Name: magnetic-, acoustic-, and optical-triple-responsive microbubbles for magnetic hyperthermia and pothotothermal combination cancer therapy
Uploaded: 2020-05-22T14:30:00.000+00:00
Duration: 9 min 1 s
Description: Watch this Scientific Journal Video about Magnetic-, Acoustic-, and Optical-Triple-Responsive Microbubbles for Magnetic Hyperthermia and Pothotothermal Combination Cancer Therapy at JoVE.com

这项工作得到了中国国家自然科学基金委员会（81601608）和NUPTSF（NY216024）的支持。

所有动物实验都是按照OG药物指南批准的动物护理和实验室动物使用指南进行的。这些议定书遵循了OG制药公司实验室动物伦理委员会的准则。
1. 纳米粒子壳微泡 （NSM） 合成
<ol><li>将磁性纳米粒子（Fe3O4，氧化铁）分散在电离水中，形成10毫克/mL的库存溶液。</li>
	<li>将含有离子溶液的管子放入超声波清洁机中 20 分钟。使用前获取均匀分散的 IONP 解决方案。</li>
	<li>添加 150 μL 的去离子水、150 μL 的 10 mM 钠硫酸钠 （SDS） 和 400 μL 的离子体库存溶液，从第 1.1 步开始。在 1.5 mL 离心机管中。</li>
	<li>在冰浴中用脚手架修复同质化器。</li>
	<li>将混合物的管子放在冰浴中，并将同质化器探针精确定位，以浸入混合物溶液中。</li>
	<li>将同质化器速度调整到 20，000 rpm，并打开同质化器 3 分钟。</li>
	<li>关闭同质化物，将管子从冰浴中取出。</li>
	<li>将管子放在管架上，在室温下稳定12小时。</li>
	<li>用磁铁将产生的 NSM 吸附到管壁上，并取出超高分子。然后补充1兆L的新鲜去离子水。</li>
	<li>重复洗涤过程三次，并在 1 mL 的新鲜蒸馏水中重新悬挂 NSM。</li>
	<li>轻微摇晃后，将 10 μL 的 NSM 悬架转移到干净的玻璃滑梯上。</li>
	<li>使用 20 倍放大倍率的荧光显微镜可视化 NSM 形态。确保在随机区域拍照。</li>
	<li>使用开放式纳米测量器 1.2 软件从照片中测量 NSM 的直径。至少计算200个微泡。<ol>
<li>单击"文件"和"打开"以选择要处理的图像文件。图像导入后，按"尺子"。绘制一条与标尺长度相同的红线。</li>
		<li>然后按"设置"|"统治者"，并进入统治者的长度。绘制与图像中单个微泡直径相同的长度的线条。完成所有测量，然后单击"报告"| 查看报告"。</li>
	</ol>
</li>
</ol>2. NSM 的声学响应
<ol><li>用 800 μL 去离子水稀释 200 μL 的 NSM，然后转移到 1.5 mL 离心机管中形成库存溶液。</li>
	<li>连接功能发生器、放大器、阻抗匹配和自制聚焦传感器。将传感器放在人工凹陷底部的中心，并将水听器与示波器连接起来，以监测输出超声波强度（图1）。将电离水加入水槽中，让传感器浸入水中。</li>
	<li>将功能发生器调整到扫描模式，调整频率范围从 10 kHz 到 900 kHz，并将振幅设置为 20 Vpp（电压峰值峰值）。通过放大器将超声波功率调整到 0.1%。每个周期的持续时间为 4s，时间间隔为 1s。</li>
	<li>在管子中准备 1 mL NSM 库存溶液样品，并在自制聚焦传感器顶部用脚手架固定管子。将磁铁连接到管子底部并吸引 NSM。</li>
	<li>打开功能发生器和放大器的电源。应用 5 个周期（25 s） 超声波后关闭功能发生器。取下磁铁并收集包含释放的 IONP 的溶液的 1 mL。在离心机管中加入 1 mL 的去离子水。</li>
	<li>重复步骤 2.5。直到管子里的 Nsm 完全崩溃</li>
	<li>通过电感耦合等离子光学发射光谱 （ICP-OES） 对所有已释放的 ION 进行量化，如前13所述。</li>
</ol>3. NSM 的光学响应
注:在这项工作中，使用了一个激光系统，包含808纳米激光功率和红外热像仪之前描述的徐等人。
<ol><li>激光系统制备<ol>
<li>打开激光的电源，让它加热几分钟。将光纤耦合 808 nm 激光二极管固定在反驳支架上。</li>
		<li>通过光纤将激光束引导到样品阶段，并聚焦在样品阶段，通过凸透镜实现 6 mm 光点（直径）。</li>
		<li>使用激光功率计测量功率输出，并将功率调整到 1 W/cm2。</li>
		<li>修复三脚架上的红外热像仪。打开相机并检查是否工作（例如，监控重点感兴趣的区域 （ROI） 温度）。关闭电源和红外热像仪。</li>
	</ol></li>
	<li>在溶液中进行光热测量<ol>
<li>在 1.5 mL 离心机管中，以不同的 IONP 浓度（1.05 毫克/毫升、1.35 毫克/mL、3.65 毫克/兆升、5 毫克/兆升）准备 1 mL 样品。</li>
		<li>将感兴趣的管子放在激光束的聚焦区域，并记录样品的基线温度。</li>
		<li>打开激光电源和红外热像仪，连续照射样品 10 分钟。同时，实时记录温度。</li>
		<li>经过 10 分钟的辐照后，关闭激光电源和红外热像仪。等待区域温度返回基线。</li>
		<li>重复 3.2.2 到 3.2.4 用于测量其他样品。 注:在 20 °C 下使用电离水作为光热测量的控制。</li>
	</ol></li>
	<li>培养细胞中的光热测量 注:Murine乳腺癌细胞（4T1）被选为研究光热治疗的抑制作用的模型。<ol>
