Nondestructive, High Resolution Imaging Platform For Tissue Regeneration Research

用于组织再生研究的无损高分辨率成像平台

• 批准号: 8620994
• 负责人: SHAY SOKER
• 金额: $ 23.68万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8620994
• 关键词: Achievement; Address; Area; Ballistics; Biological; Biological Assay; Biological Process; Biology; Biomedical Engineering; Bioreactors; Blood Vessels; Blood capillaries; Cancer Biology; Cell Communication; Cell Culture Techniques; Cell Differentiation process; Cell physiology; Cells; Coculture Techniques; Complex; Computational Technique; Confocal Microscopy; Data; Development; Developmental Biology; Disease; Disease model; Documentation; Endothelial Cells; Extracellular Matrix; Fiber; Fiber Optics; Fluorescent Probes; Functional disorder; Future; Generations; Goals; Histocompatibility Testing; Histology; Human; Image; Imagery; Imaging Device; Imaging technology; In Vitro; Label; Light; Manuscripts; Medical Research; Methods; Microscopy; Modeling; Molecular; Monitor; Motivation; Muscle; Muscle Fibers; Natural regeneration; Optics; Organism; Pathogenesis; Pathology; Penetration; Pharmaceutical Preparations; Photons; Physiology; Play; Process; Publishing; Research; Resolution; Role; Sampling; Seminal; Skeletal Muscle; Solutions; Structure; System; Techniques; Technology; Testing; Time; Tissue Engineering; Tissue Model; Tissues; United States National Institutes of Health; Validation; absorption; base; capillary; cell type; clinically relevant; fluorescence imaging; high throughput screening; image reconstruction; imaging modality; improved; in vivo; injured; instrument; interdisciplinary approach; minimally invasive; molecular imaging; new technology; novel; optical fiber; optical imaging; public health relevance; research and development; scaffold; stem cells; tissue culture; tissue regeneration; tomography

DESCRIPTION (provided by applicant): Tissue development and regeneration is highly complex and dynamic, involved with extensive remodeling of cells and the extracellular matrix surrounding them inside the developing and/or injured tissues. Despite the rapid development of tissue engineering technologies to study regeneration, a major barrier still exists in our inabilit to monitor dynamic biological processes in a minimally invasive real-time fashion, which significantly reduces the clinical relevance of these techniques. Most available assessment methods are static, requiring sacrifice of experimental samples at fixed time points. Therefore, there is an unmet need for new technologies that will provide non-destructive and dynamic monitoring of the development and regeneration processes. Optical imaging in biology can be broadly classified as either ballistic imaging or diffusive imaging. The combination of fluorescent and bioluminescent probes with optoelectronics and computing techniques has led to the development of optical molecular imaging tools that allow the visualization of biologic interactions in complex, living systems over time. However, despite the great potential of optical molecular imaging, it has not yet been harnessed as an enabling technology for tissue regeneration research, due to tissue turbidity, resulting in strong scatter and absorption of light and limited penetration depth, requiring direct view of the tissue. We have recently published several seminal manuscripts describing the development of an indirect, non-destructive, cellular-level imaging instrument through a combination of fiber optic technology and an image reconstruction approach and generation of bioengineered mature and vascularized skeletal muscle tissue using combinations of fluorescently labeled cells. These achievements serve as the motivation for the current proposal, which aims to utilize the model of bioengineered skeletal muscle to develop and validate a novel optical molecular tomography platform, which could be broadly used for tissue regeneration research. We hypothesize that 1) optical imaging, photon transport modeling, and image reconstruction will allow for the non- invasive (indirect), dynamic analysis of bioengineered muscle tissue constructs~ and 2) tomography of distinct fluorescent probes will improve the examination of developing bioengineered muscle constructs, comprised of multiple cell types. We will test these hypotheses by developing a multiwell tissue culture dish equipped with fiber-based imaging system. We will first test the capacity of the imaging system to generate optical phantoms of fluorescently labeled cells and subsequently use the imaging system to assess the organization and differentiation of muscle progenitor and endothelial cells into a multicellular skeletal muscle tissue in vitro. These studies have the potential to drive a paradigm shift from static assays of cellular function in 2D culture models towards systematic analyses of 3D tissues. Achieving the goals set forth in this proposal will establish a novel technology to construct and image 3D composite bioengineered tissues and improve our understanding of tissue development and regeneration mechanisms.

描述（由申请人提供）：组织发育和再生是高度复杂和动态的，涉及细胞及其周围的细胞外基质在发育和/或受损组织内的广泛重塑。尽管用于研究再生的组织工程技术迅速发展，但我们无法以微创实时方式监测动态生物过程仍然存在一个主要障碍，这大大降低了这些技术的临床相关性。大多数可用的评估方法都是静态的，需要在固定时间点牺牲实验样本。因此，对能够对发育和再生过程提供非破坏性和动态监测的新技术的需求尚未得到满足。 生物学中的光学成像可大致分为弹道成像或扩散成像。荧光的组合 具有光电子学和计算技术的生物发光探针促进了光学分子成像工具的发展，这些工具可以使复杂的生命系统中的生物相互作用随着时间的推移而可视化。然而，尽管光学分子成像潜力巨大，但由于组织混浊，导致光的散射和吸收强烈，穿透深度有限，需要直接观察组织，因此它尚未被用作组织再生研究的使能技术。我们最近发表了几篇开创性的手稿，描述了通过结合光纤技术和图像重建方法开发间接、非破坏性细胞级成像仪器，以及使用荧光标记细胞组合生成生物工程成熟和血管化骨骼肌组织。这些成就是当前提案的动力，该提案旨在利用生物工程骨骼肌模型来开发和验证一种新型光学分子断层扫描平台，该平台可广泛用于组织再生研究。我们假设 1) 光学成像、光子传输模型和图像重建将允许对生物工程肌肉组织结构进行非侵入性（间接）动态分析，2) 不同荧光探针的断层扫描将改进对由多种细胞类型组成的正在开发的生物工程肌肉结构的检查。我们将通过开发配备基于光纤的成像系统的多孔组织培养皿来测试这些假设。我们将首先测试成像系统生成荧光标记细胞的光学幻像的能力，然后使用成像系统评估肌肉祖细胞和内皮细胞在体外组织和分化为多细胞骨骼肌组织。 这些研究有可能推动从 2D 培养模型中的细胞功能静态分析转向 3D 组织的系统分析的范式转变。实现该提案中提出的目标将建立一种新技术来构建 3D 复合生物工程组织并对其进行成像，并提高我们对组织发育和再生机制的理解。