这项工作是由协同Ceramisys有限公司的EPSRC CASE助学金和支持也与米德求新，EPSRC中心相关于医疗器械[授权号EP / K029592 / 1]创新制造。笔者也想在谢菲尔德哈勒姆大学，感谢罗伯特·伯顿的XRF分析。

<img alt="图1" src="/files/ftp_upload/55343/55343fig1.jpg" /> 图1.仿生纳米羟基磷灰石的快速混合配制的示意图。磷酸溶液倒入氢氧化钙悬浮液中。该悬浮液定居过夜后，将羟基磷灰石是用去离子水洗涤，在60至80℃干燥之前。的羟基磷灰石然后在玛瑙研钵和杵及烧结研磨调查磷灰石产物的热稳定性。 <a href="http://ecsource.jove.com/files/ftp_upload/55343/55343fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 1.纳米羟基磷灰石的快速混合生产<ol><li>钙，磷的解决方案，准备使用钙与磷1.67摩尔比为准备5克纳米羟基磷灰石。 <ol><li>添加3.705克氢氧化钙与500ml的去离子水搅拌在磁力搅拌器板以400rpm 1小时。 </li><li>在一个单独的烧杯中，溶解3.459克的在250毫升去离子水的磷酸（85％）。 </li></ol></li><li>倾磷溶液倒入搅拌的氢氧化钙悬浮液在大约100毫升/秒的速率。用封口膜封住（Bemis公司，USA）烧杯中。 </li><li>离开该悬浮液搅拌在400rpm 1小时。 </li><li>就拿烧杯关闭搅拌器板，并留下沉降过夜。 </li><li>通过倾倒掉上清，加入500毫升去离子水并在400rpm下搅拌1分钟，洗该悬浮液。重复此步骤三次总共，每次洗涤之间2小时。 </li><li>离开纳米羟基磷灰石悬挂沉降过夜。 </li><li>倒出上层清液，然后将结算磷灰石悬浮液在烘箱中于60至80℃的干燥。 </li><li>干燥时，将干燥的羟基磷灰石到玛瑙研钵和研杵和研直到细。 </li><li>有2.5克的亲duced磷灰石粉末在氧化铝坩埚并烧结粉末在1000℃2小时，使用10℃/分钟的升温速率。热处理后，离开磷灰石在炉内冷却。 </li><li>商店的粉末在真空干燥器。 </li></ol> 2.纳米羟基磷灰石的表征<ol><li> 使用的传输模式的衍射仪的X射线衍射（XRD） <ol><li>放置聚（乙烯醇）的少量（ 即小于200微升）（PVA）胶醋酸膜和与羟基磷灰石粉末的少量（ 即小于100毫克）混合。 </li><li>用热风枪，直到干燥处理。 </li><li>装入样品到样品保持器和负载到具有Cu的Kαα辐射的传输模式X射线衍射仪。 </li><li>使用40千伏和35毫安衍射设置，以10〜70°的2θ范围内。 </li><li>分析产生的X射线衍射图案。 </li><li>使用相鉴定以下XRD卡:羟基磷灰石:9-432。 β磷酸三钙:04-014-2292。 </li></ol></li><li> 透射电子显微镜（TEM） <ol><li>放置粉末在茹少量（ 即小于10毫克）和增加大约3毫升乙醇中。 </li><li> 15超声处理样品 - 30分钟直到一个均匀的悬浮液是观察。 </li><li>吸取少量的溶液（ 即低于1毫升以下）到具有碳膜400目铜网上，并晾干。 </li><li>图像样品在80千伏的加速电压。 </li></ol></li><li> X射线荧光（XRF）的材料与工程研究所（梅里）在谢菲尔德哈勒姆大学服务 <ol><li>结合0.8克纳米羟基磷灰石粉末与8克四硼酸锂。 </li><li>熔化在使用炉设置为1200℃的铂 - 金合金坩混合物。 </li><li>在X射线荧光光谱仪分析所得的样品，以确定的元素组成样本。 </li></ol></li><li> 傅立叶变换中衰减全反射模式红外光谱（FTIR-ATR） <ol><li>从4000执行64后台扫描- 400 -1用4厘米-1的分辨率。 </li><li>放置磷灰石粉末在金刚石的顶端少量（ 即小于100毫克）在衰减全反射模式适配器和压缩到使用螺丝顶部金刚石的表面上。 </li><li>从4000执行扫描32 - 400 -1为4厘米的分辨率-1从样品扫描减去后台扫描。 </li></ol></li></ol>