<li>用罗斯韦尔公园纪念研究所-1640（RPMI-1640）中等补充10%的胎儿牛血清（FBS）和1%青霉素喂养细胞。将栽培环境设置为 37 °C 和 5% CO2。</li>
		<li>在T25烧瓶中培养4T1细胞，当达到90%的汇合时，以1:2的比例通过细胞。 注:亚栽培比例可根据不同实验室的特定细胞条件进行调整。</li>
		<li>删除并丢弃培养介质。用1x PBS溶液冲洗细胞层，去除含有试丁酶抑制剂的残留血清。</li>
		<li>在烧瓶中加入 2 mL 的 Trypsin-EDTA 溶液 （0.25%），用于分离。然后加入3 mL的1640中等和吸气细胞轻轻地管道。</li>
		<li>以 500 x g 收集 5 mL 的电池悬架和离心机，3 分钟。</li>
		<li>取出超高纳坦，加入1 mL的新鲜1640介质，形成细胞悬浮。</li>
		<li>将 100 μL 的电池悬架添加到包含 1 mL 培养介质的共聚焦盘中。确保细胞悬浮浓度为 9 x 105/mL，细胞培养共焦盘中的最终细胞浓度为 8.1 x 104/mL。</li>
		<li>将接种的细胞培养共聚焦盘放入孵化器中24小时。</li>
		<li>稀释不同浓度（1.05 毫克/mL，1.35 毫克/mL，3.65 毫克/mL，5 毫克/mL）的离子体样品，使 1 mL 溶液与无血清 1640 介质。</li>
		<li>从焦盘中吸气培养介质，并添加准备好的样品溶液。</li>
		<li>打开激光电源。将激光束聚焦在盘的中心，并将输出功率调整到 1 W/cm2。打开红外热像仪，连续照射焦盘中的细胞 10 分钟。实时记录重点区域的温度。</li>
		<li>关闭激光和红外热像仪。将辐照盘转移到孵化器再持续 24 小时。</li>
		<li>去除并丢弃培养介质，在共焦盘中加入 1 mL 的新鲜培养介质。在菜中加入 5 μL 的钙素-AM 溶液 （1 毫克/兆升）。</li>
		<li>在 37 °C 和 5% CO2 下孵育焦盘 15 分钟。用 1x PBS 溶液冲洗细胞层两次。</li>
		<li>通过共聚焦荧光显微镜观察和成像细胞，激发波长为488纳米，发射波长为500-540纳米。</li>
		<li>随机选择共聚焦图像中的 5 个区域，手动计算每个区域的活 4T1 细胞数量。通过比较所有实验组的活细胞数量与对照组，量化 4T1 细胞的生存能力。 注意:在 20 °C 下使用无血清 1640 介质作为光热测量的控制。使用无激光照射的样品作为细胞生存控制组。</li>
	</ol></li>
	<li>体内光热测量<ol>
<li>准备3只8周大的ICR雄性小鼠，平均体重为25±2克。</li>
		<li>在 20 mL 的去离子水中加入 2 克明胶粉末。将溶液加热到 40 - 50 °C，完全溶解明胶凝胶，形成透明清晰的溶液。</li>
		<li>从 3.4.2 中向解决方案添加 100 毫克离子。</li>
		<li>将凝胶加热到 40 ~ 50 °C，立即将 500 μL 的明胶溶液注入鼠标的右胸垫。</li>
		<li>打开激光电源，将激光束聚焦在兴趣区（小鼠的右胸垫）。将输出功率调整到 1 W/cm2。 打开红外热像仪，连续照射兴趣区 10 分钟。实时记录感兴趣的区域的温度。</li>
		<li>关闭激光和红外热像仪。</li>
		<li>通过CO2 窒息和颈椎脱位或经研究所动物研究委员会批准的任何方法对小鼠实施安乐死。</li>
	</ol></li>
</ol>4. 磁性体温过高测量
注:这里利用了吴等人以前描述的磁性高温系统（21）。
<ol><li>准备磁性高温系统包括交替磁场 （AFM） 发生器和红外热成像摄像机。<ol>
<li>打开冷水机组 10 分钟，然后在中等射频加热机上（即 AFM 发电机）上供电。</li>
		<li>将机器的参数设置为:频率 （f） = 415 kHz，磁场强度 = 1.8 kA/m。</li>
	</ol>
</li>
	<li>在溶液中的磁性体温过高测量<ol>
<li>在 1.5 mL 离心机管中，以不同的离子浓度（1.05 毫克/mL、1.35 毫克/mL、3.65 毫克/mL、5 毫克/兆升）准备 1 mL 的样品。</li>
		<li>将管子放在水冷磁感应铜线圈的中心。</li>
		<li>打开交替磁场 （AFM） 和红外热像仪。连续诱导样品10分钟，实时记录温度。 注:摄像机位于样品顶部，提供样品的横截面视图。</li>
		<li>关闭自动对焦和红外热像仪。等待铜线圈的温度回到基线 10 分钟。 注意:注意高温，避免直接接触手部，等待冷却后再取出样品。</li>
		<li>重复 4.2.2 到 4.2.4 用于测量其他样品。</li>
		<li>关闭中等射频加热机 （AFM） 和冷水机组。 注:在 20 °C 下使用去离子化水作为磁性体温过高测量的控制。</li>
	</ol>
</li>
	<li>体内磁性体温过高测量<ol>
<li>准备3只8周大的ICR雄性小鼠，平均体重为25±2克。</li>
		<li>准备 20 mL 的 10% 明胶凝胶，其中含有 5 毫克/mL IONP 溶液。</li>
		<li>将明胶凝胶加热到 40 - 50 °C，立即将 500 μL 的明胶溶液注入动物的右胸垫。 注:磁诱导体温过高实验使用与体外测试相同的参数在加热机上进行。</li>
		<li>打开交替磁场 （AFM） 和红外热像仪。将小鼠的右胸垫放在水冷磁感应铜线圈的中心。</li>
		<li>打开红外热像仪，连续拍摄 10 分钟的兴趣区域（小鼠的右乳房垫），实时记录兴趣区域的温度。</li>
		<li>感应 10 分钟后关闭机器的电源开关和红外热像仪。等待铜线圈的温度返回基线 10 分钟。</li>
		<li>重复 4.3.4 到 4.3.6 用于测量其他样品。</li>
		<li>关闭中等射频加热机 （AFM） 和冷水机组。 注意:注意高温，避免直接接触手部，等待冷却后再取出样品。</li>
		<li>通过CO2 窒息和颈椎脱位或经研究所动物研究委员会批准的任何方法对小鼠实施安乐死。</li>
	</ol>
</li>
</ol>

