Name: quantifying fibrillar collagen organization with curvelet transform-based tools
Uploaded: 2020-11-11T14:00:00.000+00:00
Duration: 7 min 58 s
Description: Watch this Scientific Journal Video about Quantifying Fibrillar Collagen Organization with Curvelet Transform-Based Tools at JoVE.com

我们感谢许多贡献者和用户的CT-FIRE和曲线对齐多年来，包括罗布诺瓦克博士， 卡罗琳·佩尔克博士、杰里米·布雷德费尔特博士、古内特·梅塔博士、安德鲁·莱希特博士、阿迪布·凯霍斯拉维博士、马特·康克林博士、杰恩·松鼠博士、保罗·普罗文扎诺博士、布伦达·奥格尔博士、帕特里夏·基利博士、约瑟夫·舒尔切夫斯基博士、苏珊娜·波尼克博士以及斯瓦蒂·阿南德和柯蒂斯·鲁登的其他技术贡献。这项工作得到了半导体研究公司、莫格里奇研究所的资助，国家卫生研究院向K.W.E.提供了R01CA19996、R01CA181385和U54CA210190。

作者没有什么可透露的。

此协议描述了使用 CT-FIRE 和曲线对齐进行纤维胶原蛋白量化，可应用于任何具有胶原纤维或其他线状或纤维状拉伸结构的图像，这些结构适合 CT-FIRE 或曲线对齐进行分析。例如，弹性蛋白或弹性纤维可以在此平台上以类似的方式进行处理。我们已经在计算产生的合成纤维21上测试了这两种工具。根据应用情况，用户应选择最适合其数据的分析模式。CT纤维分析模式可以直接使用CT?...

纤维胶原蛋白是突出的结构 ECM 成分。他们的组织变化影响组织功能，并可能与许多疾病的进展有关，从骨质疏松症1，心脏功能障碍2，伤口愈合3到不同类型的癌症，包括乳腺癌4，5，6，卵巢7，8，肾9，胰腺癌10。许多已建立的成像模式可用于可视化纤维胶原蛋白，如第二谐波一代显微镜11，污渍或染料与明亮的场或荧光显微镜或偏振光显微镜12，液晶基极化显微镜（LC-PolScope）13，电子显微镜14。随着纤维胶原蛋白组织的重要性越来越明显，而且这些方法的使用也越来越多，改进胶原纤维分析方法的需求也越来越大。
有许多努力来开发自动测量纤维状胶原蛋白的计算方法。免费提供的软件工具主要侧重于光纤对齐或方向的计算，采用第一个衍生或结构张力像素15，16，或傅立业基于转换的频谱分析图像磁贴17。所有这些工具都受到限制，例如手动步骤的要求、在嘈杂的背景中检测光纤边缘的不准确性或缺乏本地化特征特征。
与其他免费提供的开源免费软件工具相比，本协议中描述的方法使用CT——一种最佳、多尺度、定向的图像表示方法——去除纤维胶原蛋白图像中的噪声，增强或跟踪光纤边缘。有关位置和方向的信息可以直接从光纤提供，而不是使用间接像素或窗口信息来推断光纤组织的指标。这种基于CT的框架18、19、20、21可以在全球、ROI或光纤的基础上量化光纤组织，主要通过两个单独但相互关联的包，分别命名为"CT-FIRE"18、21和"曲线对齐"19、21。就软件的实现而言，在 CT-FIRE 中，多比例的 CT 系数可用于重建增强边缘和降低噪音的图像。然后，将单个光纤提取算法应用于 CT 重建图像中，以跟踪光纤以查找其代表性的中心点，将光纤分支从中心点延伸，并将光纤分支连接起来，形成光纤网络。在 Curve 对齐中，用户指定比例的 CT 系数可用于跟踪本地光纤方向，其中提取曲线小径的方向和位置并进行分组，以估计相应位置的光纤方向。由此产生的量化框架已经开发了十多年，在功能、用户界面和模块化等许多方面都取得了长足的发展。例如，此工具可以可视化局部光纤方向，并允许用户测量多达 30 个光纤功能，包括长度、角度、宽度和直度等单个光纤特性，以及密度和对齐等批量测量。此外，用户可以测量光纤角度相对于手动或自动分割的边界，例如，在基于图像的生物标志物发展在乳腺癌22和胰腺癌研究10发挥重要作用。此平台提供多个功能模块，包括用于投资回报率分析、自动边界创建和后处理的功能模块。投资回报率模块可用于注释不同形状的投资回报率，并进行相应的投资回报率分析。作为应用示例，自动边界创建模块可用于用第二谐波生成 （SHG） 图像注册血氧林和 eosin （H&E） 亮场图像，并从注册的 H&E 图像中生成肿瘤边界的图像掩膜。后处理模块可以帮助处理和集成来自单个图像的输出数据文件，以便进行可能的统计分析。
此量化平台不需要事先的编程或图像处理经验，可以处理包括数百或数千张图像在内的大型数据集，从而能够高效量化用于生物或生物医学应用的胶原蛋白组织。它已被包括我们在内的世界各地许多研究人员广泛应用于不同的研究领域。有四个主要出版物CT-FIRE和曲线对齐18，19，20，21，其中前三个已被引用272次（根据谷歌学者2020-05-04年）。对引用该平台（CT-FIRE 或曲线对齐）的出版物的审查表明，大约有 110 篇期刊论文直接用于分析，其中大约 35 篇出版物与我们的小组合作，其他出版物（约 75 篇）由其他组撰写。例如， 该平台用于以下研究:乳腺癌22，23，24，胰腺癌10，25，肾癌9，26，伤口愈合3，27，28，29，30，卵巢癌8，31，7，子宫韧带32，缺氧登丁3 3、基底细胞癌34例、缺氧肉瘤35例、软骨组织36例、心脏功能障碍37例、神经元38例、胶质母细胞瘤39例、淋巴收缩40例、纤维化41例、胃癌42例、微管癌43例、膀胱纤维化44例。图 1演示了 Curve 对齐的癌症成像应用，从 SHG 图像中查找乳腺癌19的肿瘤相关胶原蛋白特征。图 2描述了此平台的典型示意图工作流程。虽然这些工具已经审查技术18，19，21，和常规协议20对齐分析曲线对齐也可用，一个视觉协议，显示所有的基本功能可能是有用的。如本文所示，可视化协议将促进使用此平台的学习过程，并更有效地解决用户可能提出的问题和问题。