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作者什么都没有透露。

天然磷灰石是由具有钙10-XY的近似化学式[（HPO 4）（PO 4）] 6-X（CO 3）Y（OH）2-X型非化学计量碳酸羟磷灰石纳米颗粒。于天然存在的矿物已经报道了生产接近化学相似的生物材料，以促进最佳的生物反应。例如，在仿生缺钙碳酸磷灰石的研究已经表明，它是能刺激增殖和鼠前成骨细胞的细胞的碱性磷酸酶活性更大程度比常规磷灰石19。 在这项研究中，HA的这表明局部热分解在1000℃（ 图2）的析出建议缺钙的HA的生成。用X射线荧光光谱数据获得的P比（1.63）（T:这是由比化学计量的Ca下部支承能1）。可以理解的是一个减小的Ca:P比与较低的热稳定性20，21，22，23相关联。在该方法中，迅速加入磷酸溶液急剧降低反应悬浮液的pH以产生HPO 4离子。 HPO存在4组促进钙缺乏的HA的沉淀，用分子式为Ca 10-X（HPO 4）×（PO 4）6-X（OH）2-x，其中0 &lt;X &lt;1。 因此，迅速加入磷酸对反应的沉淀动力学有显着影响。如前所述，涉及在室温下进行的氢氧化钙和磷酸滴定反应倾向于具有高的纵横比13，以产生颗粒。对于titratioÑ涉及这些反应物的反应，有必要使用升高的温度，以产生具有较低纵横比这更类似于生物磷灰石13的颗粒。当晶体成核速率比晶体生长速率24慢的高纵横比的颗粒产生。在这项研究中开发的新方法，迅速加入磷酸溶液可能提供这导致小的圆形颗粒的存在增加，而不是更少的粒子具有较大的长宽比成核位点的更大的数字。如作者还没有充分研究的缓慢浇注磷酸成氢氧化钙悬浮液中，以达到一致的结果的影响，我们建议的磷酸在视频（大约100毫升/显示与相称的速率倾S）。 在该方法的发展，作者INVestigated一些增量变化的基础上普拉卡什等人芽制备方法。 13，包括与缓慢滴定并迅速加入磷酸溶液25的所生产的产品的比较。我们发现的磷酸的缓慢滴定入氢氧化钙悬浮液导致与氢氧化钙残基的产物。我们建议，所造成的快速加入磷酸的pH变化鼓励氢氧化钙的溶解，因此，允许对成功转化的反应物到羟基磷灰石。的产品的比较使用在室温和升高的温度（60℃）快速混合方法制备发现升高的温度导致更高的导电性的反应完成后。这表明，残留的氢氧化钙存在这很可能是由于氢氧化钙中的溶解度较低温度升高。残留的氢氧化钙的存在是不希望的，该化合物的基本性质可能危及生物相容性。 红外探测用HA（ 图3）相关联的特性磷酸酯和羟基基团的活性。有人指出，对于烧结产品的光谱显示更清晰磷酸盐和羟基峰。这些变化已经与一个更大的产品结晶度26，27提供的B型碳酸替代，其中碳酸根离子已经取代磷酸基团的证据.The烧结谱有关。这是相对于A型的取代，其中碳酸根离子可以取代的羟基17。据报道，B型碳酸盐替代在生物磷灰石3发生。然而，Tampieri 等。报告说，虽然B型取代是predomin蚂蚁在骨头青年，A型碳酸盐替代是老年人28骨头越来越呈现。碳酸盐替代已发现减小磷灰石的结晶度和热稳定性，而增加其溶解度。这些变化已经提出向碳酸取代的HA 29的增加生物活性。还已知的生物的HA含有一些记录在X射线荧光分析（ 表1）的其他元素，如镁，钠，锶30的。这些元素的存在也可能有助于提高生物效力。今后的工作应在这些纳米取代磷灰石的制备被引导，并且还制品具有增加的生物功能如掺杂银的磷灰石31。为了制备取代的羟基磷灰石，该元件可以与相应的减少预期的元素与substitut被引入e表示， 例如 ，在钙化合物的量减少时锶，镁或锌置换尝试32。可替代地，另一种方法可以是与提供"掺杂"离子是存在的羟基磷灰石的表面上，而不必有意的元件替换到HA晶格31的意图添加元素。对于这些修改的方法是可能的制备混合溶液，例如氢氧化钙和硝酸银，并作为此处所描述的进行该反应的相同方法。 总之，本文报道了仿生羟基磷灰石的制备新颖快速和基本上改进方法。对于这种方法，化学品的快速混合时间不超过5秒，这是在时间上的显着减少相比滴定反应通常需要仔细监测小时。它有在使用BIOMAT潜力巨大erial发展，由于其相对简单和低成本相比目前使用的工业磷灰石的制造方法，其中的当前商业系统的结果在长期的研究和开发时间的固有的复杂性，并大大提高了制造成本。特别是，这种新方法是优于连续流动过程或者由于显著下启动设备投资需求热液技术。