<ol>
	<li>Kievit, F. M., Zhang, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+nanotheranostics:+improving+imaging+and+therapy+by+targeted+delivery+across+biological+barriers.">Cancer nanotheranostics: improving imaging and therapy by targeted delivery across biological barriers.</a> Advanced Materials. 23 (36), 217-247 (2011).</li><li>Wu, H., 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=Fe3O4-Based+Multifunctional+Nanospheres+for+Amplified+Magnetic+Targeting+Photothermal+Therapy+and+Fenton+Reaction.">Fe3O4-Based Multifunctional Nanospheres for Amplified Magnetic Targeting Photothermal Therapy and Fenton Reaction.</a> ACS Biomaterials Science &amp; Engineering. 5 (2), 1045-1056 (2018).</li><li>Thorat, N. D., 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=Physically+stimulated+nanotheranostics+for+next+generation+cancer+therapy:+Focus+on+magnetic+and+light+stimulations.">Physically stimulated nanotheranostics for next generation cancer therapy: Focus on magnetic and light stimulations.</a> Applied Physics Reviews. 6 (4), 041306 (2019).</li><li>Sun, Q., Zhou, Z., Qiu, N., Shen, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+of+Cancer+Nanomedicine:+Nanoproperty+Integration+and+Synchronization.">Design of Cancer Nanomedicine: Nanoproperty Integration and Synchronization.</a> Advanced Materials. 29 (14), 1606628 (2017).</li><li>Minchinton, A. I., Tannock, I. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug+penetration+in+solid+tumours.">Drug penetration in solid tumours.</a> Nature Reviews Cancer. 6 (8), 583-592 (2006).</li><li>Anchordoquy, T. J., 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=Mechanisms+and+Barriers+in+Cancer+Nanomedicine:+Addressing+Challenges,+Looking+for+Solutions.">Mechanisms and Barriers in Cancer Nanomedicine: Addressing Challenges, Looking for Solutions.</a> ACS Nano. 11 (1), 12-18 (2017).</li><li>Cazares-Cortes, E., 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=Recent+insights+in+magnetic+hyperthermia:+From+the+"hot-spot"+effect+for+local+delivery+to+combined+magneto-photo-thermia+using+magneto-plasmonic+hybrids.">Recent insights in magnetic hyperthermia: From the "hot-spot" effect for local delivery to combined magneto-photo-thermia using magneto-plasmonic hybrids.</a> Advanced Drug Delivery Reviews. 138, 233-246 (2019).</li><li>Espinosa, A., 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=of+Iron+Oxide+Nanoparticles+in+Cancer+Therapy:+Amplification+of+Heating+Efficiency+by+Magnetic+Hyperthermia+and+Photothermal+Bimodal+Treatment.">of Iron Oxide Nanoparticles in Cancer Therapy: Amplification of Heating Efficiency by Magnetic Hyperthermia and Photothermal Bimodal Treatment.</a> ACS Nano. 10 (2), 2436-2446 (2016).</li><li>Rastinehad, A. 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=Gold+nanoshell-localized+photothermal+ablation+of+prostate+tumors+in+a+clinical+pilot+device+study.">Gold nanoshell-localized photothermal ablation of prostate tumors in a clinical pilot device study.</a> Proceedings of the National Academy of Sciences of the United States of America. 116 (37), 18590-18596 (2019).</li><li>Sharma, S. K., Shrivastava, N., Rossi, F., Tung, L. D., Thanh, N. T. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanoparticles-based+magnetic+and+photo+induced+hyperthermia+for+cancer+treatment.">Nanoparticles-based magnetic and photo induced hyperthermia for cancer treatment.</a> Nano Today. 29, 100795 (2019).</li><li>Das, R., Rinaldi-Montes, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Boosted+Hyperthermia+Therapy+by+Combined+AC+Magnetic+and+Photothermal+Exposures+in+Ag/Fe3O4+Nanoflowers.">Boosted Hyperthermia Therapy by Combined AC Magnetic and Photothermal Exposures in Ag/Fe3O4 Nanoflowers.</a> ACS Applied Materials &amp; Interfaces. 8 (38), 25162-25169 (2016).</li><li>Yang, Y., 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=1D+Coordination+Polymer+Nanofibers+for+Low-Temperature+Photothermal+Therapy.">1D Coordination Polymer Nanofibers for Low-Temperature Photothermal Therapy.</a> Advanced Materials. 29 (40), 1703588 (2017).</li><li>Curcio, A., 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=Iron+Oxide+Nanoflowers+CuS+Hybrids+for+Cancer+Tri-Therapy:+Interplay+of+Photothermal+Therapy,+Magnetic+Hyperthermia+and+Photodynamic+Therapy.">Iron Oxide Nanoflowers CuS Hybrids for Cancer Tri-Therapy: Interplay of Photothermal Therapy, Magnetic Hyperthermia and Photodynamic Therapy.</a> Theranostics. 9 (5), 1288-1302 (2019).</li><li>Espinosa, A., 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=Hyper)Thermia+or+Photothermia?+Progressive+Comparison+of+Iron+Oxide+and+Gold+Nanoparticles+Heating+in+Water,+in+Cells,+and+In+Vivo.">Hyper)Thermia or Photothermia? Progressive Comparison of Iron Oxide and Gold Nanoparticles Heating in Water, in Cells, and In Vivo.</a> Advanced Functional Materials. 28 (37), 1803660 (2018).</li><li>Xu, C., 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=Magnetic+Hyperthermia+Ablation+of+Tumors+Using+Injectable+Fe(3)O(4)/Calcium+Phosphate+Cement.">Magnetic Hyperthermia Ablation of Tumors Using Injectable Fe(3)O(4)/Calcium Phosphate Cement.</a> ACS Applied Materials &amp; Interfaces. 7 (3), 13866-13875 (2015).</li><li>Wilhelm, S., 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=Analysis+of+nanoparticle+delivery+to+tumours.">Analysis of nanoparticle delivery to tumours.</a> Nature Reviews Materials. 1, 16014 (2016).</li><li>Chen, H., Zhang, W., Zhu, G., Xie, J., Chen, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rethinking+cancer+nanotheranostics.">Rethinking cancer nanotheranostics.</a> Nature Reviews Materials. 2, (2017).</li><li>Rapoport, N. Y., Kennedy, A. M., Shea, J. E., Scaife, C. L., Nam, K. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+and+targeted+tumor+chemotherapy+by+ultrasound-activated+nanoemulsions/microbubbles.">Controlled and targeted tumor chemotherapy by ultrasound-activated nanoemulsions/microbubbles.</a> Journal of Controlled Release : The Official Journal of the Controlled Release Society. 138 (3), 268-276 (2009).</li><li>Gao, Y., 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=Controlled+nanoparticle+release+from+stable+magnetic+microbubble+oscillations.">Controlled nanoparticle release from stable magnetic microbubble oscillations.</a> NPG Asia Materials. 8 (4), 260 (2016).</li><li>Bao, B., 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=Mussel-inspired+functionalization+of+semiconducting+polymer+nanoparticles+for+amplified+photoacoustic+imaging+and+photothermal+therapy.">Mussel-inspired functionalization of semiconducting polymer nanoparticles for amplified photoacoustic imaging and photothermal therapy.</a> Nanoscale. 11 (31), 14727-14733 (2019).</li><li>Wu, H., 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=Enhanced+Tumor+Synergistic+Therapy+by+Injectable+Magnetic+Hydrogel+Mediated+Generation+of+Hyperthermia+and+Highly+Toxic+Reactive+Oxygen+Species.">Enhanced Tumor Synergistic Therapy by Injectable Magnetic Hydrogel Mediated Generation of Hyperthermia and Highly Toxic Reactive Oxygen Species.</a> ACS Nano. 13 (12), 14013-14023 (2019).</li><li>Alzaraa, A., 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=Targeted+microbubbles+in+the+experimental+and+clinical+setting.">Targeted microbubbles in the experimental and clinical setting.</a> American Journal of Surgery. 204 (3), 355-366 (2012).</li></ol>

