composition, architecture, and mechanics, which result from the complex interplay between cells and ECM. Correctly approaching these characteristics will be fundamental to engineer next-generation platforms for in vitro modeling of healthy and diseased tissues. Further, the capacity to miniaturize 3D microenvironments for faster composition screening and discovery will also be critical to benefit from the ever-growing toolbox of 3D hydrogels and tissue-like cues that can be integrated into engineered constructs. Within that arena, microfluidic biofabrication is a potent tool to miniaturize 3D cell-laden hydrogel constructs. Still, these are only relevant as models if important tissue characteristics such as 3D cell/ECM architectures are integrated. Additionally, the complexity of 3D models is only helpful if some tools and methodologies enable efficient and reliable quantification of ongoing biological events into measurable data. This thesis presents advances in all these critical perspectives, aiming for microfluidics to fabricate improved 3D platforms that can model, screen, and quantify biological events with unprecedented efficiency. Initially, we demonstrate how the microfluidic manipulation of hydrogel precursors can be employed to create 3D gradients with space-varying mechanics and material composition. Allied to automated single-cell screening, we show how optimal hydrogel formulations can be detected with highthroughput to elicit biological responses ranging from cell adhesion to the triggering of stem cell differentiation. Then, we explore how 3D flow-focusing microfluidics can introduce a plethora of 3D architectures within hydrogel microfibers, using a single-chip, down to sub-50μm diameters. We demonstrate how different architectures can be combined with various cells and materials to model 3D cancer invasion, vascular tissue networks, or function as all-in-one tissue engineering platforms. Finally, multi-compartment hydrogel fibers are interfaced with light-guiding in polysaccharide-based optical fibers. These fibers can detect 3D mechanical deformations, the presence of biotargets of interest, and transport live cells while simultaneously guiding light. The living optical fibers are then used to convert complex biological events into directly quantifiable optical signals, tracking the 3D growth of a cancer fiberoid and revealing inhibitory drug thresholds with unprecedented efficacy. Overall, this thesis demonstrates the wide adaptability and capabilities of microfluidic-driven hydrogel biofabrication, presenting high-throughput for miniaturizing, modeling, and direct conversion of complex biological events into quantifiable data, with high potential for the future of precision and data-driven medicine approaches. |
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| Autores principais: | Guimarães, Carlos Ferreira |
| Assunto: | Bioengineering Biomaterials Cancer Microfluidics 3D Tissue Models Bioengenharia Biomateriais Cancro Microfluídica Modelos de Tecidos 3D |
| Ano: | 2022 |
| País: | Portugal |
| Tipo de documento: | tese de doutoramento |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade do Minho |
| Idioma: | inglês |
| Origem: | RepositóriUM - Universidade do Minho |
| Resumo: | The functionality of living tissues depends entirely on their intricate 3D composition, architecture, and mechanics, which result from the complex interplay between cells and ECM. Correctly approaching these characteristics will be fundamental to engineer next-generation platforms for in vitro modeling of healthy and diseased tissues. Further, the capacity to miniaturize 3D microenvironments for faster composition screening and discovery will also be critical to benefit from the ever-growing toolbox of 3D hydrogels and tissue-like cues that can be integrated into engineered constructs. Within that arena, microfluidic biofabrication is a potent tool to miniaturize 3D cell-laden hydrogel constructs. Still, these are only relevant as models if important tissue characteristics such as 3D cell/ECM architectures are integrated. Additionally, the complexity of 3D models is only helpful if some tools and methodologies enable efficient and reliable quantification of ongoing biological events into measurable data. This thesis presents advances in all these critical perspectives, aiming for microfluidics to fabricate improved 3D platforms that can model, screen, and quantify biological events with unprecedented efficiency. Initially, we demonstrate how the microfluidic manipulation of hydrogel precursors can be employed to create 3D gradients with space-varying mechanics and material composition. Allied to automated single-cell screening, we show how optimal hydrogel formulations can be detected with highthroughput to elicit biological responses ranging from cell adhesion to the triggering of stem cell differentiation. Then, we explore how 3D flow-focusing microfluidics can introduce a plethora of 3D architectures within hydrogel microfibers, using a single-chip, down to sub-50μm diameters. We demonstrate how different architectures can be combined with various cells and materials to model 3D cancer invasion, vascular tissue networks, or function as all-in-one tissue engineering platforms. Finally, multi-compartment hydrogel fibers are interfaced with light-guiding in polysaccharide-based optical fibers. These fibers can detect 3D mechanical deformations, the presence of biotargets of interest, and transport live cells while simultaneously guiding light. The living optical fibers are then used to convert complex biological events into directly quantifiable optical signals, tracking the 3D growth of a cancer fiberoid and revealing inhibitory drug thresholds with unprecedented efficacy. Overall, this thesis demonstrates the wide adaptability and capabilities of microfluidic-driven hydrogel biofabrication, presenting high-throughput for miniaturizing, modeling, and direct conversion of complex biological events into quantifiable data, with high potential for the future of precision and data-driven medicine approaches. |
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