从威康信托基金会的资金是体谅（批准号085245/Z/08/Z），。诺丁汉纳米技术和纳米科学中心是善意的拉曼系统，为东部地区发展署确认用于资助该设备。

1。的低结垢背景的制备<ol><li>放入到50mL离心管中称出2克的聚（甲基丙烯酸羟乙酯）（PHEMA）（Sigma公司 - 细胞培养测试）。溶解在50毫升95％（体积/体积）乙醇在水中。这通常需要24小时的超声。 </li><li>浸涂环氧官能载玻片（Genetix）与PHEMA溶液。环氧基团会迅速形成共价键的PHEMA涂层。浸涂用镊子保持载玻片和浸渍到溶液中的滑动来实现。通常为5毫米的幻灯片留下未涂布，这是很有用的取向的幻灯片，并也可以作为作为粘接面的阳性对照。然后将幻灯片从PHEMA溶液排出超过1秒的期间，反相并放置干燥10分钟，然后放置在滑动架中的水平位置。 </li><li> PHEMA包被的载玻片，然后留在大气条件下，1周，以允许完全蒸发吨他的溶剂。 </li></ol> 2。单体溶液的制备<ol><li>称取120毫克的光聚合引发剂2,2 - 二甲氧基-2 - 苯基苯乙酮，并加至3毫升二甲基甲酰胺（DMF）中，使4％（重量/体积）的光引发剂的溶液。每个打印运行之前，这最好是新鲜的，所以制成的溶液的质量和体积可以变化，以适应需要多少光引发剂溶液。该溶液是稳定的长达一个月。 </li><li>单体溶液是由通过加入1光引发剂溶液的一部分，3份整齐单体。这是通过移液到384以及源板5μL的光聚合引发剂的单体溶液和15μL。总体积为20μL点形成的理想选择。产量增加需要额外，印迹均匀点之前可以生产。更小的体积，可能会导致不完整的引脚加载。 </li><li>此方法是目前使用的丙烯酸酯/甲基丙烯酸酯单体是可溶的在DMF中所vi限于rtue这些已经探索的组合，仅丙烯酰胺和其它溶剂有可能是适合于印刷过程中。为了实现建议的单体浓度（75％w / w的）液态单体也是必需的，尽管较低的单体浓度可以用于固体单体（单体浓度降低不出现不利改变所得聚合物的表面化学性质的但它是的分子量和玻璃化转变温度可能被改变）。高挥发性单体也很难使用之前由于快速蒸发的单体聚合时，UV固化步骤。将已蒸发根据一个运行可能需要长达6小时，并朝向端运行的挥发性单体的单体溶液的数量从源板。可以通过以下来实现印刷阶段冷却，和使用短打印仅运行使用的挥发性单体。 </li><li>除了少数高亲水性单体如聚（乙二醇）丙烯酸酯，溶解在水相缓冲液的所得聚合物点没有被观察到，因此，不需要的交联单体的使用，虽然还没有被排除。 </li></ol> 3。聚合物芯片的形成在图1中示意性地描绘了用于聚合物微阵列形成的，典型的过程。 <ol><li>微阵列的形成是使用接触机器人（Biodot），使用XYZ载物台（图2）实现的。使用直径为220μm的开槽上的大头针（Arrayit 96B）。在二氯甲烷中通过超声处理10分钟之前印数的，同样也应该被清洗的销保持器的所有引脚应清洗。 </li><li>引脚被装入保持器，的印迹和阵列幻灯片加载，然后用氩气填充整个腔室，以降低含氧量水平低于2000ppm，这是足够的低，以避免由氧猝灭聚合自由基。湿度就是互斗d在30％至40％之间。包括湿度允许PHEMA膨胀允许所形成的聚合物的相互渗透的PHEMA层和成为物理截留到的表面2。 </li><li>印数开始。每次运行包括: <ul><li>将样品装入源板。引脚被降低到的解决方案，在速度为25毫米/秒，2.5秒，在溶液中举行，然后在25 mm / s的（图2）的速度排出。 </li><li>在打印前删除单体溶液从外部引脚引脚必须被抹杀。随后单体交货的quilled部分的引脚，以实现一致的斑形成。干净的载玻片上使用的杂交序列包含33个触点。前四的位置联络的数是10，5，分别为4和3。接下来的四个位置2个触点，并在过去的3个位置接触。为每个联系人的总接触时间为10毫秒，方法和撤离速度为175毫米/秒。由这点，形成的斑点，应该有一个一致的形状和几何结构。在这一点上的故障指示不洁的标签或灰尘的单体溶液中的污染物。 </li><li>的单体溶液，然后印到PHEMA涂敷的载玻片（图2）。一个接触在锁销的移动为175 mm / s的每个点和接触时间为10毫秒，这取决于单体溶液的粘度和表面能量，给出了一个平均的光点直径为400μm。通常为3复制阵列印刷到每个载玻片，共10张幻灯片上印有一个单一的运行。这相当于每个周期的30点。 </li><li>脚在DMF洗涤。新鲜的DMF中的2.5 L的设置用于整个洗涤运行。销流浴搅拌洗涤。 </li><li>与洗涤的同时，照射的刚印过的幻灯片用短波UV（365 nm）的源，在洗涤期间，持续30秒的持续时间为30毫伏/厘米2的密度。 </li></ul></li><li>一归根结底单体溶液印刷阵列用UV照射（365nm）的额外的10分钟。 </li><li>刚印过的阵列被保持在低压力（&lt;50毫乇）1周，以除去未聚合的单体和溶剂。 </li></ol> 4。高通量的表面特性（HTSC） 在图3中示出的高温超导技术的一般方案。中部的自动化的，高吞吐量的方法是与表征装置的聚合物微阵列的取向。在所有的情况下，这是通过使用一个摄像头，给阵列的顶视图。最初，该阵列是与XY移动的阶段旋转对齐。然后位于阵列的拐角点和指定的特定坐标。各聚合物点的位置，然后，可以使用阵列的尺寸预测。 <ol><li>水接触角测量（WCA） <ol><li> WCA使用上躺滴法测量时自动KRUSS DSA 100仪器。 〜400皮升（pL）的体积的单个水滴被分配到各聚合物点的使用压电驱动的打印头。的聚合物点的直径一般是300微米，它的基的聚合物点上的水滴的直径通常为100微米，因此，只有一个测量可以得到每个聚合物现货。 </li><li> XY载物台允许下的打印头7的各聚合物点的自动定位。这是通过使用位于阵列的顶视图，上面的示例，提供的相机。最初，摄像机的位置必须被调整对齐水液滴分配到相机视图的中心，则该数组可以向打印头对齐。 </li><li>高速摄像机记录的液滴触及的表面，随后蒸发后的侧面轮廓。帧捕获的初始冲击的下拉是用来测量接触角。一个圆嵌合下拉利润Ie和接触角随后确定。 </li></ol></li><li>飞行时间二次离子质谱（TOF-SIMS） <ol><li>样品加载到阶段。这涉及到首先衬用干净的铝箔，这有助于电荷补偿，然后通过使用金属螺丝标签，幻灯片。 </li><li>样品载物台放置到输送室的TOF-SIMS和允许抽空至1.6×10 -6毫巴。然后将样品转移到的主要分析室，其通常具有1.0×10 -8毫巴的真空压力。 </li><li> TOF-SIMS测量是使用ION-TOF IV使用单一同位素Bi 3 +的一次离子源工作在25 kV的"揉成模式"操作的仪器进行的。在分析区域，一个10秒的100×100微米面积的芯片，每个聚合物现货收集的正面和负面的二次离子脉冲1 pA的一次离子束扫描光栅第二次采集时间。离子群众的确定，采用了高分辨率飞行时间的分析仪可以准确的质量分配。典型的质量分辨率（M / Z 41）是6000多。低能量的电子（20 eV）的使用，以补偿所造成的在该绝缘表面8带正电的一次离子束的表面充电。 </li></ol></li><li>原子力显微镜（AFM） <ol><li>一个大舞台的原子力显微镜的自动化阶段，需要分析载玻片上的阵列。一个的尺寸或ICON显微镜是用于此目的的理想选择。 </li><li>样品被放置在舞台上和舞台位置驻留右上角的数组。 </li><li>阵列的左下角的位置被发现，允许所有其它点的位置进行插值。 </li><li> A位置列表被生成并送入自动采样功能的软件。 </li><li>原子力显微镜的测量是使用奈米级3000A仪器在利用模E。约300千赫的谐振频率和力常数为40 N / m的硅尖与被使用（Tap300Al，预算传感器）。的5x5的微米区域的聚合物的量9。 </li><li>均方根均方根（RMS）粗糙度测量在这一地区的每个图像使用SPIP批处理软件。每个图像最初是平坦化之前的粗糙度测量。 </li><li>所有的图像都需要一个手动观看，以确定可能需要从粗糙度测量的成像文物。在此步骤中的图像也可以根据常用的表面特征9分类。 </li></ol></li><li> X-射线光电子能谱（XPS） <ol><li>一个微阵列玻片使用双面胶带贴在样品棒，随后装入一个奎托斯轴超XPS仪器的样品室。 </li><li>进入样品室，并等待，直到它到达一个压力低于10 -8乇。 </li><li>相应的操作作为单色器单色化的X-射线源（铝，1486.​​6 eV）的参数，例如，阳极电势和电流（10kV及15毫安），光圈尺寸（110μm），通过能量（80 eV的宽扫描和20 eV的高分辨率扫描）是选择。重点是使用两个相对的角部的微阵列和优化从样品的氧信号的X-射线源。个别点的x和y位置，然后确定和记录的位置列表。 </li><li>编写的程序图，然后运行数据的采集，包括广泛的扫描和高分辨率扫描特定元素的。 </li></ol></li><li>拉曼光谱<ol><li>拉曼光谱是使用拉曼显微镜的LabRam HR仪（Horiba Jobin Yvon公司）的各聚合物点。最初，拉曼信号被校准使用的硅晶片和在520.7厘米-1的拉曼位移的Si。 </li><li>然后将样品放置在舞台上的焦点的激光（波长= 523 nm）的调整，以最大限度地提高CH拉曼位移在2950厘米-1。 </li><li>然后，在微阵列上的左上点的中心的位置设定为原点，在地图上的微阵列是使用阵列功能的LabRam HR软件的安装。 </li><li>累积10采集频谱对每个样品的照射时间为0.5 s。 </li></ol></li></ol> 5。细菌试验该阵列可以接触到许多不同的生物测定法，包括附着和增殖的骨髓间充质干细胞，其他细胞类型和细菌3,10,4。在这里，我们描述了一种细菌附着试验，这是示意性地示出在图4中。 <ol><li>一种细菌菌株转化的GFP表达质粒培养过夜，在37℃下在10毫升的LB培养基在50毫升falcon试管中以200rpm振荡。 </li><li>的细菌培养物的OD 600测量和足够的过夜培养物接种到15毫升的RPMI-1640定义的介质，导致在最终OD 600为0.01。 </li><li>数组是在无菌蒸馏水预洗10分钟以除去任何可溶解组件从阵列（未聚合的单体，低聚物和溶剂） </li><li>洗水阵列载玻片放置在一个干净的陪替氏培养皿和UV灭菌10分钟。 </li><li>与阵列的滑动放入陪替氏培养皿和15毫升的接种的RMPI-1640培养基中加入。同时另一个幻灯片被放置在陪替氏培养皿中，并温育与非innoculatd的RPMI-1640作为对照。 </li><li>相片数是在37℃下温育72小时的振荡在60rpm。 </li><li>幻灯片用15毫升无菌PBS洗涤两次。 </li><li>幻灯片洗涤两次，用15毫升无菌蒸馏水。 </li><li>幻灯片空气干燥。 </li><li>幻灯片成像使用GenePix自动加载磁带机的4200AL扫描仪（Molecular Devices公司，美国）与488 nm的激光激发和标准的蓝光发射滤波器（510-560nm处）。具有代表性的聚合物自体荧光图像如图2中所示。这应该是尽快完成，以避免腐烂的GFP。