作者想表达他们在乌得勒支UMC从开发力学组感谢的Jeroen先生范登贝尔赫和Matthijs Wassink先生为他们的包裹骨软骨插头的过程中提供帮助。这项工作是由来自荷兰关节炎基金会的资助。

注意:这里介绍的协议是从最近的研究论文6，8，9，实验和计算程序通过。该协议是在图1中示出。 该尸体材料与乌得勒支大学的兽医学院的许可收集。 1.样品和浴的制备<ol><li>使用定制的钻头（ 图1），同时喷涂凉爽磷酸盐缓冲盐水（PBS），以防止过热和随后的软骨损伤钻出从尸体马股骨髁圆柱形骨软骨塞（8.5mm的直径）。 </li><li>用塑料收缩套筒以最小化重叠浴的横向扩散热缩骨软骨塞。 <ol><li>安装骨软骨插头与日软骨E中的塑料收缩套内侧顶最初和通过吹送热空气到它跟随。加入软骨，防止热相关的损害表面湿棉片。 </li></ol></li><li>分别准备电荷的（ 例如，650微升，420毫摩尔，碘克沙酸盐，分子量（Mw）= 1269道尔顿，电荷= -1）和中性（650μL，420毫米，碘克沙醇，MW = 1550道尔顿）溶质的有限浴。 </li><li>加载使用注射器将软骨的表面上制备的有限浴并放置一个软木塞上包裹样品，以防止在实验过程中的蒸发在RT（ 图2A）。为了研究中性溶质运移，将碘克沙醇浴，并研究带负电荷的溶质运移地方碘克酸浴到软骨表面。 </li></ol> 2.成像和图像处理<ol><li>将带有软木塞上附连到微CT的电动载物台定制的保持器中分离的包裹样品。地方Ť他取样所以覆盖有造影剂溶液朝上软骨表面。 </li><li>扫描使用微CT（40×40×40微米的3体素尺寸，2分钟的扫描时间，90千伏的管电压和180μA管电流）的视图，包括软骨，软骨下板的领域，并且finite-浴在几个时间点，直到平衡状态（48小时）达到（ 图2A）。当浓度值不随时间变化达到平衡状态。 </li><li>注册在基于初始图像上使用制造商的软件，方便利息（ROI）的区域定位不同时间点的3D图像（ 例如，分析）。 </li><li>转换3D使用制造商的软件处理它们之前重建微CT图像为二维标记图像文件格式（TIFF）栈。 </li><li>全局段（ImageJ的&gt;调整&gt;阈值）从软骨下骨和在软骨在软件躺在浴缸。 </li><li>获得在不同时间点软骨的平均灰度值使用所生成的软骨掩模在前面的步骤（ImageJ的&gt;分析&gt;测量）。 </li><li>鉴于该软骨的初始浴浓度和初始造影剂浓度，使用线性校准曲线的平均灰度值转换为溶质的实际浓度。以前的数据支持该灰度值保持与造影剂的浓度成线性关系这一事实。 </li><li>剧情中的溶质浓度与实验的时间点。 </li></ol> 3.计算建模注意:在这个问题的扩散假设采取1D位置（沿着z轴），其与实验边界条件的要求。因此，几何形状可以任意创建。 <ol><li>构建基于有限浴软骨多区域模式:1）由软骨浅表区<html> （总软骨厚度的20％），中间区（总软骨厚度的50％）和深区（总软骨厚度的30％）12和2）在FEBio 13，14（ 图2B有限浴）。 </li><li>分配软骨和沐浴在FEBio的不同区域的机械和物理性能。杨氏模量（10MPa的）被假定为足够高以抵抗由覆浴施加的渗透压，因此保护软骨免受过度的变形。 <ol><li>使用的10 -3毫米的液压渗透性4 / Ns和泊松在模拟8，0。使用的浴的实际溶质扩散系数比9。 </li></ol></li><li>生成网格（8节点六面体三线性元素）和完善它的边界附近（ 图2B）类="外部参照"&gt; 8,9。 </li><li>双相溶质模型<ol><li>在对应于它的浴和有效压力施加初始的溶质浓度。看的有效压力的说明在图9中 ，15。 </li><li>运行瞬态模式模型根据在不同的区域软骨规定扩散系数，以获得溶质浓度对时间的曲线。 </li></ol></li><li>多相模型 注意:浴和组织之间的电波动可以通过添加两个单价抗衡离子的浴，将组织两者来规避。 <ol><li>对于稳态模型:同时增加FCD到其期望值使用软骨和覆浴相同的有效流体压力和浓度。 </li><li>对于瞬态模型:通过保持溶质扩散系数在浴中足够高的创建充分搅拌的有限浴。注入溶质从浴 - 空气界面到浴达到其期望的浓度值。 </li><li>瞬态:除去在先前步骤中规定的溶质浓度的边界条件，并恢复有限浴的扩散系数其实际扩散系数。 </li><li>运行模型，以获得基于施加FCDS和扩散系数在不同的软骨区溶质浓度 - 时间曲线。 </li></ol></li><li> FEBio-MATLAB界面<ol><li>开发一个MATLAB代码在FEBio和情节浓度-时间曲线（FEBio-MATLAB接口）8，9自动执行仿真。 </li><li>改变扩散系数和FCDS使用FEBio-MATLAB界面软骨区。在FEBio运行模式和提取的溶质浓度-时间曲线8,9。 </li><li>与实验进行比较所获得的溶质浓度 - 时间曲线基于最小根扩散系数的数据，并获取集和FCDS在不同软骨区均方误差（RMSE）8,9。 </li></ol></li></ol><img alt="图1" class="xfigimg" src="/files/ftp_upload/54984/54984fig1.jpg" /> 图1: 实验设置。 A）使用一个特制的钻头样品提取过程。 B）显微CT成像程序以监测扩散过程。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54984/54984fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" class="xfigimg" src="/files/ftp_upload/54984/54984fig2.jpg" /> 图2: 示意图。A）实验设计。 B）多区域计算模型包括有限浴，软骨以及相关联的网格的浅，中，深区。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54984/54984fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contrast+agent's+transport+across+healthy+articular+cartilage+under+various+bath+conditions.">Contrast agent's transport across healthy articular cartilage under various bath conditions.</a> , (2015).</li><li>Arbabi, V., Pouran, B., Weinans, H., Zadpoor, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+a+biphasic-solute+model+in+predicting+diffusive+properties+of+osteochondral+interface.">Application of a biphasic-solute model in predicting diffusive properties of osteochondral interface.</a> , (2016).</li></ol>

