Name: software-assisted quantitative measurement of osteoarthritic subchondral bone thickness
Uploaded: 2022-03-18T14:00:00
Description: Watch this Scientific Journal Video about Software-Assisted Quantitative Measurement of Osteoarthritic Subchondral Bone Thickness at JoVE.com

这项工作得到了美国国立卫生研究院（NIH）国家关节炎和肌肉骨骼及皮肤病研究所（编号为R01 AR059088），国防部（DoD）研究奖编号为W81XWH-12-1-0304以及玛丽和保罗哈灵顿杰出教授捐赠基金的支持。

该协议中包含的所有动物程序均由堪萨斯大学医学中心的机构动物护理和使用委员会（IACUC）批准，符合所有联邦和州法律法规。
1. 小鼠膝关节OA的产生
<ol><li>通过手术破坏内侧半月板（DMM）的小鼠OA的小鼠模型，如Glasson等人描述的那样，在10-11 周龄的22只野生型BALB / c小鼠中。对具有相同背景和年龄的八只小鼠进行假手术作为对照程序。 注?...

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测量 SCB 增厚是组织学评估 OA 严重程度的重要组成部分。大多数现有的OA分级系统主要关注软骨变化15，16，17。广泛使用的小鼠骨关节炎SCB厚度分级方法将SCB增厚分为轻度，中度和重度，这在很大程度上是主观的，其可靠性尚未得到充分验证15。本研究已经开发并验证了一种新的测量方案来量化SCB厚度，其中?...

骨关节炎 （OA） 是关节炎最常见的形式，其放射学特征是关节软骨、骨赘和软骨下 （SCB） 硬化症导致关节间隙变窄1，2。虽然关节周围骨在OA病因中的作用尚不完全清楚，但骨赘形成和SCB硬化通常被认为是疾病过程的结果而不是致病因素，但关节周围骨骼结构/形状和生物学的变化可能有助于OA的发展和进展3，4.开发准确且易于执行的OA分级系统，包括SCB测量，对于研究实验室之间的比较研究以及评估旨在预防或减弱OA进展的治疗剂的功效至关重要。
SCB由薄圆顶状骨板和下层小梁骨组成。SCB板是皮质薄片，与钙化软骨平行并紧挨着钙化软骨下方。动脉和静脉血管的小分支以及神经穿透SCB板中的通道，在钙化软骨和小梁骨之间相通。软骨下小梁骨含有血管，感觉神经，骨髓，并且比SCB板更具多孔性和代谢活性。因此，SCB具有减震和支持功能，对正常关节的软骨营养供应和新陈代谢也很重要5，6，7，8。
SCB增厚（在组织学中）和硬化症（在X线照相中）是OA的主要标志和OA病理生理学的关键研究领域。测量 SCB 增厚是组织学评估 OA 严重程度的重要组成部分。先前报道的数字显微放射成像用于测量啮齿动物SCB矿物密度9，以及基于微计算机断层扫描（micro-CT）的OA10，11，12，13啮齿动物模型中的SCB定量测量，提高了我们对SCB结构和SCB变化在OA病理生理学中的作用的理解。SCB面积和厚度也已使用具有特定且昂贵的骨组织形态测量软件的复杂计算机系统通过组织学载玻片进行量化14。然而，基于视觉估计的半定量OA分级系统，包括SCB增厚分级，目前比micro-CT更广泛地使用，因为分级系统易于使用，特别是用于筛选许多组织学图像。然而，大多数现有的OA分级系统主要关注软骨变化15，16，17。广泛使用的骨关节炎SCB厚度分级方法将SCB增厚分为轻度，中度和重度，这在很大程度上是主观的，其可靠性尚未得到充分验证15。可靠且易于执行的分步骨关节炎SCB厚度测量方案要么尚未完全开发，要么未标准化。
本研究旨在开发一种可重复，灵敏且易于执行的方案，以定量测量OA小鼠模型中的SCB厚度。我们严格的测量测试和统计分析表明，这种ImageJ软件辅助的定量测量方案可以量化正常和骨关节炎膝关节的SCB厚度。新开发的方案是可重复的，并且比广泛使用的视觉分级系统对轻度SCB变化更敏感。它可用于检测早期骨关节炎SCB变化，并用于评估OA治疗与OA软骨分级的 体内 疗效。

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			<td>Safranin-O</td>
			<td>Sigma-Aldrich</td>
			<td>S8884</td>
			<td></td>
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		<tr>
			<td>Fast green</td>
			<td>Sigma-Aldrich</td>
			<td>F7252</td>
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			<td>Hematoxylin</td>
			<td>Sigma-Aldrich</td>
			<td>GHS216</td>
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			<td>Eosin</td>
			<td>Sigma-Aldrich</td>
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			<td>illustrator</td>
			<td>Adobe</td>
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software-assisted quantitative measurement of osteoarthritic subchondral bone thickness

