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 YuasaYuji Toiyama

Research output: Contribution to journalArticlepeer-review

Abstract

Objective: In living tissue, it has been difficult to make microscopic-level observations without damaging the tissue. Summary background data: We 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. Methods: In 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. Results: In 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. Conclusion: CVS-IFOM may be supportive for intraoperative pathological diagnosis during surgeries in Hirschsprung disease.

Original languageEnglish
JournalAnnals of Surgery
DOIs
Publication statusAccepted/In press - 2024

All Science Journal Classification (ASJC) codes

  • Surgery

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