作者什么都没有透露。

我们提出了一个实验方案用有限元建模程序组合来研究跨关节软骨中性和带电溶质的扩散。根据我们最近的研究，所提出的模型可以准确地描述这两种中性（双相溶质）的跨关节软骨8,9的不同区域的运输和带负电荷（多相）溶质。人们普遍认为，关节软骨变得通过其重要部件的损耗功能的限制，诸如OA进展21，22，23，24中带负电荷的大分子GAG以及胶原纤维。使用在本研究中提出的技术，可以潜在地检查关节软骨的健全性。中性溶质的运输可能在OA PRIMAR来扩充<html> ILY由于胶原的溶质之间的较低的相互作用以及聚糖和。在另一方面，带负电荷的溶质运输可以帮助获得相对于蛋白聚糖的固定收费浓度的信息，从而给OA发展水平的一些指示。 该双相溶质和多相模型基础上根据以前的研究有限浴概念的出现是被开发可以作为由可以提供关节软骨的纬向性的精确估计的平台。用造影剂，大浴相关的限制即可能射束硬化伪影，和分配单扩散系数关节软骨7，20，25，26，27，28，姑娘="外部参照"&gt; 29，30开车到开发当前研究的动机。在未来OA的研究，我们开发的车型可能会潜在地发现早期OA诊断中的应用。 有两个实验和计算机模拟所需的一些关键步骤。为了在实验过程中保持软骨完整性，一个将需要加入足够量的蛋白酶抑制剂，以防止后续的酶活性。在使用一个样本一个以上的实验的情况下，洗出时间为平衡后的渗透溶质约为48小时。洗出效率需要使用微CT进行检查。当使用我们的设置采用马软骨，上覆浴相当于无限浴的最小体积计算比软骨量的5倍。此外，为了进行扩散的有限元建模，重要的是应用实际溶质扩散coefficient以来从偏离洗澡可能会影响结果。 在底层的有限元模型，溶质尺寸效应并没有实现，因此无法进行研究。我们提出的有限浴模型提供了一定的优势，在铰接接头更恰当地理解，即扩散过程，并减少束硬化伪影。当应用带正电的造影剂我们所提出的技术相结合的实验和计算模型能使审议软骨的扩散属性。在洗澡的实际溶质扩散系数的知识似乎是关键，因为这可能影响显著跨关节软骨所获得的扩散系数的准确性。这将需要获得任一浴的扩散系数实验或容易地使用文献值。 总之，我们提出了包括实验的通用协议，计算研究带电和不带电的溶质的跨关节软骨的运输。使用协议，可以成功地获得扩散系数和固定带电密度在不同软骨层。