作者没有什么可透露的。

在这里，我们提出了一个协议，通过自组装，协同磁性，声学和光学响应性在一个纳米治疗平台制造氧化铁纳米粒子壳微泡（NSM）。IONP 被密集地包装在空气核心周围，形成磁壳，磁壳可以由外部磁场控制以进行瞄准。一旦交付，通过超声波触发器可以实现离子点的释放。释放的 IONP 可以通过 PTT 和 MHT 的 NIR 光和 AFM 激活，也可以同时激活两者的组合。
在整个协议中，NSM的合成步骤起着重要的作用，这是整个协议的基础。同时，对照放出的离子体外验证了NSM的声学反应。在溶液中光热测量和在溶液中磁性高温测量的方案也分别验证了NSM的磁性和光学响应。
为了成功准备 NSM，在使用前必须对离子体的溶液进行 20 分钟的声波处理，以确保离子在水中均匀分散。执行搅拌时，同质化器探头必须完全浸入溶液中。在研究NSM的声学反应时，样品管必须直接放置在传感器的顶部，并通过磁铁将NSM吸引到管的底部，以防止超声波强度的减弱。此外，如果在光热测量或磁性高温测量期间温度升高不显著，则可能是因为样品既不在激光束的聚焦处，也不在水冷磁感应铜线圈的中心。
应当指出，该议定书仍有一些局限性。例如，尽管预制的 NSM 的平均直径与一些临床上使用的微泡22（ 用于超声波成像）的直径相似，但 NSM 大小的均匀性需要改进。此外，应通过微泡表面的改变来改善血液中NSM的循环。除此之外，协同治疗尚未在体内得到验证，体内的疗效尚不得而知。
拟议的IONP交付策略不仅实现了离子的按需释放，而且促进了离子的渗透到肿瘤组织中。目前，肿瘤间质液压力的增大和肿瘤的致密性极强，极大地限制了纳米粒子的输送效率。因此，本协议为纳米医学的提供提供了组织穿透策略，对癌症治疗非常感兴趣。
我们还证明，以IONPs为媒介的PTT和MHT在体外和体内都有效。结果表明，离子体具有良好的光热转换和磁热转换能力，能有效消融肿瘤。PTT 和 MHT 的组合足以确保癌细胞完全死亡并提高抗癌效果。今后，NSM的双热疗法（即PTT和MHT）将为诊所治疗深层次固体肿瘤提供新的选择。

癌症是最致命的疾病之一，每年在全世界造成数百万人死亡和巨大的经济损失。在诊所，传统的抗癌疗法，如手术切除，放疗和化疗仍然不能提供令人满意的治疗效果2。这些疗法的局限性是高毒副作用，高复发率和高转移率3。例如，化疗是遭受化疗药物的低交付效率正是肿瘤部位4。药物无法深入肿瘤组织跨越生物屏障，包括细胞外基质和高肿瘤间质液压，也是导致疗效低的原因。此外，肿瘤耐药性通常发生在接受单次化疗治疗的患者中。因此，发生肿瘤热消融的技术，如光热疗法（PTT）和磁性高温治疗（MHT），已经显示出了降低肿瘤抵抗力的可喜结果，并已在临床试验7，8，9中出现。
PTT通过光热转换剂在激光能量照射下的作用触发癌细胞的热消融。产生的高温（高于50°C）诱导完整的细胞坏死10。最近，氧化铁纳米粒子（IONPs）被证明是一种光热转换剂，可以通过近红外（NIR）光11激活。 尽管近红外区域的摩尔吸收系数较低，但IONP是低温（43°C）光热疗法的候选者，这是一种经过改良的疗法，旨在减少热暴露对正常组织造成的损害，并启动肿瘤转移的抗肿瘤免疫力。PTT的局限性之一是激光的低穿透深度。对于深座肿瘤，交替磁场（AFM）诱导加热的氧化铁纳米粒子，也被称为磁性高温，是PTT13，14的替代疗法。MHT的主要优点是磁场15的高渗透率。然而，需要相对较高的IONP浓度仍然是其临床应用的主要劣势。由于循环、积累和渗透等一系列障碍，纳米医学（或纳米粒子）对动物实体肿瘤的输送效率一直为1-10%。因此，具有高组织渗透能力的受控和有针对性的IONP输送策略对癌症治疗非常感兴趣。
超声介质纳米粒子的输送已经显示出它的能力，以促进纳米粒子深入肿瘤组织，由于现象称为微泡腔18，19。在本研究中，我们通过在一个纳米治疗平台中自组装、协同磁性、声学和光学响应性来制造离子体壳化微气泡（NSM）。NSM 包含空气核心和氧化铁纳米粒子壳，直径约为 5.4 μm。NSM 可以磁引导到肿瘤位点。然后，IONP 的释放由超声波触发，并伴有微泡空腔和微流。从微流获得的势头有助于离子体渗透到肿瘤组织中。PTT 和 MHT 可以通过 NIR 激光照射或 AFM 应用或两者的组合实现。