软骨下骨增厚和硬化是骨关节炎（OA）的主要标志，无论是在动物模型中还是在人类中。目前，组织学软骨下骨增厚的严重程度主要由基于视觉估计的半定量分级系统确定。本文提出了一种可重复且易于执行的方案，用于定量测量由内侧半月板（DMM）不稳定诱导的膝关节OA小鼠模型中的软骨下骨厚度。该协议利用ImageJ软件在定义股骨内侧髁和内侧胫骨平台的感兴趣区域后，量化组织学图像上的软骨下骨厚度，其中软骨下骨增厚通常发生在DMM诱导的膝关节OA中。使用假手术的膝关节的组织学图像作为对照。统计分析表明，新开发的定量软骨下骨测量系统具有高度的可重复性，观察者和观察者之间的变异性较低。结果表明，与广泛使用的视觉分级系统相比，新方案对细微或轻度软骨下骨增厚更敏感。该协议适用于检测早期和进展的骨关节炎软骨下骨变化，并用于评估OA治疗与OA软骨分级的 体内 疗效。

视觉估计分级和 ImageJ 辅助定量测量之间的再现性比较： 48个感兴趣区域（ROI）（24 MFC和24 MTP）的SCB厚度，由三个独立个体使用文献中描述的现有0-3视觉评分方案进行评分，其中0 =正常（无SCB增厚），1 =轻度，2 =中度，3 =重度SCB增厚。这些图像是从DMM或假手术后2，8和16周的三个不同的术后时间点中选择的。通常，采用DMM手术的小鼠在术后...

Watch this Scientific Journal Video about Software-Assisted Quantitative Measurement of Osteoarthritic Subchondral Bone Thickness at JoVE.com

Software-Assisted Quantitative Measurement of Osteoarthritic Subchondral Bone Thickness

本方法论文章提出了一种软件辅助定量测量方案，用于量化小鼠骨关节炎膝关节和正常膝关节的组织学软骨下骨厚度作为对照。该方案对细微增厚高度敏感，适用于检测早期骨关节炎软骨下骨变化。

软件辅助骨关节炎软骨下骨厚度的定量测量

Subchondral bone thickening and sclerosis are the major hallmarks of osteoarthritis (OA), both in animal models and in humans. Currently, the severity of ...

软骨下骨增厚或硬化症是骨关节炎的标志之一，无论是在动物模型中还是在人类中。目前，组织学软骨下骨增厚的严重程度主要通过基于半定量分级系统的视觉估计来确定。本研究旨在开发一种可重复且易于执行的方案，以定量测量由内侧半月板不稳定诱导的膝关节OA小鼠模型中的软骨下骨厚度。我们通过手术破坏内侧半月板的稳定性来创建膝关节骨关节炎的小鼠模型，该模型在手术显微镜和无菌条件下进行。小鼠膝关节的组织学过程包括组织固定、脱钙、修剪、加工、包埋、切片和染色，然后进行显微镜成像。屏幕上的图像显示了小鼠膝关节骨关节炎伴关节软骨病变，骨赘形成和软骨下增厚使用ImageJ软件定量测量骨关节炎软骨下骨。从 imagej.nih.gov/ij 下载图像J。打开图像J。单击"文件"选项卡，然后单击"打开"选项以打开组织学图像。找到文件目录地址，选择图片文件，然后单击"打开"。使用直线工具绘制一个长度单位（一千分尺），然后单击"分析"，然后单击"设置比例"。将已知距离和像素长宽比设置为 1，然后单击 Ok.ImageJ 可以将像素长度转换为单位长度，即千分尺。将测量因子设置为面积。单击分析，然后单击设置测量，然后在新窗口下选中面积和阈值限制框。此步骤将 ImageJ 设置为测量所选阈值内的参数区域。定义的总软骨下感兴趣区域，如无线电所示，覆盖软骨下皮质板和毗邻皮质板的底层小梁骨的一部分。使用图像主窗口下的选择工具绘制总软骨下骨面积的轮廓。选择工具为系统提供一个阈值，用于在选择阈值后限制测量。单击分析，然后单击测量。将打开一个带有面积测量结果的窗口。单击"编辑"，然后单击"清除外部"以排除总软骨下骨区域之外的区域。单击"清除外部"选项后，仅可见总软骨下骨面积。单击"图像"，然后单击"调整"和"颜色阈值"以打开阈值颜色窗口。单击阈值颜色窗口底部的"原始"，将图片恢复到原始状态。使用选择工具在骨物质区域绘制一个小盒子。单击窗口底部的"样本选项"以定义骨物质区域。选定的骨质区域将变为红色。单击阈值颜色窗口底部的选择以创建面积测量阈值。单击分析，然后在 ImageJ 主菜单中测量。骨物质面积测量结果将显示在结果窗口中。保存总软骨下骨面积和骨质面积的数据。观测者之间和观测者内部的变异性和再现性均采用皮尔逊相关系数分析确定。研究组之间差异的显著性是使用学生t检验或单因素方差分析确定的，然后是事后检验。此图显示了上面板和下面板。上图显示了来自 Sham 组和 DMM 组的组织学图像的显微照片，用于基于视觉估计的软骨下骨分级。下图显示了来自 Sham 组和 DMM 组的组织学图像的显微照片，用于 ImageJ 辅助的定量软骨下骨测量。用Adobe Illustrator制作的黄色虚线勾勒出的框定义了感兴趣的总软骨下骨区域。盒子内的骨物质区域以橙色突出显示。使用ImageJ软件可以量化股骨髁内侧和胫骨平台内侧软骨下骨的显着增厚。在该图中，观察者间变异测试表明，对于内侧胫平台和内侧股骨髁中软骨厚度的第一次和第二次测量，观察者之间具有很高的再现性。在该图中，观察者变异试验表明，在内侧胫平台和内侧股骨髁区域，第一和第二软骨下骨厚度测量之间的重复性很高。表1显示了视觉分级和ImageJ辅助的软骨下骨厚度定量测量之间再现性的比较分析。相关系数检验表明，定量测量相对上比视觉分级系统更具可重复性。该图显示了视觉分级的比较灵敏度分析和ImageJ辅助的内侧股骨髁和内侧胫骨平台软骨下骨厚度的定量测量。将用于视觉估计分级的组织学图像分为三组。对于视觉评分为零的DMM图像，所有三个观察者的定量软骨下骨厚度值均显著高于视觉评分为零的Sham图像，表明定量测量比视觉分级对轻度软骨下骨增厚更敏感。新开发的用于定量测量骨关节炎软骨下骨厚度的方案比广泛使用的视觉分级系统对轻度软骨下骨变化更敏感。该协议可用于检测早期骨关节炎软骨下骨的变化，并用于评估骨关节炎治疗与骨关节炎软骨分级的体内疗效。