<html>分子运输在铰接接头的内稳态中起着至关重要的作用，治疗剂递送至关节软骨和对比度增强软骨成像1，2，3。因素，如软骨整合和完整性，溶质电荷和大小以及重量渗透摩尔浓度，并与软骨可能影响传输速率4，5，6接触浴的浓度。溶质，无论是中性的或带电荷的传输，可以是关节软骨区域之间不同，因为每个区由不同浓度的和主要的细胞外基质分子的取向，即蛋白聚糖素（PGs）和II型胶原蛋白1，7，8，9，小姑娘="外部参照"&gt; 10,11。更重要的是，带电荷的溶质运输可以是高度依赖于包括横跨关节软骨8,9增加了的细胞外基质内负的固定电荷的蛋白聚糖的浓度。这些参数尤其是固定电荷密度（FCD），胶原原纤维和穿过软骨含水量变化的取向可经历改变为骨关节炎（OA）的进行，从而表示跨越软骨研究扩散的重要性。 在目前的研究中，协议基于先前建立的实验和计算研究6，8，9，提出了准确调查使用中性和带电溶质扩散的有限浴模型中的各种边界条件下扩散。 Ť他提出的方法是由包括软骨和由先进的双相 - 溶质和多相有限元模型支持的有限浴的系统的微型计算机断层扫描成像（微CT）的。这些模型使跨关节软骨的不同区域获得中性和带电分子以及FCDS的扩散系数。利用这些模型，可以更好地了解可用于研究软骨和覆盖有限浴之间的相互作用的扩散中性和带电分子的行为。

<table border="0" cellpadding="0" cellspacing="0" width="688">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Hexabrix</td>
			<td style="width:149px;">Guerbet</td>
			<td style="width:123px;">15HX005D</td>
			<td style="width:201px;">Negatively charged contrast agent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Visipaque</td>
			<td style="width:149px;">GE healthcare</td>
			<td align="right" style="width:123px;">12570511</td>
			<td>Nuetral contrast agent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">PBS</td>
			<td style="width:149px;">Life technologies</td>
			<td align="right" style="width:123px;">10010023</td>
			<td>Medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">micro-CT</td>
			<td style="width:149px;">Perkin Elmer</td>
			<td style="width:123px;"></td>
			<td>Monitoring diffusion</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Freezing-point osmometer</td>
			<td style="width:149px;">Advanced instruments</td>
			<td style="width:123px;"></td>
			<td>Measuring solution osmolality</td>
		</tr>
	</tbody>
</table>

an experimental and finite element protocol to investigate the transport of neutral and charged solutes across articular cartilage

<html>骨关节炎（OA）是与关节软骨和软骨下骨的退化相关联的令人衰弱的疾病。关节软骨的退化基本上损害其承载功能，因为它经历了巨大的化学降解， 即 ，蛋白多糖的损失和胶原纤维的破坏。 OA期间，调查化学损伤机制中的一种有前途的方法是将试样软骨暴露到外部溶质和监控分子的扩散。软骨损伤（ 即浓度和基本大分子的构型）的程度与外部溶质的碰撞能量损失，同时跨越关节软骨移动相比健康软骨产生不同的扩散特性相关联。在这项研究中，我们引入一个协议，该协议包括几个步骤，并且基于先前开发的试验微-C14px的;"&gt; omputedŤomography（微CT）和有限元建模使用微CT，随后是通过施加双相-溶质和多相有限元模型，以获得扩散系数首先被记录带电和不带电碘化分子的运输。固定跨软骨区的电荷密度。

