TR&D-3: Bio-instructive Bioreactors

项目概要/摘要： 生物反应器系统对于粘性工程组织从细胞和支架成熟至关重要。许多 生物反应器与基本环境参数的周期性、非侵入性测量兼容， 溶解氧、pH值和温度以及其他组织特异性过程中和最终产品质量属性。 然而，（1）针对一系列重新确定的， 组织特异性生物和机械质量属性，以及（2）提供生物和机械质量属性 对结构组织的刺激是不存在的。此外，封闭/密封系统可减轻污染 需要非侵入式测量技术，特别是用于测量 自体/个人化组织，其仅可存在单个单元。我们将开发一个封闭的，生物教学 生物反应器平台，适合于各种结构组织的培养，并结合传感和致动 用于过程参数的反馈控制，使用以下特定目的。目标1：发展和 集成传感器以监测生理化学细胞状态/成熟度和组织机械 特性.我们将首先研究将光学传感器集成到生物反应器容器中，重点是 优化的信号采集和分析。我们将开发（然后也整合）超声评估， 组织构建体的机械性能，例如一维模量、组织不均匀性和表面 缺陷连同关键代谢物和营养物的离线分析（例如，氨基酸，生长因子）， 这些输入将提供关于工程组织的表型和健康的详细信息。目标二： 模块化生物反应器开发，包括用于机械刺激、连续生长的致动器 因子调节和注入系统。将处理来自嵌入式、在线和离线传感器的输入 并输出到两种类型的可编程致动器：1）用于化学刺激的流体分配器，和2）机械 通过屈曲、剪切和/或压缩传递的刺激。生物反应器平台将开发为 强调与Tissue Foundry模块的集成，以及与其自动化和 数据管理骨干网。生物反应器的内部配置将特定于组织， 由我们的CP开发，并将设计用于细胞接种，培养基交换和 通过PLC控制的执行器和阀门进行营养添加。对于需要机械刺激的组织， 模块化反应器将具有集成的可变形聚合物壁，组织可以通过该可变形聚合物壁机械地 以无菌的方式操作。无菌端口和光纤端口将嵌入生物反应器壁中， 生物效应物的递送并使O2摄取或其他分析物测量成为可能。反馈控制 将开发用于感测和致动的算法，用于单一组织类型的机械质量控制。

Project Summary/Abstract: Bioreactor systems are critical for cohesive engineered tissues to mature from cells and scaffolds. Many bioreactors are compatible with the periodic, non-invasive measurement of basic environmental parameters, like dissolved O2, pH and temperature and other, tissue-specific in-process and final product quality attributes. However, the ability to (1) non-invasively measure cell and tissue maturation against a series of redetermined, tissue-specific biological and mechanical quality attributes and (2) to provide biological and mechanical stimulation to structural tissues, does not exist. Furthermore, closed/sealed systems to mitigate contamination require non-invasive measurement technologies, especially for measuring quality attributes of autologous/personalized tissues of which only a single unit may exist. We will develop a closed, bio-instructive bioreactor platform, amenable to the culture of various structural tissues, and incorporating sensing and actuation for feedback control of process parameters using the following specific aims. Aim 1: Development and integration of sensors to monitor physio-chemical cell status/maturity, and tissue mechanical properties. We will first investigate integration of optical sensors into bioreactor vessels with a focus on optimized signal acquisition and analysis. We will develop (and then also integrate) ultrasonic assessment of tissue construct mechanical properties such as one-dimensional modulus, and tissue inhomogeneity and surface defects. Together with the off-line analysis of key metabolites and nutrients (e.g., amino acids, growth factors), these inputs will provide detailed information on the phenotype and health of the engineered tissue. Aim 2: Modular bioreactor development, including actuators for mechanical stimulation, sequential growth factor regulation, and injection systems. Inputs from embedded, in-line, and off-line sensors will be processed and exported to two types of programmable actuators:1) fluid dispensers for chemical stimuli, and 2) mechanical stimuli delivered via flexion, sheer, and/or compression. The bioreactor platforms will be developed with emphasis on integration with Tissue Foundry modules, and on downstream compatibility with its automation and data management backbone. The internal configuration of the bioreactor will be specific to the tissues in development by our CPs and will be designed for optimal fluid flow paths for cell seeding, media exchanges and nutrient addition via PLC-controlled actuators and valves. For tissues requiring mechanical stimulation, the modular reactor will have integrated deformable polymeric walls, through which the tissue can be mechanically manipulated in a sterile manner. Sterile ports and fiberoptic ports will be embedded into the bioreactor walls for the delivery of biological effectors and to enable O2 uptake or other analyte measurements. Feedback control algorithms will be developed for sensing and actuation for mechanical quality control of a single tissue type.