陆军部 #W81XWH-08-1-0587, #W81XWH-15-1-0006;球州立大学2010立志补助金。

<ol>
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作者声明他们没有竞争的财政利益。

本研究的目的是发展一种非侵入性的方法, 以确定胫骨的变形在高影响活动。对胫骨负荷的影响进行量化, 将有助于更好地理解胫骨应力性骨折。在本研究中, 开发了一个特定于主题的肌肉骨骼模型, 并运行计算机模拟, 以复制在实验室环境中进行的降落运动。研究了降落地高度对胫骨应变的影响。在本研究中, 我们观察到, 随着降落高度的增加, 峰值最大的主要菌株也是如此。此外, 在三个着陆条件?...

骨应激损伤, 如应力性骨折, 是严重的过度使用性损伤, 需要长时间的恢复, 并招致重大医疗费用1,2。应力性骨折在运动和军事人口中都很常见。在所有运动相关的损伤中, 应力性骨折占总3的10%。特别是, 田径运动员在 20%4时面临较高的伤害率。士兵也经历了高压力骨折的发病率。例如, 据报道, 美国陆军的6% 受伤率为1 , 在以色列军队的5中报告了31% 的伤害率。在所有报告的应力性骨折中, 胫骨应力骨折是最常见的一个6,7,8。
运动和体育训练, 有较高的胫骨应力骨折风险通常与高地面影响 (例如, 跳跃, 着陆和切割)。在运动过程中, 当脚接触地面时, 地面撞击力会被应用到身体上。这种撞击力被肌肉骨骼系统和鞋类所消散。骨骼系统作为一系列杠杆, 允许肌肉施加力量来吸收地面撞击9。当腿部肌肉不能充分减少地面撞击时, 下肢骨骼必须吸收残余力。骨结构在这个过程中会经历变形。残余冲击力的重复吸收可能导致骨骼中的 microdamages, 这将积聚并成为应力性骨折。到目前为止, 与外部地面撞击力有关的骨骼反应的信息是有限的。研究胫骨骨在动态运动过程中如何响应高冲击力引入的机械载荷是很重要的。在高撞击活动中检查胫骨骨的变形, 可以更好地了解胫骨应力性骨折的发生机制。
用于测量骨骼变形的常规技术在体内依赖于仪器应变表10,11,12,13,14,15。在骨表面植入应变片需要手术治疗。由于侵入性的性质,在体内研究受志愿者的小样本的限制。此外, 应变计只能监测骨骼表面的一个小区域。本文介绍了一种利用计算机模拟分析骨应变的非侵入性方法1617。这种方法可以将肌肉骨骼模型和计算模拟结合起来, 研究人体运动过程中的骨骼应变。
肌肉骨骼模型由骨骼和骨骼肌肉代表。骨架由骨段组成, 它们是刚性或非变形体。骨骼肌肉被建模为控制器使用渐进积分-导数 (PID) 算法。三期 PID 控制使用估计中的错误来提高输出精度18。从本质上讲, 代表肌肉的 PID 控制器试图通过发展必要的力量来复制身体运动, 从而在一段时间内产生肌肉的长度变化。PID 控制器使用长度/时间曲线中的误差来修改复制运动的力。这个模拟过程创造了一个可行的解决方案, 协调所有的肌肉一起工作, 以移动骨骼和产生身体运动。
肌肉骨骼模型骨架中的一个或多个段可以被建模为柔性体, 以允许测量变形。例如, 胫骨骨可以分解成有限数量的元素, 由数以千计的元素和节点组成。通过有限元分析, 可以研究机械载荷对挠性胫骨的影响。有限元分析计算了各个元素随时间推移的加载响应。随着骨元素和节点数量的增加, 有限元分析的计算时间将显著增加。
为降低计算成本, 并对柔性体的变形进行精确评价, 采用模态有限元分析方法, 在汽车和航天工业中应用了19,20。该过程不是分析单个 FE 元素对时间域内机械载荷的响应, 而是根据频率域中不同的振动频率来评估物体的机械响应。此方法使计算时间大大减少, 同时提供了对变形20的精确测量。虽然模态有限元分析已广泛应用于汽车和航天领域的机械疲劳研究, 但这种方法在人类运动科学中的应用却十分有限。Al Nazer et, 使用一种模态有限元分析, 检查人体步态中的胫骨变形, 并报告令人鼓舞的结果16,17。然而, 只有利用有限的运动学数据进行计算机仿真, 才对其方法产生很大的影响;没有真正的地面撞击力用于辅助模拟。这种方法可能是合理的研究低冲击慢动作, 如步行, 但它不是一个可行的解决方案, 研究高地面撞击运动。因此, 为了检查在动态高影响活动期间的下肢骨反应, 必须开发一种创新的方法来解决与以前报告的方法相关的限制。具体来说, 必须开发一种利用精确的实验运动数据和实际地面冲击力的方法。因此, 本研究的目的是建立一个特定主题的肌肉骨骼模型, 进行多体动力学模拟, 用模态有限元分析, 检查胫骨应变在高冲击活动。选择了由不同高度的降落所表示的动态高冲击运动来测试该方法。

