This research was supported by the Technology Innovation Program (Contract grant No. 0037915, WPM Biomedical Materials-Implant Materials) and Industrial Strategic Technology Development Program (Contract grant No. 10045329, Development of customized implant with porous structure for bone replacement), funded by the Ministry of Trade, industry &#38; Energy (MI, Korea), and BK21 PLUS SNU Materials Division for Educating Creative Global Leaders (Contract grant No. 21A20131912052).

1.制备多孔金属支架的<ol><li>通过混合市售的Ti粉末，莰烯，和KD-4 如表1中描述的多孔钛支架具有四个初始孔隙率（40，50，60，和70）称量材料的适当的量后制备的Ti-莰泥浆。倾浆料到500ml聚乙烯（PE）瓶和旋转瓶在55℃下30分钟在球磨机烘箱在30rpm。 </li><li>倾从PE瓶浆料成圆柱形的铝（Al）的模具，其直径为60毫米和60毫米的高度。密封每个铝模具与相应的A1盖玻片并旋转在球磨机烘箱模具以30rpm的速度在55℃下进行10分钟。 <ol><li>随后，降低球磨机烘箱的温度升高到44℃，并连续旋转模具以30rpm的速度在44℃进行12小时的恒温。 </li></ol></li><li>取出模具从球 -磨炉之后附加地旋转所述模具在RT 1小时的冷却过程。从使用Al柱塞铝模具中取出固化的钛/莰生坯。 </li><li>将固化的钛/莰生坯在一橡胶袋通过手和完全由捆扎袋的口用字符串密封橡胶袋。将橡胶袋在冷等静压（CIP），机的水槽中，并应用​​为200MPa的等静压力10分钟。从橡胶袋中取出压缩坯体。 </li><li>上手工传送的Ti-莰生坯氧化铝坩埚放置在坩埚中的冷冻干燥机。冷冻干燥该生坯升华莰相在生坯在 - 40℃下放置24小时。 </li><li>接着，关闭该坩埚有氧化铝盖玻片并将封闭坩埚在真空炉中（低于10 -6托 ）于RT。然后，增加了炉的温度升高到1300℃，在一个加热ř吃了5℃/分钟并保持温度在1300℃进行2小时。 </li><li>热处理后，保持烧结多孔钛在炉为6-7小时，直至炉完全冷却至室温。 注意：在6小时的冷却过程中，400℃以上的熔炉的平均冷却速度为〜15℃/分钟和400℃以下的熔炉的平均冷却速度为2〜℃/分钟。 </li><li>如果需要的话，切割烧结的多孔钛的块到盘形样品，直径为通过放电加工16个毫米（EDM）。27 注意：根据所述Al模具的大小，需要通过加工过程（图2A）进行修改的烧结多孔的Ti的大小。 </li><li>放置在高压釜中的多孔钛样品的玻璃烧杯中，并灭菌样品在121℃下进行15分钟。从高压釜中取出样品。洗多孔钛样品用蒸馏水洗涤两次，然后用70％乙醇两次。最后，离开多孔钛到培养皿并空气干燥在RT样品上在UV光下净化台。 </li></ol>支架的2浸涂与生物活性剂<ol><li>通过混合1毫升的GFP用9ml的Dulbecco磷酸盐缓冲盐水（DPBS，pH7.4）中的溶液在10毫升灭菌的从1毫克/毫升，以在清洁台上100微克/毫升稀释商用绿色荧光蛋白（GFP）的聚苯乙烯（PS）管，如表1所示。 </li><li>通过将钛样品到PS管在RT将GFP溶液和放置在净化台浸没灭菌致密或多孔钛在10毫升稀释的GFP溶液（100微克/毫升）。 </li><li>放置在PS管在真空干燥器并撤离干燥器10分钟，以确保将GFP溶液更有效地渗入多孔钛的孔中。 </li><li>除去使用镊子将PS管上的多孔钛。将GFP-涂覆的多孔钛成直径为10cm的聚乙烯三道菜和风干O / N在室温在一个干净的长椅上。 </li><li>用在玻璃烧杯中10个​​ml的Dulbecco氏磷酸盐缓冲盐水（DPBS）中冲洗多孔钛两次，并使用镊子和风干移动多孔钛成直径为10cm的培养皿在RT上的净化台。 </li></ol> 3.致密多孔支架的<ol><li>放置在一个圆柱形钢模具的GFP-涂覆的多孔钛的样品不同的高度，并插入了一组冲头进入钢模具（图3A）的顶部和底部的孔。 </li><li>用压制机以0.05〜0.1秒的中间应变率压缩所述多孔钛在RT中的样品（图3A）的z方向上的钢模具组件内-1针对在表2所示的预定的施加应变。握持压力卸货前1分钟。 </li><li>从钢模具去除致密的Ti样品。用10毫升的DPBS洗涤两次致密样品在一个干净的长椅的烧杯中，空气干燥O / N在室温。 </li></ol>绿色荧光蛋白涂层支架4释放测试<ol><li>沉浸三种试样（GFP涂覆致密的Ti（后步2），绿色荧光蛋白涂覆的多孔钛（后步骤1和2）和GFP-涂覆致密多孔的Ti（后步骤1-3））的5ml DPBS（pH值7.4）在37℃下在清洁台上所载10ml的灭菌的PS管溶液。 </li><li>吸出所有来自每个PS管的DPBS溶液与GFP-涂覆的样品和补充与使用根据预定的次数的1，2，3，5，8，12的吸移管新5 ml的DPBS溶液（pH 7.4），浸泡后15，22和29天。 </li><li>取GFP涂覆样品的荧光图像浸渍（第0天）前，用共聚焦激光扫描光谱（CLSM）后22天，浸泡。 </li><li>测量所释放的GFP在1ml溶液中的荧光信号强度从总共5毫升DPBS溶液从每个试管的PS在节4.2绘制使用紫外光谱在215纳米的波长。的强度值转换成利用标准曲线将GFP溶液的浓度。 注意：在测量之前，通过测量在0纳克/毫升的浓度范围将GFP溶液的荧光信号强度绘制的GFP溶液的标准曲线 - 10微克/毫升。 </li></ol> 5.制造分级多孔钛支架的<ol><li>通过重复步骤1.1至步骤1.7产生烧结多孔Ti的块。 </li><li>机根据预定结构设计 （如， 图5a和 5d）通过电火花烧结多孔的Ti块。 </li><li>放置加工的Ti样品高度分布在一个钢模其中多孔钛的直径为〜小于模具的直径0.1毫米小，并插入了一组冲头进入钢模具的顶部和底部的孔。 </li><li>执行步骤3.2和3.3。 </li></ol> 6.孔隙率我钛支架的asurement <ol><li>衡量的Ti支架的质量（m 多个）。 </li><li>计算的表观容积（V S），通过测量钛支架长度，宽度和高度。 </li><li>使用下列公式计算孔隙率： <img alt="式（1）" src="/files/ftp_upload/53279/53279eq1.jpg" /> 其中P是总孔隙率，ρTi为钛和米S的理论密度/ V S是样品的测量的密度。 注意：钛样品的孔隙率可以从显微图像来直接检索后显微成像是使用微计算机断层扫描仪。 </li></ol>

