New Experimental and Computational Tools for Tissue Engineering

描述(申请人提供)：组织工程学的目标是通过将分离的细胞悬浮到可生物降解的聚合物支架中，在其上形成新的组织来创建功能性生物假体。迫切需要更好的手段来比较工程组织，特别是软骨和其他软组织与正常组织。组织学是缓慢的、繁琐的和破坏性的，因为它需要首先切开组织，然后对其进行染色。此外，由于染色强度和颜色的高度变异性，组织学的定量可靠性微乎其微。这一第一阶段的SBIR应用旨在实现组织评估的一次量子飞跃，这将通过将时间分辨荧光光谱的独特实现与模拟组织中光传播的蒙特卡罗代码相结合来实现。使用关节软骨结构作为模型系统，我们的团队将展示对软骨细胞中胶原表达的定量跟踪，软骨细胞的增殖，以及在整个生长过程中与细胞外基质中软骨细胞相邻形成的胶原交联物的空间分布。特异性目的1是追踪作为软骨细胞胶原合成标志的GFP的活性，特异性目的2是追踪胶原交联物在软骨细胞周围区域形成时的活性。一旦成功演示，这些能力将使测试大量可能影响组织生长和结构最终功能的变量成为可能。第二阶段的高通量组织工程测试最终可能导致假体性能更好，植入后持续更长时间。在第一阶段工作中获得的数据和科学知识在实时医疗诊断和伤口愈合方面也有潜在的应用。组织工程通过在可生物降解的聚合物支架上培养细胞形成新组织来创造功能性生物假体。这一第一阶段SBIR应用的目标是改进技术，在植入实验室培养的组织之前对其进行测试，从而产生性能更好、寿命更长的假体。

DESCRIPTION (provided by applicant): Tissue engineering aims to create functional biologic prostheses by suspending dissociated cells into a biodegradable polymer scaffold upon which new tissue forms. Better means are urgently needed to compare engineered tissues, particularly cartilage and other soft tissues, with normal tissue. Histology is slow, cumbersome, and destructive because it requires first sectioning the tissue, then staining it. In addition, the quantitative reliability of histology is marginal owing to high variability of the stain intensity and color. This Phase I SBIR application aims for a quantum leap forward in tissue assessment, which will be realized by combining a unique implementation of time-resolved fluorescence spectroscopy with Monte-Carlo code that models light propagation in tissue. Using articular cartilage constructs as a model system, our team will demonstrate quantitative tracking of collagen expression in the chondrocytes, chondrocyte proliferation, and the spatial distribution of the collagen cross-links forming adjacent to the chondrocytes in the extracellular matrix over the entire growth process. Specific Aim 1 is to track the activity of GFP that serves as a marker of collagen synthesis in chondrocytes and Specific Aim 2 is to track the activity of collagen cross-linker formation as it develops in the region around the chondrocytes. Once successfully demonstrated, these capabilities will make it possible to test a vastly expanded number of variables that potentially affect tissue growth and the ultimate function of the construct. Tests of high throughput tissue engineering in Phase II could ultimately lead to prostheses that perform better and for a longer period after implantation. The data and scientific knowledge gained during the Phase I effort also has potential applications in real-time medical diagnostics and wound healing. Tissue engineering creates functional biologic prostheses by growing cells that form new tissue on a biodegradable polymer scaffold. The goal of this Phase I SBIR application is improved techniques to test the laboratory grown tissues before they are implanted, resulting in prostheses that perform better and last longer.