TR&D-2: Sensor Enabled Scaffolds

项目概要/摘要： 在制造工程化组织构建体的过程中，能够评估初始细胞是至关重要的。 接种和随后的细胞功能在真实的时间。本次研发将重点关注生物传感器技术的发展， 将利用组织工程中支架的广泛使用来生产传感器启用的scaf的技术， 折叠在不同的材料和配置，以服务于我们的CP/SP。我们的关键创新是将传感 功能，可用于动态监测多种特定的生物学特性， 纵向地，从而使得能够控制和改变细胞接种、营养交换、代谢废物 去除和分化的线索。我们将在两个具体目标中实现这一目标： 具体目标1：生成传感支架以监测和自动化细胞接种。在自动化 在TE构建体的制造过程中，关键是能够实时评估细胞附着，以便 验证播种过程或在需要时采取补救措施。因为不同的支架生物材料和 合成为细胞提供不同的微环境线索，评估细胞的附着和分布 在TEMP组装过程中在支架内的应用可以帮助优化该过程。我们将设计信标型适体 其结合细胞全细胞或细胞表面分子并与之反应，因此可用作短- 传感器在本申请中适体将具有分子信标类型的报告化学， 在结合它们的靶细胞时被构象变化激活 信号与接种到支架上的细胞数量成比例。我们的适体传感器可以附着在 整合到支架/生物材料/生物墨水中。该技术的变化将包括光学光导aug- 用于评估厚结构的内部。 具体目标2：用于测量O2、葡萄糖和乳酸的传感器平台。O2、葡萄糖和乳酸是3个关键指标 代谢活动。在组织的分化和/或成熟过程中，监测这些是必不可少的，以确定 挖掘它们的功能和生化质量，并再次在必要时提供补救行动的信号。 我们将开发O2，葡萄糖和乳酸光电极使用新的化学，并将它们与支架整合在一起。 多种特定于应用的方法，用于测量组织内部的O2、葡萄糖和乳酸水平， 组织表面，并最终在本体介质中。我们还将开发一个测量氧气摄取率的系统 在工程结构的表面。传感器将使用生物相容性材料， 植入支架中或使用新型微锚可逆地连接到支架上。传感器将进行稳定性测试 以及在组织分化和成熟过程中的生物相容性和稳定性。

Project Summary/Abstract: During the process of fabricating an engineered tissue construct, it is critical to be able to assess the initial cell seeding and subsequent cell functions in real time. This TR&D will focus on the development of biosensor tech- nology that will leverage the widespread use of scaffolds in tissue engineering to produce sensor-enabled scaf- folds in different materials and configurations to serve our CPs/SPs. Our key innovation is to incorporate sensing functions into the scaffolds that can be used to monitor multiple specific biological properties dynamically and longitudinally, thus enabling the potential to control and modify cell seeding, nutrient exchange, metabolic waste removal and differentiation cues in real-time. We will accomplish this in two Specific Aims: Specific Aim 1: Generation of sensing scaffolds to monitor and automate cellular seeding. During the automated fabrication process of a TE construct, it is critical to be able to assess cell attachment in real-time in order to validate the seeding process or take remedial action if called for. Because different scaffold biomaterials and syntheses provide different microenvironmental cues for cells, assessing the attachment and distribution of cells within the scaffold during TEMP assembly can help optimize the process. We will design beacon-type aptamers that bind and react to cell whole cells or alternately to cell surface molecules and are therefore usable as short- term sensors in this application. The aptamers will have a molecular beacon-type of reporter chemistry that will be activated by conformational changes upon binding their target cell and will provide a fluorescent feedback signal proportional to the number of cells seeded to the scaffold. Our aptamer sensor that can be attached to or integrated into scaffolds/biomaterials/bioinks. Variations of the technology will include optic light guide aug- mented scaffolds to assess the interior of thick structures. Specific Aim 2: Sensor platforms to measure O2, glucose and lactate. O2, glucose and lactate are 3 key markers of metabolic activity. Monitoring of these is essential during differentiation and/or maturation of tissues to deter- mine both their functionality and biochemical quality, and to, again, provide signals for remedial action if needed. We will develop O2, glucose, and lactate optodes using novel chemistries, and integrate them with scaffolds in a variety of application-specific ways to measure levels of O2, glucose, and lactate in the tissue interior, at the tissue surface and eventually in the bulk medium. We will also develop a system to measure oxygen uptake rate at the surface of the engineered construct. The sensors will use biocompatible materials and will be integrated into the scaffolds or reversibly attached to them using novel micro-anchors. The sensors will be tested for stability during scaffold storage, and for biocompatibility and stability during tissue differentiation and maturation.