有为了提高患者的生活质量，并减少全球人口老龄化的医疗负担很大的临床需要与增强功能先进的生物材料。羟基磷灰石已广泛，由于其良好的生物相容性应用于多年医疗应用。最近，已在使用纳米级的羟基磷灰石（羟基磷灰石）的兴趣增加，特别是用于医学和牙科矿化组织再生。在骨骼和牙齿的釉质中发现的矿物是缺钙，多取代的，纳米级羟基磷灰石。生物纳米羟基磷灰石血小板的大小估计报告的50纳米×30纳米×2纳米的1维，在不成熟的骨2所述更小的结构。与此相反，在牙齿珐琅质的矿物比在长度和宽度3,4骨组织中发现的大10至100倍。合成纳米羟基磷灰石<html>可能会更好地称为仿生，而不是仿生，因为我们正在寻求翻译关于自然材料的特性与性能改进医疗技术的意见。它已被提出，仿生羟基磷灰石可以是骨骼和牙齿的组织再生应用更有利的，因为它相似于天然存在的矿物5。 有已报道以制备羟基磷灰石包括热液6，喷雾干燥7和溶胶-凝胶8技术的各种方法。这些中，湿式沉淀法被认为是用于制备羟基磷灰石的相对方便的方法。公布的羟基磷灰石湿沉淀方法一般包括一个滴定步骤混合的钙和磷的化学前体9，10，11时，裁判"&gt; 12，13，14，但是，这些方法都具有许多缺点，包括冗长和复杂的过程结合在某些情况下，与需要昂贵的设备相关联。商业生产可能更复杂，专利描述用于复杂的反应器高品质的医疗级磷灰石15的制造。尽管这样，氢氧化钙和磷酸之间的中和反应是有利的，由于副产物的缺乏有毒化学品。 加工条件和磷灰石产物的形态之间的关系已被报道用于缓慢滴定反应。具体地，对于涉及氢氧化钙和磷酸滴定方法中，升高的温度似乎赞成粒子具有低纵横比13的制备。这项工作得到了大大代扩展瓷砖等。 16谁表现出对羟基磷灰石的产品从广泛的方法的质量温度等加工条件的关系。他的结论是，普拉卡什13的湿化学沉淀法制造的高质量的产品，但应注意的是，结果是依赖于技术上的挑战和慢/混合过程。原始普拉卡什滴定步骤接管一小时。然而，更长的滴定时间可能需要要制备大批量。 为了总结，而几个因素，包括温度的影响现在已经广泛地在减少复杂性的研究，几乎没有关注已指示和相关联的执行基于滴定的方法所需要的时间。因此，这项研究的目的是探讨应用快速混合方式，以仿生纳米羟基磷灰石制造的影响，并充分characteri泽所得到的材料。如果成功的话，简单的快速混合的方式将对实验室的研究人员和业界的一致好评，其中制造成本可以不包括质量大幅减少很大的好处。