如果这是不可能的细胞应接受固定。此外，列入适当的阳性对照，用已知的细菌反应是必须要能够正常化的结果，并允许在不同的日子进行定量比较的实验。聚合物的荧光强度从点获得使用GenePix Pro 6的软件（Molecular Devices公司，美国）。这是为幻灯片培养细菌和媒体的。要查找的荧光由于细菌生长的介质上的控制的荧光中减去从与细菌孵育在幻灯片上的荧光测定。 </li><li>幻灯片还可以染色20μMSYTO17核酸染料（Invitrogen公司，英国）在室温下30分钟，使用的是卡尔蔡司LSM 700激光扫描显微镜拍摄的2009年与ZEN成像软件（卡尔蔡司，德国）。细菌的覆盖范围的表面上使用开源图像J 1.44软件（国立卫生研究院，美国）。 </li></ol> 6。代表性的成果印刷的条件进行了优化，打印最高质量的聚合物微阵列。湿度应保持在30％至40％之间。剥离的聚合物在含水环境中经常观察到的斑点印在低于30％的湿度的阵列，这表明此湿度不足以溶胀PHEMA层和允许的物理截留的聚合物基板。改变的聚合物点的直径，可以进一步增加的湿度，但，这是依赖于的单体化学。例如，对于聚合溶液的等体积的印刷和湿度上升至40％至80％的光点直径减小从430微米至370微米的单体含有亲水性的乙烯乙二醇部分等体积，而为莫nomer含有疏水性的脂族碳环结构的光点直径的增加从290微米至350微米（图5）。 的聚合度可以监视使用拉曼光谱测量的C = C的拉曼位移被检测到的在1640厘米-1，与在1720cm -1的C = O喇曼位移应被标准化。的拉曼光谱测定的聚合物聚合多样化的暴露于紫外线（图6）的点。 C = C:C = O比减少紫外线照射增加，从0到50秒，于是没有进一步下降的C = C:C = O比率进一步紫外线照射（图6）。拉曼光谱进行测定聚合物斑点在多样化的O 2的水平和C =Ç拉曼位移被观察到的O 2的水平降低至2000ppm，但是没有进一步降低，观察到的O 2低于这个水平（图7A聚合）。拉曼光谱的真空抽气S也展示了能力TEP，以除去未聚合的单体。到抽真空之前的C = C的拉曼位移是更大的聚合的聚合物用2000ppm的（图7A）相比，在3300 ppm的，然而，抽真空后的拉曼位移的高度是没有区别（图7B），这表明所有的未聚合的单体真空提取步骤期间已被移除。来概括，聚合条件包括30-40％的湿度，紫外线曝光大于50秒的O 2的水平低于2000ppm与印刷后7天的真空提取步骤。 印刷和真空萃取后的成功聚合物的聚合点可以通过简单的光学显微镜的识别和异常点形态评估。通常情况下，斑点出现圆形和均匀的，在左侧上的图8中所示。几何形状的变化，可能的原因是损坏或不洁针。对于一个小数量的单体的组合，我们已经观察到畸形斑点，为电子xample一个大的顶部有一个中央点的中央位置，如图8所示，在右侧的小斑点，或一个煎鸡蛋形状的卫星，平坦的位置。这可能是由于通过相分离有关的单体的粘度，亲水性，波动或表面张力的差异，并建议该单体的组合是不兼容这种格式打印之前。的技术（如TOF-SIMS）的聚合物点的附加化学映射也是一个重要的和有时是必要的质量控制步骤，以确定材料的化学成分的分布跨越的斑点和阵列。该技术可以识别一些不可见的光镜的材料的过度蔓延，并确定在个别的聚合物景点相分离。 <img alt="图1" src="/files/ftp_upload/3636/3636fig1.jpg" /> 图1原理图描绘涉及的各个步骤中形成的口服lymer点。 <img alt="图2" src="/files/ftp_upload/3636/3636fig2.jpg" /> 图2。引脚的印刷方法，包括初始加载的引脚与单体在源板，然后在基板上沉积单体接触的示意图。 XYZ机器人手臂的控制引脚。插图示出了典型的图像从生产后的数组中的自发荧光。 <img alt="图3" src="/files/ftp_upload/3636/3636fig3.jpg" /> 图3。示意图HTSC生物测定施加到聚合物微阵列研究与突出的技术。 <img alt="图4" src="/files/ftp_upload/3636/3636fig4.jpg" /> 图4的细菌附着试验的示意图。 <img alt="图5" src="/files/ftp_upload/3636/3636fig5.jpg" /> 图5，P印刷olymer光斑直径在多样化的湿度为两种不同的单体:4 - 叔丁基环己基丙烯酸酯，二（乙二醇）乙基醚甲基丙烯酸酯。 <img alt="图6" src="/files/ftp_upload/3636/3636fig6.jpg" /> 图6的拉曼强度的比例的C = C的拉曼位移1640cm-1 处的C = O从4 -叔丁基环己基丙烯酸酯的聚合物点具有不同的紫外线暴露在1720cm -1的喇曼位移。误差棒相当于一个标准偏差（n = 3）。 <img alt="图7" src="/files/ftp_upload/3636/3636fig7.jpg" /> 图7的 Raman光谱测定聚合物斑点印在多样化的O 2的水平的4 -叔丁基环己基丙烯酸酯，左侧的各频谱表示，（A）的前和（B）后，抽真空。的C = C的拉曼位移1640cm-1 处和C = O的喇曼位移在1720cm -1的拉曼强度的比率</su对&gt;所示的各频谱的权利。 <img alt="图8" src="/files/ftp_upload/3636/3636fig8.jpg" /> 图8。光镜图像的两种聚合物的斑点。左边示出了形成良好的点上的光斑，而右侧上的光斑是含有单体的分布很不均匀的点的一个例子。比例尺为500μm。