<table><tbody><tr><td>CT-FIRE</td><td>Univerity of Wisconsin-Madison</td><td>N/A</td><td>open source software available from https://eliceirilab.org/software/ctfire/</td></tr><tr><td>CurveAlign</td><td>University of Wisconsin-Madison</td><td>N/A</td><td>open source software available from https://eliceirilab.org/software/curvealign/</td></tr></tbody></table>

quantifying fibrillar collagen organization with curvelet transform-based tools

纤维状胶原蛋白是突出的细胞外基质 （ECM） 成分，其拓扑变化已证明与乳腺癌、卵巢癌、肾脏癌和胰腺癌等多种疾病的进展有关。免费提供的光纤量化软件工具主要侧重于光纤对齐或方向的计算，它们受到手动步骤要求、嘈杂背景中光纤边缘检测不准确或缺乏本地化特征特征等限制。本文中描述的胶原纤维定量工具的特点是使用曲线变换 （CT） 启用的最佳多尺度图像表示。这种算法方法允许去除纤维胶原蛋白图像中的噪声，并增强光纤边缘，直接从光纤中提供位置和方向信息，而不是使用从其他工具获得的间接像素或窗口信息。此基于 CT 的框架包含两个单独但链接的包，名为"CT-FIRE"和"曲线对齐"，可在全球、感兴趣区域 （ROI） 或单个光纤基础上量化光纤组织。这个量化框架已经开发了十多年，现在已经发展成为一个全面和用户驱动的胶原蛋白量化平台。使用此平台，可以测量多达 30 种光纤功能，包括长度、角度、宽度和直度等单个纤维特性，以及密度和对齐度等批量测量。此外，用户可以测量光纤角度相对于手动或自动分割的边界。该平台还提供了多个附加模块，包括用于投资回报率分析、自动边界创建和后处理的模块。使用此平台不需要事先的编程或图像处理经验，它可以处理包括数百或数千张图像在内的大型数据集，从而能够高效量化用于生物或生物医学应用的胶原纤维组织。