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The authors declare that they have no competing financial interests.

而biometal系统已被广泛地用于生物医学应用，特别是，作为承重材料，高刚度和金属的低生物活性已被视为重大的挑战。在这项研究中，我们建立了一个新的金属系统，致密多孔金属支架具有仿生机械性能以及生物活性表面与可持续释放行为的制造方法。我们的制作方法的主要优点包括：1）在前面的动态冷冻的铸造方法没有变化，我们已经开发，28 2）中的一个参数度的控制致密化-实现机...

而金属生物材料已被广泛用作由于其优异的机械强度和韧性，1-3承重植入物和内固定装置，他们涉及两个关键的挑战：1）机械失配，因为金属是比生物体组织硬得多，从而导致不希望的损害到周围组织和2）低的生物活性，往往导致界面差与生物组织，往往挑起异物反应 （如炎症或血栓形成）。已经提出了促进骨向内生长的结构4-6多孔金属支架，改善骨-植入物接触，而应力屏蔽效果，因为它们降低的刚性抑制7-9此外，各种表面改性已经应用于提高金属植入物的生物活性;这些修改包括涂层将金属表面与生物活性分子（例如，生长的fac器）或药物 （如万古霉素，四环素）10-12但是，存在的问题，如多孔金属支架降低机械性能，降低的硬度和快速释放的生物活性涂层的层仍然没有得到解决。13-16 特别是，钛（Ti）和Ti的合金是一体的，因为它们的优异的机械性能，化学稳定性的最流行​​biometal系统，以及良好的生物相容性。13,17-19其泡沫形的应用也引起越来越多的关注，因为在3D多孔网络促进除了骨样的机械性能的骨向内生长。20-22已作出努力通过开发新的制造技术，包括聚合海绵的复制，金属粒子，快速成型（RP）的方法的烧结来提高机械性能，并为了控制孔的各种特征空间保持器的方法（例如，孔隙分数，形状，大小，分布，和连接）和材料性质（例如，金属相和杂质）23-25 ​​最近，水性金属浆料的冻结铸造已经获得了相当大的注意，以产生机械增强的Ti形式具有良好对准孔结构通过利用在凝固过程中的单向冰枝晶生长;然而，所造成的金属粉末与水接触氧污染需要特别小心，以尽量减少钛支架的脆化。14,15 因此，我们开发了对制造生物活性和机械可调多孔钛支架的新方法。25支架最初有超过50％的孔隙率多孔结构。所制造的多孔支架涂覆有生物活性分子，然后进行压缩使用期间的最后的孔隙率，机械性能和药物释放行为由APPLI分别控制一个机械压力机编辑应变。致密多孔钛种植体，尽管低刚度比得上骨（3-20 GPA）2由于被覆层的一者所示低孔隙率具有良好的强度，致密多孔钛的生物活性显著改善。因为引起的致密化过程中的独特扁平孔结构。此外，涂布的生物活性分子被视为被逐渐从支架释放时，保持其功效为长时间。 在这项研究中，我们介绍了我们建立的方法来制作致密多孔钛支架的生物医学应用的潜在用途。该协议包括动态凝固的铸造​​金属浆料和多孔支架的致密化。首先，为了制造多孔钛支架具有良好的延展性的动态冷冻铸造方法被引入，如图1A。钛粉末分散在液体莰;然后，通过降低温度，液相固化，从而在钛粉末网络和固体莰晶体之间的相分离。接着，将固化的Ti-莰生坯进行烧结，其中的Ti粉末缩合的连续钛支柱，和莰相完全除去，得到多孔结构。该涂层和致密化过程用所获得的多孔支架受雇于，变化的致密化和初始孔隙的程度。被覆层和它的释放行为进行观察和使用绿色荧光蛋白（GFP）包被的多孔钛具有和不致密化相比，GFP-涂覆致密的Ti定量。最后，提出并通过改变多孔支架的内层和外层部分的致密化的程度表现出功能梯度的Ti支架具有两个不同的多孔结构。

