Microfabricated bioactive hydrogel platform as in vitro models to understand the mechanobiology of cell-matrix interaction in human tissue

体外模型是研究人类组织发育和衰老的有价值的工具；然而，简单易用的概括细胞外微环境信号的体外模型尚未开发出来。拟议的研究计划旨在应用微制造和生物活性水凝胶创建一个生物仿生平台，用于在健康微环境和年龄或应力重塑的微环境中对角膜和血管组织中细胞-基质相互作用的机制进行研究。细胞-基质反应是复杂的，细胞反应也受物理和化学微环境的影响。微环境中的物理线索，如刚性和地形，已被证明显著影响细胞的行为。这个项目的首要假设是，细胞外微环境的机械生物学可以影响细胞行为和细胞功能，进而影响细胞的功能。我们小组一直在研究细胞-材料相互作用的机械生物学，用于组织工程应用。利用微加工技术，我们开发了在3D非平面表面形成水凝胶图案化的各种技术，并证明了应用这些平台开发体外组织模型的可行性。该计划的关键是使用微加工技术和水凝胶来构建生物材料平台，以概括仿生微环境。主要目标是：O1利用水凝胶微加工技术建立具有仿生形态和力学特性的角膜后弹力膜(DM)体外模型，研究健康和增龄后角膜微环境中细胞的迁移、细胞与细胞和细胞与基质的相互作用；O2开发集成仿生形貌、力学特性和机械刺激的3D平台，研究复杂血管几何结构和老化血管中细胞的响应；以及03建立具有仿生机械特性和外部机械刺激的二维体外模型，以了解机械刺激在揉眼睛中的作用。这项研究计划将使使用工程平台作为衰老研究工具的体外模型的开发成为可能，并允许对影响潜在干预成功的细胞信号和生物力学因素的基础科学有重要的新理解。该计划还将通过影响新产品的设计和开发可申请专利的概念和技术来产生经济影响。体外模型可能导致可应用于生物医学工程领域的新设备，最终应用于药物筛选和干预衰老。此外，该计划还将为生物技术、生物医学工程、材料科学和制药工业等领域提供一批训练有素的科学家和工程师。

In vitro models are valuable tools to study human tissue development and aging; however, simple-to-use in vitro models that recapitulate extracellular microenvironmental signals are yet to be developed. The proposed research program aims to apply microfabrication and bioactive hydrogel to create a biomimicking platform for mechanistic studies of cell-matrix interactions in corneal and vascular tissues, in both the healthy microenvironment and age- or stress-remodeled microenvironment. Cell-matrix responses are complex and cell responses are also influenced by the physical and chemical microenvironment. Physical cues in the microenvironment, such as rigidity and topography, have been shown to significantly affect cell behaviours. The overarching hypothesis of this program is that the mechanobiology of the extracellular microenvironment could affect cell behaviors and cell functions, in turn affecting the functionality of cells. Our group has been studying the mechanobiology of cell-material interactions for tissue engineering applications. Using microfabrication technologies, we have developed various techniques for patterning hydrogel in 3D non-planar surfaces and have demonstrated the feasibility of applying these platforms to develop in vitro tissue models. The key approach of the proposed program is to use microfabrication techniques and hydrogel to build biomaterial platforms to recapitulate biomimicking microenvironments. The main objectives are: O1 to develop a 2D in vitro model with biomimicking topographical and mechanical properties of corneal Descemet's membrane (DM) with microfabrication of hydrogel, to study cell migration, cell-cell and cell-matrix interactions in healthy and age-remodeled corneal microenvironment; O2 to develop a 3D platform with integrated biomimicking topography, mechanical properties and mechanical stimulation, to study cell responses in complex vascular geometry and in aging vasculature, and O3 to develop a 2D in vitro model with biomimicking mechanical properties with external mechanical stimulation to understand the role of mechanical stimuli in eye-rubbing. This research program will enable the development of in vitro models using engineering platforms as tools for aging research and allow for significant new understanding of the fundamental science in cell signalling and biomechanical factors that affect the success of potential interventions. The program will also provide economic impact by influencing the design of new products and developing concepts and technologies that can be patented. The in vitro model may lead to new devices applicable in biomedical engineering areas, with eventual applications in drug-screening and interventions in aging. In addition, the program will provide a pool of highly trained scientists and engineers for the fields of biotechnology, biomedical engineering, materials science and pharmaceutical industry.