Research

JoVE Journal

Medicine

Quantitative Autonomic Testing

定量自主测试

Disorders associated with dysfunction of autonomic nervous system are quite common yet frequently unrecognized. Quantitative autonomic testing can be ...

科研

医学

Models of Bone Metastasis

骨转移模型

Bone metastases are a common occurrence in several malignancies, including breast, prostate, and lung. Once established in bone, tumors are responsible ...

Quantitative Analysis of Chromatin Proteomes in Disease

在疾病的染色质蛋白质组的定量分析

In the nucleus reside the proteomes whose functions are most intimately linked with gene regulation. Adult mammalian cardiomyocyte nuclei are unique due ...

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

采用实时定量PCR检测供体细胞植入一个具有竞争力的小鼠骨髓移植模型

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

Pre-clinical Evaluation of Tyrosine Kinase Inhibitors for Treatment of Acute Leukemia

酪氨酸激酶抑制剂的临床前评价急性白血病的治疗

Receptor tyrosine kinases have been implicated in the development and progression of many cancers, including both leukemia and solid tumors, and are ...

Using a Knee Arthrometer to Evaluate Tissue-specific Contributions to Knee Flexion Contracture in the Rat

用膝关节关节仪评价大鼠膝关节屈曲收缩的特定组织贡献

Normal knee range of motion (ROM) is critical to well-being and allows one to perform basic activities such as walking, climbing stairs and sitting. Lost ...

Quantitative [18F]-Naf-PET-MRI Analysis for the Evaluation of Dynamic Bone Turnover in a Patient with Facetogenic Low Back Pain

Quantitative [18F]-Naf-PET-MRI Analysis for the Evaluation of Dynamic Bone Turnover in a Patient with Facetogenic Low Back Pain

定量 =18F_-Naf-PET-MRI 分析,用于评估面部腰痛患者的动态骨质周转率

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. Bone is a dynamic tissue undergoing ...

Flow Cytometry Analysis of Immune Cell Subsets within the Murine Spleen, Bone Marrow, Lymph Nodes and Synovial Tissue in an Osteoarthritis Model

骨关节炎模型中骨髓、骨髓、淋巴结和合成组织内免疫细胞子集的流动细胞测量分析

Osteoarthritis (OA) is one of the most prevalent musculoskeletal diseases, affecting patients suffering from pain and physical limitations. Recent ...

Evaluation of Amino Acid Consumption in Cultured Bone Cells and Isolated Bone Shafts

Bone development and homeostasis is dependent upon the differentiation and activity of bone forming osteoblasts. Osteoblast differentiation is ...

Precision of In Vivo Quantitative Tooth Wear Measurement Using Intra-Oral Scans

Precision of In Vivo Quantitative Tooth Wear Measurement Using Intra-Oral Scans

使用口腔内扫描进行 体内 定量牙齿磨损测量的精度

Quantitative wear measurement is of increasing interest for measuring tooth wear progression. However, most research on quantitative wear measurement has ...