<table><tbody><tr><td>808 nm laser power</td><td>Changchun New Industries Optoelectronics Tech</td><td>MDL-F-808-5W-18017023</td><td></td></tr><tr><td>Calcein-AM</td><td>Thermo Fisher SCIENTIFIC</td><td>C3099</td><td></td></tr><tr><td>Fetal bovine serum</td><td>Invitrogen</td><td>16000-044</td><td></td></tr><tr><td>Fluorescence Microscope</td><td>Olympus</td><td>IX71</td><td></td></tr><tr><td>Function generator</td><td>Keysight</td><td>33500B series</td><td>20 MHz, 2 channels with arbitrary waveform generation capability</td></tr><tr><td>Gelatin gel</td><td>Sigma</td><td>9000-70-8</td><td></td></tr><tr><td>Heating machine</td><td>Shuangping</td><td>SPG-06- II</td><td></td></tr><tr><td>Homemade focused transducer</td><td></td><td></td><td>Frequency=855, R-X=36.2W+5.8W, |Z|-&amp;theta;=37W+8&amp;deg;</td></tr><tr><td>Homogenizer</td><td>SCILOGEX</td><td>D-160</td><td>8000-30000 rpm</td></tr><tr><td>Hydrophone</td><td>T&amp;amp;C</td><td>NH1000</td><td></td></tr><tr><td>ICR male mice</td><td>OG Pharmaceutical. Co. Ltd</td><td></td><td>8-week-old</td></tr><tr><td>Inductively coupled plasma optical emission spectrometry</td><td>PerkinElmer</td><td></td><td></td></tr><tr><td>Infrared thermal imaging camera.</td><td>FLIR</td><td>E50</td><td></td></tr><tr><td>Iron(II,III) oxide</td><td>Alfa Aesar</td><td>1317-61-9</td><td>50-100nm APS Powder</td></tr><tr><td>Laser power meter</td><td>Changchun New Industries Optoelectronics Tech</td><td></td><td></td></tr><tr><td>Oscilloscope</td><td>Keysight</td><td>DSOX3054T</td><td>Bandwidth 500 MHz, Sampling Rate 5 GS/S, 4 channels</td></tr><tr><td>RF Power Amplifier</td><td>T&amp;amp;C</td><td>AG1020</td><td>The signal source can also be connected to an external signal source. The gain can be adjusted from 0 to 100%. It has multiple functions such as frequency sweep, pulse, and triangle.</td></tr><tr><td>Roswell Park Memorial Institute-1640</td><td>KeyGEN BioTECH</td><td>KGM31800</td><td></td></tr><tr><td>Sodium dodecyl sulfate</td><td>Sigma</td><td>151-21-3</td><td></td></tr></tbody></table>

magnetic-, acoustic-, and optical-triple-responsive microbubbles for magnetic hyperthermia and pothotothermal combination cancer therapy

抗癌剂的精确输送，旨在有针对性和深度渗透的分娩，以及在肿瘤部位的对照释放，已受到挑战。在这里，我们通过自组装、协同磁性、声学和光学响应性，在一个纳米治疗平台上制造氧化铁纳米粒子壳化微泡 （NSM）。氧化铁纳米粒子既可用作磁性剂，又可用作光热剂。静脉注射后，NSM可磁性引导至肿瘤位点。超声波触发氧化铁纳米粒子的释放，由于微泡的腔作用，促进纳米粒子深入肿瘤。此后，磁性高温和光热疗法可以在肿瘤上进行合并癌症治疗，这是治疗肿瘤异质性对癌症的抵抗力的一种解决方案。在本协议中，对NSM进行了合成和定性，包括结构、化学、磁性和声学特性。此外，还利用体外细胞培养物对热疗法的抗癌疗效进行了调查。拟议的分娩策略和联合治疗在癌症治疗方面有很大的希望，以提高分娩和抗癌的疗效。

本研究中使用的三响应纳米粒子壳微泡 （NSM） 是通过搅拌表面活性剂和离子体的混合物而制备的。IONP （50 nm） 在液体和气体核心的接口上自行组装，形成密集的磁壳。NSM 的形态显示在 图 1A 中。结果NSM呈现球形，平均直径为5.41±1.78μm（图1B）。结果表明，NSM的准备工作是成功的。当储存在水中时，微泡保持完好无损超过1年，并在缓冲器和细胞培养介质中稳定至少10天19。如图 1D所示，Fe的逐步释放是随着应用超声波周期的增加而实现的。经过10个周期后，大约20%的Fe被释放。直到50个超声波周期，释放Fe的量达到80%左右的高原。这些结果表明，IONP通过外部超声波触发器按需释放。
在射电溶液中，IONP 介介质的光热测量如图2所示。IONP的温度随着辐照时间的增加而迅速增加，如图2A，B所示。暴露在NIR激光器（808 nm，1 W/cm2）下10分钟，在5毫克/毫升的Fe浓度下，温度可升高30°C。
PTT产生的热量可以杀死癌细胞。通过NIR激光（808纳米，1 W/cm2）治疗10分钟，对PTT4T1细胞的生存能力进行了评估。如图3A，B，与对照组相比，在高浓度Fe（5mg/mL）孵育时，形态和活细胞数没有差异，这表明IONP具有良好的生物可用性。一旦受到NIR激光的照射，细胞就变成圆形，表明细胞凋亡。活细胞数的量化，即细胞的生存能力在图3C中显示。在NIR辐照下孵育的具有高离子浓度（3.65毫克/毫升和5毫克/毫升）的细胞死亡率最高，分别为80%和100%左右。低离子浓度（1.025毫克/mL和1.35毫克/mL）治疗组表现出类似的杀伤效率约40%。结果表明，NSMS的光热效应可以有效治疗癌症。
如图4A，B所示，在5分钟的NIR辐照后，明胶注射区的温度迅速上升了约20°C。小鼠感兴趣区域的实际表面温度可达到 57 °C 左右。 如图5所示，当暴露在AFM下时，红外热像仪（图5A）监测不同浓度的离子体（1.05毫克/兆升、1.35毫克/兆升、3.65毫克/兆升、5毫克/兆升）的热成像，并在不同时间间隔（图5B）记录和绘制温度曲线。其中，1.35 毫克/mL、3.65 毫克/mL 和 5 毫克/mL IONP 可在 10 分钟感应后快速加热溶液并增加温度（分别为 20 °C、30 °C、40°C）。结果表明NSMS的磁场响应特性。
在体内磁性高温实验中，小鼠暴露在415 kHz的频率和1.8千卡/米的磁振幅10分钟。加热过程由红外热像仪实时监控（图6A，6B）。观察到感兴趣的区域的显著温度变化（图6）。温度随时间而迅速升高，10 分钟的感应增量为 50 °C。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/61208/61208fig1.jpg"> 图1:NSM.（A）代表NSM的亮场显微镜图像的特征和受控离子体释放。（B） NSM 的直径分布，n = 200。（C）实验中使用的超声波设备图。（D）在超声波刺激下，从 NSM 中累积释放 IONP 配置文件。<a href="https://www.jove.com/files/ftp_upload/61208/61208fig1large.jpg" target="_blank">请单击此处查看此图的较大版本。</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/61208/61208fig2.jpg"> 图2:以离子体为媒介的射电热测量。（A） 10分钟激光照射后，10分钟激光照射1W/cm2不同浓度的红外热图像。（B）不同浓度的离子体（808 nm，1 W/cm2，10分钟）的典型温度高程曲线。<a href="https://www.jove.com/files/ftp_upload/61208/61208fig2large.jpg" target="_blank">请单击此处查看此图的较大版本。</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/61208/61208fig3v2.jpg"> 图3:4T1细胞中IONP介质光热测量。（A）活4T1细胞在24小时孵育后，具有不同浓度的离子蛋白（沾染有钙素-AM，绿色）的聚焦荧光显微镜图像。NIR处理过的细胞暴露在808纳米激光下10分钟（1 W/cm2）。秤吧:50μm。（B） IONP 不同浓度的典型温度高程曲线处理 4T1 细胞 10 分钟的 NIR 辐照 （1 W/cm2）。（C）量化与离子体一起孵育的 4T1 细胞的生存能力，无论是否进行 NIR 治疗。<a href="https://www.jove.com/files/ftp_upload/61208/61208fig3v2large.jpg" target="_blank">请单击此处查看此图的较大版本。</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/61208/61208fig4v2.jpg"> 图4:活体中IONP介质的光热测量。（A）在不同时间间隔捕获的暴露在NIR激光下的小鼠兴趣区域的红外热图像（808 nm，1 W/cm2，10分钟）。（B） NIR激光（808 nm，1 W/cm2，10分钟）治疗后，不同时间间隔温度曲线升高。<a href="https://www.jove.com/files/ftp_upload/61208/61208fig4v2large.jpg" target="_blank">请单击此处查看此图的较大版本。</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/61208/61208fig5.jpg"> 图5:在无声中由离子体介质的磁性高温测量。（A）在自动对焦器下不同浓度的离子体溶液的红外热图像，频率为415 kHz，磁振幅为1.8 kA/m，为10分钟。 （B）以415 kHz的频率和1.8千卡/米的磁振幅在自动对焦下不同浓度的IONP溶液的典型温度<a href="https://www.jove.com/files/ftp_upload/61208/61208fig5large.jpg" target="_blank">曲线。</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/61208/61208fig6.jpg"> 图6:活体中IONPs介质的磁性高温。（A）在自动对焦下以415 kHz的频率和1.8千卡/米的磁振幅在10分钟内在自动对焦下不同时间间隔捕获的小鼠兴趣区域的红外热图像。 （B）在 AFM 下不同时间间隔的温度曲线升高，频率为 415 kHz，磁振幅为 1.8 kA/m。<a href="https://www.jove.com/files/ftp_upload/61208/61208fig6large.jpg" target="_blank">请单击此处查看此图的较大版本。</a>