<table>
	<tbody>
		<tr>
			<td>Titanium powder</td>
			<td>Alfa Aesar</td>
			<td>#42624</td>
			<td>-325 mesh, 99.5% (metals basis)</td>
		</tr>
		<tr>
			<td>Camphene</td>
			<td>SigmaAldrich</td>
			<td>#456055</td>
			<td>95%, C10H16</td>
		</tr>
		<tr>
			<td>KD-4</td>
			<td>Croda</td>
			<td>&#173;</td>
			<td>Hypermer, polymeric dispersant</td>
		</tr>
		<tr>
			<td>Phosphate Buffer Solution (PBS)</td>
			<td>Welgene</td>
			<td>ML 008-01</td>
			<td>&#173;</td>
		</tr>
		<tr>
			<td>Green Fluorescent Protein (GFP)</td>
			<td>Genoss Co.</td>
			<td>-</td>
			<td>&#62;98% purity, 1mg/ml</td>
		</tr>
		<tr>
			<td>Ball mill oven</td>
			<td>SAMHENUG ENERGY</td>
			<td>SH-BDO150</td>
			<td>&#173;</td>
		</tr>
		<tr>
			<td>Freeze dryer</td>
			<td>Ilshin Lab.</td>
			<td>PVTFD50A</td>
			<td>&#173;</td>
		</tr>
		<tr>
			<td>Cold isostatic pressing (CIP) machine</td>
			<td>SONGWON SYSTEMS</td>
			<td>CIP 42260</td>
			<td>&#173;</td>
		</tr>
		<tr>
			<td>Vaccum furnace</td>
			<td>JEONG MIN INDUSTRIAL</td>
			<td>JM-HP20</td>
			<td>&#173;</td>
		</tr>
		<tr>
			<td>electical chaege machine</td>
			<td>FANUC robocut</td>
			<td>0iB</td>
			<td>External use</td>
		</tr>
		<tr>
			<td>Press machine</td>
			<td>CG&#38;S</td>
			<td>AJP-200</td>
			<td>&#173;</td>
		</tr>
		<tr>
			<td>Confocal laser scanning spectroscopy (CLSM)</td>
			<td>Olympus</td>
			<td>FluoView FV1000</td>
			<td>External use</td>
		</tr>
	</tbody>
</table>

fabrication of mechanically tunable and bioactive metal scaffolds for biomedical applications

Biometal systems have been widely used for biomedical applications, in particular, as load-bearing materials. However, major challenges are high stiffness and low bioactivity of metals. In this study, we have developed a new method towards fabricating a new type of bioactive and mechanically reliable porous metal scaffolds-densified porous Ti scaffolds. The method consists of two fabrication processes, 1) the fabrication of porous Ti scaffolds by dynamic freeze casting, and 2) coating and densification of the porous scaffolds. The dynamic freeze casting method to fabricate porous Ti scaffolds allowed the densification of porous scaffolds by minimizing the chemical contamination and structural defects. The densification process is distinctive for three reasons. First, the densification process is simple, because it requires a control of only one parameter (degree of densification). Second, it is effective, as it achieves mechanical enhancement and sustainable release of biomolecules from porous scaffolds. Third, it has broad applications, as it is also applicable to the fabrication of functionally graded porous scaffolds by spatially varied strain during densification.

用于生产多孔钛的支架的制造方法示于图1A。钛粉末被保持在莰均匀分散由容器的连续旋转在44℃下12小时和，而液体莰被完全固化，相对较重的Ti粉末的任何沉积物最小化。其结果是，将均匀的Ti-莰生坯使用动态冷冻浇铸方法生产，如图1B所示，在其中三维地相互连接的大莰孔隙由钛粉末相（图1C）所包围。然而，不正确的容器的转动...

Watch this Scientific Journal Video about Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications at JoVE.com

Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications

Bioactive and mechanically reliable metal scaffolds have been fabricated through a method which consists of two processes, dynamic freeze casting for the fabrication of porous Ti, and coating and densification of the Ti scaffolds. The densification process is simple, effective and applicable to the fabrication of functionally graded scaffolds.

加工机械可调和生物活性金属支架的生物医学应用

Biometal systems have been widely used for biomedical applications, in particular, as load-bearing materials. However, major challenges are high stiffness ...

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Transformations of Stress and Strain

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10.6K 次观看.  Korea Institute of Industrial Technology. Bioactive and mechanically reliable metal scaffolds have been fabricated through a method which consists of two processes, dynamic freeze casting for the fabrication of porous Ti, and coating and densification of the Ti scaffolds. The densification process is simple, effective and applicable to the fabrication of functionally graded scaffolds.

