New Experimental and Computational Tools for Tissue Engineering

描述（由申请人提供）：组织工程旨在通过将解离的细胞悬浮到可生物降解的聚合物支架中，在该支架上形成新的组织来创建功能性生物假体。我们迫切需要更好的方法来比较工程组织，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，破坏性的，因为它需要首先将组织切片，然后染色。此外，由于染色强度和颜色的高度可变性，组织学的定量可靠性是边缘的。第一阶段SBIR应用旨在实现组织评估的飞跃，这将通过将时间分辨荧光光谱的独特实现与对组织中的光传播进行建模的蒙特-卡罗代码相结合来实现。使用关节软骨结构作为模型系统，我们的团队将展示软骨细胞中胶原蛋白表达的定量跟踪，软骨细胞增殖，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蛋白交联的空间分布。具体目标1是跟踪GFP的活性，GFP用作软骨细胞中胶原合成的标志物，具体目标2是跟踪胶原交联剂形成的活性，因为它在软骨细胞周围的区域中发展。一旦成功证明，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会导致假体在植入后表现更好和更长时间。在第一阶段工作中获得的数据和科学知识在实时医学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生长形成新组织的细胞来制造功能性生物假体。第一阶段SBIR应用的目标是改进技术，在植入前测试实验室生长的组织，从而使假体性能更好，寿命更长。

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.