Watch this Scientific Journal Video about Magnetic-, Acoustic-, and Optical-Triple-Responsive Microbubbles for Magnetic Hyperthermia and Pothotothermal Combination Cancer Therapy at JoVE.com

Magnetic-, Acoustic-, and Optical-Triple-Responsive Microbubbles for Magnetic Hyperthermia and Pothotothermal Combination Cancer Therapy

这里介绍了一个协议，通过自组装，协同磁性，声学和光学响应性在一个纳米治疗平台的磁性高温和光热组合癌症治疗制造氧化铁纳米粒子壳微泡（NSMs）。

磁性、声学和光学三响应性微泡，用于磁性高温和波托托热组合癌症治疗

The precision delivery of anti-cancer agents which aim for targeted and deep-penetrated delivery as well as a controlled release at the tumor site has ...

magnetic-acoustic-optical-triple-responsive-microbubbles-for-magnetic

Nanyang Technological University

Research

JoVE Journal

Bioengineering

3.0K 次观看.  Nanjing University of Posts & Telecommunications. 这里介绍了一个协议，通过自组装，协同磁性，声学和光学响应性在一个纳米治疗平台的磁性高温和光热组合癌症治疗制造氧化铁纳米粒子壳微泡（NSMs）。

视频: Magnetic-, Acoustic-, and Optical-Triple-Responsive Microbubbles for Magnetic Hyperthermia and Pothotothermal Combination Cancer Therapy

Contrast Ultrasound Targeted Treatment of Gliomas in Mice via Drug-Bearing Nanoparticle Delivery and Microvascular Ablation

We are developing minimally-invasive contrast agent microbubble based therapeutic approaches in which the permeabilization and/or ablation of the microvasculature are controlled by varying ultrasound pulsing parameters.  Specifically, we are testing whether such approaches may be used to treat malignant brain tumors through drug delivery and microvascular ablation.  Preliminary studies have been performed to determine whether targeted drug-bearing nanoparticle delivery can be facilitated by the ultrasound mediated destruction of "composite" delivery agents comprised of 100nm poly(lactide-co-glycolide) (PLAGA) nanoparticles that are adhered to albumin shelled microbubbles. We denote these agents as microbubble-nanoparticle composite agents (MNCAs). When targeted to subcutaneous C6 gliomas with ultrasound, we observed an immediate 4.6-fold increase in nanoparticle delivery in MNCA treated tumors over tumors treated with microbubbles co-administered with nanoparticles and a 8.5 fold increase over non-treated tumors. Furthermore, in many cancer applications, we believe it may be desirable to perform targeted drug delivery in conjunction with ablation of the tumor microcirculation, which will lead to tumor hypoxia and apoptosis. To this end, we have tested the efficacy of non-theramal cavitation-induced microvascular ablation, showing that this approach elicits tumor perfusion reduction, apoptosis, significant growth inhibition, and necrosis. Taken together, these results indicate that our ultrasound-targeted approach has the potential to increase therapeutic efficiency by creating tumor necrosis through microvascular ablation and/or simultaneously enhancing the drug payload in gliomas.

Insonation of microbubbles is a promising strategy for tumor ablation at reduced time-averaged acoustic powers, as well as for the targeted delivery of therapeutics. The purpose of the present study is to develop low duty cycle ultrasound pulsing strategies and nanocarriers to maximize non-thermal microvascular ablation and payload delivery to subcutaneous C6 gliomas.

超声造影有针对性的治疗胶质瘤的小鼠通过药物轴承纳米粒子交付和微血管消融

We are developing minimally-invasive contrast agent microbubble based therapeutic approaches in which the permeabilization and/or ablation of the ...

科研

生物工程

Magnetic Tweezers for the Measurement of Twist and Torque

Single-molecule techniques make it possible to investigate the behavior of individual biological molecules in solution in real time. These techniques include so-called force spectroscopy approaches such as atomic force microscopy, optical tweezers, flow stretching, and magnetic tweezers. Amongst these approaches, magnetic tweezers have distinguished themselves by their ability to apply torque while maintaining a constant stretching force. Here, it is illustrated how such a &#8220;conventional&#8221; magnetic tweezers experimental configuration can, through a straightforward modification of its field configuration to minimize the magnitude of the transverse field, be adapted to measure the degree of twist in a biological molecule. The resulting configuration is termed the freely-orbiting magnetic tweezers. Additionally, it is shown how further modification of the field configuration can yield a transverse field with a magnitude intermediate between that of the &#8220;conventional&#8221; magnetic tweezers and the freely-orbiting magnetic tweezers, which makes it possible to directly measure the torque stored in a biological molecule. This configuration is termed the magnetic torque tweezers. The accompanying video explains in detail how the conversion of conventional magnetic tweezers into freely-orbiting magnetic tweezers and magnetic torque tweezers can be accomplished, and demonstrates the use of these techniques. These adaptations maintain all the strengths of conventional magnetic tweezers while greatly expanding the versatility of this powerful instrument.

