Development of nonlinear endomicroscopy: toward assessing articular cartilage repair In vivo

非线性内窥镜的发展：评估体内关节软骨修复

• 批准号: 10244921
• 负责人: Tong Ye
• 金额: $ 25.41万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10244921
• 关键词: 3-Dimensional; Affect; Age; Animals; Architecture; Arthroscopy; Assessment tool; Biopsy; Cartilage; Cavia; Cell Survival; Cells; Centers of Research Excellence; Chondrocytes; Clinic; Clinical; Clinical Research; Collagen; Complex; Data; Defect; Degenerative polyarthritis; Dermatologic; Development; Diagnostic radiologic examination; Dyes; Elastin Fiber; Endoscopy; Evaluation; Evaluation Studies; Fluorescence; Future; Generations; Goals; Gold; Health; Histologic; Histology; Human; Image; Imaging Device; Imaging Techniques; Imaging technology; Knee joint; Label; Lasers; Lesion; Magnetic Resonance Imaging; Medical Imaging; Microscopy; Modeling; Monitor; Morphology; Musculoskeletal; Natural regeneration; Needles; Operative Surgical Procedures; Optical Coherence Tomography; Optics; Outcome; Pathology; Performance; Preparation; Problem Solving; Procedures; Process; Property; Research; Resolution; Rod; Roentgen Rays; Scanning; Signal Transduction; Source; Speed; Stains; Structure; System; Techniques; Technology; Testing; Thick; Thinness; Time; Tissue Engineering; Tissue imaging; Tissues; Translating; Translational Research; Ultrasonography; Visualization; X-Ray Computed Tomography; articular cartilage; base; cartilage repair; cell dimension; clinical application; clinical examination; clinical imaging; design; disability; follow-up; healing; imaging modality; imaging probe; imaging study; imaging system; improved; in vivo; in vivo imaging; intravital imaging; method development; microendoscopy; microscopic imaging; minimally invasive; models and simulation; novel; optical imaging; quantitative imaging; repair model; repaired; research clinical testing; second harmonic; success; two-photon

SUMMARY Repair and regeneration of articular cartilage remains a clinical and scientific challenge. Reliable assessment tools for evaluating outcomes of cartilage repair are critical for both refinement of existing methods and development of new techniques. Histological analysis of biopsies is the gold standard for repair assessment; however, biopsies are invasive procedures and therefore limited in clinical evaluation and studies of the cartilage repair. The common medical imaging methods, such as x-ray radiography, computed tomography (CT), magnetic resonance imaging (MRI), and ultrasound, can perform imaging non-destructively; however, their spatial resolutions are not sufficient to reveal the complex cell and matrix architecture of articular cartilage. Though some imaging techniques are non-destructive and can image tissue, such as arthroscopy, laser scanning confocal arthroscopy (LSCA) and optical coherence tomography (OCT), they are all performed as surgical procedures because of using thick endoscopy probes. The inability to perform clinical in vivo imaging on cartilage tissue with high spatial resolution remains a problem. To solve the problem, we propose to develop a nonlinear optical microscopy (NLOM) based endomicroscopy system for assessment of cartilage repair in vivo. In NLOM imaging of cartilage tissue, second harmonic generation (SHG) signal provides high- resolution information of fibril organization of collagen while two-photon excited fluorescence (TPEF) enables visualization of chondrocytes and elastin fibers. However, the current cartilage NLOM imaging devices all use tabletop systems that are too bulky to be used directly in clinical applications. Thus, our long-term goal is to translate this technology into a clinical imaging tool for assessment of articular cartilage repair and treatment at the cellular level. In this application, we will focus on three specific aims as follows. (1) We will determine the efficacy of using NLOM to evaluate morphological changes of articular cartilage. Using spontaneous OA guinea pigs as an articular cartilage pathology model, we will test if NLOM imaging can detect the quantitative differences among the early stages of OA cartilage tissues. (2) We will design and build a compact and high- speed NLOM imaging system with a thin rod objective as the imaging probe. A numerical simulation model will be developed to help optimize the system design. (3) With the developed endomicroscope, we will first evaluate its performance by performing a similar quantitative imaging study as described in Aim1 on excised cartilage tissues from guinea pigs with OA. We will then use the endomicroscope and tabletop system to perform a quantitative imaging study on a cartilage repair model to test if the endomicroscope can detect morphological differences between tissues in non-treated and microfracture treated defects. With the success of this study, we will be able to determine the usefulness and limitation for using NLOM to assess cartilage repair and to perform further in vivo animal study in preparation for future clinical studies on cartilage repair with the developed endomicroscope system.

概括 关节软骨的修复和再生仍然是临床和科学挑战。可靠的评估 评估软骨修复结果的工具对于现有方法的完善和 开发新技术。活检的组织学分析是维修评估的黄金标准。 但是，活检是侵入性程序，因此在临床评估中受到限制和研究 软骨维修。常见的医学成像方法，例如X射线射线照相，计算机断层扫描 （CT），磁共振成像（MRI）和超声可以进行非破坏性成像。然而， 它们的空间分辨率不足以揭示关节软骨的复杂细胞和基质结构。 尽管某些成像技术是无损的，并且可以成像组织，例如关节镜检查，激光 扫描共聚焦关节镜检查（LSCA）和光学相干断层扫描（OCT），它们均以作为 手术程序是因为使用了厚的内窥镜探针。无法在体内成像进行临床 在具有高空间分辨率的软骨组织上仍然是一个问题。为了解决问题，我们建议 开发基于非线性光学显微镜（NLOM）的内在显微镜系统，以评估软骨 在体内维修。在软骨组织的NLOM成像中，第二次谐波产生（SHG）信号提供了高 胶原蛋白原纤维组织的分辨率信息，而两光激发荧光（TPEF）可以实现 软骨细胞和弹性蛋白纤维的可视化。但是，当前的软骨NLOM成像设备都使用 桌面系统太大，无法直接用于临床应用中。因此，我们的长期目标是 将该技术转换为一种临床成像工具，以评估关节软骨修复和治疗 细胞水平。在此应用程序中，我们将重点关注以下三个特定目标。 （1）我们将确定 使用NLOM评估关节软骨的形态变化的功效。使用自发的OA 豚鼠作为关节软骨病理模型，我们将测试NLOM成像是否可以检测到定量 OA软骨组织的早期阶段之间的差异。 （2）我们将设计和建造一个紧凑而高的 用薄杆物镜作为成像探针的速度NLOM成像系统。数值模拟模型将 开发以帮助优化系统设计。 （3）使用已发达的内分体显微镜，我们将首先 通过进行AIM1中所述的类似定量成像研究来评估其性能 带有OA的豚鼠的软骨组织。然后，我们将使用内分体显微镜和桌面系统进行 对软骨维修模型进行定量成像研究，以测试内分物显微镜是否可以检测到 未处理和微裂纹处理的缺陷中组织之间的形态差异。取得成功 在这项研究中，我们将能够确定使用NLOM评估软骨的有用性和限制 修复并进行进一步的体内动物研究，以准备未来的软骨修复临床研究 随着开发的内部分子显微镜系统。