这些方法已成功地应用于许多研究。一些典型应用包括:1） Conklin 等人22 使用曲线对齐来计算肿瘤相关的胶原蛋白特征，并发现胶原纤维更频繁地垂直于原位导管癌 （DCIS） 病变中的导管周长:2） Drifka等人使用曲线 对齐中的CT-FIRE模式来量化胰腺导管腺癌和正常/慢性胰腺炎组织的频闪胶原对齐，发现与正常/慢性组织相比，癌组织中的对齐度有所提高:3） Alkmin<sup...

Watch this Scientific Journal Video about Quantifying Fibrillar Collagen Organization with Curvelet Transform-Based Tools at JoVE.com

Quantifying Fibrillar Collagen Organization with Curvelet Transform-Based Tools

在这里，我们提出了一个协议，使用基于曲线转换的开源 MATLAB 软件工具，用于量化正常组织和患病组织的细胞外基质中的纤维状胶原蛋白组织。此工具可应用于胶原纤维或其他类型的线状结构的图像。

使用基于曲线的转换工具量化纤维胶原蛋白组织

Fibrillar collagens are prominent extracellular matrix (ECM) components, and their topology changes have been shown to be associated with the progression ...

quantifying-fibrillar-collagen-organization-with-curvelet-transform

Research

JoVE Journal

Bioengineering

6.0K 次观看.  University of Wisconsin-Madison. 在这里，我们提出了一个协议，使用基于曲线转换的开源 MATLAB 软件工具，用于量化正常组织和患病组织的细胞外基质中的纤维状胶原蛋白组织。此工具可应用于胶原纤维或其他类型的线状结构的图像。

视频: Quantifying Fibrillar Collagen Organization with Curvelet Transform-Based Tools

Observing and Quantifying Fibroblast-mediated Fibrin Gel Compaction

Cells embedded in collagen and fibrin gels attach and exert traction forces on the fibers of the gel. These forces can lead to local and global reorganization and realignment of the gel microstructure. This process proceeds in a complex manner that is dependent in part on the interplay between the location of the cells, the geometry of the gel, and the mechanical constraints on the gel. To better understand how these variables produce global fiber alignment patterns, we use time-lapse differential interference contrast (DIC) microscopy coupled with an environmentally controlled bioreactor to observe the compaction process between geometrically spaced explants (clusters of fibroblasts). The images are then analyzed with a custom image processing algorithm to obtain maps of the strain. The information obtained from this technique can be used to probe the mechanobiology of various cell-matrix interactions, which has important implications for understanding processes in wound healing, disease development, and tissue engineering applications.

Time-lapse microscopy and image processing techniques were used to observe and analyze fibroblast-mediated gel compaction and fibrin fiber realignment in an environmentally controlled bioreactor over a 48 hr&#160;period.

观察和量化纤维细胞介导的纤维素凝胶压实

Cells embedded in collagen and fibrin gels attach and exert traction forces on the fibers of the gel. These forces can lead to local and global ...

科研

生物工程

Engineering Fibrin-based Tissue Constructs from Myofibroblasts and Application of Constraints and Strain to Induce Cell and Collagen Reorganization

Collagen content and organization in developing collagenous tissues can be influenced by local tissue strains and tissue constraint. Tissue engineers aim to use these principles to create tissues with predefined collagen architectures. A full understanding of the exact underlying processes of collagen remodeling to control the final tissue architecture, however, is lacking. In particular, little is known about the (re)orientation of collagen fibers in response to changes in tissue mechanical loading conditions. We developed an in&#160;vitro model system, consisting of biaxially-constrained myofibroblast-seeded fibrin constructs, to further elucidate collagen (re)orientation in response to i) reverting biaxial to uniaxial static loading conditions and ii) cyclic uniaxial loading of the biaxially-constrained constructs before and after a change in loading direction, with use of the Flexcell FX4000T loading device. Time-lapse confocal imaging is used to visualize collagen (re)orientation in a nondestructive manner.
Cell and collagen organization in the constructs can be visualized in real-time, and an internal reference system allows us to relocate cells and collagen structures for time-lapse analysis. Various aspects of the model system can be adjusted, like cell source or use of healthy and diseased cells. Additives can be used to further elucidate mechanisms underlying collagen remodeling, by for example adding MMPs or blocking integrins. Shape and size of the construct can be easily adapted to specific needs, resulting in a highly tunable model system to study cell and collagen (re)organization.