这里提供的代表性的结果是从先前的研究论文6，8，9，16日通过。 在OA中，关节软骨发生显著变化最重要的GAG损失，以及胶原纤维17的损坏，18个 ，19。这些变化可通过关节软骨20,21影响溶质的扩散行为。我们研究了两种碘化造影剂的轴向扩散， 即 ，碘克沙醇（电荷= 0）和碘克沙酸盐（电荷= -1），在使用显微CT尸体马骨软骨塞。为了定量的中性溶质（碘克沙醇），双相-溶质模型和带电SOLU的扩散过程<html> TE（碘克酸）多相模型中在FEBio所考虑软骨的纬向结构发达。将两相-溶质和多相模型可以预测碘克沙醇的扩散和整个关节软骨碘克沙酸盐（ 图3）。启用这些模型在不同的软骨区8获得的碘克沙醇（双相-溶质）和扩散系数的扩散系数以及FCD（碘克沙酸盐），9。 <img alt="图3" class="xfigimg" src="/files/ftp_upload/54984/54984fig3.jpg" /> 图3: 计算上曲线拟合数据。 A）多区域双相-溶质（虚线）与实验数据和B）多相模型拟合（虚线）与实验数据（符号）8,9。://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54984/54984fig3large.jpg"目标=‘_空白’&gt;点击此处查看该图的放大版本。

Watch this Scientific Journal Video about An Experimental and Finite Element Protocol to Investigate the Transport of Neutral and Charged Solutes across Articular Cartilage at JoVE.com

An Experimental and Finite Element Protocol to Investigate the Transport of Neutral and Charged Solutes across Articular Cartilage

我们提出了一个协议，以调查带电和不带电分子的横跨关节软骨与最近开发的实验和数值方法的帮助下运输。

实验和有限元议定书调查中性和带电溶质的跨关节软骨运输

Osteoarthritis (OA) is a debilitating disease that is associated with degeneration of articular cartilage and subchondral bone. Degeneration of articular ...

an-experimental-finite-element-protocol-to-investigate-transport

Nanyang Technological University

Research

JoVE Journal

Bioengineering

6.2K 次观看.  Delft University of Technology (TU Delft). 我们提出了一个协议，以调查带电和不带电分子的横跨关节软骨与最近开发的实验和数值方法的帮助下运输。 

视频: An Experimental and Finite Element Protocol to Investigate the Transport of Neutral and Charged Solutes across Articular Cartilage

Patterned Photostimulation with Digital Micromirror Devices to Investigate Dendritic Integration Across Branch Points

Light is a versatile and precise means to control neuronal excitability. The recent introduction of light sensitive effectors such as channel-rhodopsin and caged neurotransmitters have led to interests in developing better means to control patterns of light in space and time that are useful for experimental neuroscience. One conventional strategy, employed in confocal and 2-photon microscopy, is to focus light to a diffraction limited spot and then scan that single spot sequentially over the region of interest. This approach becomes problematic if large areas have to be stimulated within a brief time window, a problem more applicable to photostimulation than for imaging. An alternate strategy is to project the complete spatial pattern on the target with the aid of a digital micromirror device (DMD). The DMD approach is appealing because the hardware components are relatively inexpensive and is supported by commercial interests. Because such a system is not available for upright microscopes, we will discuss the critical issues in the construction and operations of such a DMD system. Even though we will be primarily describing the construction of the system for UV photolysis, the modifications for building the much simpler visible light system for optogenetic experiments will also be provided. The UV photolysis system was used to carryout experiments to study a fundamental question in neuroscience, how are spatially distributed inputs integrated across distal dendritic branch points. The results suggest that integration can be non-linear across branch points and the supralinearity is largely mediated by NMDA receptors.

Digital micromirror devices (DMD) can generate complex patterns in time and space with which to control neuronal excitability. Issues relevant to the design, construction, and operation of DMD systems are discussed. Such a system enabled the demonstration of non-linear integration across distal dendritic branch points.

图案与数字微镜装置Photostimulation调查跨分支点的树突状融合

Light is a versatile and precise means to control neuronal excitability. The recent introduction of light sensitive effectors such as channel-rhodopsin ...