视频: Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications

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

Preparation of 3D Fibrin Scaffolds for Stem Cell Culture Applications

Stem cells are found in naturally occurring 3D microenvironments in vivo, which are often referred to as the stem cell niche 1. Culturing stem cells inside of 3D biomaterial scaffolds provides a way to accurately mimic these microenvironments, providing an advantage over traditional 2D culture methods using polystyrene as well as a method for engineering replacement tissues 2. While 2D tissue culture polystrene has been used for the majority of cell culture experiments, 3D biomaterial scaffolds can more closely replicate the microenvironments found in vivo by enabling more accurate establishment of cell polarity in the environment and possessing biochemical and mechanical properties similar to soft tissue.3 A variety of naturally derived and synthetic biomaterial scaffolds have been investigated as 3D environments for supporting stem cell growth. While synthetic scaffolds can be synthesized to have a greater range of mechanical and chemical properties and often have greater reproducibility, natural biomaterials are often composed of proteins and polysaccharides found in the extracelluar matrix and as a result contain binding sites for cell adhesion and readily support cell culture. Fibrin scaffolds, produced by polymerizing the protein fibrinogen obtained from plasma, have been widely investigated for a variety of tissue engineering applications both in vitro and in vivo 4. Such scaffolds can be modified using a variety of methods to incorporate controlled release systems for delivering therapeutic factors 5. Previous work has shown that such scaffolds can be used to successfully culture embryonic stem cells and this scaffold-based culture system can be used to screen the effects of various growth factors on the differentiation of the stem cells seeded inside 6,7.
 This protocol details the process of polymerizing fibrin scaffolds from fibrinogen solutions using the enzymatic activity of thrombin. The process takes 2 days to complete, including an overnight dialysis step for the fibrinogen solution to remove citrates that inhibit polymerization. These detailed methods rely on fibrinogen concentrations determined to be optimal for embryonic and induced pluripotent stem cell culture. Other groups have further investigated fibrin scaffolds for a wide range of cell types and applications - demonstrating the versatility of this approach 8-12.

This work details the preparation of 3D fibrin scaffolds for culturing and differentiating plutipotent stem cells. Such scaffolds can be used to screen the effects of various biological compounds on stem cell behavior as well as modified to contain drug delivery systems.

制备纤维蛋白三维支架干细胞培养应用

Stem cells are found in naturally occurring 3D microenvironments in vivo, which are often referred to as the stem cell niche 1. Culturing stem cells ...

Designing Silk-silk Protein Alloy Materials for Biomedical Applications

Fibrous proteins display different sequences and structures that have been used for various applications in biomedical fields such as biosensors, nanomedicine, tissue regeneration, and drug delivery. Designing materials based on the molecular-scale interactions between these proteins will help generate new multifunctional protein alloy biomaterials with tunable properties. Such alloy material systems also provide advantages in comparison to traditional synthetic polymers due to the materials biodegradability, biocompatibility, and tenability in the body. This article used the protein blends of wild tussah silk (Antheraea pernyi) and domestic mulberry silk (Bombyx mori) as an example to provide useful protocols regarding these topics, including how to predict protein-protein interactions by computational methods, how to produce protein alloy solutions, how to verify alloy systems by thermal analysis, and how to fabricate variable alloy materials including optical materials with diffraction gratings, electric materials with circuits coatings, and pharmaceutical materials for drug release and delivery. These methods can provide important information for designing the next generation multifunctional biomaterials based on different protein alloys.

Blending is an efficient approach to generate biomaterials with a broad range of properties and combined features. By predicting the molecular interactions between different natural silk proteins, new silk-silk protein alloy platforms with tunable mechanical resiliency, electrical response, optical transparency, chemical processability, biodegradability, or thermal stability can be designed.

设计丝绸蚕丝蛋白合金生物医用材料

Fibrous proteins display different sequences and structures that have been used for various applications in biomedical fields such as biosensors, ...