<ol>
	<li>Hook, A. L., Anderson, D. G., Langer, R., Williams, P., Davies, M. C., Alexander, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+throughput+methods+applied+in+biomaterial+development+and+discovery.">High throughput methods applied in biomaterial development and discovery.</a> Biomaterials. 31, 187-198 (2010).</li><li>Anderson, D. G., Levenberg, S., Langer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanoliter-scale+synthesis+of+arrayed+biomaterials+and+application+to+human+embryonic+stem+cells.">Nanoliter-scale synthesis of arrayed biomaterials and application to human embryonic stem cells.</a> Nature Biotechnology. 22, 863-866 (2004).</li><li>Mei, Y., Saha, K., Bogatyrev, S. R., Yang, J., Hook, A. L., Kalcioglu, Z. I., Cho, S. W., Mitalipova, M., Pyzocha, N., Rojas, F., Van Vliet, K. J., Davies, M. C., Alexander, M. R., Langer, R., Jaenisch, R., Anderson, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combinatorial+development+of+biomaterials+for+clonal+growth+of+human+pluripotent+stem+cells.">Combinatorial development of biomaterials for clonal growth of human pluripotent stem cells.</a> Nature Materials. 9, 768-778 (2010).</li><li>Pernagallo, S., Wu, M., Gallagher, M. P., Bradley, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Colonising+new+frontiers-microarrays+reveal+biofilm+modulating+polymers.">Colonising new frontiers-microarrays reveal biofilm modulating polymers.</a> Journal of Materials Chemistry. 21, 96-101 (2011).</li><li>Yang, J., Mei, Y., Hook, A. L., Taylor, M., Urquhart, A. J., Bogatyrev, S. R., Langer, R., Anderson, D. G., Davies, M. C., Alexander, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polymer+surface+functionalities+that+control+human+embryoid+body+cell+adhesion+revealed+by+high+throughput+surface+characterization+of+combinatorial+material+microarrays.">Polymer surface functionalities that control human embryoid body cell adhesion revealed by high throughput surface characterization of combinatorial material microarrays.</a> Biomaterials. 31, 8827-8838 (2010).</li><li>Urquhart, A. J., Anderson, D. G., Taylor, M., Alexander, M. R., Langer, R., Davies, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+throughput+surface+characterisation+of+a+combinatorial+material+library.">High throughput surface characterisation of a combinatorial material library.</a> Advanced Materials. 19, 2486-2491 (2007).</li><li>Taylor, M., Urquhart, A. J., Zelzer, M., Davies, M. C., Alexander, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Picoliter+water+contact+angle+measurement+on+polymers.">Picoliter water contact angle measurement on polymers.</a> Langmuir. 23, 6875-6878 (2007).</li><li>Urquhart, A. J., Taylor, M., Anderson, D. G., Langer, R., Davies, M. C., Alexander, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TOF-SIMS+analysis+of+a+576+micropatterned+copolymer+array+to+reveal+surface+moieties+that+control+wettability.">TOF-SIMS analysis of a 576 micropatterned copolymer array to reveal surface moieties that control wettability.</a> Analytical Chemistry. 80, 135-142 (2008).</li><li>Hook, A. L., Yang, J., Chen, X., Roberts, C. J., Mei, Y., Anderson, D. G., Langer, R., Alexander, M. R., Davies, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acrylate+polymers+with+hydro-responsive+topography.">Acrylate polymers with hydro-responsive topography.</a> Soft Matter. 7, 7194-9197 (2011).</li><li>Pernagallo, S., Unciti-Broceta, A., Diaz-Mochon, J. J., Bradley, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deciphering+cellular+morphology+and+biocompatibility+using+polymer+microarrays.">Deciphering cellular morphology and biocompatibility using polymer microarrays.</a> Biomedical Materials. 3, (2008).</li></ol>