<table border="0" cellpadding="0" cellspacing="0" width="1375">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:399px;">Calcium hydroxide (purity of &#8805; 96%)</td>
			<td style="width:168px;">Sigma Aldrich UK</td>
			<td align="right" style="width:115px;">31219</td>
			<td style="width:695px;">Good laboratory practise should be used at all times including the use of appropriate personal protective equipment.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Phosphoric acid (85 %)</td>
			<td>Sigma Aldrich UK</td>
			<td align="right">345245</td>
			<td>Safety goggles and a faceshield should be used when handling this product (see safety data sheet from Sigma Aldrich for further information).</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">STOE IP x-ray diffractometer</td>
			<td>Phillips</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">International centre for diffraction data (ICDD) PDF4+ database</td>
			<td colspan="2">International Centre for Diffraction Data</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Holey carbon films on 300 mesh grids</td>
			<td>Agar Scientific</td>
			<td>S147-3H&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tecnai G2 Spirit transmission electron microscope</td>
			<td>FEI</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Lithium tetraborate</td>
			<td>ICPH, Malz&#233;ville, France</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">PW2440 XRF spectrometer&#160;</td>
			<td>Philips</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">ThermoScientific Nikolett Spectrometer</td>
			<td>Unicam Ltd</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

rapid mix preparation of bioinspired nanoscale hydroxyapatite for biomedical applications

羟基磷灰石（HA）已被广泛地用作医疗陶瓷由于其良好的生物相容性和骨传导性。最近有关于使用仿生纳米羟基磷灰石（NHA）的兴趣。然而，生物磷灰石是已知的钙缺乏和碳酸取代的纳米级片状的形态。仿生羟基磷灰石具有刺激最佳骨组织再生的潜力，因为它相似骨和牙齿珐琅质矿物。许多方法目前用来制造磷灰石在实验室和商业上，涉及冗长过程和复杂的设备。因此，本研究的目的是开发一种快速，可靠的方法来制备高品质的仿生纳米羟基磷灰石。开发的快速混合方法是基于涉及氢氧化钙和磷酸的酸 - 碱反应。简言之，磷酸溶液倒入氢氧化钙溶液中，接着搅拌，洗涤和干燥阶段。批料的一部分混合物在1000℃在为了研究产品的高温稳定性烧结2小时。 X射线衍射分析显示成功形成的HA，其显示热分解为β-磷酸三钙的高温处理中，这是典型的钙缺乏HA之后磷酸盐。傅里叶变换红外光谱表明碳酸酯基团的在沉淀出的产品的存在。芽颗粒与50×30纳米的大致尺寸，接近生物磷灰石的尺寸低纵横比。该材料也是钙缺乏与钙:1.63 P摩尔比，这样生物学磷灰石比1.67的化学计量的HA比率低。因此，这种新方法是用于制造仿生羟基磷灰石的，克服了冗长滴定和复杂的设备需要一个可靠的和更方便的过程。所得仿生HA产品是适用于各种用途医疗和消费保健应用。