科研

生物工程

Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the ex vivo Human Placental Perfusion Model

Decades ago the human placenta was thought to be an impenetrable barrier between mother and unborn child. However, the discovery of thalidomide-induced birth defects and many later studies afterwards proved the opposite. Today several harmful xenobiotics like nicotine, heroin, methadone or drugs as well as environmental pollutants were described to overcome this barrier. With the growing use of nanotechnology, the placenta is likely to come into contact with novel nanoparticles either accidentally through exposure or intentionally in the case of potential nanomedical applications. Data from animal experiments cannot be extrapolated to humans because the placenta is the most species-specific mammalian organ 1. Therefore, the ex vivo dual recirculating human placental perfusion, developed by Panigel et al. in 1967 2 and continuously modified by Schneider et al. in 1972 3, can serve as an excellent model to study the transfer of xenobiotics or particles.
Here, we focus on the ex vivo dual recirculating human placental perfusion protocol and its further development to acquire reproducible results.
The placentae were obtained after informed consent of the mothers from uncomplicated term pregnancies undergoing caesarean delivery. The fetal and maternal vessels of an intact cotyledon were cannulated and perfused at least for five hours. As a model particle fluorescently labelled polystyrene particles with sizes of 80 and 500 nm in diameter were added to the maternal circuit. The 80 nm particles were able to cross the placental barrier and provide a perfect example for a substance which is transferred across the placenta to the fetus while the 500 nm particles were retained in the placental tissue or maternal circuit. The ex vivo human placental perfusion model is one of few models providing reliable information about the transport behavior of xenobiotics at an important tissue barrier which delivers predictive and clinical relevant data.

Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the ex vivo Human Placental Perfusion Model

The ex vivo dual recirculating human placental perfusion model can be used to investigate the transfer of xenobiotics and nanoparticles across the human placenta. In this video protocol we describe the equipment and techniques required for a successful execution of a placenta perfusion.

通过胎盘的外来物质和纳米材料的运输率测定<em&gt;体外</em&gt;人胎盘灌注型号

Decades ago the human placenta was thought to be an impenetrable barrier between mother and unborn child. However, the discovery of thalidomide-induced ...

Models and Methods to Evaluate Transport of Drug Delivery Systems Across Cellular Barriers

Sub-micrometer carriers (nanocarriers; NCs) enhance efficacy of drugs by improving solubility, stability, circulation time, targeting, and release. Additionally, traversing cellular barriers in the body is crucial for both oral delivery of therapeutic NCs into the circulation and transport from the blood into tissues, where intervention is needed. NC transport across cellular barriers is achieved by: (i) the paracellular route, via transient disruption of the junctions that interlock adjacent cells, or (ii) the transcellular route, where materials are internalized by endocytosis, transported across the cell body, and secreted at the opposite cell surface (transyctosis). Delivery across cellular barriers can be facilitated by coupling therapeutics or their carriers with targeting agents that bind specifically to cell-surface markers involved in transport. Here, we provide methods to measure the extent and mechanism of NC transport across a model cell barrier, which consists of a monolayer of gastrointestinal (GI) epithelial cells grown on a porous membrane located in a transwell insert. Formation of a permeability barrier is confirmed by measuring transepithelial electrical resistance (TEER), transepithelial transport of a control substance, and immunostaining of tight junctions. As an example, ~200&#160;nm polymer NCs are used, which carry a therapeutic cargo and are coated with an antibody that targets a cell-surface determinant. The antibody or therapeutic cargo is labeled with 125I for radioisotope tracing and labeled NCs are added to the upper chamber over the cell monolayer for varying periods of time. NCs associated to the cells and/or transported to the underlying chamber can be detected. Measurement of free 125I allows subtraction of the degraded fraction. The paracellular route is assessed by determining potential changes caused by NC transport to the barrier parameters described above. Transcellular transport is determined by addressing the effect of modulating endocytosis and transcytosis pathways.

Many therapeutic applications require safe and efficient transport of drug carriers and their cargoes across cellular barriers in the body. This article describes an adaptation of established methods to evaluate the rate and mechanism of transport of drug nanocarriers (NCs) across cellular barriers, such as the gastrointestinal (GI) epithelium.

模型和方法来评价药物输送系统在整个细胞屏障的运输

Sub-micrometer carriers (nanocarriers; NCs) enhance efficacy of drugs by improving solubility, stability, circulation time, targeting, and release. ...