<table border="0" cellpadding="0" cellspacing="0" width="575">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:143px;">CT Scanner</td>
			<td style="width:123px;">GE Medical System</td>
			<td style="width:107px;">N/A</td>
			<td style="width:203px;">Light Speed VCT. For performing tibia CT scan.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Motion Capture System</td>
			<td>Vicon Inc</td>
			<td>N/A</td>
			<td>Vicon FX40 high speed cameras. For performing 3D motion capture.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Force plates</td>
			<td>AMTI Inc</td>
			<td>N/A</td>
			<td>Collecting 3D ground reaction forces</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Vicon Nexus</td>
			<td>Vicon Inc</td>
			<td>N/A</td>
			<td>Motion capture software program. For processing visual marker trajectory data.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Visual 3D</td>
			<td>C-Motion Inc</td>
			<td>N/A</td>
			<td>Biomechanics analysis software. For computing 3D kinematics and kinetics of human movements.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">MATLAB</td>
			<td>Mathworks Inc</td>
			<td>N/A</td>
			<td>Computer programming software. For performing raw data filtering, data conversion, and data processing.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">ADAMS 2012</td>
			<td>MSC Software Inc</td>
			<td>N/A</td>
			<td>Multibody dynamic computer simulation program.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">LifeMOD</td>
			<td>Lifemodeler Inc</td>
			<td>N/A</td>
			<td>A software Plug-in in ADAMS. For building human body musculo-skeletal models.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">MIMICS 13</td>
			<td>Materialise Inc</td>
			<td>N/A</td>
			<td>Image processing program. A 3D modeling tool to process imaging data. For creating 3D tibia model from CT scans.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">MARC 2012</td>
			<td>MSC Software Inc</td>
			<td>N/A</td>
			<td>Finite element analysis software. For performing volumn meshing, generating tibia FE model, and running modal FE analysis.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">SPSS 19</td>
			<td>IBM Inc</td>
			<td>N/A</td>
			<td>Statistical analysis software.</td>
		</tr>
	</tbody>
</table>

subject-specific musculoskeletal model for studying bone strain during dynamic motion