我们什么都没有透露。

生物测定中的新型聚合物通过筛选数百聚合物芯片已被成功地用于新材料的发现和识别"点击"材料随后可以被按比例放大的有用设备。在这种情况下，可以采用描述的表面特征可能随后的生物测定和专门的"命中"的材料，更详细地研究这些材料。 HTSC是不提供给实验者采用这种方法，这种策略可能会感兴趣的。但是，为了充分利用聚合物微阵列研究的生物物质的相互作用，整个阵列的数百材料应进行分析之前，生物测定中使用的HTSC的方法，可在随后被用来观测一般结构与功能的趋势。 接触印刷依赖于向上和向下滑动销保持器内自由的金属销。引脚与引脚保持清洁是最重要的是确保printing成功，应严格进行。开始前一个印刷运行适当的运动的销保持器内的销可以通过执行干运行测试，没有单体本。应继续，直到锁销的移动来实现再现的清洗工序。 可观的想法应该进入的单体混合物的设计。为了容易地产生一个组合文库的聚合物，通过混合数按不同比例的单体的共聚物形成的数百个。通常情况下，我们生产的576个成员库，形成了一个24×24的阵列，这是适用于载玻片上的几何形状。为了产生一个组合库，探索最组合的空间最简单的方法是将24个单体，成对地以2:1的比率混合。可替换地，在阵列内的组成梯度的纳入用于使观察的趋势，它允许为d的最佳的单体组合物是有用的etermined。作为一个例子，可以使用这22单体作为第一组分依次稀释1 6第二组分中的共聚单体混合物。如果5稀释使用，例如在90:10，75:25，50:50，25:75和10:90的比率混合的第一和第二部件，这将导致在488独特的共聚物溶液。为了使总数达到576个，复制所用的单体可以推出，这往往是一个重要的参考样品的均聚物。 576个单体的解决方案，应分配到2 384孔板。对于编程的机器人，它是更容易在单体的位置上有两个相同的板，因此，单体溶液应均匀地拆分在两个板之间。 一个显着的时间内，可以保存在源板的准备，通过使用多道移液器，源板的设计应确定在利用多道移液器使用。&lt;/ P&gt; 为了实现自动化的阵列的高温超导点的位置必须成功地将与表征设备。丙烯酸酯阵列的间距通常为500-1000微米的聚合物的光点直径为300μm。大多数XY工作台具有低于10微米的分辨率，从而有足够的耐受性的表面特性评价装置，以可靠地访问的阵列的位置，一旦正确的尺寸已被输入到样品定位软件。到自动定位的限制实际上是在阵列的准确的印刷。重要的是要防止在基板上的印刷阶段的运动，以确保准确的打印使用真空抽吸或弹簧夹连同适当的幻灯片尺寸（注意，一个美国和欧盟标准的幻灯片大小存在）。 TOF-SIMS是一个极其表面的敏感技术，将遵守任何污染的样本。因此，必须小心至上以避免与该表面接触。样品应仅被处理，但感兴趣的表面不接触，用干净的手套（优选聚乙烯）与新鲜清洁的镊子。我们通常用氯仿和己烷。最好是在测量之前样品储存的样品支架，保持相隔的幻灯片，例如5滑动架或20滑动架。 该阵列是专门设计用于与许多生物测定格式和读数兼容，即，所使用的基板是一种非常适合于荧光扫描仪和光学显微镜的显微镜载玻片。这意味着探索多种生物材料的相互作用是非常适合的格式。此外，该格式允许平行数百材料进行筛选。这使得更多的材料进行筛选，比传统的方法，使每个单独进行筛选新材料化学。的范围扩大生物材料的相互作用使s的生物表面的相互作用，以及对于一个给定的应用程序中找到的最佳材料是澄清的机制。

<table border="1"><tbody><tr><td> 的试剂/设备名称 </td><td> 公司 </td><td> 目录/型号 </td></tr><tr><td>环氧幻灯片</td><td> Genetix </td><td> K2652 </td></tr><tr><td>联系打印机</td><td> Biodot </td><td> XYZ3060平台</td></tr><tr><td>金属针</td><td> ArrayIt </td><td> 946MP6B </td></tr><tr><td> TOF-SIMS仪的</td><td> ION-TOF </td><td></td></tr><tr><td> XPS仪器</td><td>奎托斯</td><td></td></tr><tr><td> WCA设备</td><td> KRUSS </td><td> DSA 100 </td></tr><tr><td> AFM </td><td>布鲁克</td><td>尺寸图标</td></tr><tr><td>的RPMI-1640细胞培养基</td><td> Sigma-Aldrich公司</td><td> R0883 </td></tr><tr><td> SYTO17 </td><td> Invitrogen公司</td><td> S-7579 </td></tr></tbody></table>

polymer microarrays for high throughput discovery of biomaterials

混合是一个单元操作成均匀的混合物，结合两个或更多个组件。此工作包括使用在线静态混合器混合两种粘性液体流。混频器是一个分裂和重新结合的设计，采用剪切和拉伸流增加的部件之间的界面接触。原型分割和重组（SAR）的混频器，构建通过对齐了一系列薄的激光切割的聚（甲基丙烯酸甲酯）甲基丙烯酸甲酯（PMMA）板保持就位在PVC管。在此设备中的混合的照片在图中示出。 1。红染料添加到测试流体的一部分，并用作被混入主要组件（未染色）的次要组分。在混频器的入口，注入层分成两层的示踪流体流经它的混合部分。在每个后续的混合部分，水平层的数目是重复的。最终，单个数据流被均匀地分散的染料苏氨酸oughout设备的横截面。 使用一种非牛顿型的测试流体的0.2％的卡波姆和掺杂的示踪剂的类似组合物的流体，混合中的单元被可视化，使用磁共振成像（MRI）。 MRI是一个非常强大的实验探针分子的化学和物理环境以及样品的结构尺度从微米到厘米的长度。在这些技术的广泛应用，这种敏感性导致物理，化学和/或生物性质，从人类的食物至多孔介质中的1，2的材料的特征。这里所使用的设备和条件，是适合用于成像含有大量的NMR移动1 H，如普通的水和有机液体，包括油的液体。传统MRI利用超导磁铁有不适合用于工业环境的实验室内（ 图2），不便于携带。最新进展我Ň磁体技术，已批准建设大容量，符合工业磁铁，适用于成像处理流程。在这里，MRI提供空间分辨的成分浓度在不同的轴向位置，在混合过程中。这项工作的文件实时高粘度流体的混合，通过分布混合应用到个人护理产品。

Watch this Scientific Journal Video about Polymer Microarrays for High Throughput Discovery of Biomaterials at JoVE.com

Polymer Microarrays for High Throughput Discovery of Biomaterials

形成的聚合物的微阵列，使用一个芯片上的光聚合技术的描述。使用原子力显微镜（AFM），水的接触角的测量，X-射线光电子能谱法和时间飞行二次离子质谱法和细胞附着试验的高吞吐量表面表征也被描述。

聚合物芯片高通量发现的生物材料

The discovery of novel biomaterials that are optimized for a specific biological application is readily achieved using polymer microarrays, which allows a ...

polymer-microarrays-for-high-throughput-discovery-of-biomaterials

Research

JoVE Journal

Bioengineering

14.4K 次观看.  University of Nottingham. 形成的聚合物的微阵列，使用一个芯片上的光聚合技术的描述。使用原子力显微镜（AFM），水的接触角的测量，X-射线光电子能谱法和时间飞行二次离子质谱法和细胞附着试验的高吞吐量表面表征也被描述。

