Name: semiautomated longitudinal microcomputed tomography-based quantitative structural analysis of a nude rat osteoporosis-related vertebral fracture model
Uploaded: 2017-09-28T13:00:00
Description: Watch this Scientific Journal Video about Semiautomated Longitudinal Microcomputed Tomography-based Quantitative Structural Analysis of a Nude Rat Osteoporosis-related Vertebral Fracture Model at JoVE

这项研究得到了加利福尼亚再生医学研究所 (CIRM) (TR2-01780) 的资助。

所有动物实验都是根据由雪松-西奈医疗中心的机构动物保育和使用委员会 (IACUC) 批准的协议执行的 (3609 号议定书)。麻醉是为所有的成像和手术程序。所有动物都按照批准的 IACUC 协议安置. 
 注意: 该协议的实验设计如图 1 所示。购买六周大的老鼠与他们的卵巢手术切除和饲料他们的 LCD 包括0.01% 钙和0.77% 磷酸盐。经过4月的 LCD, 钻 critical-size 脊椎缺损在第四和第五腰椎...

骨质疏松是脊椎压迫性骨折的最常见原因, 脊柱的负荷增加, 导致椎体塌陷。然而, 在真正复制类似椎体塌陷的啮齿动物中, 几乎不可能产生损伤。相反, 研究人员在椎体中心创建一个圆柱形空洞以模拟 OVCFs16,17,18,19,20,21,24<sup...

全世界有超过2亿人患有骨质疏松症1。骨密度 (BMD) 和改变的骨微结构的潜在病理降低增加了骨脆性, 因此, 骨折的相对风险为2。骨质疏松症是如此普遍和有害健康, 世界卫生组织已确定它是一个重大的公共卫生问题。此外, 随着世界人口的老龄化, 骨质疏松症有望变得更加普遍。
骨质疏松性椎体压缩骨折是最常见的脆性骨折, 估计在美国每年超过75万。它们与显著的发病率和九倍高的死亡率3有关。在临床试验中, 目前可获得的手术干预, 如椎体成形术和成形, 被发现没有比假治疗更有效4,5, 只留下疼痛管理可供这些患者使用。由于目前的 OVCF 治疗是有限的, 所以必须开发一种能够复制紊乱的动物模型6,7,8。这种动物模型可以促进对目前治疗方法的研究和新疗法的发展, 将转化为临床实践。骨质疏松症已诱导和持续的模型动物通过管理的低饮食 (LCD) 与卵巢切除联合1,9,10,11,12,13,14,15. 为进一步模型骨质疏松与 OVCFs, 椎体骨缺损建立了免疫大鼠的骨骼损伤 16,17,18,19, 20,21,22,23,24。在这项工作中, 一个模型的免疫损伤大鼠骨质疏松症提出。这种新的模型可以用来评估细胞疗法涉及干细胞从各种来源和物种的修复有挑战性的骨折, 如 OVCFs。
骨显像是评价骨折和骨骼疾病的重要组成部分。为了准确评估结构骨的变化和再生策略, 开发了先进的成像方法25。其中, µCT 成像已经成为一种非侵入性的、易用的、廉价的提供高分辨率3D 图像的方法。µCT 成像在评估骨质疏松症患者方面优于其他模式, 因为它提供高分辨率的3D 骨微体系结构26 , 然后可以对其进行定量分析。后者可以用来比较治疗效果的建议治疗。事实上,在体内µCT 成像是一个金标准的脊椎缺损再生监测1,16,27。但是, 很少有出版物28、29、30、31使用了自动注册工具来最小化µCT 的用户依赖性、插值偏差和精度错误imaging-based 分析。最近, 我们是第一个使用注册程序, 以改善骨再生的标准化骨空洞的分析, 如本协议32中所述。
本文所描述的方法可以用来研究新的细胞治疗 OVCFs 的效果, 不受宿主 T 细胞的反应, 可能拒绝异种或同种异体细胞。通过卵巢切除 (卵巢) 和4月的液晶显示器, 诱导幼鼠骨质疏松。年轻的卵巢鼠, 结合 LCD, 我们可以达到一个低峰值骨量, 模仿绝经后骨质疏松, 导致不可逆转的骨质流失。这可以解释部分原因是, 在 LCD 和大约3月的年龄, 大鼠从骨骼模型过渡到重建腰椎椎体的阶段33, 从而增加了维持骨质疏松症的可能性时间.使用幼兽使这种模型更具成本效益, 因为它们成本更低。然而, 它是有限的内在不核算的生物变化的衰老动物。

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semiautomated longitudinal microcomputed tomography-based quantitative structural analysis of a nude rat osteoporosis-related vertebral fracture model