<html> X射线衍射图案（ 图2）表现出纯的HA相具有宽峰的沉淀，指示相对小的晶粒尺寸和/或无定形性质。高温烧结后，β磷酸三钙（β-TCP）检测，沿着HA的主相。的衍射峰的锐化， 即在全宽度半最大值的降低，烧结后所指示的微晶尺寸的增加。 <img alt="图2" src="/files/ftp_upload/55343/55343fig2.jpg" /> 产品图2.晶相分析。 X射线衍射（XRD）在1000℃烧结2小时未烧结纳米级的羟基磷灰石（羟基磷灰石）粉末和磷灰石粉末的图案。峰标签:▼羟基磷灰石峰，β■磷酸三钙峰。ge.jpg"目标="_空白"&gt;点击此处查看该图的放大版本。 FTIR-ATR光谱（ 图3）确认一个的HA相由特性磷酸盐和羟基频带17,18的形成。详细频带被分配如下:3750厘米-1（OH -拉伸νOH）; 1086和1022厘米-1（PO 4 3-ν3）; 962厘米-1（PO 4 3-ν1）; 630厘米-1（OH -振动δOH）; 600和570 -1（PO 4 3-ν4）。在未烧结样品的附加的峰被分配如下:约有3,400 -1（吸附水分子）为中心的宽峰; 1455和1410厘米-1（CO 3 2-ν3）; 880厘米<sup&gt; -1（CO 3 2-ν2）。在未烧结粉末观察到的吸收水和碳酸酯基团在高温烧结阶段被拆除。烧结过程也削尖这是由一个更大的高峰表现到谷底的距离羟基和磷酸盐带。 <img alt="图3" src="/files/ftp_upload/55343/55343fig3.jpg" /> 图3的产品的红外光谱。在未烧结纳米级的羟基磷灰石（羟基磷灰石）粉末和磷灰石粉末在1000℃烧结2小时的衰减全反射模式（FTIR-ATR）光谱傅里叶变换红外。 <a href="http://ecsource.jove.com/files/ftp_upload/55343/55343fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 透射电镜照片（ 图4），显示出纳米级颗粒机智的形成通过30纳米50纳米ħ近似尺寸。所述颗粒具有约1.7的低纵横比（颗粒长度/颗粒宽度）。纳米产品的大小和形状相似尺寸的生物磷灰石1。 <img alt="图4" src="/files/ftp_upload/55343/55343fig4.jpg" /> 图4.产品的纳米形貌。纳米羟基磷灰石（NHA）的透射电子显微照片（TEM）采用快速混合方法在两个倍数，准备。 <a href="http://ecsource.jove.com/files/ftp_upload/55343/55343fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 通过XRF（ 表1）的羟基磷灰石粉末的定量化学分析允许的钙:磷比率被计算为1.63，这比在化学计量的HA略低whicH有钙:1.67磷比例。 XRF也表现出了纳米羟基磷灰石产品的高纯度记录其他元素的只有微量。 <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> 复合 </td><td> 重量％ </td></tr><tr><td>氧化钙</td><td> 51.52 </td></tr><tr><td> P 2 O 5 </td><td> 39.89 </td></tr><tr><td>氧化镁</td><td> 0.46 </td></tr><tr><td>娜2 O </td><td> 0.13 </td></tr><tr><td> Y 2 O 3 </td><td> 0.07 </td></tr><tr><td> 的 Al 2 O 3 </td><td> 0.03 </td></tr><tr><td> SiO 2的</td><td> 0.03 </td></tr><tr><td>锰3 O 4 </td><td> 0.03 </td></tr><tr><td>的SrO </td><td> 0.02 </td></tr><tr><td> 二氧化钛</td><td> 0.01 </td></tr></tbody></table>表1.定量化学品的分析。 X射线荧光（XRF）的结果为未烧结羟基磷灰石粉末显示&gt; 99％纯度（重量）。

Watch this Scientific Journal Video about Rapid Mix Preparation of Bioinspired Nanoscale Hydroxyapatite for Biomedical Applications at JoVE.com

Rapid Mix Preparation of Bioinspired Nanoscale Hydroxyapatite for Biomedical Applications

本文介绍了高品质的仿生纳米羟基磷灰石的快速制造的新方法。该生物材料是在一个广泛的骨科，颅面外科和牙科临床应用提供创新的医疗设备制造具有重要意义。

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 ...

rapid-mix-preparation-bioinspired-nanoscale-hydroxyapatite-for

Research

JoVE Journal

Bioengineering

19.1K 次观看.  University of Sheffield. 本文介绍了高品质的仿生纳米羟基磷灰石的快速制造的新方法。该生物材料是在一个广泛的骨科，颅面外科和牙科临床应用提供创新的医疗设备制造具有重要意义。 

视频: Rapid Mix Preparation of Bioinspired Nanoscale Hydroxyapatite for Biomedical Applications

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 ...

科研

生物工程

Graphene Coatings for Biomedical Implants

Atomically smooth graphene as a surface coating has potential to improve implant properties. This demonstrates a method for coating nitinol alloys with nanometer thick layers of graphene for applications as a stent material. Graphene was grown on copper substrates via chemical vapor deposition and then transferred onto nitinol substrates. In order to understand how the graphene coating could change biological response, cell viability of rat aortic endothelial cells and rat aortic smooth muscle cells was investigated. Moreover, the effect of graphene-coatings on cell adhesion and morphology was examined with fluorescent confocal microscopy. Cells were stained for actin and nuclei, and there were noticeable differences between pristine nitinol samples compared to graphene-coated samples. Total actin expression from rat aortic smooth muscle cells was found using western blot. Protein adsorption characteristics, an indicator for potential thrombogenicity, were determined for serum albumin and fibrinogen with gel electrophoresis. Moreover, the transfer of charge from fibrinogen to substrate was deduced using Raman spectroscopy. It was found that graphene coating on nitinol substrates met the functional requirements for a stent material and improved the biological response compared to uncoated nitinol. Thus, graphene-coated nitinol is a viable candidate for a stent material.

