This work was partially funded through the Richard Barber Foundation and a Thomas Rumble Fellowship (TJW).

1，水/油/水双重乳液微球生产<ol><li>首先准备2％和0.5％重量/在去离子水中的聚乙烯醇（PVA）诉的解决方案。搅拌溶液，在50℃，直到清楚，这可能需要几个小时。制备的2％体积/体积异丙醇在去离子水中的溶液。 </li><li>制备所需的亲水性蛋白的水溶液。下表提供了示例制剂。 </li></ol><img alt="表1" src="/files/ftp_upload/51517/51517table1highres.jpg" /> 表1：实施例的蛋白质解下列蛋白的解决方案已经成功地封装和使用该协议静电。根据需要，其它亲水蛋白溶液可被使用。 <ol start="3"><li>放置40毫升的0.5％PVA溶液到50ml离心管中，放在一边。 </li><li>在15毫升离心管中溶解300毫克65:35聚在3ml二氯甲烷中（乳酸 - 共 - 乙醇酸）（PLGA）。一种涡流混合器可用于加速的PLGA溶解。 </li><li>结合200微升的蛋白质溶液和4微升2％的PVA溶液。倒入蛋白混合物进入PLGA溶液（步骤1.4）。该解决方案将保持大部分独立。 </li><li>将管放入冰水中的烧杯中。用魔杖超声仪在约10瓦（RMS），搅拌几秒钟（5-10）溶液，直至形成均匀的乳白色乳液。 </li><li>倾乳液进50毫升的试管含0.5％的PVA（步骤1.3）。混合溶液在高速涡流混合器〜20秒。该解决方案将制定一个浑浊的外观。 </li><li>传送乳液到200ml烧杯中并置于搅拌板上，在350 rpm离心2分钟。加入50毫升2％的异丙醇的搅拌板的烧杯中。使混合物继续搅拌至少1小时，以允许DCM蒸发，PLGA硬化。 </li><li>转让吨他微球溶液到离心管中。 </li><li>离心机在425×g离心3分钟。微球将收集在试管的底部并出现白色。小心地从管除去上清液，将微球上，并储存在500ml的瓶子。 </li><li>通过填充管四分之三满并摇动它以重新分布的微球体在液体中用去离子水冲洗该微球。 </li><li>重复步骤1.10和1.11的四倍。 </li><li>以下最后的漂洗，与其它样品再次和地方除去在500ml瓶中的上清液。冻结收集在离心管中的微球，然后冷冻干燥至少24小时。 </li><li>可视化在光学显微镜下，或用扫描电子显微镜，微球。微不超过60微米的大都是有效的静电。如果微球太大，再超声处理或涡旋倍，可能需要在步骤1.6或1.7。 </li> &lt;LI&gt;存放干燥的微球在-20℃的冰箱中。 </li><li>可选：使用蛋白质测定中，按照制造商的说明，从步骤1.10 30测试在500毫升瓶的蛋白质的量。这被用来计算百分数蛋白包封在微球中，减去的量在从什么在生产过程中所用的溶液。 </li></ol>注意：要在可视化微球蛋白质位置添加罗丹明2微克/毫升的PLGA溶液31和封装一FITC标记的蛋白质图1显示了一个例子。 2，静电与微<ol><li>之前制备电纺丝溶液，通过溶解在37℃下创建0.5％w / v的溶液的光引发剂，在去离子水中。这个过程可能需要几个小时。 </li><li>创建一个2％w / v的甲基丙烯酸酯化的透明质酸（的MeHA）（见的Burdick 等人的合成）29，3％w / v的900 kD的聚（环氧乙烷）（PEO）和0.05％的重量/在去离子水中v光引发剂溶液。 <ol><li>计算的MeHA和PEO的正确量为所需的体积。例如，将10毫升静电纺丝溶液需要200毫克的MeHA和300毫克的PEO。 </li><li>溶解于去离子水中的PEO的90％所需的最终体积（9毫升这个例子）。这可能需要几个小时，在37℃下加热搅拌盘或水浴可以加速这一进程。 </li><li>接下来添加的MeHA并用旋涡混合器搅拌溶液，直到清楚。这将只需要几分钟的时间。 </li><li>最后加入0.5％的光引发剂溶液，以填补剩下的10％的体积（1毫升本示例）。 </li></ol></li><li>添加微球体在所需要的浓度高达400毫克/毫升。混在涡旋混合器上的溶液，直到微球均匀地分布在该溶液中。 </li><li>将溶液转移到一个注射器，并附加一个6英寸的18号BLUNT尖的针。 </li><li>发生在注射泵的注射器，并把它设置为免除1.2毫升/小时。 </li><li>胶带铝箔上收集板或型芯的层。这允许容易清理成品支架和存储。一种旋转心轴被用来创建排列的纤维。的平板或固定芯棒将导致随机排列的纤维。 </li><li>从高电压电源的收集装置连接的接地线。正极导线连接到针。 </li><li>调整注射器泵和收集表面，以便有针尖和收集表面的15公分。 </li><li>启动该聚合物泵送，当溶液是在注射器的端部可见，接通电压源和设置电压24千伏注意 ：一旦电压接通时不要触摸系统中的任何金属部分。负责人也可以跳很短的距离，从带电部位的皮肤。 </li><li>运行解决方案，直到ðesired脚手架厚度。当完全关闭电压源和注射器泵。 </li><li>删除箔支架连接。含蛋白质已完成支架储存在-20℃的冰箱中。 </li></ol> 3，蛋白质的生物活性测试<ol><li>制备的细胞培养介质。加入10％体积/体积的胎牛血清，1％体积/体积L-谷氨酰胺和1％体积/体积的青霉素 - 链霉素，以Dulbecco改良的Eagle培养基。 </li><li>选择盖玻片适合完全陷入了孔板。 </li><li>用3（三甲氧基硅基）丙基甲基丙烯酸酯如制造商所述治疗的盖玻片。 Methacrylation增强脚手架坚持盖玻片。 </li><li>静电纺丝之前，请安装甲基丙烯酸盖玻片与可拆卸的双面胶带electrospinner的收集区。纺纱到盖玻片简化了操作和观看。 </li><li>静电纺丝到所需的厚度，如上所述。 </li><li> á压脚提升静电纺丝小心地从心轴上取下盖玻片。放置在支架涂覆的盖玻片成清晰氮气室，并确保所有的氧气被吹洗。 </li><li>放置在10毫瓦/厘米2 365纳米的光的腔室和支架15分钟。后交联处成适当大小的孔板。确保棚架的一面朝上。 </li><li>在杀菌灯30分钟的地方支架进行消毒。如果需要，纤连蛋白或其它蛋白质被用作涂层以增强细胞附着。按照制造商的说明大衣支架。 </li><li>嘉实背根神经节（DRG）由霍伦贝克32如前所述。一DRG将需要为每个支架覆盖盖玻片进行测试。 </li><li>将100-200微升媒体在孔板每个支架。小心将一个背根神经节的媒体液滴每个支架。对于厚的支架可能需要更多的媒体; DRG需要被完全浸没，而不是FLOA婷。 </li><li>孵育所述支架和背根神经节，在37℃下搅拌4小时，以使细胞附着在支架上。 </li><li>培养基灌装到适当的水平井，并将进入孵化器。继续温育4-6天。 </li><li>潜伏期后，小心地从每一个媒体好，用PBS轻轻洗一次。固定的细胞用4％w / v的多聚甲醛固定30分钟。 </li><li>用抗体染色为神经丝染色的细胞。这将允许可视化神经突向外生长的定量。 DAPI也可以用来查看细胞核。一个例子染色协议是由Sundararaghavan和他的同事14描述。 </li><li>使用可视化荧光显微镜细胞。 <ol><li>放置孔平板上的显微镜的阶段。 </li><li>找到了用DAPI过滤器和励磁设置的细胞团。 </li><li>一旦该细胞是切换滤波器来FITC以可视化的扩展的突起。 ü唱在显微镜的线圈功能收集和组合一样多的图像，必要时看到整个结构。重复DAPI，FITC和亮场。 </li></ol></li></ol>