Magnetic tweezers, a powerful single-molecule manipulation technique, can be adapted for the direct measurements of the twist (using a configuration called freely-orbiting magnetic tweezers) and torque (using a configuration termed magnetic torque tweezers) in biological macromolecules. Guidelines for performing such measurements are given, including applications to the study of DNA and associated nucleo-protein filaments.

磁性镊子的扭曲和扭矩的测量

Single-molecule techniques make it possible to investigate the behavior of individual biological molecules in solution in real time. These techniques ...

Fabrication of a Functionalized Magnetic Bacterial Nanocellulose with Iron Oxide Nanoparticles

In this study, bacterial nanocellulose (BNC) produced by the bacteria Gluconacetobacter xylinus is synthesized and impregnated in situ with iron oxide nanoparticles (IONP) (Fe3O4) to yield a magnetic bacterial nanocellulose (MBNC). The synthesis of MBNC is a precise and specifically designed multi-step process. Briefly, bacterial nanocellulose (BNC) pellicles are formed from preserved G. xylinus strain according to our experimental requirements of size and morphology. A solution of iron(III) chloride hexahydrate (FeCl3&#183;6H2O) and iron(II) chloride tetrahydrate (FeCl2&#183;4H2O) with a 2:1 molar ratio is prepared and diluted in deoxygenated high purity water. A BNC pellicle is then introduced in the vessel with the reactants. This mixture is stirred and heated at 80 &#176;C in a silicon oil bath and ammonium hydroxide (14%) is then added by dropping to precipitate the ferrous ions into the BNC mesh. This last step allows forming in situ magnetite nanoparticles (Fe3O4) inside the bacterial nanocellulose mesh to confer magnetic properties to BNC pellicle. A toxicological assay was used to evaluate the biocompatibility of the BNC-IONP pellicle. Polyethylene glycol (PEG) was used to cover the IONPs in order to improve their biocompatibility. Scanning electron microscopy (SEM) images showed that the IONP were located preferentially in the fibril interlacing spaces of the BNC matrix, but some of them were also found along the BNC ribbons. Magnetic force microscope measurements performed on the MBNC detected the presence magnetic domains with high and weak intensity magnetic field, confirming the magnetic nature of the MBNC pellicle. Young&#39;s modulus values obtained in this work are also in a reasonable agreement with those reported for several blood vessels in previous studies.

Here, we present a protocol to make a bacterial nanocellulose (BNC) magnetic for applications in damaged blood vessel reconstruction. The BNC was synthesized by G. xylinus strain. On the other hand, magnetization of the BNC was realized through in situ precipitation of Fe2+ and Fe3+ ferrous ions inside the BNC mesh.

一机能磁性纳米纤维素的细菌与氧化铁纳米粒子的制备

In this study, bacterial nanocellulose (BNC) produced by the bacteria Gluconacetobacter xylinus is synthesized and impregnated in situ with iron oxide ...

Custom-designed Laser-based Heating Apparatus for Triggered Release of Cisplatin from Thermosensitive Liposomes with Magnetic Resonance Image Guidance

Liposomes have been employed as drug delivery systems to target solid tumors through exploitation of the enhanced permeability and retention (EPR) effect resulting in significant reductions in systemic toxicity. Nonetheless, insufficient release of encapsulated drug from liposomes has limited their clinical efficacy. Temperature-sensitive liposomes have been engineered to provide site-specific release of drug in order to overcome the problem of limited tumor drug bioavailability. Our lab has designed and developed a heat-activated thermosensitive liposome formulation of cisplatin (CDDP), known as HTLC, to provide triggered release of CDDP at solid tumors. Heat-activated delivery in vivo was achieved in murine models using a custom-built laser-based heating apparatus that provides a conformal heating pattern at the tumor site as confirmed by MR thermometry (MRT). A fiber optic temperature monitoring device was used to measure the temperature in real-time during the entire heating period with online adjustment of heat delivery by alternating the laser power. Drug delivery was optimized under magnetic resonance (MR) image guidance by co-encapsulation of an MR contrast agent (i.e.,&#160;gadoteridol) along with CDDP into the thermosensitive liposomes as a means to validate the heating protocol and to assess tumor accumulation. The heating protocol consisted of a preheating period of 5 min prior to administration of HTLC and 20 min heating post-injection. This heating protocol resulted in effective release of the encapsulated agents with the highest MR signal change observed in the heated tumor in comparison to the unheated tumor and muscle. This study demonstrated the successful application of the laser-based heating apparatus for preclinical thermosensitive liposome development and the importance of MR-guided validation of the heating protocol for optimization of drug delivery.

AN MRI-compatible&#160;custom-designed laser-based heating apparatus has been developed to provide local heating of subcutaneous tumors in order to activate release of agents from thermosensitive liposomes specifically at the tumor region.

定制设计的基于激光的加热装置顺铂触发释放的热敏脂质体与磁共振成像制导

Liposomes have been employed as drug delivery systems to target solid tumors through exploitation of the enhanced permeability and retention (EPR) effect ...

Magnetic Resonance-Guided High Intensity Focused Ultrasound Generated Hyperthermia: A Feasible Treatment Method in a Murine Rhabdomyosarcoma Model

Magnetic resonance-guided high intensity focused ultrasound (MRgHIFU) is an established method for producing localized hyperthermia. Given the real-time imaging and acoustic energy modulation, this modality enables precise temperature control within a defined area. Many thermal applications are being explored with this noninvasive, nonionizing technology, such as hyperthermia generation, to release drugs from thermosensitive liposomal carriers. These drugs can include chemotherapies such as doxorubicin, for which targeted release is desired due to the dose-limiting systemic side effects, namely cardiotoxicity. Doxorubicin is a mainstay for treating a variety of malignant tumors and is commonly used in relapsed or recurrent rhabdomyosarcoma (RMS). RMS is the most common solid soft tissue extracranial tumor in children and young adults. Despite aggressive, multimodal therapy, RMS survival rates have remained the same for the past 30 years. To explore a solution for addressing this unmet need, an experimental protocol was developed to evaluate the release of thermosensitive liposomal doxorubicin (TLD) in an immunocompetent, syngeneic RMS mouse model using MRgHIFU as the source of hyperthermia for drug release.