This model system starts from a myofibroblast-populated fibrin gel that can be used to study endogenous collagen (re)organization real-time in a nondestructive manner. The model system is very tunable, as it can be used with different cell sources, medium additives, and can be adapted easily to specific needs.

肌成纤维细胞和约束的应用工程纤维蛋白的​​组织结构和应变诱导细胞和胶原蛋白（RE）组织

Collagen content and organization in developing collagenous tissues can be influenced by local tissue strains and tissue constraint. Tissue engineers aim ...

Examining the Dynamics of Cellular Adhesion and Spreading of Epithelial Cells on Fibronectin During Oxidative Stress

The adhesion and spreading of cells onto the extracellular matrix (ECM) are essential cellular processes during organismal development and for the homeostasis of adult tissues. Interestingly, oxidative stress can alter these processes, thus contributing to the pathophysiology of diseases such as metastatic cancer. Therefore, understanding the mechanism(s) of how cells attach and spread on the ECM during perturbations in redox status can provide insight into normal and disease states. Described below is a step-wise protocol that utilizes an immunofluorescence-based assay to specifically quantify cell adhesion and spreading of immortalized fibroblast cells on fibronectin (FN) in vitro. Briefly, anchorage-dependent cells are held in suspension and exposed to the ATM kinase inhibitor Ku55933 to induce oxidative stress. Cells are then plated on FN-coated surface and allowed to attach for predetermined periods of time. Cells that remain attached are fixed and labeled with fluorescence-based antibody markers of adhesion (e.g., paxillin) and spreading (e.g., F-actin). Data acquisition and analysis are performed using commonly available laboratory equipment, including an epifluorescence microscope and freely available Fiji software. This procedure is highly versatile and can be modified for a variety of cell lines, ECM proteins, or inhibitors in order to examine a broad range of biological questions.

This method is useful for quantifying the early dynamics of cellular adhesion and spreading of anchorage-dependent cells onto the fibronectin. Furthermore, this assay can be used to investigate the effects of altered redox homeostasis on cell spreading and/or cell adhesion-related intracellular signaling pathways.

在氧化应激期间细胞粘附和纤维蛋白扩散动力学

The adhesion and spreading of cells onto the extracellular matrix (ECM) are essential cellular processes during organismal development and for the ...

生物化学

Revealing the Cytoskeletal Organization of Invasive Cancer Cells in 3D

Cell migration has traditionally been studied in 2D substrates. However, it has become increasingly evident that there is a need to study cell migration in more appropriate 3D environments, which better resemble the dimensionality of the physiological processes in question. Migratory cells can substantially differ in their morphology and mode of migration depending on whether they are moving on 2D or 3D substrates. Due to technical difficulties and incompatibilities with most standard protocols, structural and functional analysis of cells embedded within 3D matrices still remains uncommon. This article describes methods for preparation and imaging of 3D cancer cell cultures, either as single cells or spheroids. As an appropriate ECM substrate for cancer cell migration, we use nonpepsinized rat tail collagen I polymerized at room-temperature and fluorescently labeled to facilitate visualization using standard confocal microscopes. This work also includes a protocol for 3D immunofluorescent labeling of endogenous cell cytoskeleton. Using these protocols we hope to contribute to a better description of the molecular composition, localization, and functions of cellular structures in 3D.

This article presents a method to fluorescently label collagen that can be further used for both fix and live imaging of 3D cell cultures. We also provide an optimized protocol to visualize endogenous cytoskeletal proteins of cells cultured in 3D environments.

在3D揭示侵入性癌症细胞骨架组织

Cell migration has traditionally been studied in 2D substrates. However, it has become increasingly evident that there is a need to study cell migration ...