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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Neurons: Methods and Applications for the Cell Biologist. , (2003).</li><li>Boer, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+sciatic+nerve+repair+with+a+poly-lactic-co-glycolic+acid+scaffold+and+nerve+growth+factor+releasing+microspheres.">Rat sciatic nerve repair with a poly-lactic-co-glycolic acid scaffold and nerve growth factor releasing microspheres.</a> Microsurgery. 31, 293-302 (2011).</li><li>Pujic, Z., Goodhill, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+dual+compartment+diffusion+chamber+for+studying+axonal+chemotaxis+in+3D+collagen.">A dual compartment diffusion chamber for studying axonal chemotaxis in 3D collagen.</a> Journal of Neuroscience Methods. 215, 53-59 (2013).</li><li>Madduri, S., di Summa, P., Papalo&#239;zos, M., Kalbermatten, D., Gander, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+controlled+co-delivery+of+synergistic+neurotrophic+factors+on+early+nerve+regeneration+in+rats.">Effect of controlled co-delivery of synergistic neurotrophic factors on early nerve regeneration in rats.</a> Biomaterials. 31, 8402-8409 (2010).</li><li>Xu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polyphosphoester+microspheres+for+sustained+release+of+biologically+active+nerve+growth+factor.">Polyphosphoester microspheres for sustained release of biologically active nerve growth factor.</a> Biomaterials. 23, 3765-3772 (2002).</li><li>Yan, Q., Yin, Y., Li, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+new+PLGL-RGD-NGF+nerve+conduits+for+promoting+peripheral+nerve+regeneration.">Use new PLGL-RGD-NGF nerve conduits for promoting peripheral nerve regeneration.</a> Biomed Eng Online. 11, (2012).</li><li>Gungor-Ozkerim, P. S., Balkan, T., Kose, G. T., Sarac, A. S., Kok, F. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incorporation+of+growth+factor+loaded+microspheres+into+polymeric+electrospun+nanofibers+for+tissue+engineering+applications.">Incorporation of growth factor loaded microspheres into polymeric electrospun nanofibers for tissue engineering applications.</a> J Biomed Mater Res A. , (2013).</li><li>Li, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Encapsulation+of+proteins+in+poly(L-lactide-co-caprolactone)+fibers+by+emulsion+electrospinning.">Encapsulation of proteins in poly(L-lactide-co-caprolactone) fibers by emulsion electrospinning.</a> Colloids Surf B Biointerfaces. 75, 418-424 (2010).</li><li>Wang, C. -. 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The authors have nothing to disclose.