骨应激损伤在运动和军事训练中普遍存在。在训练过程中, 重复的大地面撞击力可能是原因。重要的是要确定高地冲击力对下肢骨变形的影响, 以更好地了解骨应激损伤的机制。传统的应变测量方法用于研究体内胫骨变形。这种方法与限制, 包括程序的侵袭性, 少数人类的参与, 和有限的应变数据从小骨表面积。本研究旨在介绍一种新的方法来研究胫骨骨应变在高冲击载荷条件下。一个主题特定的肌肉骨骼模型被创建来代表一个健康的男性 (19 岁, 80 公斤, 1800 毫米)。基于 ct 扫描的对象右胫骨, 建立了一种灵活的有限元胫骨模型。进行了实验室运动捕获, 以获得不同高度 (26、39、52厘米) 降落的运动学和地面反应力。采用多体动态计算机模拟方法, 结合弹性胫骨的模态分析, 对下落着陆时胫骨应变进行量化。计算出的胫骨应变数据与先前的体内研究吻合良好。很明显, 这种非侵入性的方法可以应用于研究大型队列的高冲击活动胫骨骨应变, 这将有助于更好地理解胫骨应力性骨折的损伤机制。

一个健康的高加索男性 (19 岁, 身高1800毫米, 质量80公斤) 自愿为研究。在收集数据之前, 该主题审查并签署了大学机构审查委员会批准的同意表格, 然后才参加研究。这项试验是根据《赫尔辛基宣言》进行的。实验是根据下面的协议进行的。
为了验证正向动态仿真的正确性, 将模拟中的低体关节角与由生物力学分析程序处理的运...

Watch this Scientific Journal Video about Subject-specific Musculoskeletal Model for Studying Bone Strain During Dynamic Motion at JoVE.com

用于研究动态运动期间骨应变的特定主题肌肉骨骼模型 |生物工程|视频

Subject-specific Musculoskeletal Model for Studying Bone Strain During Dynamic Motion

在着陆期间, 下半身的骨骼经历了巨大的机械负荷, 并且变形了。测量骨的变形以更好地了解与撞击有关的骨应激损伤的机制是非常必要的。一种新的方法, 结合学科特定的肌肉骨骼模型和有限元分析是用来测量胫骨应变在动态运动。

动态运动过程中骨应变的学科特异性肌肉模型研究

Bone stress injuries are common in sports and military trainings. Repetitive large ground impact forces during training could be the cause. It is ...

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Universidad de Cartagena UDEC

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Unit 1

Transformations of Stress and Strain

Musculoskeletal System

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9.6K 次观看.  Ball State University. 在着陆期间, 下半身的骨骼经历了巨大的机械负荷, 并且变形了。测量骨的变形以更好地了解与撞击有关的骨应激损伤的机制是非常必要的。一种新的方法, 结合学科特定的肌肉骨骼模型和有限元分析是用来测量胫骨应变在动态运动。

视频: Subject-specific Musculoskeletal Model for Studying Bone Strain During Dynamic Motion

Microfabricated Platforms for Mechanically Dynamic Cell Culture

The ability to systematically probe in vitro cellular response to combinations of mechanobiological stimuli for tissue engineering, drug discovery or fundamental cell biology studies is limited by current bioreactor technologies, which cannot simultaneously apply a variety of mechanical stimuli to cultured cells. In order to address this issue, we have developed a series of microfabricated platforms designed to screen for the effects of mechanical stimuli in a high-throughput format. In this protocol, we demonstrate the fabrication of a microactuator array of vertically displaced posts on which the technology is based, and further demonstrate how this base technology can be modified to conduct high-throughput mechanically dynamic cell culture in both two-dimensional and three-dimensional culture paradigms.

In this protocol, we demonstrate the fabrication of a microactuator array of vertically displaced posts on which the technology is based, and how this base technology can be modified to conduct high-throughput mechanically dynamic cell culture in both two-dimensional and three-dimensional culture paradigms.

微机械动态细胞培养平台

The ability to systematically probe in vitro cellular response to combinations of mechanobiological stimuli for tissue engineering, drug discovery or ...