视频: Polymer Microarrays for High Throughput Discovery of Biomaterials

Candida albicans Biofilm Chip (CaBChip) for High-throughput Antifungal Drug Screening

Candida albicans remains the main etiological agent of candidiasis, which currently represents the fourth most common nosocomial bloodstream infection in US hospitals1. These opportunistic infections pose a growing threat for an increasing number of compromised individuals, and carry unacceptably high mortality rates. This is in part due to the limited arsenal of antifungal drugs, but also to the emergence of resistance against the most commonly used antifungal agents. Further complicating treatment is the fact that a majority of manifestations of candidiasis are associated with the formation of biofilms, and cells within these biofilms show increased levels of resistance to most clinically-used antifungal agents2. Here we describe the development of a high-density microarray that consists of C. albicans nano-biofilms, which we have named CaBChip3. Briefly, a robotic microarrayer is used to print yeast cells of C. albicans onto a solid substrate. During printing, the yeast cells are enclosed in a three dimensional matrix using a volume as low as 50 nL and immobilized on a glass substrate with a suitable coating. After initial printing, the slides are incubated at 37 &deg;C for 24 hours to allow for biofilm development. During this period the spots grow into fully developed "nano-biofilms" that display typical structural and phenotypic characteristics associated with mature C. albicans biofilms (i.e. morphological complexity, three dimensional architecture and drug resistance)4. Overall, the CaBChip is composed of ~750 equivalent and spatially distinct biofilms; with the additional advantage that multiple chips can be printed and processed simultaneously. Cell viability is estimated by measuring the fluorescent intensity of FUN1 metabolic stain using a microarray scanner. This fungal chip is ideally suited for use in true high-throughput screening for antifungal drug discovery. Compared to current standards (i.e. the 96-well microtiter plate model of biofilm formation5), the main advantages of the fungal biofilm chip are automation, miniaturization, savings in amount and cost of reagents and analyses time, as well as the elimination of labor intensive steps. We believe that such chip will significantly speed up the antifungal drug discovery process.

Candida albicans Biofilm Chip CaBChip for High-throughput Antifungal Drug Screening

We have developed a high-density microarray platform consisting of 3D nano-biofilms of C. albicans called CaBChip. The susceptibility profile of drugs tested on a CaBChip is comparable to the conventional 96-well plate model, suggesting that the fungal chip is ideally suited for true high-throughput screening of antifungal drugs.

白色念珠菌生物膜芯片（CA BChip）为高通量的抗真菌药物筛选

Candida  albicans remains the main etiological agent of candidiasis, which  currently represents the fourth most common nosocomial bloodstream infection ...

科研

生物工程

Combinatorial Synthesis of and High-throughput Protein Release from Polymer Film and Nanoparticle Libraries

Polyanhydrides are a class of biomaterials with excellent biocompatibility and drug delivery capabilities. While they have been studied extensively with conventional one-sample-at-a-time synthesis techniques, a more recent high-throughput approach has been developed enabling the synthesis and testing of large libraries of polyanhydrides1. This will facilitate more efficient optimization and design process of these biomaterials for drug and vaccine delivery applications. The method in this work describes the combinatorial synthesis of biodegradable polyanhydride film and nanoparticle libraries and the high-throughput detection of protein release from these libraries. In this robotically operated method (Figure 1), linear actuators and syringe pumps are controlled by LabVIEW, which enables a hands-free automated protocol, eliminating user error. Furthermore, this method enables the rapid fabrication of micro-scale polymer libraries, reducing the batch size while resulting in the creation of multivariant polymer systems. This combinatorial approach to polymer synthesis facilitates the synthesis of up to 15 different polymers in an equivalent amount of time it would take to synthesize one polymer conventionally. In addition, the combinatorial polymer library can be fabricated into blank or protein-loaded geometries including films or nanoparticles upon dissolution of the polymer library in a solvent and precipitation into a non-solvent (for nanoparticles) or by vacuum drying (for films). Upon loading a fluorochrome-conjugated protein into the polymer libraries, protein release kinetics can be assessed at high-throughput using a fluorescence-based detection method (Figures 2 and 3) as described previously1. This combinatorial platform has been validated with conventional methods2 and the polyanhydride film and nanoparticle libraries have been characterized with 1H NMR and FTIR. The libraries have been screened for protein release kinetics, stability and antigenicity; in vitro cellular toxicity, cytokine production, surface marker expression, adhesion, proliferation and differentiation; and in vivo biodistribution and mucoadhesion1-11. The combinatorial method developed herein enables high-throughput polymer synthesis and fabrication of protein-loaded nanoparticle and film libraries, which can, in turn, be screened in vitro and in vivo for optimization of biomaterial performance.

This method describes the combinatorial synthesis of biodegradable polyanhydride film and nanoparticle libraries and the high-throughput detection of protein release from these libraries.

组合合成和高通量的蛋白质释放的聚合物薄膜和纳米粒子库

Polyanhydrides are a class of biomaterials with excellent biocompatibility and drug delivery capabilities. While they have been studied extensively with ...