Fabrication of a Bioactive, PCL-based "Self-fitting" Shape Memory Polymer Scaffold

Tissue engineering has been explored as an alternative strategy for the treatment of critical-sized cranio-maxillofacial (CMF) bone defects. Essential to the success of this approach is a scaffold that is able to conformally fit within an irregular defect while also having the requisite biodegradability, pore interconnectivity and bioactivity. By nature of their shape recovery and fixity properties, shape memory polymer (SMP) scaffolds could achieve defect &#8220;self-fitting.&#8221; In this way, following exposure to warm saline (~60 &#186;C), the SMP scaffold would become malleable, permitting it to be hand-pressed into an irregular defect. Subsequent cooling (~37 &#186;C) would return the scaffold to its relatively rigid state within the defect. To meet these requirements, this protocol describes the preparation of SMP scaffolds prepared via the photochemical cure of biodegradable polycaprolactone diacrylate (PCL-DA) using a solvent-casting particulate-leaching (SCPL) method. A fused salt template is utilized to achieve pore interconnectivity. To realize bioactivity, a polydopamine coating is applied to the surface of the scaffold pore walls. Characterization of self-fitting and shape memory behaviors, pore interconnectivity and in vitro bioactivity are also described.

Fabrication of a Bioactive, PCL-based &quot;Self-fitting&quot; Shape Memory Polymer Scaffold

Scaffolds capable of fitting within cranio-maxillofacial (CMF) bone defects while exhibiting osteoconductivity and bioactivity are of interest. This protocol describes the preparation of a shape memory scaffold based on polycaprolactone diacrylate (PCL-DA) using a solvent-casting particulate-leaching (SCPL) method employing a fused salt template and application of a bioactive polydopamine coating.

生物活性，PCL为基础的“自我装配”形状记忆聚合物支架的制备

Tissue engineering has been explored as an alternative strategy for the treatment of critical-sized cranio-maxillofacial (CMF) bone defects. Essential to ...

Fabricating Superhydrophobic Polymeric Materials for Biomedical Applications

Superhydrophobic materials, with surfaces possessing permanent or metastable non-wetted states, are of interest for a number of biomedical and industrial applications. Here we describe how electrospinning or electrospraying a polymer mixture containing a biodegradable, biocompatible aliphatic polyester (e.g., polycaprolactone and poly(lactide-co-glycolide)), as the major component, doped with a hydrophobic copolymer composed of the polyester and a stearate-modified poly(glycerol carbonate) affords a superhydrophobic biomaterial. The fabrication techniques of electrospinning or electrospraying provide the enhanced surface roughness and porosity on and within the fibers or the particles, respectively. The use of a low surface energy copolymer dopant that blends with the polyester and can be stably electrospun or electrosprayed affords these superhydrophobic materials. Important parameters such as fiber size, copolymer dopant composition and/or concentration, and their effects on wettability are discussed. This combination of polymer chemistry and process engineering affords a versatile approach to develop application-specific materials using scalable techniques, which are likely generalizable to a wider class of polymers for a variety of applications.

Two- and three-dimensional superhydrophobic polymeric materials are prepared by electrospinning or electrospraying biodegradable polymers blended with a lower surface energy polymer of similar composition.

制造超疏水高分子材料生物医学应用

Superhydrophobic materials, with surfaces possessing permanent or metastable non-wetted states, are of interest for a number of biomedical and industrial ...

Fabrication of a Dipole-assisted Solid Phase Extraction Microchip for Trace Metal Analysis in Water Samples

This paper describes a fabrication protocol for a dipole-assisted solid phase extraction (SPE) microchip available for trace metal analysis in water samples. A brief overview of the evolution of chip-based SPE techniques is provided. This is followed by an introduction to specific polymeric materials and their role in SPE. To develop an innovative dipole-assisted SPE technique, a chlorine (Cl)-containing SPE functionality was implanted into a poly(methyl methacrylate) (PMMA) microchip. Herein, diverse analytical techniques including contact angle analysis, Raman spectroscopic analysis, and laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) analysis were employed to validate the utility of the implantation protocol of the C-Cl moieties on the PMMA. The analytical results of the X-ray absorption near-edge structure (XANES) analysis also demonstrated the feasibility of the Cl-containing PMMA used as an extraction medium by virtue of the dipole-ion interactions between the highly electronegative C-Cl moieties and the positively charged metal ions.