科研

生物工程

Microfluidic Mixers for Studying Protein Folding

The process by which a protein folds into its native conformation is highly relevant to biology and human health yet still poorly understood. One reason for this is that folding takes place over a wide range of timescales, from nanoseconds to seconds or longer, depending on the protein1. Conventional stopped-flow mixers have allowed measurement of folding kinetics starting at about 1 ms. We have recently developed a microfluidic mixer that dilutes denaturant ~100-fold in ~8 &mu;s2. Unlike a stopped-flow mixer, this mixer operates in the laminar flow regime in which turbulence does not occur. The absence of turbulence allows precise numeric simulation of all flows within the mixer with excellent agreement to experiment3-4.
Laminar flow is achieved for Reynolds numbers Re &le;100. For aqueous solutions, this requires micron scale geometries. We use a hard substrate, such as silicon or fused silica, to make channels 5-10 &mu;m wide and 10 &mu;m deep (See Figure 1). The smallest dimensions, at the entrance to the mixing region, are on the order of 1 &mu;m in size. The chip is sealed with a thin glass or fused silica coverslip for optical access. Typical total linear flow rates are ~1 m/s, yielding Re~10, but the protein consumption is only ~0.5 nL/s or 1.8 &mu;L/hr. Protein concentration depends on the detection method: For tryptophan fluorescence the typical concentration is 100 &mu;M (for 1 Trp/protein) and for FRET the typical concentration is ~100 nM.
The folding process is initiated by rapid dilution of denaturant from 6 M to 0.06 M guanidine hydrochloride. The protein in high denaturant flows down a central channel and is met on either side at the mixing region by buffer without denaturant moving ~100 times faster (see Figure 2). This geometry causes rapid constriction of the protein flow into a narrow jet ~100 nm wide. Diffusion of the light denaturant molecules is very rapid, while diffusion of the heavy protein molecules is much slower, diffusing less than 1 &mu;m in 1 ms. The difference in diffusion constant of the denaturant and the protein results in rapid dilution of the denaturant from the protein stream, reducing the effective concentration of the denaturant around the protein. The protein jet flows at a constant rate down the observation channel and fluorescence of the protein during folding can be observed using a scanning confocal microscope5.

In this work we explain the fabrication and use of a microfluidic mixer capable of mixing two solutions in ~8 &mu;s. We also demonstrate the use of these mixers with spectroscopic detection using UV fluorescence and fluorescence resonance energy transfer (FRET).

研究蛋白质折叠的微流体混合器

The process by which a protein folds into its native conformation is highly relevant to biology and human health yet still poorly understood. One reason ...

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, direct visualization of individual viral complexes in their host cellular environment with cryoET is challenging2, due to the infrequent and dynamic nature of viral entry, particularly in the case of HIV-1. While time-lapse live-cell imaging has yielded a great deal of information about many aspects of the life cycle of HIV-13-7, the resolution afforded by live-cell microscopy is limited (~ 200 nm). Our work was aimed at developing a correlation method that permits direct visualization of early events of HIV-1 infection by combining live-cell fluorescent light microscopy, cryo-fluorescent microscopy, and cryoET. In this manner, live-cell and cryo-fluorescent signals can be used to accurately guide the sampling in cryoET. Furthermore, structural information obtained from cryoET can be complemented with the dynamic functional data gained through live-cell imaging of fluorescent labeled target.
In this video article, we provide detailed methods and protocols for structural investigation of HIV-1 and host-cell interactions using 3D correlative high-speed live-cell imaging and high-resolution cryoET structural analysis. HeLa cells infected with HIV-1 particles were characterized first by confocal live-cell microscopy, and the region containing the same viral particle was then analyzed by cryo-electron tomography for 3D structural details. The correlation between two sets of imaging data, optical imaging and electron imaging, was achieved using a home-built cryo-fluorescence light microscopy stage. The approach detailed here will be valuable, not only for study of virus-host cell interactions, but also for broader applications in cell biology, such as cell signaling, membrane receptor trafficking, and many other dynamic cellular processes.

We describe a correlative microscopy method that combines high-speed 3D live-cell fluorescent light microscopy and high-resolution cryo-electron tomography. We demonstrate the capability of the correlative method by imaging dynamic, small HIV-1 particles interacting with host HeLa cells.