许多研究表明，神经细胞可以由沟槽宽度（纤维取向）和化学信号（生长因子）1,2,10,11,35被引导。静电是一种简便的方法来创建排列的纤维。生长因子促进神经生长，但以它们包括成神经管（NC），需要持续释放的方法。要创建一个更强大的系统，这两个线索，这两个信号应该合并。几种方法已经被先前研究的静电纤维支架用于神经再生中提供蛋白的延长释放，但没有迄今一直能持续释放...

其中在神经组织​​工程的持续挑战是创建一个神经导管（NC），模仿的细胞外基质，其中神经自然生长。有研究表明，细胞多种因素在其环境中，包括机械，地形，粘合剂和化学信号1-3响应。一个在该领域的主要挑战是确定的信号的适当组合，并制造一个系统，可以维持线索长时间，以支持细胞生长4。外周神经元已知喜欢的软衬底，由排列的纤维定向，并且对神经生长因子（NGF）5-7响应。奈米可为周提供化学线索已经示出，以提供改进的功能恢复接近该同种异体移植，目前金标准神经修复8,9。 各种材料和生产方法可用于生产机械和地形人的线索10-13。机械线索是内在所选择的材质，使得选择该应用程序的关键1,13合适的材料。的生产方法，以控制沟槽宽度包括相分离，自组装和电纺丝1,13。对于微型应用程序，微流体，光图案化，蚀刻，盐滤出，或气体的泡沫也可以使用14-17。静电已成为最流行 ​​的方式来设计的纤维基材进行组织培养，由于其灵活性和易于生产13,18-23的。电纺纳米纤维是通过施加高电压的聚合物溶液使其排斥本身并横跨一个短间隙放电24制成。对准支架可以通过收集在纤维上接地的旋转心轴被创建和不结盟支架被收集在固定板25上。粘附信号可以通过涂覆纤维支架机智来实现ħ纤连蛋白或静电26日前缀粘附肽，如RGD，房委会。 化学信号，例如生长因子，是最难以保持在延长的时期，因为它们需要用于控制释放的来源。许多系统已经试图控制释放添加到静电纺丝纤维网具有不同程度的成功。这些方法包括共混静电纺丝，静电纺丝乳液，芯壳电纺和蛋白质结合27。另外，电纺丝是在挥发性溶剂中，可以影响蛋白28的生存能力传统上完成的，因此保持了蛋白质的生物活性必须加以考虑。 这种方法专门针对机械​​相结合，地形，化学和胶粘剂信号来创建一个可调谐的支架周围神经生长。脚手架力学正是通过合成来控制甲基丙烯酸酯化透明质酸（HA）。该methacrylation站点用来连接光反应的交联剂。交联的材料不再是水溶性的，并且是专门分解通过酶29。交联的量的变化的降解速率，机械和材料的其它物理性能。使用HA以30％methacrylation，其具有〜500 Pa的拉伸弹性模量，创建一个软基质中靠近神经组织的天然机制，并且通常优选的由神经元26,29。静电纺丝在旋转心轴被用来创建排列的纤维的地形线索。使用静电与微球提供了支架在延长期限内的化学信号。支持含有NGF用来建立化学信号的神经突生长的微球体。与大多数静电材料医管局易溶于水，使神经生长因子不会在生产过程中遇到的恶劣的溶剂。要添加粘合剂信号时，SCAffold涂覆有纤连蛋白。完成后的系统包含了所有四种类型的上述信号：软（机械）对齐（地形）纤维释放神经生长因子微球（化学）涂有纤维连接蛋白（胶粘剂）。生产与本系统的测试在此协议中描述。 该过程开始于生产用与水包油包水双乳液的微球。该乳液是稳定用表面活性剂，聚乙烯醇（PVA）。的内水相含有蛋白质。因为它被加入到油相中，含有PLGA壳材料溶解在二氯甲烷（DCM），所述表面活性剂产生保护从DCM中的蛋白质相之间的屏障。此乳液是比分散在含有PVA的制造微球体的外表面上的另一水相。稳定的乳化液进行搅拌，以使DCM蒸发。冲洗和冷冻干燥后，只剩下干微球续癌宁的蛋白质。 后微球完成他们准备静电纺丝成支架。首先，你准备静电解决方案。该溶液的粘度为适当的纤维形成的关键。纯HA的解决方案并不能满足这一要求; PEO中加入作为载体的聚合物，以允许静电。将微球加入到溶液中，并静电纺丝所得的纤维支架与整个分布的微球。 一旦生产完成后，将蛋白进行测试，以验证其可行性。要做到这一点，它响应NGF原代细胞可以被使用。该协议采用背根神经节（DRG）从8-10日龄的鸡胚。单元束被接种到含微球填充有NGF或那些是空的支架。如果NGF仍然是可行的，你应该看到在NGF含支架增强轴突的生长。如果NGF不再可行它将不能促进轴突延伸，应该出现类似的控制。 本文所描述的确切过程但是集中在神经支撑，以简单的修改的材料，​​静电纺丝法，以及蛋白的系统可以为各种细胞和组织类型进行优化。