Presented here is a protocol to use controlled hyperthermia, generated by magnetic resonance-guided high intensity focused ultrasound, to trigger drug release from temperature-sensitive liposomes in a rhabdomyosarcoma mouse model.

磁共振引导的高强度聚焦超声产生热疗:小鼠横纹肌肉瘤模型中的可行治疗方法

Magnetic resonance-guided high intensity focused ultrasound (MRgHIFU) is an established method for producing localized hyperthermia. Given the real-time ...

癌症研究

Fabricating and Labeling Microbubbles with Fluorescent and Radioactive Tracers

Microbubbles are lipid-shelled, gas-filled particles that have evolved from vascular ultrasound contrast agents into revolutionary cancer therapy platforms. When combined with therapeutic focused ultrasound (FUS), they can safely and locally overcome physiological barriers (e.g., blood-brain barrier), deliver drugs to otherwise inaccessible cancers (e.g., glioblastoma and pancreatic cancer), and treat neurodegenerative diseases. The therapeutic arsenal of microbubble-FUS is advancing in new directions, including synergistic combination radiotherapy, multimodal imaging, and all-in-one drug loading and delivery from microbubble shells.
Labeling microbubbles with radiotracers is key to establishing these expanded theranostic capabilities. However, existing microbubble radiolabeling strategies rely on purification methodologies known to perturb microbubble physicochemical properties, use short-lived radioisotopes, and do not always yield stable chelation. Collectively, this creates ambiguity surrounding the accuracy of microbubble radioimaging and the efficiency of tumor radioisotope delivery.
This protocol describes a new one-pot, purification-free microbubble labeling methodology that preserves microbubble physicochemical properties while achieving &#62;95% radioisotope chelation efficiency. It is versatile and can be applied successfully across custom and commercial microbubble formulations with differing acyl lipid chain length, charge, and chelator/probe (porphyrin, DTPA, DiI) composition. It can be adaptively applied during ground-up microbubble fabrication and to pre-made microbubble formulations with modular customizability of fluorescence and multimodal fluorescence/radioactive properties. Accordingly, this flexible method enables the production of tailored, traceable (radio, fluorescent, or radio/fluorescent active) multimodal microbubbles that are useful for advancing mechanistic, imaging, and therapeutic microbubble-FUS applications.

This protocol outlines the fabrication of lipid microbubbles and a compatible one-pot microbubble radiolabeling method with purification-free &gt;95% labeling efficiency that conserves microbubble physicochemical properties. This method is effective across diverse lipid microbubble formulations and can be tailored to generate radioactive and/or fluorescent microbubbles.

使用荧光和放射性示踪剂制造和标记微气泡

Microbubbles are lipid-shelled, gas-filled particles that have evolved from vascular ultrasound contrast agents into revolutionary cancer therapy ...

Studying Cavitation Enhanced Therapy

Interest in the therapeutic applications of ultrasound is significant and growing, with potential clinical targets ranging from cancer to Alzheimer&#8217;s disease. Cavitation - the formation and subsequent motion of bubbles within an ultrasound field - represents a key phenomenon underpinning many of these treatments. There remains, however, considerable uncertainty regarding the detailed mechanisms of action by which cavitation promotes therapeutic effects and there is a need to develop reliable monitoring techniques that can be implemented clinically. In particular, there is significant variation between studies in the exposure parameters reported as successfully delivering therapeutic effects and the corresponding acoustic emissions. The aim of this paper is to provide design guidelines and an experimental protocol using widely available components for performing studies of cavitation-mediated bioeffects, and&#160;include real-time acoustic monitoring. It is hoped that the protocol will enable more widespread incorporation of acoustic monitoring into therapeutic ultrasound experiments and facilitate easier comparison across studies of exposure conditions and their correlation to relevant bio-effects.

The presented experimental protocol can be used to perform real time measurements of cavitation activity in a cell culture device with the aim of enabling investigation of the conditions required for successful drug delivery and/or other bioeffects.

研究屈化强化疗法

Interest in the therapeutic applications of ultrasound is significant and growing, with potential clinical targets ranging from cancer to ...

模拟双极射频热疗系统对肿瘤的影响

A Computational Modeling Approach to Investigate the Influence of Hyperthermia on the Tumor Microenvironment

The biophysical properties of the tumor microenvironment differ substantially from normal tissues. A constellation of features, including decreased vascularity, lack of lymphatic drainage, and elevated interstitial pressure, diminishes the penetration of therapeutics into tumors. Local&#160;hyperthermia within the tumor can alter microenvironmental properties, such as interstitial fluid pressure, potentially leading to improvements in drug penetration. In this context, multi-physics computational models can provide insight into the interplay between the biophysical parameters within the tumor microenvironment and can guide the design and interpretation of experiments that test the bioeffects of local hyperthermia.
This paper describes a step-by-step workflow for a computational model coupling partial differential equations describing electrical current distribution, bioheat transfer, and fluid dynamics. The main objective is to study the effects of hyperthermia delivered by a bipolar radiofrequency device on the interstitial fluid pressure within the tumor.&#160;The system of mathematical expressions linking electrical current distribution, bioheat transfer, and interstitial fluid pressure is presented, emphasizing the changes in the distribution of the interstitial fluid pressure that could be induced by the thermal intervention.

作者聚焦:计算局部射频热疗干预对肿瘤生物力学的影响

The article describes a protocol to simulate the transient temperature profiles and the coupled spatiotemporal variation of the interstitial fluid pressure following the heating delivered by a dipolar radiofrequency hyperthermia system. The protocol can be used to assess the response of biophysical parameters characterizing the tumor microenvironment to interventional hyperthermia techniques.

一种计算建模方法研究热疗对肿瘤微环境的影响

The biophysical properties of the tumor microenvironment differ substantially from normal tissues. A constellation of features, including decreased ...

磁性、声学和光学三响应性微泡，用于磁性高温和波托托热组合癌症治疗

摘要

探索更多视频

此视频中的章节

超声造影有针对性的治疗胶质瘤的小鼠通过药物轴承纳米粒子交付和微血管消融

磁性镊子的扭曲和扭矩的测量

一机能磁性纳米纤维素的细菌与氧化铁纳米粒子的制备

定制设计的基于激光的加热装置顺铂触发释放的热敏脂质体与磁共振成像制导

磁共振引导的高强度聚焦超声产生热疗:小鼠横纹肌肉瘤模型中的可行治疗方法

使用荧光和放射性示踪剂制造和标记微气泡

研究屈化强化疗法

一种计算建模方法研究热疗对肿瘤微环境的影响

一种计算建模方法研究热疗对肿瘤微环境的影响

一种计算建模方法研究热疗对肿瘤微环境的影响