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

Home-Based Monitor for Gait and Activity Analysis

Current outcomes in neuromuscular disorder clinical trials include motor function scales, timed tests, and strength measures performed by trained clinical evaluators. These measures are slightly subjective and are performed during a visit to a clinic or hospital and constitute therefore a point assessment. Point assessments can be influenced by daily patient condition or factors such as fatigue, motivation, and intercurrent illness. To enable home-based monitoring of gait and activity, a wearable magneto-inertial sensor (WMIS) has been developed. This device is a movement monitor composed of two very light watch-like sensors and a docking station. Each sensor contains a tri-axial accelerometer, gyroscope, magnetometer, and a barometer that record linear acceleration, angular velocity, the magnetic field of the movement in all directions, and barometric altitude, respectively. The sensors can be worn on the wrist, ankle, or wheelchair to record the subject&#8217;s movements during the day. The docking station enables data uploading and recharging of sensor batteries during the night. Data are analyzed using proprietary algorithms to compute parameters representative of the type and intensity of the performed movement. This WMIS can record a set of digital biomarkers, including cumulative variables, such as total number of meters walked, and descriptive gait variables, such as the percentage of the most rapid or longest stride that represents the top performance of patient over a predefined period of time.

This innovative device uses magneto-inertial sensors to permit gait and activity analysis in uncontrolled environments. Currently in the qualification process as an outcome measure in the European Medical Agency, one of the applications will be to serve as a clinical endpoint in clinical trials in neuromuscular diseases.

基于家庭的盖特和活动分析监视器

Current outcomes in neuromuscular disorder clinical trials include motor function scales, timed tests, and strength measures performed by trained clinical ...

Calvarial Model of Bone Augmentation in Rabbit for Assessment of Bone Growth and Neovascularization in Bone Substitution Materials

The basic principle of the rabbit calvarial model is to grow new bone tissue vertically on top of the cortical part of the skull. This model allows assessment of bone substitution materials for oral and craniofacial bone regeneration in terms of bone growth and neovascularization support. Once animals are anesthetized and ventilated (endotracheal intubation), four cylinders made of polyether ether ketone (PEEK) are screwed onto the skull, on both sides of the median and coronal sutures. Five intramedullary holes are drilled within the bone area delimited by each cylinder, allowing influx of bone marrow cells. The material samples are placed into the cylinders which are then closed. Finally, the surgical site is sutured, and animals are awaken. Bone growth may be assessed on live animals by using microtomography. Once animals are euthanized, bone growth and neovascularization may be evaluated by using microtomography, immune-histology and immunofluorescence. As the evaluation of a material requires maximum standardization and calibration, the calvarial model appears ideal. Access is very easy, calibration and standardization are facilitated by the use of defined cylinders and four samples may be assessed simultaneously. Furthermore, live tomography may be used and ultimately a large decrease in animals to be euthanized may be anticipated.

Here we present a surgical protocol in rabbits with the aim to assess bone substitution materials in terms of bone regeneration capacities. By using PEEK cylinders fixed onto rabbit skulls, osteoconduction, osteoinduction, osteogenesis and vasculogenesis induced by the materials may be evaluated either on live or euthanized animals.

兔子骨增增的千叶模型,用于骨代材料骨生长和新血管化的评估

The basic principle of the rabbit calvarial model is to grow new bone tissue vertically on top of the cortical part of the skull. This model allows ...