医学

Preparation of 3D Collagen Gels and Microchannels for the Study of 3D Interactions In Vivo

Historically, most cellular processes have been studied in only 2 dimensions. While these studies have been informative about general cell signaling mechanisms, they neglect important cellular cues received from the structural and mechanical properties of the local microenvironment and extracellular matrix (ECM). To understand how cells interact within a physiological ECM, it is important to study them in the context of 3 dimensional assays. Cell migration, cell differentiation, and cell proliferation are only a few processes that have been shown to be impacted by local changes in the mechanical properties of a 3-dimensional ECM. Collagen I, a core fibrillar component of the ECM, is more than a simple structural element of a tissue. Under normal conditions, mechanical cues from the collagen network direct morphogenesis and maintain cellular structures. In diseased microenvironments, such as the tumor microenvironment, the collagen network is often dramatically remodeled, demonstrating altered composition, enhanced deposition and altered fiber organization. In breast cancer, the degree of fiber alignment is important, as an increase in aligned fibers perpendicular to the tumor boundary has been correlated to poorer patient prognosis1. Aligned collagen matrices result in increased dissemination of tumor cells via persistent migration2,3. The following is a simple protocol for embedding cells within a 3-dimensional, fibrillar collagen hydrogel. This protocol is readily adaptable to many platforms, and can reproducibly generate both aligned and random collagen matrices for investigation of cell migration, cell division, and other cellular processes in a tunable, 3-dimensional, physiological microenvironment.

Preparation of 3D Collagen Gels and Microchannels for the Study of 3D Interactions In Vivo

Collagen is a core component of the ECM, and provides essential cues for several cellular processes ranging from migration to differentiation and proliferation. Provided here is a protocol for embedding cells within 3D collagen hydrogels, and a more advanced technique for generating randomized or aligned collagen matrices using PDMS microchannels.

3D胶原凝胶和微通道3D相互作用的研究准备<em&gt;在体内</em

Historically, most cellular processes have been studied in only 2 dimensions.  While these studies have been informative about general cell signaling ...

Assessing Collagen and Elastin Pressure-dependent Microarchitectures in Live, Human Resistance Arteries by Label-free Fluorescence Microscopy

The pathogenic contribution of resistance artery remodeling is documented in essential hypertension, diabetes and the metabolic syndrome. Investigations and development of microstructurally motivated mathematical models for understanding the mechanical properties of human resistance arteries in health and disease have the potential to aid understanding how disease and medical treatments affect the human microcirculation. To develop these mathematical models, it is essential to decipher the relationship between the mechanical and microarchitectural properties of the microvascular wall. In this work, we describe an ex vivo method for passive mechanical testing and simultaneous label-free three-dimensional imaging of the microarchitecture of elastin and collagen in the arterial wall of isolated human resistance arteries. The imaging protocol can be applied to resistance arteries of any species of interest. Image analyses are described for quantifying i) pressure-induced changes in internal elastic lamina branching angles and adventitial collagen straightness using Fiji and ii) collagen and elastin volume densities determined using Ilastik software. Preferably all mechanical and imaging measurements are performed on live, perfused arteries, however, an alternative approach using standard video-microscopy pressure myography in combination with post-fixation imaging of re-pressurized vessels is discussed. This alternative method provides users with different options for analysis approaches. The inclusion of the mechanical and imaging data in mathematical models of the arterial wall mechanics is discussed, and future development and additions to the protocol are proposed.

We describe simultaneous mechanical testing and 3D-imaging of the arterial wall of isolated, live human resistance arteries, and Fiji and Ilastik image analyses for the quantification of elastin and collagen spatial organization and volume densities. We discuss the use of these data in mathematical models of arterial wall mechanics.

用无标签荧光显微镜评价活体、人体阻力动脉中胶原蛋白和弹性蛋白的压力依赖性 Microarchitectures

The pathogenic contribution of resistance artery remodeling is documented in essential hypertension, diabetes and the metabolic syndrome. Investigations ...

