Bimodal platform for nondestructive analysis of engineered vascular biomaterials

 描述(申请人提供)：本次资助申请的目标是研究、测试和验证结合光学和超声成像技术的双模式诊断平台，用于实时、无损地对用于血管组织工程的细胞外基质(ECM)支架的成分、结构功能和特定部位的细胞再繁殖进行体内和体外分析。该方法有可能极大地推进血管组织工程领域，促进工程血管材料向临床应用的转化。建议的平台的非破坏性的性质使得能够在体外和体内重复评估细胞外基质支架和再细胞结构的结构-功能关系。因此，拟议的技术减轻了对跨多个时间点的破坏性分析方法的需求，这些方法成本高、耗时长，而且往往不切实际。此外，拟议的技术将有助于(A)支架生产方法的体外快速筛选和批次质量的非破坏性评估；以及(B)跨多个时间点的非终止性体内评估，从而为工程血管组织再生过程提供机械洞察力。拟议的双模式平台将集成两种用于无标记组织分析的非电离辐射技术：(1)用于评估血管生物材料的ECM成分和生化异质性的多光谱时间分辨荧光光谱(TRFs)系统；以及(2)用于评估血管生物材料的结构特性和形态的高频超声(US)成像。这可以通过超声背向散射显微镜(UBM)进行平面扫描或常规血管内超声(IVUS)进行旋转扫描来实现。将解决四个具体目标。目标1专注于开发一套用于血管支架和结构的体外和体内评估的定制工具(仪器和数据分析方法)。目的2重点论证双模式平台作为评估血管支架性能的非破坏性工具的可行性。目的3重点展示双模式平台作为体外研究和监测血管组织结构形成的非破坏性工具的能力。目的4着重于证明双模式技术作为一种非破坏性工具用于监测体内植入后血管构筑物成熟的可行性。总之，本赠款申请中建议开发和验证的技术为评估与血管生物材料的成熟度和功能相关的许多重要特征(成分、结构和功能)提供了一种非破坏性的解决方案。这可能会提高我们生产工程化血管组织的能力 在实验室进行体内植入，可以加快植入物的整合时间 与周围的宿主组织结合，从而恢复患者所需的生活质量。重点将放在工程血管组织的评估上，但如果成功，这种非破坏性技术可以应用于评估各种工程组织。

 DESCRIPTION (provided by applicant): The objective of this grant application is to research, test and validate a bi-modal diagnostic platform combining optical and ultrasound imaging technologies for real-time, non-destructive in-vitro and in-vivo analysis of composition, structure function and site specific cellular repopulation of extracellular matrix (ECM) scaffolds utilized fr vascular tissue engineering. The proposed approach has the potential to significantly advance the field of vascular tissue engineering and facilitate translation of engineered vascular material to clinical application. The non-destructive nature of the proposed platform enables repeated assessment of ECM scaffold and recellularized construct structure-function relationships both in-vitro and in-vivo. The proposed technology therefore alleviates the need for destructive analysis methods across multiple time points, which are costly, time consuming and frequently impractical. Moreover, the proposed technology will facilitate (a) in-vitro rapid screening of scaffold production methods and non-destructive assessment of batch quality; and (b) non- terminal in-vivo assessment across multiple time points, thereby providing mechanistic insights into engineered vascular tissue regenerative processes. The proposed bi-modal platform will integrate two non-ionizing radiation techniques for label-free tissue analysis: (1) Multispectral Time-Resolved Fluorescence Spectroscopy (TRFS) system for evaluation of ECM composition and biochemical heterogeneities of vascular biomaterials; and (2) High-frequency Ultrasound (US) imaging for evaluation of structural properties and morphology in vascular biomaterials. This is enabled by either Ultrasound Backscatter Microscopy (UBM) for planar scanning or conventional Intravascular Ultrasound (IVUS) for rotational scanning. Four specific aims will be addressed. Aim 1 is focused on developing a set of customized tools (instrumentation and data analysis methods) for in- vitro and in-vivo assessment of vascular scaffolds and constructs. Aim 2 in focused on demonstrating the feasibility of the bi-modal platform as a non-destructive tool for assessment of vascular scaffold properties. Aim 3 is focused on demonstrating the bi-modal platform's ability as a non-destructively tool for in-vitro studying and monitoring of vascular tissue construct formation. Aim 4 is focused on demonstrating the feasibility of the bi-modal technique as a non-destructive tool for monitoring the maturation of vascular constructs in-vivo post- implantation. In summary, the technology proposed for development and validation in this grant application offers a non-destructive solution for the evaluation of many important features (compositional, structural and functional) associated with the maturity and functionality of vascular biomaterials. This is likely to improve our ability to produce engineered vascular tissues in the laboratory for in-vivo implantation which can accelerate the integration time of the implant with the surrounding host tissue, thus restoring the desired quality of life to the patient. Emphasis will be placed on the evaluation of engineered vascular tissue, though, if successful, this non-destructive technique can be applied to assess a variety of engineered tissues.