TY - JOUR
T1 - DYNAMIC PATHOLOGY OF ENTERIC NEURAL NETWORK USING CURCUMIN-ASSISTED MULTIPHOTON LASER IMAGING IN HIRSCHSPRUNG DISEASE
AU - Koike, Yuhki
AU - Mizoguchi, Akira
AU - Uchida, Keiichi
AU - Sato, Yuki
AU - Higashi, Koki
AU - Nagano, Yuka
AU - Matsushita, Kohei
AU - Sai, Kousyoku
AU - Kaito-Yamagishi, Aika
AU - Wang, Shujie
AU - Kayahara, Tetsuro
AU - Okugawa, Yoshinaga
AU - Tanaka, Kyosuke
AU - Inoue, Mikihiro
AU - Funabiki, Kazuo
AU - Kimura, Kazushi
AU - Goto, Hidemasa
AU - Yuge, Mizuki
AU - Nishimura, Yuhei
AU - Yuasa, Hiroto
AU - Toiyama, Yuji
N1 - Publisher Copyright:
Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.
PY - 2024
Y1 - 2024
N2 - 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.
AB - 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.
KW - curcumin vital staining
KW - green fluorescent protein
KW - Hirschsprung disease
KW - intravital fluorescence
KW - multi-photon laser-scanning microscopy
UR - http://www.scopus.com/inward/record.url?scp=85204031135&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85204031135&partnerID=8YFLogxK
U2 - 10.1097/SLA.0000000000006528
DO - 10.1097/SLA.0000000000006528
M3 - Article
C2 - 39263745
AN - SCOPUS:85204031135
SN - 0003-4932
JO - Annals of Surgery
JF - Annals of Surgery
ER -