Graphene offers potential as a coating material for biomedical implants. In this study we demonstrate a method for coating nitinol alloys with nanometer thick layers of graphene and determine how graphene may influence implant response.

生物医学植入物的石墨涂料

Atomically smooth graphene as a surface coating has potential to improve implant properties. This demonstrates a method for coating nitinol alloys with ...

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, ...

Nanomanipulation of Single RNA Molecules by Optical Tweezers

A large portion of the human genome is transcribed but not translated. In this post genomic era, regulatory functions of RNA have been shown to be increasingly important. As RNA function often depends on its ability to adopt alternative structures, it is difficult to predict RNA three-dimensional structures directly from sequence. Single-molecule approaches show potentials to solve the problem of RNA structural polymorphism by monitoring molecular structures one molecule at a time. This work presents a method to precisely manipulate the folding and structure of single RNA molecules using optical tweezers. First, methods to synthesize molecules suitable for single-molecule mechanical work are described. Next, various calibration procedures to ensure the proper operations of the optical tweezers are discussed. Next, various experiments are explained. To demonstrate the utility of the technique, results of mechanically unfolding RNA hairpins and a single RNA kissing complex are used as evidence. In these examples, the nanomanipulation technique was used to study folding of each structural domain, including secondary and tertiary, independently. Lastly, the limitations and future applications of the method are discussed.

Optical tweezers have been used to study RNA folding by stretching individual molecules from their 5&#8217; and 3&#8217; ends. Here common procedures are described to synthesize RNA molecules for tweezing, calibration of the instrument, and methods to manipulate single molecules.

单个RNA分子的光镊纳米操作

A large portion of the human genome is transcribed but not translated. In this post genomic era, regulatory functions of RNA have been shown to be ...

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 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.

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 ...

Synthesis of Keratin-based Nanofiber for Biomedical Engineering

Electrospinning, due to its versatility and potential for applications in various fields, is being frequently used to fabricate nanofibers. Production of these porous nanofibers is of great interest due to their unique physiochemical properties. Here we elaborate on the fabrication of keratin containing poly (&#949;-caprolactone) (PCL) nanofibers (i.e., PCL/keratin composite fiber). Water soluble keratin was first extracted from human hair and mixed with PCL in different ratios. The blended solution of PCL/keratin was transformed into nanofibrous membranes using a laboratory designed electrospinning set up. Fiber morphology and mechanical properties of the obtained nanofiber were observed and measured using scanning electron microscopy and tensile tester. Furthermore, degradability and chemical properties of the nanofiber were studied by FTIR. SEM images showed uniform surface morphology for PCL/keratin fibers of different compositions. These PCL/keratin fibers also showed excellent mechanical properties such as Young&#39;s modulus and failure point. Fibroblast cells were able to attach and proliferate thus proving good cell viability. Based on the characteristics discussed above, we can strongly argue that the blended nanofibers of natural and synthetic polymers can represent an excellent development of composite materials that can be used for different biomedical applications.

Electrospun nanofibers have a high surface area to weight ratio, excellent mechanical integrity, and support cell growth and proliferation. These nanofibers have a wide range of biomedical applications. Here we fabricate keratin/ PCL nanofibers, using the electrospinning technique, and characterize the fibers for possible applications in tissue engineering.

基于角蛋白纳米纤维生物医学工程合成

Electrospinning, due to its versatility and potential for applications in various fields, is being frequently used to fabricate nanofibers. Production of ...

Preparation of Silicon Nanowire Field-effect Transistor for Chemical and Biosensing Applications

Surveillance using biomarkers is critical for the early detection, rapid intervention, and reduction in the incidence of diseases. In this study, we describe the preparation of polycrystalline silicon nanowire field-effect transistors (pSNWFETs) that serve as biosensing devices for biomarker detection. A protocol for chemical and biomolecular sensing by using pSNWFETs is presented. The pSNWFET device was demonstrated to be a promising transducer for real-time, label-free, and ultra-high-sensitivity biosensing applications. The source/drain channel conductivity of a pSNWFET is sensitive to changes in the environment around its silicon nanowire (SNW) surface. Thus, by immobilizing probes on the SNW surface, the pSNWFET can be used to detect various biotargets ranging from small molecules (dopamine) to macromolecules (DNA and proteins). Immobilizing a bioprobe on the SNW surface, which is a multistep procedure, is vital for determining the specificity of the biosensor. It is essential that every step of the immobilization procedure is correctly performed. We verified surface modifications by directly observing the shift in the electric properties of the pSNWFET following each modification step. Additionally, X-ray photoelectron spectroscopy was used to examine the surface composition following each modification. Finally, we demonstrated DNA sensing on the pSNWFET. This protocol provides step-by-step procedures for verifying bioprobe immobilization and subsequent DNA biosensing application.