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and regenerative medicine. With digital control&#160;cells, scaffolds, and growth factors can be precisely deposited to the desired two-dimensional (2D) and three-dimensional (3D) locations rapidly. Therefore, this technology is an ideal approach to fabricate tissues mimicking their native anatomic structures. In order to engineer cartilage with native zonal organization, extracellular matrix composition (ECM), and mechanical properties, we developed a bioprinting platform using a commercial inkjet printer with simultaneous photopolymerization capable for 3D cartilage tissue engineering. Human chondrocytes suspended in poly(ethylene glycol) diacrylate (PEGDA) were printed for 3D neocartilage construction via layer-by-layer assembly. The printed cells were fixed at their original deposited positions, supported by the surrounding scaffold in simultaneous photopolymerization. The mechanical properties of the printed tissue were similar to the native cartilage. Compared to conventional tissue fabrication, which requires longer UV exposure, the viability of the printed cells with simultaneous photopolymerization was significantly higher. Printed neocartilage demonstrated excellent glycosaminoglycan (GAG) and collagen type II production, which was consistent with gene expression. Therefore, this platform is ideal for accurate cell distribution and arrangement for anatomic tissue engineering.

The methods described in this paper show how to convert a commercial inkjet printer into a bioprinter with simultaneous UV polymerization. The printer is capable of constructing 3D tissue structure with cells and biomaterials. The study demonstrated here constructed a 3D neocartilage.

人体软骨组织制作采用三维喷墨打印技术

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and ...

A Coupled Experiment-finite Element Modeling Methodology for Assessing High Strain Rate Mechanical Response of Soft Biomaterials

This study offers a combined experimental and finite element (FE) simulation approach for examining the mechanical behavior of soft biomaterials (e.g. brain, liver, tendon, fat, etc.) when exposed to high strain rates. This study utilized a Split-Hopkinson Pressure Bar (SHPB) to generate strain rates of 100-1,500 sec-1. The SHPB employed a striker bar consisting of a viscoelastic material (polycarbonate). A sample of the biomaterial was obtained shortly postmortem and prepared for SHPB testing. The specimen was interposed between the incident and transmitted bars, and the pneumatic components of the SHPB were activated to drive the striker bar toward the incident bar. The resulting impact generated a compressive stress wave (i.e. incident wave) that traveled through the incident bar. When the compressive stress wave reached the end of the incident bar, a portion continued forward through the sample and transmitted bar (i.e. transmitted wave) while another portion reversed through the incident bar as a tensile wave (i.e. reflected wave). These waves were measured using strain gages mounted on the incident and transmitted bars. The true stress-strain behavior of the sample was determined from equations based on wave propagation and dynamic force equilibrium. The experimental stress-strain response was three dimensional in nature because the specimen bulged. As such, the hydrostatic stress (first invariant) was used to generate the stress-strain response. In order to extract the uniaxial (one-dimensional) mechanical response of the tissue, an iterative coupled optimization was performed using experimental results and Finite Element Analysis (FEA), which contained an Internal State Variable (ISV) material model used for the tissue. The ISV material model used in the FE simulations of the experimental setup was iteratively calibrated (i.e. optimized) to the experimental data such that the experiment and FEA strain gage values and first invariant of stresses were in good agreement.

The current study prescribes a coupled experiment-finite element simulation methodology to obtain the uniaxial dynamic mechanical response of soft biomaterials (brain, liver, tendon, fat, etc.). The multiaxial experimental results that arose because of specimen bulging obtained from Split-Hopkinson Pressure Bar testing were rendered to a uniaxial true stress-strain behavior when simulated through iterative optimization of the finite element analysis of the biomaterial.

一个耦合试验，有限元建模方法以评估生物材料软的高应变率力学响应

This study offers a combined experimental and finite element (FE) simulation approach for examining the mechanical behavior of soft biomaterials (e.g. ...

