• Annals of surgery · Sep 2024

    Dynamic Pathology of Enteric Neural Network using Curcumin-assisted Multiphoton Laser Imaging in Hirschsprung Disease.

    • Yuhki Koike, Akira Mizoguchi, Keiichi Uchida, Yuki Sato, Koki Higashi, Yuka Nagano, Kohei Matsushita, Kousyoku Sai, Aika Kaito-Yamagishi, Shujie Wang, Tetsuro Kayahara, Yoshinaga Okugawa, Kyosuke Tanaka, Mikihiro Inoue, Kazuo Funabiki, Kazushi Kimura, Hidemasa Goto, Mizuki Yuge, Yuhei Nishimura, Hiroto Yuasa, and Yuji Toiyama.
    • Departments of Gastrointestinal and Pediatric Surgery, Mie University Graduate School of Medicine; Tsu, Japan.
    • Ann. Surg. 2024 Sep 12.

    ObjectiveIn living tissue, it has been difficult to make microscopic-level observations without damaging the tissue.Summary Background DataWe have invented a novel intravital fluorescent observation method (IFOM) for real-time tissue observation, combining multi-photon laser scanning microscopy (MPLSM) with curcumin vital staining (CVS-IFOM). The aim of this study was to use CVS-IFOM to analyze the enteric nervous system (ENS) in mice and human patients with hypoganglionosis and Hirschsprung disease.MethodsIn an initial viability study, we compared live ENS images from non-fluorescent C57BL6 mice stained with curcumin (n=5) and GFP mice (n=5) using MPLSM. We then explored CVS-IFOM for the live examination of resected colon tissues from one hypoganglionosis and three Hirschsprung disease patients.ResultsIn the viability study, detailed ENS histological features were only observed in the curcumin-stained mice. In the hypoganglionosis patient, CVS-IFOM provided ENS details that were not visualized under H&E staining or calretinin immunohistochemistry, allowing the analysis of ENS size, neural bundle number, and neural cell number per plexus. In Hirschsprung disease patients, CVS-IFOM showed a gradual hypoplastic change in the ENS from the oral wedge to the anal wedge, detecting disproportionate changes in the ENS within the same intestinal level, supporting a circumferentially uneven distribution of the intestinal ENS.ConclusionCVS-IFOM may be supportive for intraoperative pathological diagnosis during surgeries in Hirschsprung disease.Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.

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