Semi-automated Analysis of Mouse Skeletal Muscle Morphology and Fiber-type Composition

For years, distinctions between skeletal muscle fiber types were best visualized by myosin-ATPase staining. More recently, immunohistochemical staining of myosin heavy chain (MyHC) isoforms has emerged as a finer discriminator of fiber-type. Type I, type IIA, type IIX and type IIB fibers can now be identified with precision based on their MyHC profile; however, manual analysis of these data can be slow and down-right tedious. In this regard, rapid, accurate assessment of fiber-type composition and morphology is a very desirable tool. Here, we present a protocol for state-of-the-art immunohistochemical staining of MyHCs in frozen sections obtained from mouse hindlimb muscle in concert with a novel semi-automated algorithm that accelerates analysis of fiber-type and fiber morphology. As expected, the soleus muscle displayed staining for type I and type IIA fibers, but not for type IIX or type IIB fibers. On the other hand, the tibialis anterior muscle was composed predominantly of type IIX and type IIB fibers, a small fraction of type IIA fibers and little or no type I fibers. Several image transformations were used to generate probability maps for the purpose of measuring different aspects of fiber morphology (i.e., cross-sectional area (CSA), maximal and minimal Feret diameter). The values obtained for these parameters were then compared with manually-obtained values. No significant differences were observed between either mode of analysis with regards to CSA, maximal or minimal Feret diameter (all p &#62; 0.05), indicating the accuracy of our method. Thus, our immunostaining analysis protocol may be applied to the investigation of effects on muscle composition in many models of aging and myopathy.

Immunohistochemical staining of myosin heavy chain isoforms has emerged as the state-of-the-art discriminator of skeletal muscle fiber-type (i.e., type I, type IIA, type IIX, type IIB). Here, we present a staining protocol along with a novel semi-automated algorithm that facilitates rapid assessment of fiber-type and fiber morphology.

小鼠骨骼肌形态及纤维型成分的半自动化分析

For years, distinctions between skeletal muscle fiber types were best visualized by myosin-ATPase staining. More recently, immunohistochemical staining of ...

A Method to Study the Correlation Between Local Collagen Structure and Mechanical Properties of Atherosclerotic Plaque Fibrous Tissue

The rupture of atherosclerotic plaques in coronary and carotid arteries is the primary cause of fatal cardiovascular events. However, the rupture mechanics of the heterogeneous, highly collagenous plaque tissue, and how this is related to the tissue's fibrous structure, are not known yet. Existing pipelines to study plaque mechanics are limited to obtaining only gross mechanical characteristics of the plaque tissue, based on the assumption of structural homogeneity of the tissue. However, fibrous plaque tissue is structurally heterogeneous, arguably mainly due to local variation in the collagen fiber architecture.
The mechano-imaging pipeline described here has been developed to study the heterogeneous structural and mechanical plaque properties. In this pipeline, the tissue's local collagen architecture is characterized using multiphoton microscopy (MPM) with second-harmonic generation (SHG), and the tissue's failure behavior is characterized under uniaxial tensile testing conditions using digital image correlation (DIC) analysis. This experimental pipeline enables correlation of the local predominant angle and dispersion of collagen fiber orientation, the rupture behavior, and the strain fingerprints of the fibrous plaque tissue. The obtained knowledge is key to better understand, predict, and prevent atherosclerotic plaque rupture events.

We have developed a mechano-imaging pipeline to study the heterogeneous structural and mechanical atherosclerotic plaque properties. This pipeline enables correlation of the local predominant angle and dispersion of collagen fiber orientation, the rupture behavior, and the strain fingerprints of the fibrous plaque tissue.

一种研究动脉粥样硬化斑块纤维组织局部胶原结构与力学性能相关性的方法

The rupture of atherosclerotic plaques in coronary and carotid arteries is the primary cause of fatal cardiovascular events. However, the rupture ...