A Human 3D Extracellular Matrix-Adipocyte Culture Model for Studying Matrix-Cell Metabolic Crosstalk

The extracellular matrix (ECM) plays a central role in regulating tissue homeostasis, engaging in crosstalk with cells and regulating multiple aspects of cellular function. The ECM plays a particularly important role in adipose tissue function in obesity, and alterations in adipose tissue ECM deposition and composition are associated with metabolic disease in mice and humans. Tractable in vitro models that permit dissection of the roles of the ECM and cells in contributing to global tissue phenotype are sparse. We describe a novel 3D in vitro model of human ECM-adipocyte culture that permits study of the specific roles of the ECM and adipocytes in regulating adipose tissue metabolic phenotype. Human adipose tissue is decellularized to isolate ECM, which is subsequently repopulated with preadipocytes that are then differentiated within the ECM into mature adipocytes. This method creates ECM-adipocyte constructs that are metabolically active and retain characteristics of the tissues and patients from which they are derived. We have used this system to demonstrate disease-specific ECM-adipocyte crosstalk in human adipose tissue. This culture model provides a tool for dissecting the roles of the ECM and adipocytes in contributing to global adipose tissue metabolic phenotype and permits study of the role of the ECM in regulating adipose tissue homeostasis.

We describe a 3D human extracellular matrix-adipocyte in vitro culture system that permits dissection of the roles of the matrix and adipocytes in contributing to adipose tissue metabolic phenotype.

用于研究基质细胞代谢串扰的人类3D细胞外基质-脂肪培养模型

The extracellular matrix (ECM) plays a central role in regulating tissue homeostasis, engaging in crosstalk with cells and regulating multiple aspects of ...

Controlled Strain of 3D Hydrogels under Live Microscopy Imaging

External forces are an important factor in tissue formation, development, and maintenance. The effects of these forces are often studied using specialized in vitro stretching methods. Various available systems use 2D substrate-based stretchers, while the accessibility of 3D techniques to strain soft hydrogels, is more restricted. Here, we describe a method that allows external stretching of soft hydrogels from their circumference, using an elastic silicone strip as the sample carrier. The stretching system utilized in this protocol is constructed from 3D-printed parts and low-cost electronics, making it simple and easy to replicate in other labs. The experimental process begins with polymerizing thick (&#62;100 &#956;m) soft fibrin hydrogels (Elastic Modulus of ~100 Pa) in a cut-out at the center of a silicone strip. Silicone-gel constructs are then attached to the printed-stretching device and placed on the confocal microscope stage. Under live microscopy the stretching device is activated, and the gels are imaged at various stretch magnitudes. Image processing is then used to quantify the resulting gel deformations, demonstrating relatively homogenous strains and fiber alignment throughout the gel&#8217;s 3D thickness (Z-axis). Advantages of this method include the ability to strain extremely soft hydrogels in 3D while executing in situ microscopy, and the freedom to manipulate the geometry and size of the sample according to the user&#8217;s needs. Additionally, with proper adaptation, this method can be used to stretch other types of hydrogels (e.g., collagen, polyacrylamide or polyethylene glycol) and can allow for analysis of cells and tissue response to external forces under more biomimetic 3D conditions.

The presented method involves uniaxial stretching of 3D soft hydrogels embedded in silicone rubber while allowing live confocal microscopy. Characterization of the external and internal hydrogel strains as well as fiber alignment are demonstrated. The device and protocol developed can assess the response of cells to various strain regimes.

实时显微镜成像下的 3D 水凝胶控制应变

External forces are an important factor in tissue formation, development, and maintenance. The effects of these forces are often studied using specialized ...

Measuring 3D In-vivo Shoulder Kinematics using Biplanar Videoradiography

The shoulder is one of the human body's most complex joint systems, with motion occurring through the coordinated actions of four individual joints, multiple ligaments, and approximately 20 muscles. Unfortunately, shoulder pathologies (e.g., rotator cuff tears, joint dislocations, arthritis) are common, resulting in substantial pain, disability, and decreased quality of life. The specific etiology for many of these pathologic conditions is not fully understood, but it is generally accepted that shoulder pathology is often associated with altered joint motion. Unfortunately, measuring shoulder motion with the necessary level of accuracy to investigate motion-based hypotheses is not trivial. However, radiographic-based motion measurement techniques have provided the advancement necessary to investigate motion-based hypotheses and provide a mechanistic understanding of shoulder function. Thus, the purpose of this article is to describe the approaches for measuring shoulder motion using a custom biplanar videoradiography system. The specific objectives of this article are to describe the protocols to acquire biplanar videoradiographic images of the shoulder complex, acquire CT scans, develop 3D bone models, locate anatomical landmarks, track the position and orientation of the humerus, scapula, and torso from the biplanar radiographic images, and calculate the kinematic outcome measures. In addition, the article will describe special considerations unique to the shoulder when measuring joint kinematics using this approach.