<table border="0" cellpadding="0" cellspacing="0" width="906">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">DAPI</td>
			<td style="width:208px;">Invitrogen</td>
			<td style="width:139px;">D1306</td>
			<td style="width:291px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Irgacure 2959</td>
			<td style="width:208px;">BASF</td>
			<td style="width:139px;">24650-42-8</td>
			<td style="width:291px;">Protect from light</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">PEO 900 kDa</td>
			<td style="width:208px;">Sigma-Aldrich</td>
			<td align="right" style="width:139px;">189456</td>
			<td style="width:291px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:269px;">Methacryloxethyl thiocarbamoyl rhodamine B</td>
			<td style="width:208px;">Polysciences, Inc.</td>
			<td style="width:139px;">23591-100</td>
			<td style="width:291px;">Prepare stock solution in DMSO</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Syringe Pump</td>
			<td style="width:208px;">KD Scientific</td>
			<td style="width:139px;">KDS100</td>
			<td style="width:291px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Power Source</td>
			<td style="width:208px;">Gamma High Voltage</td>
			<td style="width:139px;">ES30P-5W</td>
			<td style="width:291px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Motor</td>
			<td style="width:208px;">Triem Electric Motors, Inc</td>
			<td style="width:139px;">0132022-15</td>
			<td style="width:291px;">Must attach to a custom built mandrel</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Tachometer</td>
			<td style="width:208px;">Network Tool Warehouse</td>
			<td style="width:139px;">ESI-330</td>
			<td style="width:291px;">Use to monitor mandrel speed</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:269px;">Omnicure UV Spot Cure System with collimating adapter</td>
			<td style="width:208px;">EXFO</td>
			<td style="width:139px;">S1000</td>
			<td style="width:291px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Needles</td>
			<td style="width:208px;">Fisher Scientific</td>
			<td style="width:139px;">14-825-16H</td>
			<td style="width:291px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Coverslips</td>
			<td style="width:208px;">Fisher Scientific</td>
			<td style="width:139px;">12-545-81</td>
			<td style="width:291px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Polyvinyl Alcohol</td>
			<td style="width:208px;">Sigma-Aldrich</td>
			<td style="width:139px;">P8136-250G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Isoporopyl Alcohol</td>
			<td style="width:208px;">Sigma-Aldrich</td>
			<td style="width:139px;">I9030-500mL</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Bovine Serum Albumin (BSA)</td>
			<td style="width:208px;">Fisher Scientific</td>
			<td style="width:139px;">BP9703-100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">BSA-FITC</td>
			<td style="width:208px;">Sigma-Aldrich</td>
			<td style="width:139px;">080M7400</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">&#946;-Nerve Growth Factor (NGF)</td>
			<td style="width:208px;">R&#38;D Systems</td>
			<td style="width:139px;">1156-NG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">65:35 Poly-Lactic-Glycolic-Acid (PLGA)</td>
			<td style="width:208px;">Sigma-Aldrich</td>
			<td align="right" style="width:139px;">1001554270</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Dichloromethane</td>
			<td style="width:208px;">Sigma-Aldrich</td>
			<td style="width:139px;">34856-2L</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Coomassie (Bradford) Protein Assay</td>
			<td style="width:208px;">Thermo Scientific</td>
			<td align="right" style="width:139px;">1856209</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">3-(Trimethoxysilyl)propyl methacrylate</td>
			<td style="width:208px;">Sigma-Aldrich</td>
			<td align="right" style="width:139px;">1001558456</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Fibronectin</td>
			<td style="width:208px;">Sigma-Aldrich</td>
			<td style="width:139px;">F2006</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">DMEM</td>
			<td style="width:208px;">Lonza</td>
			<td style="width:139px;">12-604F</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">FBS</td>
			<td style="width:208px;">Atlanta Biologicals</td>
			<td style="width:139px;">S11150</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">PBS</td>
			<td style="width:208px;">Hyclone</td>
			<td style="width:139px;">SH30256.01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Glutamine</td>
			<td style="width:208px;">Fisher Scientific</td>
			<td>G7513</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Pen-Strep</td>
			<td style="width:208px;">Sigma-Aldrich</td>
			<td style="width:139px;">P4333</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Paraformaldehyde</td>
			<td style="width:208px;">Alfa Aesar</td>
			<td style="width:139px;">A11313&#160;</td>
			<td></td>
		</tr>
	</tbody>
</table>