The fabrication protocol of a dipole-assisted solid phase extraction microchip for the trace metal analysis is presented.

水样中偶极辅助固相萃取Microchip的痕量金属分析的制作

This paper describes a fabrication protocol for a dipole-assisted solid phase extraction (SPE) microchip available for trace metal analysis in water ...

Rapid Mix Preparation of Bioinspired Nanoscale Hydroxyapatite for Biomedical Applications

Hydroxyapatite (HA) has been widely used as a medical ceramic due to its good biocompatibility and osteoconductivity. Recently there has been interest regarding the use of bioinspired nanoscale hydroxyapatite (nHA). However, biological apatite is known to be calcium-deficient and carbonate-substituted with a nanoscale platelet-like morphology. Bioinspired nHA has the potential to stimulate optimal bone tissue regeneration due to its similarity to bone and tooth enamel mineral. Many of the methods currently used to fabricate nHA both in the laboratory and commercially, involve lengthy processes and complex equipment. Therefore, the aim of this study was to develop a rapid and reliable method to prepare high quality bioinspired nHA. The rapid mixing method developed was based upon an acid-base reaction involving calcium hydroxide and phosphoric acid. Briefly, a phosphoric acid solution was poured into a calcium hydroxide solution followed by stirring, washing and drying stages. Part of the batch was sintered at 1,000 &#176;C for 2 h in order to investigate the products&#39; high temperature stability. X-ray diffraction analysis showed the successful formation of HA, which showed thermal decomposition to &#946;-tricalcium phosphate after high temperature processing, which is typical for calcium-deficient HA. Fourier transform infrared spectroscopy showed the presence of carbonate groups in the precipitated product. The nHA particles had a low aspect ratio with approximate dimensions of 50 x 30 nm, close to the dimensions of biological apatite. The material was also calcium deficient with a Ca:P molar ratio of 1.63, which like biological apatite is lower than the stoichiometric HA ratio of 1.67. This new method is therefore a reliable and far more convenient process for the manufacture of bioinspired nHA, overcoming the need for lengthy titrations and complex equipment. The resulting bioinspired HA product is suitable for use in a wide variety of medical and consumer health applications.

This paper describes a novel method for the rapid manufacture of high quality bioinspired nanoscale hydroxyapatite. This biomaterial is of great significance in the manufacture of a wide range of innovative medical devices for clinical applications in orthopedics, craniofacial surgery and dentistry.

仿生纳米羟基磷灰石的快速混合制备生物医学应用

Hydroxyapatite (HA) has been widely used as a medical ceramic due to its good biocompatibility and osteoconductivity. Recently there has been interest ...

Patterning Bioactive Proteins or Peptides on Hydrogel Using Photochemistry for Biological Applications

There are many biological stimuli that can influence cell behavior and stem cell differentiation. General cell culture approaches rely on soluble factors within the medium to control cell behavior. However, soluble additions cannot mimic certain signaling motifs, such as matrix-bound growth factors, cell-cell signaling, and spatial biochemical cues, which are common influences on cells. Furthermore, biophysical properties of the matrix, such as substrate stiffness, play important roles in cell fate, which is not easily manipulated using conventional cell culturing practices. In this method, we describe a straightforward protocol to provide patterned bioactive proteins on synthetic polyethylene glycol (PEG) hydrogels using photochemistry. This platform allows for the independent control of substrate stiffness and spatial biochemical cues. These hydrogels can achieve a large range of physiologically relevant stiffness values. Additionally, the surfaces of these hydrogels can be photopatterned with bioactive peptides or proteins via thiol-ene click chemistry reactions. These methods have been optimized to retain protein function after surface immobilization. This is a versatile protocol that can be applied to any protein or peptide of interest to create a variety of patterns. Finally, cells seeded onto the surfaces of these bioactive hydrogels can be monitored over time as they respond to spatially specific signals.