3D Hydrogel Scaffolds for Articular Chondrocyte Culture and Cartilage Generation

Human articular cartilage is highly susceptible to damage and has limited self-repair and regeneration potential. Cell-based strategies to engineer cartilage tissue offer a promising solution to repair articular cartilage. To select the optimal cell source for tissue repair, it is important to develop an appropriate culture platform to systematically examine the biological and biomechanical differences in the tissue-engineered cartilage by different cell sources. Here we applied a three-dimensional (3D) biomimetic hydrogel culture platform to systematically examine cartilage regeneration potential of juvenile, adult, and osteoarthritic (OA) chondrocytes. The 3D biomimetic hydrogel consisted of synthetic component poly(ethylene glycol) and bioactive component chondroitin sulfate, which provides a physiologically relevant microenvironment for in vitro culture of chondrocytes. In addition, the scaffold may be potentially used for cell delivery for cartilage repair in vivo. Cartilage tissue engineered in the scaffold can be evaluated using quantitative gene expression, immunofluorescence staining, biochemical assays, and mechanical testing. Utilizing these outcomes, we were able to characterize the differential regenerative potential of chondrocytes of varying age, both at the gene expression level and in the biochemical and biomechanical properties of the engineered cartilage tissue. The 3D culture model could be applied to investigate the molecular and functional differences among chondrocytes and progenitor cells from different stages of normal or aberrant development.

Cartilage repair represents an unmet medical challenge and cell-based approaches to engineer human articular cartilage are a promising solution. Here, we describe three-dimensional (3D) biomimetic hydrogels as an ideal tool for the expansion and maturation of human articular chondrocytes.

3D水凝胶支架的关节软骨细胞培养及软骨生成

Human articular cartilage is highly susceptible to damage and has limited self-repair and regeneration potential. Cell-based strategies to engineer ...

An Experimental Protocol for Assessing the Performance of New Ultrasound Probes Based on CMUT Technology in Application to Brain Imaging

The possibility to perform an early and repeatable assessment of imaging performance is fundamental in the design and development process of new ultrasound (US) probes. Particularly, a more realistic analysis with application-specific imaging targets can be extremely valuable to assess the expected performance of US probes in their potential clinical field of application.
The experimental protocol presented in this work was purposely designed to provide an application-specific assessment procedure for newly-developed US probe prototypes based on Capacitive Micromachined Ultrasonic Transducer (CMUT) technology in relation to brain imaging.
The protocol combines the use of a bovine brain fixed in formalin as the imaging target, which ensures both realism and repeatability of the described procedures, and of neuronavigation techniques borrowed from neurosurgery. The US probe is in fact connected to a motion tracking system which acquires position data and enables the superposition of US images to reference Magnetic Resonance (MR) images of the brain. This provides a means for human experts to perform a visual qualitative assessment of the US probe imaging performance and to compare acquisitions made with different probes. Moreover, the protocol relies on the use of a complete and open research and development system for US image acquisition, i.e. the Ultrasound Advanced Open Platform (ULA-OP) scanner.
The manuscript describes in detail the instruments and procedures involved in the protocol, in particular for&#160;the calibration, image acquisition and registration of US and MR images. The obtained results prove the effectiveness of the overall protocol presented, which is entirely open (within the limits of the instrumentation involved), repeatable, and covers the entire set of acquisition and processing activities for US images.

The development of new ultrasound (US) probes based on Capacitive Micromachined Ultrasonic Transducer (CMUT) technology requires an early realistic assessment of imaging capabilities. We describe a repeatable experimental protocol for US image acquisition and comparison with magnetic resonance images, using an ex vivo bovine brain as an imaging target.

基于 CMUT 技术的新型超声探头在脑成像中的应用研究

The possibility to perform an early and repeatable assessment of imaging performance is fundamental in the design and development process of new ...

Creating a Structurally Realistic Finite Element Geometric Model of a Cardiomyocyte to Study the Role of Cellular Architecture in Cardiomyocyte Systems Biology