electrospinning growth factor releasing microspheres into fibrous scaffolds

This procedure describes a method to fabricate a multifaceted substrate to direct nerve cell growth. This system incorporates mechanical, topographical, adhesive and chemical signals. Mechanical properties are controlled by the type of material used to fabricate the electrospun fibers. In this protocol we use 30% methacrylated Hyaluronic Acid (HA), which has a tensile modulus of ~500 Pa, to produce a soft fibrous scaffold. Electrospinning on to a rotating mandrel produces aligned fibers to create a topographical cue. Adhesion is achieved by coating the scaffold with fibronectin. The primary challenge addressed herein is providing a chemical signal throughout the depth of the scaffold for extended periods. This procedure describes fabricating&#160;poly(lactic-co-glycolic acid) (PLGA) microspheres that contain Nerve Growth Factor (NGF) and directly impregnating the scaffold with these microspheres during the electrospinning process. Due to the harsh production environment, including high sheer forces and electrical charges, protein viability is measured after production. The system provides protein release for over 60 days and has been shown to promote primary nerve cell growth.

微球50±14微米直径的85％以上的蛋白质封装一贯生产，静电成支架。大小是由成像微球的样本来自三个不同的生产批次确定。 图1，其中捕获在哪里使用商业实验室软件测定在光学显微镜和长度的图像。表示粒度分布的直方图。包封率也由三个单独的微球的批次进行测试，通过量化在生产过程中逃脱的蛋白质。 图2示出了代表性的微球与罗丹明（2微克...

Watch this Scientific Journal Video about Electrospinning Growth Factor Releasing Microspheres into Fibrous Scaffolds at JoVE.com

Electrospinning Growth Factor Releasing Microspheres into Fibrous Scaffolds

This protocol combines electrospinning and microspheres to develop tissue engineered scaffolds to direct neurons. Nerve growth factor was encapsulated within PLGA microspheres and electrospun into Hyaluronic Acid (HA) fibrous scaffolds. The protein bioactivity was tested by seeding the scaffolds with primary chick Dorsal Root Ganglia and culturing for 4-6 days.

静电生长因子释放到微纤维支架

This procedure describes a method to fabricate a multifaceted substrate to direct nerve cell growth.  This system incorporates mechanical, topographical, ...

electrospinning-growth-factor-releasing-microspheres-into-fibrous

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Bioengineering

12.2K 次观看.  Wayne State University. This protocol combines electrospinning and microspheres to develop tissue engineered scaffolds to direct neurons. Nerve growth factor was encapsulated within PLGA microspheres and electrospun into Hyaluronic Acid (HA) fibrous scaffolds. The protein bioactivity was tested by seeding the scaffolds with primary chick Dorsal Root Ganglia and culturing for 4-6 days.