A fiber-coupled multimodal imaging platform for in vitro assessment of engineering tissue

项目概要/摘要 心血管疾病是全球主要的死亡原因。替代血管的体外工程 已经成为一个有前途的研究领域，有可能为患者提供挽救生命的治疗。 自体旁路移植是不可能的。血管组织工程的挑战需要 开发定期监测的样本，但传统的评估技术很慢， 费力和破坏性。本提案的目标是实现和验证多模态光学成像 使用单一灵活光纤接口按需获取非破坏性测量的平台 从生物反应器内发育的血管结构的腔表面。该平台设计为 三种独立但互补的成像模式将能够并行操作，这些模式包括 荧光寿命成像（FLIm），用于监测结构生物化学的变化;光学相干 断层扫描（OCT），用于监测结构形态和微观结构;以及稳态荧光 用荧光成像（SSFI）跟踪绿色荧光蛋白标记的内皮细胞在管腔内的增殖情况。 面本课题的具体目标如下：（1）实现多模态成像平台兼容 对生物反应器内血管结构的管腔表面区域进行非破坏性体外评估; (2)为了使用组织体模和工程化组织来验证多模式成像平台的性能， 血管结构。这一提议的重要性在于，它提供了一个机会， 工程组织的评估，其中传统的破坏性技术被更快，非破坏性的技术所取代。 破坏性的。该提案的创新之处在于，它将允许FLIm、OCT和SSFI并行运行， 使用相同的双包层光纤接口将光引导到样品和从样品引导光。圆满完成 这项工作有可能在再生医学领域产生重大影响， 非破坏性测量的破坏性测量将降低成本，并允许更快的原型设计， 生物材料。

Project summary/Abstract Cardiovascular disease is a leading cause of death globally. In vitro engineering of replacement blood vessels has emerged as a promising area of research that has the potential to provide lifesaving therapy in patients where autologous bypass grafts are not possible. The challenges of vascular tissue engineering require the developing samples to be monitored at regular intervals, however conventional assessment techniques are slow, laborious and destructive. The goal of this proposal is to realize and validate a multimodal optical imaging platform that uses a single flexible fiber optic interface to acquire non-destructive measurements on-demand from the luminal surface of vascular constructs developing inside a bioreactor. The platform is designed such that three independent but complimentary imaging modalities will be able to operate in parallel, these include fluorescence lifetime imaging (FLIm) for monitoring changes in construct biochemistry; optical coherence tomography (OCT) for monitoring construct morphology and microstructure; and steady state fluorescence imaging (SSFI) to track the proliferation of green fluorescent protein labeled endothelial cells across the luminal surface. The specific aims of this proposal are as follows: (1) To realize a multimodal imaging platform compatible with non-destructive in vitro assessment of the luminal surface regions of vascular constructs inside a bioreactor; (2) To validate the performance of the multimodal imaging platform using tissue phantoms and engineered vascular constructs. The significance of this proposal is it presents an opportunity for a paradigm shift in the assessment of engineered tissue, where conventional destructive techniques are supplanted with faster, non- destructive ones. The innovation of this proposal is that it will allow FLIm, OCT and SSFI to operate in parallel, using the same double-clad fiber interface to guide light to and from the sample. The successful completion of this work has the potential for significant impact in the field of regenerative medicine where supplanting destructive measurements for non-destructive ones will lower costs and allow for faster prototyping of novel biomaterials.