With the advent of three-dimensional (3D) imaging technologies such as electron tomography, serial-block-face scanning electron microscopy and confocal microscopy, the scientific community has unprecedented access to large datasets at sub-micrometer resolution that characterize the architectural remodeling that accompanies changes in cardiomyocyte function in health and disease. However, these datasets have been under-utilized for&#160;investigating the role of cellular architecture remodeling in cardiomyocyte function. The purpose of this protocol is to outline how to create an accurate finite element model of a cardiomyocyte using high resolution electron microscopy and confocal microscopy images. A detailed and accurate model of cellular architecture has significant potential to provide new insights into cardiomyocyte biology, more than experiments alone can garner. The power of this method lies in its ability to computationally fuse information from two disparate imaging modalities of cardiomyocyte ultrastructure to develop one unified and detailed model of the cardiomyocyte. This protocol outlines steps to integrate electron tomography and confocal microscopy images of adult male Wistar (name for a specific breed of albino rat) rat cardiomyocytes to develop a half-sarcomere finite element model of the cardiomyocyte. The procedure generates a 3D finite element model that contains an accurate, high-resolution depiction (on the order of ~35 nm) of the distribution of mitochondria, myofibrils and ryanodine receptor clusters that release the necessary calcium for cardiomyocyte contraction from the sarcoplasmic reticular network (SR) into the myofibril and cytosolic compartment. The model generated here as an illustration does not incorporate details of the transverse-tubule architecture or the sarcoplasmic reticular network and is therefore a minimal model of the cardiomyocyte. Nevertheless, the model can already be applied in simulation-based investigations into the role of cell structure in calcium signaling and mitochondrial bioenergetics, which is illustrated and discussed using two case studies that are presented following the detailed protocol.

This protocol outlines a novel method to create a spatially detailed finite element model of the intracellular architecture of cardiomyocytes from electron microscopy and confocal microscopy images. The power of this spatially detailed model is demonstrated using case studies in calcium signaling and bioenergetics.

建立一种结构逼真的心肌细胞的有限元几何模型研究蜂窝结构在心肌系统生物学中的作用

With the advent of three-dimensional (3D) imaging technologies such as electron tomography, serial-block-face scanning electron microscopy and confocal ...

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver engineering, including in vivo organ decellularization followed by repopulation, has emerged as a promising approach over ex vivo liver engineering. However, postoperative survival was not achieved. The aim of this study is to develop a novel surgical technique of in vivo selective liver lobe perfusion in rats as a prerequisite for in vivo liver engineering. We generate a circuit bypass only through the left lateral lobe. Then, the left lateral lobe is perfused with heparinized saline. The experiment is performed with 4 groups (n = 3 rats per group) based on different perfusion times of 20 min, 2 h, 3 h, and 4 h. Survival, as well as the macroscopically visible change of color and the histologically determined absence of blood cells in the portal triad and the sinusoids, is taken as an indicator for a successful model establishment. After selective perfusion of the left lateral lobe, we observe that the left lateral lobe, indeed, turned from red to faint yellow. In a histological assessment, no blood cells are visible in the branch of the portal vein, the central vein, and the sinusoids. The left lateral lobe turns red after reopening the blocked vessels. 12/12 rats survived the procedure for more than one week. We are the first to report a surgical model for in vivo single liver lobe perfusion with a long survival period of more than one week. In contrast to the previously published report, the most important advantage of the technique presented here is that perfusion of 70% of the liver is maintained throughout the whole procedure. The establishment of this technique provides a foundation for in vivo partial liver engineering in rats, including decellularization and recellularization.

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

We establish a novel surgical technique for an in vivo single liver lobe perfusion model in rat as a prerequisite for further studying in vivo partial liver engineering in the future.

大鼠体内部分肝工程的外科技术基础

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver ...

Fabrication of Decellularized Cartilage-derived Matrix Scaffolds

Osteochondral defects lack sufficient intrinsic repair capacity to regenerate functionally sound bone and cartilage tissue. To this extent, cartilage research has focused on the development of regenerative scaffolds. This article describes the development of scaffolds that are completely derived from natural cartilage extracellular matrix, coming from an equine donor. Potential applications of the scaffolds include producing allografts for cartilage repair, serving as a scaffold for osteochondral tissue engineering, and providing in vitro models to study tissue formation. By decellularizing the tissue, the donor cells are removed, but many of the natural bioactive cues are thought to be retained. The main advantage of using such a natural scaffold in comparison to a synthetically produced scaffold is that no further functionalization of polymers is required to drive osteochondral tissue regeneration. The cartilage-derived matrix scaffolds can be used for bone and cartilage tissue regeneration in both in vivo and in vitro settings.

Decellularized cartilage-derived scaffolds can be used as a scaffold to guide cartilage repair and as a means to regenerate osteochondral tissue. This paper describes the decellularization process in detail and provides suggestions to use these scaffolds in in vitro settings.

脱碳软骨衍生基质支架的制备

Osteochondral defects lack sufficient intrinsic repair capacity to regenerate functionally sound bone and cartilage tissue. To this extent, cartilage ...