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，以监测体外和体内工程组织的功能发育。本申请的目的是开发基于光学对比度的内源性来源的光学生物标志物，该光学生物标志物避免使用外源性染色剂，并且可以用于定量报告工程化组织的生化和结构组成。拟议的研究集中在表征的脂肪和骨组织工程化的蚕丝支架与人类间充质干细胞接种。本申请的中心假设是，可以开发基于工程组织的细胞和基质组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。拟议研究的基本原理是，建立非侵入性方法，允许监测工程组织内发生的动态变化，将在开发和优化创新的功能性工程组织结构中发挥重要作用。为了实现我们的目标，我们将表征来自体外开发的工程化组织的不同细胞和基质组分的内源性荧光和光散射信号（目的1）。我们将开发一种系统，该系统将优化这些来自动物的光学信号的采集（目标2），并且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体内植入乳房脂肪垫或空腔骨缺损小鼠模型后的整合（目标3）。这将是第一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。这一建议与改善公众健康高度相关，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，数以百万计的患者接受外科手术修复或重建这些组织将最终受益于这项工作。 公共卫生相关性声明（由申请人提供）：该提案与改善公共卫生高度相关，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，数以百万计的患者接受外科手术修复或重建这些组织将最终受益于这项工作。

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.