We describe key steps for biosensing by using polysilicon nanowire field-effect transistors, including the preparation of the device and the immobilization and confirmation of a DNA molecular probe on the nanowire surface, as well as conditions for DNA sensing.

硅纳米线场效应晶体管的研制化学生物传感与应用

Surveillance using biomarkers is critical for the early detection, rapid intervention, and reduction in the incidence of diseases. In this study, we ...

Extraction of Plant-based Capsules for Microencapsulation Applications

Microcapsules derived from plant-based spores or pollen provide a robust platform for a diverse range of microencapsulation applications. Sporopollenin exine capsules (SECs) are obtained when spores or pollen are processed so as to remove the internal sporoplasmic contents. The resulting hollow microcapsules exhibit a high degree of micromeritic uniformity and retain intricate microstructural features related to the particular plant species. Herein, we demonstrate a streamlined process for the production of SECs from Lycopodium clavatum spores and for the loading of hydrophilic compounds into these SECs. The current SEC isolation procedure has been recently optimized to significantly reduce the processing requirements which are conventionally used in SEC isolation, and to ensure the production of intact microcapsules. Natural L. clavatum spores are defatted with acetone, treated with phosphoric acid, and extensively washed to remove sporoplasmic contents. After acetone defatting, a single processing step using 85% phosphoric acid has been shown to remove all sporoplasmic contents. By limiting the acid processing time to 30 hr, it is possible to isolate clean SECs and avoid SEC fracturing, which has been shown to occur with prolonged processing time. Extensive washing with water, dilute acids, dilute bases, and solvents ensures that all sporoplasmic material and chemical residues are adequately removed. The vacuum loading technique is utilized to load a model protein (Bovine Serum Albumin) as a representative hydrophilic compound. Vacuum loading provides a simple technique to load various compounds without the need for harsh solvents or undesirable chemicals which are often required in other microencapsulation protocols. Based on these isolation and loading protocols, SECs provide a promising material for use in a diverse range of microencapsulation applications, such as, therapeutics, foods, cosmetics, and personal care products.

This protocol/manuscript describes a streamlined process for the production of sporopollenin exine capsules (SECs) from Lycopodium clavatum spores and for the loading of hydrophilic compounds into these SECs.

基于植物的提取胶囊微胶囊化应用

Microcapsules derived from plant-based spores or pollen provide a robust platform for a diverse range of microencapsulation applications. Sporopollenin ...

Two Methods for Decellularization of Plant Tissues for Tissue Engineering Applications

The autologous, synthetic, and animal-derived grafts currently used as scaffolds for tissue replacement have limitations due to low availability, poor biocompatibility, and cost. Plant tissues have favorable characteristics that make them uniquely suited for use as scaffolds, such as high surface area, excellent water transport and retention, interconnected porosity, preexisting vascular networks, and a wide range of mechanical properties. Two successful methods of plant decellularization for tissue engineering applications are described here. The first method is based on detergent baths to remove cellular matter, which is similar to previously established methods used to clear mammalian tissues. The second is a detergent-free method adapted from a protocol that isolates leaf vasculature and involves the use of a heated bleach and salt bath to clear the leaves and stems. Both methods yield scaffolds with comparable mechanical properties and low cellular metabolic impact, thus allowing the user to select the protocol which better suits their intended application.

Here we present, and contrast two protocols used to decellularize plant tissues: a detergent-based approach and a detergent-free approach. Both methods leave behind the extracellular matrix of the plant tissues used, which can then be utilized as scaffolds for tissue engineering applications.

组织工程应用中植物组织去细胞的两种方法

The autologous, synthetic, and animal-derived grafts currently used as scaffolds for tissue replacement have limitations due to low availability, poor ...