Evaluation of Biomaterials for Bladder Augmentation using Cystometric Analyses in Various Rodent Models

Renal function and continence of urine are critically dependent on the proper function of the urinary bladder, which stores urine at low pressure and expels it with a precisely orchestrated contraction. A number of congenital and acquired urological anomalies including posterior urethral valves, benign prostatic hyperplasia, and neurogenic bladder secondary to spina bifida/spinal cord injury can result in pathologic tissue remodeling leading to impaired compliance and reduced capacity1. Functional or anatomical obstruction of the urinary tract is frequently associated with these conditions, and can lead to urinary incontinence and kidney damage from increased storage and voiding pressures2. Surgical implantation of gastrointestinal segments to expand organ capacity and reduce intravesical pressures represents the primary surgical treatment option for these disorders when medical management fails3. However, this approach is hampered by the limitation of available donor tissue, and is associated with significant complications including chronic urinary tract infection, metabolic perturbation, urinary stone formation, and secondary malignancy4,5.
Current research in bladder tissue engineering is heavily focused on identifying biomaterial configurations which can support regeneration of tissues at defect sites. Conventional 3-D scaffolds derived from natural and synthetic polymers such as small intestinal submucosa and poly-glycolic acid have shown some short-term success in supporting urothelial and smooth muscle regeneration as well as facilitating increased organ storage capacity in both animal models and in the clinic6,7. However, deficiencies in scaffold mechanical integrity and biocompatibility often result in deleterious fibrosis8, graft contracture9, and calcification10, thus increasing the risk of implant failure and need for secondary surgical procedures. In addition, restoration of normal voiding characteristics utilizing standard biomaterial constructs for augmentation cystoplasty has yet to be achieved, and therefore research and development of novel matrices which can fulfill this role is needed.
In order to successfully develop and evaluate optimal biomaterials for clinical bladder augmentation, efficacy research must first be performed in standardized animal models using detailed surgical methods and functional outcome assessments. We have previously reported the use of a bladder augmentation model in mice to determine the potential of silk fibroin-based scaffolds to mediate tissue regeneration and functional voiding characteristics.11,12 Cystometric analyses of this model have shown that variations in structural and mechanical implant properties can influence the resulting urodynamic features of the tissue engineered bladders11,12. Positive correlations between the degree of matrix-mediated tissue regeneration determined histologically and functional compliance and capacity evaluated by cystometry were demonstrated in this model11,12. These results therefore suggest that functional evaluations of biomaterial configurations in rodent bladder augmentation systems may be a useful format for assessing scaffold properties and establishing in vivo feasibility prior to large animal studies and clinical deployment. In the current study, we will present various surgical stages of bladder augmentation in both mice and rats using silk scaffolds and demonstrate techniques for awake and anesthetized cystometry.

Surgical stages of bladder augmentation are described using 3-D scaffolds in murine and rat models. To test the efficacy of biomaterial configurations for use in bladder augmentation, techniques for both awake and anesthetized cystometry are presented.

膀胱膀胱分析各种啮齿类动物模型中使用的增广的生物材料评价

Renal function and continence of urine are critically dependent on the proper function of the urinary bladder, which stores urine at low pressure and ...

A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo

Insufficient vascularization is considered to be one of the main factors limiting the clinical success of tissue-engineered constructs. In order to evaluate new strategies that aim at improving vascularization, reliable methods are required to make the in-growth of new blood vessels into bio-artificial scaffolds visible and quantify the results. Over the past couple of years, our group has introduced a full skin defect model that enables the direct visualization of blood vessels by transillumination and provides the possibility of quantification through digital segmentation. In this model, one surgically creates full skin defects in the back of mice and replaces them with the material tested. Molecules or cells of interest can also be incorporated in such materials to study their potential effect. After an observation time of one&#8217;s own choice, materials are explanted for evaluation. Bilateral wounds provide the possibility of making internal comparisons that minimize artifacts among individuals as well as of decreasing the number of animals needed for the study. In comparison to other approaches, our method offers a simple, reliable and cost effective analysis. We have implemented this model as a routine tool to perform high-resolution screening when testing vascularization of different biomaterials and bio-activation approaches.

A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo

Vascularization is key to approaches in successful tissue engineering. Therefore, reliable technologies are required to evaluate the development of vascular networks in tissue-constructs. Here we present a simple and cost-effective method to visualize and quantify vascularization in vivo.

一个完整的皮肤缺损模型，以评估生物材料的血管化<em&gt;体内</em

Insufficient vascularization is considered to be one of the main factors limiting the clinical success of tissue-engineered constructs. In order to ...

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. ...

Using Tomoauto: A Protocol for High-throughput Automated Cryo-electron Tomography

Cryo-electron tomography (Cryo-ET) is a powerful three-dimensional (3-D) imaging technique for visualizing macromolecular complexes in their native context at a molecular level. The technique involves initially preserving the sample in its native state by rapidly freezing the specimen in vitreous ice, then collecting a series of micrographs from different angles at high magnification, and finally computationally reconstructing a 3-D density map. The frozen-hydrated specimen is extremely sensitive to the electron beam and so micrographs are collected at very low electron doses to limit the radiation damage. As a result, the raw cryo-tomogram has a very low signal to noise ratio characterized by an intrinsically noisy image. To better visualize subjects of interest, conventional imaging analysis and sub-tomogram averaging in which sub-tomograms of the subject are extracted from the initial tomogram and aligned and averaged are utilized to improve both contrast and resolution. Large datasets of tilt-series are essential to understanding and resolving the complexes at different states, conditions, or mutations as well as obtaining a large enough collection of sub-tomograms for averaging and classification. Collecting and processing this data can be a major obstacle preventing further analysis. Here we describe a high-throughput cryo-ET protocol based on a computer-controlled 300kV cryo-electron microscope, a direct detection device (DDD) camera and a highly effective, semi-automated image-processing pipeline software wrapper library tomoauto developed in-house. This protocol has been effectively utilized to visualize the intact type III secretion system (T3SS) in Shigella flexneri minicells. It can be applicable to any project suitable for cryo-ET.

We present a protocol on how to utilize high-throughput cryo-electron tomography to determine high resolution in situ structures of molecular machines. The protocol permits large amounts of data to be processed, avoids common bottlenecks and reduces resource downtime, allowing the user to focus on important biological questions.

使用Tomoauto:一个协议，用于高通量自动化低温电子断层扫描

Cryo-electron tomography (Cryo-ET) is a powerful three-dimensional (3-D) imaging technique for visualizing macromolecular complexes in their native ...

