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.

总结 关节软骨的修复和再生仍然是临床和科学的挑战。可靠的评估 评估软骨修复结果的工具对于现有方法的改进和 开发新技术。活检的组织学分析是修复评估的金标准; 然而，活组织检查是侵入性程序，因此在临床评价和研究中受到限制。 软骨修复常用的医学成像方法，如X射线摄影、计算机断层扫描 (CT)磁共振成像（MRI）和超声可以非破坏性地执行成像;然而， 它们的空间分辨率不足以显示关节软骨的复杂细胞和基质结构。 虽然一些成像技术是非破坏性的，并且可以对组织成像，例如关节镜、激光、激光等。 扫描共焦关节镜（LSCA）和光学相干断层扫描（OCT），它们都是作为 因为使用了较厚的内窥镜探头。无法进行临床体内成像 在软骨组织上具有高空间分辨率仍然是一个问题。为了解决这个问题，我们建议 开发基于非线性光学显微镜（NLOM）的显微内镜系统，用于评估软骨 在体内修复。在软骨组织的NLOM成像中，二次谐波产生（SHG）信号提供高强度 胶原纤维组织的分辨率信息，而双光子激发荧光（TPEF）使 软骨细胞和弹性蛋白纤维的可视化。然而，目前的软骨NLOM成像设备都使用 桌面系统过于庞大，无法直接用于临床应用。因此，我们的长期目标是 将这项技术转化为临床成像工具，用于评估关节软骨修复和治疗， 细胞水平。在本申请中，我们将重点关注以下三个具体目标。(1)康贝特人将以 NLOM评价关节软骨形态学改变的有效性。使用自发性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和高- 高速NLOM成像系统与薄棒物镜作为成像探头。数值模拟模型将 有助于优化系统设计。(3)随着发展的显微内镜，我们将首先 通过进行与Aim 1中所述类似的定量成像研究，对切除的 来自患有OA的豚鼠的软骨组织。然后，我们将使用内窥镜和桌面系统， 在软骨修复模型上进行定量成像研究，以测试内窥镜是否可以检测到 未处理和微骨折处理的缺损中组织之间的形态学差异。的成功 通过本研究，我们将能够确定使用NLOM评估软骨的有用性和局限性 修复，并进行进一步的体内动物研究，为将来的软骨修复临床研究做准备 用先进的内窥镜系统