Biplane videoradiography can quantify shoulder kinematics with a high degree of accuracy. The protocol described herein was specifically designed to track the scapula, humerus, and the ribs during planar humeral elevation, and outlines the procedures for data collection, processing, and analysis. Unique considerations for data collection are also described.

使用双平面影像学测量 3D 体内肩部运动学

The shoulder is one of the human body's most complex joint systems, with motion occurring through the coordinated actions of four individual joints, ...

Cantilever Bending of Murine Femoral Necks

Fractures in the femoral neck are a common occurrence in individuals with osteoporosis. Many mouse models have been developed to assess disease states and therapies, with biomechanical testing as a primary outcome measure. However, traditional biomechanical testing focuses on torsion or bending tests applied to the midshaft of the long bones. This is not typically the site of high-risk fractures in osteoporotic individuals. Therefore, a biomechanical testing protocol was developed that tests the femoral necks of murine femurs in cantilever bending loading to replicate better the types of fractures experienced by osteoporosis patients. Since the biomechanical outcomes are highly dependent on the flexural loading direction relative to the femoral neck, 3D printed guides were created to maintain a femoral shaft at an angle of 20&#176; relative to the loading direction. The new protocol streamlined the testing by reducing variability in alignment (21.6&#176; &plusmn; 1.5&#176;, COV = 7.1%, n = 20) and improved reproducibility in the measured biomechanical outcomes (average COV = 26.7%). The new approach using the 3D printed guides for reliable specimen alignment improves rigor and reproducibility by reducing the measurement errors due to specimen misalignment, which should minimize sample sizes in mouse studies of osteoporosis.

The present protocol describes the development of a reproducible testing platform for murine femoral necks in a cantilever bending set-up. Custom 3D printed guides were used to consistently and rigidly fix the femurs in optimal alignment.

鼠股颈悬臂弯曲

Fractures in the femoral neck are a common occurrence in individuals with osteoporosis. Many mouse models have been developed to assess disease states and ...

A GMP-Compliant Procedure for the Generation of Gene-Modified T cells

The field of Adoptive Cell Therapy (ACT) has been revolutionized by the development of genetically modified cells, specifically Chimeric Antigen Receptor (CAR)-T cells. These modified cells have shown remarkable clinical responses in patients with hematologic malignancies. However, the high cost of producing these therapies and conducting extensive quality control assessments has limited their accessibility to a broader range of patients. To address this issue, many academic institutions are exploring the feasibility of in-house manufacturing of genetically modified cells, while adhering to guidelines set by national and international regulatory agencies.
Manufacturing genetically modified T cell products on a large scale presents several challenges, particularly in terms of the institution's production capabilities and the need to meet infusion quantity requirements. One major challenge involves producing large-scale viral vectors under Good Manufacturing Practice (GMP) guidelines, which is often outsourced to external companies. Additionally, simplifying the T cell transduction process can help minimize variability between production batches, reduce costs, and facilitate personnel training. In this study, we outline a streamlined process for lentiviral transduction of primary human T cells with a fluorescent marker as the gene of interest. The entire process adheres to GMP-compliant standards and is implemented within our academic institution.

This protocol outlines the process of transducing primary human T cells with a gene of interest, ensuring compatibility with Good Manufacturing Practice (GMP) standards.

符合GMP的基因修饰T细胞生成程序

The field of Adoptive Cell Therapy (ACT) has been revolutionized by the development of genetically modified cells, specifically Chimeric Antigen Receptor ...