A Microfluidic Platform for High-throughput Single-cell Isolation and Culture

Studying the heterogeneity of single cells is crucial for many biological questions, but is technically difficult. Thus, there is a need for a simple, yet high-throughput, method to perform single-cell culture experiments. Here, we report a microfluidic chip-based strategy for high-efficiency single-cell isolation (~77%) and demonstrate its capability of performing long-term single-cell culture (up to 7 d) and cellular heterogeneity analysis using clonogenic assay. These applications were demonstrated with KT98 mouse neural stem cells, and A549 and MDA-MB-435 human cancer cells. High single-cell isolation efficiency and long-term culture capability are achieved by using different sizes of microwells on the top and bottom of the microfluidic channel. The small microwell array is designed for precisely isolating single-cells, and the large microwell array is used for single-cell clonal culture in the microfluidic chip. This microfluidic platform constitutes an attractive approach for single-cell culture applications, due to its flexibility of adjustable cell culture spaces for different culture strategies, without decreasing isolation efficiency.

Here, we present a protocol for isolating and culturing single cells with a microfluidic platform, which utilizes a new microwell design concept to allow for high-efficiency single cell isolation and long-term clonal culture.

高通量单细胞的分离和文化的微流体平台

Studying the heterogeneity of single cells is crucial for many biological questions, but is technically difficult. Thus, there is a need for a simple, yet ...

High-throughput Identification of Bacteria Repellent Polymers for Medical Devices

Medical devices are often associated with hospital-acquired infections, which place enormous strain on patients and the healthcare system as well as contributing to antimicrobial resistance. One possible avenue for the reduction of device-associated infections is the identification of bacteria-repellent polymer coatings for these devices, which would prevent bacterial binding at the initial attachment step. A method for the identification of such repellent polymers, based on the parallel screening of hundreds of polymers using a microarray, is described here. This high-throughput method resulted in the identification of a range of promising polymers that resisted binding of various clinically relevant bacterial species individually and also as multi-species communities. One polymer, PA13 (poly(methylmethacrylate-co-dimethylacrylamide)), demonstrated significant reduction in attachment of a number of hospital isolates when coated onto two commercially available central venous catheters. The method described could be applied to identify polymers for a wide range of applications in which modification of bacterial attachment is important.

A high-throughput microarray method for the identification of polymers which reduce bacterial surface binding on medical devices is described.

医疗器械细菌高通量鉴定驱蚊聚合物

Medical devices are often associated with hospital-acquired infections, which place enormous strain on patients and the healthcare system as well as ...

3D Microtissues for Injectable Regenerative Therapy and High-throughput Drug Screening

To upgrade traditional 2D cell culture to 3D cell culture, we have integrated microfabrication with cryogelation technology to produce macroporous microscale cryogels (microcryogels), which can be loaded with a variety of cell types to form 3D microtissues. Herein, we present the protocol to fabricate versatile 3D microtissues and their applications in regenerative therapy and drug screening. Size and shape-controllable microcryogels can be fabricated on an array chip, which can be harvested off-chip as individual cell-loaded carriers for injectable regenerative therapy or be further assembled on-chip into 3D microtissue arrays for high-throughput drug screening. Due to the high elastic nature of these microscale cryogels, the 3D microtissues exhibit great injectability for minimally invasive cell therapy by protecting cells from mechanical shear force during injection. This ensures enhanced cell survival and therapeutic effect in the mouse limb ischemia model. Meanwhile, assembly of 3D microtissue arrays in a standard 384-multi-well format facilitates the use of common laboratory facilities and equipment, enabling high-throughput drug screening on this versatile 3D cell culture platform.

This protocol describes the fabrication of elastic 3D macroporous microcryogels by integrating microfabrication with cryogelation technology. Upon loading with cells, 3D microtissues are generated, which can be readily injected in vivo to facilitate regenerative therapy or assembled into arrays for in vitro high-throughput drug screening.

3D Microtissues 用于可注射再生疗法和高通量药物筛选

To upgrade traditional 2D cell culture to 3D cell culture, we have integrated microfabrication with cryogelation technology to produce macroporous ...

Biological Compatibility Profile on Biomaterials for Bone Regeneration

Large non-union bone fractures are a significant challenge in orthopedic surgery. Although auto- and allogeneic bone grafts are excellent for healing such lesions, there are potential complications with their use. Thus, material scientists are developing synthetic, biocompatible biomaterials to overcome these problems. In this study, we present a multidisciplinary platform for evaluating biomaterials for bone repair. We combined expertise from bone biology and immunology to develop a platform including in vitro osteoclast (OC) and osteoblast (OB) assays and in vivo mouse models of bone repair, immunogenicity, and allergenicity. We demonstrate how to perform the experiments, summarize the results, and report on biomaterial biocompatibility. In particular, we tested OB viability, differentiation, and mineralization and OC viability and differentiation in the context of &#946;-tricalcium phosphate (&#946;-TCP) disks. We also tested a &#946;-TCP/Collagen (&#946;-TCP/C) foam which is a commercially available material used clinically for bone repair in a critical-sized calvarial bone defect mouse model to determine the effects on the early phase of bone healing. In parallel experiments, we evaluated immune and allergic responses in mice. Our approach generates a biological compatibility profile of a bone biomaterial with a range of parameters necessary for predicting the biocompatibility of biomaterials used for bone healing and repair in patients.

The number of novel biomaterials engineered for repairing large bone lesions is continuously expanding with the aim to enhance bone healing and overcome the complications associated with bone transplantation. Here, we present a multidisciplinary strategy for pre-clinical biocompatibility testing of biomaterials for bone repair.

骨再生生物材料的生物相容性曲线

Large non-union bone fractures are a significant challenge in orthopedic surgery. Although auto- and allogeneic bone grafts are excellent for healing such ...