Optical monitoring of engineered tissues

总结 我们的长期目标是开发非侵入性的光学技术，以监测功能发育。 体外和体内工程组织。本申请的目的是开发基于光学生物标志物的生物标志物。 关于内源性光学对比度的来源，它限制了外源性染色剂的使用，并可用于报告 定量地对工程组织的生化和结构组成进行分析。拟议的研究重点 对从接种有以下物质的丝支架开发的脂肪和骨工程化组织的表征 人骨髓间充质干细胞该应用程序的中心假设是线性和非线性深度- 基于细胞和基质的自然光散射和荧光特征的分辨成像方法 可以开发工程化组织的组分以报告在移植前和移植后发生的动态变化。 在植入工程组织之后。我们的假设是基于从1999年获得的初步证据。 体外样品，这表明内源性光信号可用于监测细胞内的变化。 分化干细胞、丝支架和沉积胶原的生物化学和形态学。的理由 对于拟议的研究是，建立非侵入性的方法，允许监测的 在工程化组织内发生的动态变化将在发育中起重要作用， 优化创新的、功能性的工程化组织构建体。为了实现我们的目标，我们将描述 来自工程化的细胞的不同细胞和基质组分的内源性荧光和光散射信号 体外发育的组织（目标1）。我们将开发一个系统，将优化这些光学采集 来自动物的信号（目的2），我们将使用这些生物标志物来非侵入性地表征整合 这些工程化组织在体内植入乳房脂肪垫或腔静脉骨内后 缺陷小鼠模型（Aim 3）。这将是第一次，动态监测的生化和 植入的工程化组织的结构功能使用基于以下的非侵入性手段在体内实现： 内源性光信号 这一建议与改善公共卫生密切相关，因为它将促进有效发展 功能性骨和脂肪工程组织。因此，数百万接受外科手术的患者 因为这些组织的修复或重建将最终受益于这项工作。项目叙述 这一建议与改善公共卫生密切相关，因为它将促进有效发展 功能性骨和脂肪工程组织。因此，数百万接受外科手术的患者 因为这些组织的修复或重建将最终受益于这项工作。

Summary Our long-term goal is to develop non-invasive, optical technologies to monitor the functional development of engineered tissues in vitro and in vivo. The objective of this application is to develop optical biomarkers based on endogenous sources of optical contrast that obviate the use of exogenous stains and can be used to report quantitatively on the biochemical and structural composition of engineered tissues. The proposed studies focus on the characterization of adipose and bone engineered tissues developed from silk scaffolds seeded with human mesenchymal stem cells. The central hypothesis of the application is that linear and non-linear depth- resolved imaging methods based on the natural light scattering and fluorescence signatures of cell and matrix components of engineered tissues can be developed to report on the dynamic changes that occur prior to and following implantation of engineered tissues. Our hypothesis is based on preliminary evidence acquired from in vitro samples, which indicate that endogenous optical signals can be used to monitor changes in the biochemistry and morphology of differentiating stem cells, silk scaffolds and deposited collagen. The rationale for the proposed research is that the establishment of non-invasive methods that allow monitoring of the dynamic changes that occur within engineered tissues will play an essential role in the development and optimization of innovative, functional engineered tissue constructs. To achieve our goal we will characterize the endogenous fluorescence and light scattering signals from different cell and matrix components of engineered tissues developed in vitro (Aim 1). We will develop a system that will optimize acquisition of these optical signals from animals (Aim 2) and we will use these biomarkers to characterize non-invasively the integration of these engineered tissues in vivo following implantation either within a mammary fat pad or a cavarial bone defect mouse model (Aim 3). This will be the first time that dynamic monitoring of the biochemical and structural function of implanted engineered tissues is achieved in vivo using non-invasive means based on endogenous optical signals. This proposal is highly relevant to the improvement of public health as it will enable the efficient development of functional bone and adipose engineered tissues. Thus, millions of patients that undergo surgical procedures for the repair or reconstruction of such tissues will ultimately benefit from this work. Project Narrative This proposal is highly relevant to the improvement of public health as it will enable the efficient development of functional bone and adipose engineered tissues. Thus, millions of patients that undergo surgical procedures for the repair or reconstruction of such tissues will ultimately benefit from this work.