In this method, we use photopolymerization and click chemistry techniques to create protein or peptide patterns on the surface of polyethylene glycol (PEG) hydrogels, providing immobilized bioactive signals for the study of cellular responses in vitro.

生物活性蛋白或多肽在水凝胶上的应用

There are many biological stimuli that can influence cell behavior and stem cell differentiation. General cell culture approaches rely on soluble factors ...

Melt Electrospinning Writing of Three-dimensional Poly(&#949;-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology with great potential for biomedical applications. The technique facilitates the direct deposition of biocompatible polymer fibers to fabricate well-ordered scaffolds in the sub-micron to micro scale range. The establishment of a stable, viscoelastic, polymer jet between a spinneret and a collector is achieved using an applied voltage and can be direct-written. A significant benefit of a typical porous scaffold is a high surface-to-volume ratio which provides increased effective adhesion sites for cell attachment and growth. Controlling the printing process by fine-tuning the system parameters enables high reproducibility in the quality of the printed scaffolds. It also provides a flexible manufacturing platform for users to tailor the morphological structures of the scaffolds to their specific requirements. For this purpose, we present a protocol to obtain different fiber diameters using melt electrospinning writing (MEW) with a guided amendment of the parameters, including flow rate, voltage and collection speed. Furthermore, we demonstrate how to optimize the jet, discuss often experienced technical challenges, explain troubleshooting techniques and showcase a wide range of printable scaffold architectures.

Melt Electrospinning Writing of Three-dimensional Poly(&amp;#949;-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

This protocol serves as a comprehensive guideline to fabricate scaffolds via electrospinning with polymer melts in a direct writing mode. We systematically outline the process and define the appropriate parameter settings for achieving targeted scaffold architectures.

用于组织工程应用的三维聚 (ε-内) 支架的熔体静电纺丝

This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology ...

Fabrication and Characterization of Griffithsin-modified Fiber Scaffolds for Prevention of Sexually Transmitted Infections

Electrospun fibers (EFs) have been widely used in a variety of therapeutic applications; however, they have only recently been applied as a technology to prevent and treat sexually transmitted infections (STIs). Moreover, many EF technologies focus on encapsulating the active agent, relative to utilizing the surface to impart biofunctionality. Here we describe a method to fabricate and surface-modify poly(lactic-co-glycolic) acid (PLGA) electrospun fibers, with the potent antiviral lectin Griffithsin (GRFT). PLGA is an FDA-approved polymer that has been widely used in drug delivery due to its outstanding chemical and biocompatible properties. GRFT is a natural, potent, and safe lectin that possesses broad activity against numerous viruses including human immunodeficiency virus type 1 (HIV-1). When combined, GRFT-modified fibers have demonstrated potent inactivation of HIV-1 in vitro. This manuscript describes the methods to fabricate and characterize GRFT-modified EFs. First, PLGA is electrospun to create a fiber scaffold. Fibers are subsequently surface-modified with GRFT using 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS)chemistry. Scanning electron microscopy (SEM) was used to assess the size and morphology of surface-modified formulations. Additionally, a gp120 or hemagglutinin (HA)-based ELISA may be used to quantify the amount of GRFT conjugated to, as well as GRFT desorption from the fiber surface. This protocol can be more widely applied to fabricate fibers that are surface-modified with a variety of different proteins.

This manuscript describes the procedure to fabricate and characterize Griffithsin-modified poly(lactic-co-glycolic acid) electrospun fibers that demonstrate potent adhesive and antiviral activity against human immunodeficiency virus type 1 infection in vitro. Methods used to synthesize, surface-modify, and characterize the resulting morphology, conjugation, and desorption of Griffithsin from surface-modified fibers are described.

Griffithsin 修饰纤维支架预防性传播感染的制备与表征

Electrospun fibers (EFs) have been widely used in a variety of therapeutic applications; however, they have only recently been applied as a technology to ...