了解肌腱撞击 – 用于研究机械生物学的小鼠外植体模型

Murine Hind Limb Explant Model for Studying the Mechanobiology of Achilles Tendon Impingement

Tendon impingement upon bone generates a multiaxial mechanical strain environment with markedly elevated transverse compressive strain, which elicits a localized fibrocartilage phenotype characterized by accumulation of glycosaminoglycan (GAG)-rich matrix and remodeling of the collagen network. While fibrocartilage is a normal feature in impinged regions of healthy tendons, excess GAG deposition and disorganization of the collagen network are hallmark features of tendinopathy. Accordingly, impingement is clinically recognized as an important extrinsic factor in the initiation and progression of tendinopathy. Nevertheless, the mechanobiology underlying tendon impingement remains understudied. Prior efforts to elucidate the cellular response to tendon impingement have applied uniaxial compression to cells and excised tendon explants in vitro. However, isolated cells lack a three-dimensional extracellular environment crucial to mechanoresponse, and both in vitro and excised explant studies fail to recapitulate the multiaxial strain environment generated by tendon impingement in vivo, which depends on anatomical features of the impinged region. Moreover, in vivo models of tendon impingement lack control over the mechanical strain environment. To overcome these limitations, we present a novel murine hind limb explant model suitable for studying the mechanobiology of Achilles tendon impingement. This model maintains the Achilles tendon in situ to preserve local anatomy and reproduces the multiaxial strain environment generated by impingement of the Achilles tendon insertion upon the calcaneus during passively applied ankle dorsiflexion while retaining cells within their native environment. We describe a tissue culture protocol integral to this model and present data establishing sustained explant viability over 7 days. The representative results demonstrate enhanced histological GAG staining and decreased collagen fiber alignment secondary to impingement, suggesting elevated fibrocartilage formation. This model can easily be adapted to investigate different mechanical loading regimens and allows for the manipulation of molecular pathways of interest to identify mechanisms mediating phenotypic change in the Achilles tendon in response to impingement.

作者聚焦：揭开肌腱撞击的力学生物学 – 多轴鼠后肢外植体模型 

We present a custom experimental platform and tissue culture protocol that recreates fibrocartilaginous change driven by impingement of the Achilles tendon insertion in murine hind limb explants with sustained cell viability, providing a model suitable for exploring the mechanobiology of tendon impingement.

小鼠后肢外植体研究跟腱撞击力学生物学的模型

Tendon impingement upon bone generates a multiaxial mechanical strain environment with markedly elevated transverse compressive strain, which elicits a ...

使用基于曲线的转换工具量化纤维胶原蛋白组织

摘要

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观察和量化纤维细胞介导的纤维素凝胶压实

肌成纤维细胞和约束的应用工程纤维蛋白的组织结构和应变诱导细胞和胶原蛋白（RE）组织

在氧化应激期间细胞粘附和纤维蛋白扩散动力学

在3D揭示侵入性癌症细胞骨架组织

3D胶原凝胶和微通道3D相互作用的研究准备

用无标签荧光显微镜评价活体、人体阻力动脉中胶原蛋白和弹性蛋白的压力依赖性 Microarchitectures

小鼠骨骼肌形态及纤维型成分的半自动化分析

一种研究动脉粥样硬化斑块纤维组织局部胶原结构与力学性能相关性的方法

小鼠后肢外植体研究跟腱撞击力学生物学的模型

小鼠后肢外植体研究跟腱撞击力学生物学的模型

使用基于曲线的转换工具量化纤维胶原蛋白组织

摘要

探索更多视频

此视频中的章节

观察和量化纤维细胞介导的纤维素凝胶压实

肌成纤维细胞和约束的应用工程纤维蛋白的​​组织结构和应变诱导细胞和胶原蛋白（RE）组织

在氧化应激期间细胞粘附和纤维蛋白扩散动力学

在3D揭示侵入性癌症细胞骨架组织

3D胶原凝胶和微通道3D相互作用的研究准备

用无标签荧光显微镜评价活体、人体阻力动脉中胶原蛋白和弹性蛋白的压力依赖性 Microarchitectures

小鼠骨骼肌形态及纤维型成分的半自动化分析

一种研究动脉粥样硬化斑块纤维组织局部胶原结构与力学性能相关性的方法

小鼠后肢外植体研究跟腱撞击力学生物学的模型

小鼠后肢外植体研究跟腱撞击力学生物学的模型

肌成纤维细胞和约束的应用工程纤维蛋白的组织结构和应变诱导细胞和胶原蛋白（RE）组织