Kobayashi Masakuni, Sumiyama Kazuki, Shimojima Naoki, Ieiri Satoshi, Okano Hideyuki, Kamba Shunsuke, Fujimura Takumi, Hirobe Seiichi, Kuroda Tatsuo, Takahashi-Fujigasaki Junko
Department of Endoscopy and Division of Gastroenterology and Hepatology, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan.
Department of Endoscopy, The Jikei University School of Medicine, Tokyo, Japan.
J Gastroenterol Hepatol. 2017 Sep;32(9):1604-1610. doi: 10.1111/jgh.13754.
In preceding studies, we identified that the myenteric plexus (MP) could be visualized with confocal laser endomicroscopy (CLE) by applying neural fluorescent probes lacking clinical safety profiling data from the submucosal side. In this study, we evaluated the technical feasibility of MP visualization using probe-based CLE (pCLE) from the serosal side with cresyl violet (CV), which has been used clinically for chromoendoscopy.
The dye affinity of CV for MP was first explored in an in vivo transgenic mouse model using neural crest derivatives labeled with green fluorescent protein. We also tested the feasibility of CV-assisted visualization of MP in human surgical specimens, wherein the tissue dying and pCLE observation were performed from the serosal side. In the human study, rate of MP visualization by pCLE was evaluated as the primary outcome. We also evaluated the sensitivity and specificity of MP visualization by pCLE, using pathological presence/absence of MP as the gold standard.
We confirmed the dye affinity of CV to MP in all tested models. The MP appeared as brightly stained ladder-like structures with pCLE, and in the human study, MP was visualized in 12/14 (85.7%) samples, with 92.3% sensitivity and 100% specificity. In positive cases showing the ladder-like structure of MP by pCLE, the mean maximum and minimum widths of nerve strands were 54.3 (± 23.6) and 19.7 (± 6.0) μm, respectively. A ganglion was detected by pCLE in 10 cases (10/12, 83.3%).
This study demonstrated the technical feasibility of visualizing the MP in real time by CV-assisted pCLE (UMIN-CTR number, UMIN000015056).
在之前的研究中，我们发现通过从黏膜下层应用缺乏临床安全性分析数据的神经荧光探针，肌间神经丛（MP）可以用共聚焦激光内镜显微镜（CLE）进行可视化。在本研究中，我们评估了使用基于探针的CLE（pCLE）从浆膜侧用甲酚紫（CV）可视化MP的技术可行性，CV已在临床上用于色素内镜检查。
首先在体内转基因小鼠模型中利用绿色荧光蛋白标记的神经嵴衍生物探索CV对MP的染料亲和力。我们还测试了CV辅助在人类手术标本中可视化MP的可行性，其中从浆膜侧进行组织染色和pCLE观察。在人体研究中，将通过pCLE可视化MP的比率作为主要结果进行评估。我们还以MP的病理存在与否作为金标准，评估了通过pCLE可视化MP的敏感性和特异性。
我们在所有测试模型中均证实了CV对MP的染料亲和力。通过pCLE，MP呈现为染色明亮的梯状结构，在人体研究中，14个样本中有12个（85.7%）可视化了MP，敏感性为92.3%，特异性为100%。在通过pCLE显示MP梯状结构的阳性病例中，神经束的平均最大宽度和最小宽度分别为54.3（±23.6）μm和19.7（±6.0）μm。通过pCLE在10例（10/12，83.3%）中检测到神经节。
本研究证明了通过CV辅助pCLE实时可视化MP的技术可行性（UMIN-CTR编号，UMIN000015056）。
