导管内送货到兔乳腺
Summary
Here, we describe a technique for the localized delivery of reagents to the rabbit mammary gland via an intraductal injection. In addition, we describe a protocol for visualization and the confirmation of delivery by high-resolution ultrasound imaging of contrast agents.
Abstract
对于乳腺癌的局部导管的治疗提供了潜在的优势，包括有效递送至肿瘤，并降低全身毒性和副作用1，2，3，4，5，6，7。然而，一些挑战依然存在之前这些处理可以更广泛地应用。在合适的动物模型的发展和导管治疗的有效性促进导管内治疗策略对患者的发展。而小鼠乳腺已被广泛地用作乳腺发育和肿瘤的模型系统中，解剖来自人腺不同。较大的动物模型，如兔，可作为乳腺结构和管内治疗开发一个更好的模型。在Contrast小鼠，在十个由导管树木在空间沿体轴分布，在一个单独的乳头每个终止，兔子乳腺更类似于人类腺，具有多个重叠导管系统该出口通过在一个乳头分开的开口。这里，我们提出了试剂的递送微创方法直接进入兔乳腺导管和用于递送本身具有高分辨率超声波成像的可视化。
Introduction
治疗剂的导管内输送已经研究了在啮齿动物模型中和在早期的人体试验3，4，5，6，11，12。最近的I期临床试验证明导管内卡铂或女性乳房等待为浸润癌2治疗导管内聚乙二醇化脂质体阿霉素的安全性和可行性。
对导管内输送以前的协议已被开发用于小鼠和大鼠乳腺6，7，8，9。出于研究目的，导管内的肿瘤细胞注射和癌基因的慢病毒载体递送也已在啮齿动物模型中进行的裁判“> 13，14，15，16，但是， 在导管内输送过程的体内模型的理想应允许新的类治疗化合物的发展和促进临床前评估。啮齿动物和人类之间的解剖学差异有复杂的这些翻译学习。
不像小鼠，其中每个管道在一个单独的乳头结束时，人乳腺癌由5至9个独立的导管系统中，每个与奶嘴结束一个单独的开口。兔乳腺怀有四个独立导管系统中，每个分别通过在一个单一的奶头4孔可接触。兔模型更接近人体解剖，并允许导管内给药的研究更相关的情境。
在这里，我们用两种方法来评估导管内交付。的一个共同给予活体染料允许可视化通过皮肤，并提供该方法的一个简单的和快速的确认。对于一些应用，导管的更高分辨率映射可以是优选的。我们在座的管道超声成像协议通过非靶向造影剂的导管内交付。
Protocol
Representative Results
Discussion
可用于超声对比试剂和许多其他的水溶液，包括重要的染料和治疗剂导管内输送到兔的乳腺的此方法。以前的研究已经表明激素17，18，19的导管内交付。在啮齿动物模型中，核酸8的导管内输送，化学治疗6,7和纳米颗粒的载体8，20<…
Divulgations
The authors have nothing to disclose.
Acknowledgements
The authors acknowledge support from a Translational Breast Cancer Research Grant (14-60-26-BROC to AB) from the Breast Cancer Research Foundation and the American Association for Cancer Research.
Materials
| MicroMarker non-targeted contrast reagent | VisualSonics | VS-11694 | |
| Luer Lock 1mL Syringes | BD | 309628 | |
| Glycopyrrolate 0.2mg/mL | Wedgewood Compounding Pharmacy | GLYCOP-INJ013VC | 6 month shelf life, supply may be limited. |
| Atropine Sulfate 0.5 mg/mL | Animal Health International | 15320764 | If glycopyrrolate is unavailable. Not to be combined with glycopyrrolate. |
| Ketamine HCL 100mg/mL | Animal Health International | 21250699 | http://www.animalhealthinternational.com/ |
| Acepromazine 10mg/mL | Animal Health International | 17640541 | |
| Xylazine 20mg/mL | Animal Health International | 20101547 | |
| Yohimbine 0.2mg/mL | Animal Health International | 14588965 | |
| Hair Removing Cream | Veet | Sensitive skin solution. Available through local retailers. | |
| Blunt tip infusion needles | Sai Infusion Technology | B14-50 | http://www.sai-infusion.com/collections/blunt-needles |
| Veterinary Pulse Oximeter | EdanUSA | VE-H100B | http://www.edanusa.com/Product/VE-H100B-Veterinary-Pulse-Oximeter.html |
| Warm Water Pump | Gaymar | TP700 | |
| Warm Water Blanket | Animal Health International | 21232696 | Maxi-Therm Lite Warming Pads |
| Ultrasound system | VisualSonics | Vevo 2100 |
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