Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of California San Diego, La Jolla, California, USA.
Department of Nanoengineering, University of California San Diego, La Jolla, California, USA.
Otolaryngol Head Neck Surg. 2019 Nov;161(5):814-822. doi: 10.1177/0194599819866407. Epub 2019 Sep 24.
Human papillomavirus (HPV)-associated oropharyngeal cancer (OPC) is a lethal disease with increasing incidence; however, technologies for early detection are limited. Nanomotors are synthetic nanostructures that can be powered by different mechanisms and functionalized for specific applications, such as biosensing. The objective of this investigation was to demonstrate an in vitro proof of concept for a novel nanomotor-based cancer detection approach toward in vivo detection of HPV-OPC.
In vitro cell line incubated with ultrasound-propelled nanomotors.
Basic science and engineering laboratories.
Ultrasound-powered gold nanowire nanomotors were functionalized with graphene oxide and dye-labeled single-stranded DNA for the specific intracellular detection of HPV16 mRNA transcripts. Nanomotors were incubated with HPV-positive or HPV-negative human OPC cells under static conditions or with an applied ultrasound field for 15 minutes. The resulting intracellular fluorescence was assessed with fluorescence microscopy and analysis software.
Nanomotors incubated with RNA extracted from HPV-positive OPC cells resulted in 60.7% of maximal fluorescence recovery, while incubation with RNA extracted from HPV-negative cells produced negligible fluorescence. Nanomotor incubation with intact HPV-negative cells produced minimal fluorescence (0.01 au), while incubation with HPV-positive cells produced a detectable signal (0.43 au) under static conditions and had 2.3-times greater intensity when powered with ultrasound.
Acoustically powered nanomotors can successfully identify HPV16 mRNA transcripts extracellularly and within intact cells. This work represents the first step toward a novel, practical approach to address the challenge of visually detecting HPV-OPC in real time.
人乳头瘤病毒（HPV）相关的口咽癌（OPC）是一种致命疾病，发病率不断上升；然而，早期检测技术有限。纳米马达是一种可以通过不同机制驱动并针对特定应用（如生物传感）进行功能化的合成纳米结构。本研究的目的是展示一种基于新型纳米马达的癌症检测方法的体外概念验证，以实现 HPV-OPC 的体内检测。
体外细胞系与超声驱动纳米马达孵育。
基础科学和工程实验室。
超声驱动的金纳米线纳米马达与氧化石墨烯和染料标记的单链 DNA 功能化，用于 HPV16 mRNA 转录本的特定细胞内检测。在静态条件下或施加超声场 15 分钟的情况下，将纳米马达与 HPV 阳性或 HPV 阴性的人 OPC 细胞孵育。用荧光显微镜和分析软件评估所得细胞内荧光。
与从 HPV 阳性 OPC 细胞中提取的 RNA 孵育的纳米马达导致最大荧光恢复的 60.7%，而与从 HPV 阴性细胞中提取的 RNA 孵育的则产生可忽略的荧光。纳米马达与完整的 HPV 阴性细胞孵育产生最小的荧光（0.01 au），而在静态条件下与 HPV 阳性细胞孵育则产生可检测的信号（0.43 au），并且在超声驱动时强度增加 2.3 倍。
声驱动纳米马达可以成功地识别 HPV16 mRNA 转录本的细胞外和完整细胞内。这项工作代表了一种新的实用方法的第一步，用于实时解决视觉检测 HPV-OPC 的挑战。
