Three-Dimensional Image-Guided Development and Optimization of Molecular Regulating Bone Regenerative Scaffolds

三维图像引导分子调控骨再生支架的开发与优化

• 批准号: 9063060
• 负责人: Yu Chen
• 金额: $ 32.28万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-10 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9063060
• 关键词: Acetals; Algorithms; Animal Model; Architecture; Behavior; Binding; Biocompatible Materials; Bone Marrow; Bone Regeneration; Bone Tissue; Cell Survival; Cells; Cellular Structures; Cephalic; Characteristics; Clinical Medicine; Confocal Microscopy; Cues; Defect; Development; Feedback; Fluorescence; Growth; Health; Human; Hydrogels; Image; Image Analysis; Imaging Techniques; Imaging technology; In Situ; Infiltration; Investigation; Knowledge; Laboratories; Lead; Longitudinal Studies; Mesenchymal Stem Cells; Methods; Microscopy; Modality; Modeling; Molecular; Morphology; Motivation; Natural regeneration; Optical Coherence Tomography; Optical Tomography; Oryctolagus cuniculus; Osteogenesis; Outcome; Penetration; Phenotype; Population; Porosity; Principal Investigator; Property; Regenerative Medicine; Regulation; Research Personnel; Role; Signal Transduction; Stem cells; Stromal Cells; Structure; Surface; System; Techniques; Three-Dimensional Image; Time; Tissue Engineering; Tissue Grafts; Tissue imaging; Tissues; Work; base; bone; bone morphogenetic protein 2; cell behavior; clinically relevant; craniofacial; design; environmental chemical; in vivo; migration; novel; optical imaging; osteoblast differentiation; osteogenic; pre-clinical; regenerative; response; scaffold; tissue support frame; tool; two-photon

 DESCRIPTION (provided by applicant): Three-dimensional (3D) cell-based tissue grafts have been increasingly useful in tissue engineering and regenerative medicine. A critical building block in tissue engineering is the scaffold which can act as the supporting medium to deliver cell populations and induce the ingrowth of vessels and surrounding tissues. Therefore, it is necessary to develop tools to characterize the architecture of the scaffold. In addition, to study cell-scaffold interaction, namely, cell viability, migration, proliferation, and signaling within te scaffold, a non- destructive technique that can quantitatively image 3D cell behavior is required. The emerging multi-modality systems such as the combination of optical coherence tomography (OCT) with fluorescence confocal microscopy (FCM) and two-photon microscopy (TPM) that provide co-registered images of structural and functional properties of scaffolds and cells have had significant impact upon the field. However, both FCM and TPM have limited penetration depths thus precluding their characterization of cells deep inside the scaffolds. Therefore, there is a critical need for developing new methods that can analyze the engineered tissue structure in a non-destructive manner and with the ability to image deeper than microscopy. We propose a new platform of OCT and fluorescence laminar optical tomography (FLOT) for characterization cell-scaffold interaction. Two key findings from our lab provide the major motivation for this work: 1) We have demonstrated that OCT can image macroporous scaffolds and quantify their structural parameters including pore sizes, porosity, and interconnectivity; and the co-registered FCM can quantitatively describe cell composition on the scaffold surface. 2) We have developed a combined OCT and FLOT system for depth-resolved imaging of tissue morphology and molecular information in vivo up to 1-2 mm deep. The combined use of OCT and FLOT is a promising approach to characterize both structural and cellular information simultaneously to investigate cell-scaffold interaction, which enables longitudinal studies of cell viability, migraton, proliferation, and differentiation temporally as well as spatially within the scaffolds. We hypothesize that OCT and FLOT will enable 3D description of a scaffold's structure and cellular/molecular distribution, therefore elucidating cellular interactions within scaffolds. We propose to achieve our objective through three specific aims: 1) Establish the Capability of OCT/FLOT on Imaging Morphological and Functional Parameters in Engineered Tissues. 2) Image Stem-Cell-Laden Tissue Scaffolds and Correlate with Bone Formation In Vivo. 3) Prospectively Validate the Hypothesis that OCT/FLOT Imaging Can Predict Regenerative Outcomes. This project will result in a new non-destructive imaging technology for quantitative characterization of cell-scaffold interactions, which is essential to enable optimized design and materials of tissue engineering scaffolds, cell-seeding methods, and chemical/environmental cues. These studies will define a novel path towards advancing bone tissue engineering.

 描述（由申请人提供）：基于三维（3D）细胞的组织移植物在组织工程和再生医学中越来越有用。组织工程中的一个关键构件是支架，其可以作为支持介质来递送细胞群并诱导血管和周围组织的向内生长。因此，有必要开发工具来表征支架的结构。此外，为了研究细胞-支架相互作用，即支架内的细胞活力、迁移、增殖和信号传导，需要能够对3D细胞行为进行定量成像的非破坏性技术。新兴的多模态系统，如光学相干断层扫描（OCT）与荧光共聚焦显微镜（FCM）和双光子显微镜（TPM）的组合，提供支架和细胞的结构和功能特性的共配准图像，对该领域产生了重大影响。然而，FCM和TPM都具有有限的穿透深度，因此排除了它们对支架内部深处的细胞的表征。因此，迫切需要开发新的方法，可以以非破坏性的方式分析工程组织结构，并具有比显微镜更深的成像能力。我们提出了一个新的OCT和荧光层光学层析成像（FLOT）的表征细胞-支架相互作用的平台。我们实验室的两个关键发现提供了这项工作的主要动机：1）我们已经证明OCT可以对大孔支架进行成像并量化其结构参数，包括孔径，孔隙率和互连性;并且共配准FCM可以定量描述支架表面的细胞组成。2)我们已经开发了一种组合的OCT和FLOT系统，用于深度分辨成像的组织形态和分子信息在体内高达1-2毫米深。OCT和FLOT的组合使用是一种有前途的方法，同时表征结构和细胞信息，以研究细胞-支架相互作用，这使得纵向研究细胞活力，迁移，增殖和分化的时间以及空间内的支架。我们假设OCT和FLOT将能够对支架的结构和细胞/分子分布进行3D描述，从而阐明支架内的细胞相互作用。我们建议通过三个具体目标来实现我们的目标：1）建立OCT/FLOT对工程组织中成像形态和功能参数的能力。2)成像干细胞负载的组织支架和与体内骨形成的相关性。3)证实OCT/FLOT成像可以预测再生结果的假设。该项目将产生一种新的非破坏性成像技术，用于定量表征细胞-支架相互作用，这对于优化组织工程支架的设计和材料，细胞接种方法以及化学/环境线索至关重要。这些研究将为推进骨组织工程开辟一条新的道路。