Optical monitoring of engineered tissues

描述(由申请人提供)： 我们的长期目标是开发非侵入性的光学技术，以监测工程组织在体外和体内的功能发展。这项应用的目的是开发基于内源性光学对比来源的光学生物标记物，避免使用外源染色，并可用于定量报告工程组织的生化和结构组成。拟议的研究集中在脂肪和骨工程组织的特征，这些组织是由种植了人骨髓间充质干细胞的丝质支架发展而来的。应用的中心假设是，可以开发基于工程组织的细胞和基质成分的自然光散射和荧光特征的线性和非线性深度分辨成像方法，以报告在工程组织植入之前和之后发生的动态变化。我们的假设是基于从In 体外样本，这表明内源性光学信号可以用来监测 分化干细胞、丝素支架和沉积的胶原的生化和形态。这项拟议研究的基本原理是，建立能够监测工程组织内发生的动态变化的非侵入性方法，将在创新的、功能工程组织结构的开发和优化中发挥重要作用。为了实现我们的目标，我们将表征来自体外培养的工程组织的不同细胞和基质成分的内源荧光和光散射信号(目标1)。我们将开发一种系统，优化从动物身上获取这些光学信号(目标2)，我们将使用这些生物标志物来非侵入性地表征这些工程组织在植入乳房脂肪垫或腔静脉骨缺损小鼠模型后在体内的整合情况(目标3)。这将是首次使用基于内源性光学信号的非侵入性手段在体内实现对植入的工程组织的生化和结构功能的动态监测。这项建议与改善公共健康高度相关，因为它将使功能性骨和脂肪工程组织的有效开发成为可能。因此，数百万接受手术修复或重建这类组织的患者最终将从这项工作中受益。 公共健康相关声明(由申请者提供)：这项建议与改善公共健康高度相关，因为它将使功能性骨和脂肪工程组织的有效开发成为可能。因此，数百万接受手术修复或重建这类组织的患者最终将从这项工作中受益。

DESCRIPTION (provided by applicant): 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 depthresolved 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. Public Health Relevance Statement (provided by applicant): 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.