Samarasena Jason B, Tarnawski Andrzej S, Ahluwalia Amrita, Shinoura Susumu, Choi Kee Don, Lee John G, Chang Kenneth J
Division of Gastroenterology and Hepatology, Department of Medicine, University of California-Irvine, Orange, California, USA.
Division of Gastroenterology and Hepatology, Department of Medicine, University of California-Irvine, Orange, California, USA; Division of Gastroenterology and Hepatology, Department of Medicine, Long Beach Veterans Affairs Healthcare System, Long Beach, California, USA.
Gastrointest Endosc. 2015 Dec;82(6):1116-20. doi: 10.1016/j.gie.2015.06.048. Epub 2015 Aug 28.
The GI tract is innervated by the autonomic enteric nervous system, mainly composed of submucosal Meissner's plexus and myenteric Auerbach's plexus, which is essential for motility, blood flow regulation, and secretory functions. In vivo visualization of the esophageal enteric nervous system (EENS) during endoscopy has not been possible without invasive mucosal resection. This study aimed to visualize the EENS without mucosal resection, in vivo by using the novel probe, needle-based confocal laser-induced endomicroscopy (nCLE) with a fluorescence neuronal probe, NeuroTrace, under EUS guidance and to evaluate the feasibility of ex vivo imaging of the neuronal network in submucosal biopsy samples acquired at endoscopy.
Four Yorkshire pigs were anesthetized and examined. In vivo experiment: During endoscopy, NeuroTrace was injected into the submucosa and muscularis propria of the middle and distal esophagus under EUS guidance, and nCLE imaging was performed. Ex vivo experiment: Submucosal tissue biopsy specimens from the porcine esophagus were obtained for ex vivo evaluation by using a "through-the-needle" forceps technique. After incubation of the samples in NeuroTrace solution, pCLE was used to visualize the EENS elements in the tissue.
Imaging of the EENS network by using EUS-guided nCLE was successful, both within the submucosa and the muscularis propria, and clearly visualized neuronal cells, glial cells, nerve bundles, and nerve fibers provided distinctive image patterns with excellent imaging quality. The use of the "through-the-needle" forceps technique achieved ex vivo images similar to those acquired in vivo.
EUS-guided in vivo imaging of the enteric nervous system is feasible without mucosal resection and provides a novel ex vivo imaging alternative for human application. These novel, minimally invasive imaging approaches could be of tremendous diagnostic value to better characterize and explore the EENS of the GI tract.
胃肠道由自主肠神经系统支配，主要由黏膜下的迈斯纳神经丛和肌间的奥尔巴赫神经丛组成，这对胃肠蠕动、血流调节及分泌功能至关重要。在内镜检查过程中，若不进行侵入性黏膜切除，就无法对食管肠神经系统（EENS）进行体内可视化观察。本研究旨在通过使用新型探头——基于针的共聚焦激光诱导内镜显微镜（nCLE）及荧光神经元探针NeuroTrace，在超声内镜（EUS）引导下，在不进行黏膜切除的情况下对EENS进行体内可视化观察，并评估在内镜检查时获取的黏膜下活检样本中神经元网络的体外成像可行性。
对4只约克夏猪进行麻醉并检查。体内实验：在内镜检查期间，在EUS引导下将NeuroTrace注入食管中下段的黏膜下层和固有肌层，并进行nCLE成像。体外实验：采用“针穿”钳取技术获取猪食管的黏膜下组织活检标本进行体外评估。将样本在NeuroTrace溶液中孵育后，使用pCLE对组织中的EENS成分进行可视化观察。
利用EUS引导的nCLE成功实现了EENS网络在黏膜下层和固有肌层内的成像，清晰地观察到神经元细胞、神经胶质细胞、神经束和神经纤维，提供了具有优异成像质量的独特图像模式。“针穿”钳取技术获得的体外图像与体内获取的图像相似。
在不进行黏膜切除的情况下，EUS引导的肠神经系统体内成像可行，并为人体应用提供了一种新的体外成像方法。这些新颖的微创成像方法对更好地表征和探索胃肠道的EENS可能具有巨大的诊断价值。