与骨质疏松相关的椎体压缩骨折 (OVCFs) 是一种常见的和临床未满足的需要随着世界人口年龄的增加而患病率。动物 OVCF 模型对转化组织工程策略的临床前发展至关重要。虽然目前有许多模型存在, 该协议描述了一个优化的方法, 诱导多高度重现性脊柱缺损在一个单一的裸鼠。本文还详细介绍了一种基于纵向半自动 microcomputed 断层扫描 (µCT) 的椎体缺损定量结构分析方法。简单地, 老鼠在术后多时间点被成像。1天扫描被重新定向到标准位置, 定义了标准的兴趣量。随后对每只老鼠的µCT 扫描自动登记到1天扫描, 因此, 同样的兴趣量被分析, 以评估新的骨骼形成。这种通用的方法可以适应各种其他模型, 纵向 imaging-based 分析可以受益于精确的3D 半自动对准。在一起, 本议定书描述了一个容易量化和容易重现的系统骨质疏松和骨骼研究。建议的协议需要4月来诱导裸卵巢大鼠和2.7 和 4 h 之间的骨质疏松症的产生, 图像, 并分析两个椎体缺损, 取决于组织大小和设备。

使用这个协议, 你可以图像和量化的再生 n = 8 模型骨质疏松椎体缺损不同时间点。通过注册程序获得的解剖匹配, 可以在所有时间点对同一 VOI 进行分析。这导致了高度准确的纵向3D 计量分析, 即使原始缺陷的边缘不再是可识别的。我们使用五时间点 (天 1, 星期 2, 星期 4, 星期8和星期 12) 作为一个例子为骨骼再生的纵向评估 (图 7)。可通过对2D 剖面和3D ...

Watch this Scientific Journal Video about Semiautomated Longitudinal Microcomputed Tomography-based Quantitative Structural Analysis of a Nude Rat Osteoporosis-related Vertebral Fracture Model at JoVE

Semiautomated Longitudinal Microcomputed Tomography-based Quantitative Structural Analysis of a Nude Rat Osteoporosis-related Vertebral Fracture Model

半自动纵 Microcomputed Tomography-based 裸鼠骨质疏松症相关椎体骨折模型的定量结构分析

该协议的目的是产生一个裸鼠骨质疏松症相关的椎体压缩骨折模型, 可以纵向评价在体内使用半自动 microcomputed tomography-based 定量结构分析。

Osteoporosis-related vertebral compression fractures (OVCFs) are a common and clinically unmet need with increasing prevalence as the world population ...

Research

JoVE Journal

Bioengineering

Improved Visualization and Quantitative Analysis of Drug Effects Using Micropatterned Cells

改进的可视化和定量分析，药物使用Micropatterned细胞的影响

To date, most HCA (High Content Analysis) studies are carried out with adherent cell lines grown on a homogenous substrate in tissue-culture treated ...

科研

生物工程

Quantitative FRET (F&#246;rster Resonance Energy Transfer) Analysis for SENP1 Protease Kinetics Determination

Quantitative FRET FÃƒÆ’Ã‚Â¶rster Resonance Energy Transfer Analysis for SENP1 Protease Kinetics Determination

定量荧光共振能量转移（福斯特共振能量转移）SENP1蛋白酶的动力学测定分析

Reversible posttranslational modifications of proteins with ubiquitin or ubiquitin-like proteins (Ubls) are widely used to dynamically regulate protein ...

Microfluidic-based Electrotaxis for On-demand Quantitative Analysis of Caenorhabditis elegans' Locomotion

Microfluidic-based Electrotaxis for On-demand Quantitative Analysis of Caenorhabditis elegans' Locomotion

基于微流体Electrotaxis的，为按需定量分析<em&gt;线虫</em&gt;&#39;LOCOMOTION的

The nematode Caenorhabditis elegans is a versatile model organism for biomedical research because of its conservation of disease-related genes and ...

Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions

Cryo-electron tomography (cryoET) allows 3D visualization of cellular structures at molecular resolution in a close-to-physiological state1. However, ...

Athymic Rat Model for Evaluation of Engineered Anterior Cruciate Ligament Grafts

无胸​​腺大鼠模型工程前交叉韧带移植评估

Anterior cruciate ligament (ACL) rupture is a common ligamentous injury that often requires surgery because the ACL does not heal well without ...

Use of a Rat Model to Study Ventral Abdominal Hernia Repair

大鼠模型在腹腹疝修补术中的应用研究

Ventral abdominal hernia is a relatively common clinical condition that sometimes requires herniorraphy (surgical repair). The repair of ventral abdominal ...

A Method to Estimate Cadaveric Femur Cortical Strains During Fracture Testing Using Digital Image Correlation

基于数字图像相关的骨折检测中股骨皮质应变的估算方法

This protocol describes the method using digital image correlation to estimate cortical strain from high speed video images of the cadaveric femoral ...

An Efficient and Reproducible Protocol for Distraction Osteogenesis in a Rat Model Leading to a Functional Regenerated Femur

大鼠牵张成骨的高效可再生性协议

This protocol describes the use of a newly developed external fixator for distraction osteogenesis in a rat femoral model. Distraction osteogenesis (DO) ...

A Microfluidic Platform for Longitudinal Imaging in Caenorhabditis elegans

A Microfluidic Platform for Longitudinal Imaging in Caenorhabditis elegans

秀丽线虫纵向成像的微流控平台

In the last decade, microfluidic techniques have been applied to study small animals, including the nematode Caenorhabditis elegans, and have proved ...

Rat Model of Adhesive Capsulitis of the Shoulder

肩上粘 Capsulitis 大鼠模型的建立

This proposal aims to create an in vivo rat model of adhesive capsulitis for researching potential treatment options for this condition and other ...