Abramson Research Center, Children's Hospital of Philadelphia Research Institute, Philadelphia, Pennsylvania; Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania.
Abramson Research Center, Children's Hospital of Philadelphia Research Institute, Philadelphia, Pennsylvania.
Gastroenterology. 2024 Aug;167(3):547-559. doi: 10.1053/j.gastro.2024.02.045. Epub 2024 Mar 16.
BACKGROUND & AIMS: Hirschsprung's disease is defined by the absence of the enteric nervous system (ENS) from the distal bowel. Primary treatment is "pull-through" surgery to remove bowel that lacks ENS, with reanastomosis of "normal" bowel near the anal verge. Problems after pull-through are common, and some may be due to retained hypoganglionic bowel (ie, low ENS density). Testing this hypothesis has been difficult because counting enteric neurons in tissue sections is unreliable, even for experts. Tissue clearing and 3-dimensional imaging provide better data about ENS structure than sectioning.
Regions from 11 human colons and 1 ileal specimen resected during Hirschsprung's disease pull-through surgery were cleared, stained with antibodies to visualize the ENS, and imaged by confocal microscopy. Control distal colon from people with no known bowel problems were similarly cleared, stained, and imaged.
Quantitative analyses of human colon, ranging from 3 days to 60 years old, suggest age-dependent changes in the myenteric plexus area, ENS ganglion area, percentage of myenteric plexus occupied by ganglia, neurons/mm, and neuron Feret's diameter. Neuron counting using 3-dimensional images was highly reproducible. High ENS density in neonatal colon allowed reliable neuron counts using 500-μm × 500-μm regions (36-fold smaller than in adults). Hirschsprung's samples varied 8-fold in proximal margin enteric neuron density and had diverse ENS architecture in resected bowel.
Tissue clearing and 3-dimensional imaging provide more reliable information about ENS structure than tissue sections. ENS structure changes during childhood. Three-dimensional ENS anatomy may provide new insight into human bowel motility disorders, including Hirschsprung's disease.
先天性巨结肠症的定义是远端肠道缺乏肠神经系统（ENS）。主要治疗方法是“拖出”手术，切除缺乏 ENS 的肠道，并在肛门边缘附近重新吻合“正常”肠道。拖出后出现问题很常见，其中一些可能是由于保留了低神经节细胞肠（即低 ENS 密度）所致。由于即使是专家在对组织切片进行肠神经元计数时也不可靠，因此验证这一假说一直具有挑战性。组织透明化和 3 维成像比切片提供了更好的 ENS 结构数据。
对 11 个人类结肠和 1 个回肠标本进行清除，这些标本是在先天性巨结肠症拖出手术中切除的，并用抗体染色以可视化 ENS，并通过共聚焦显微镜进行成像。同样对无已知肠道问题的人的远端结肠进行了清除、染色和成像。
对从 3 天到 60 岁的人类结肠进行的定量分析表明，肠肌丛面积、ENS 神经节面积、神经节占肠肌丛的百分比、神经元/mm 和神经元 Feret 直径存在年龄依赖性变化。使用 3 维图像进行神经元计数具有高度可重复性。新生儿结肠的 ENS 密度高，使用 500-μm×500-μm 区域（比成人小 36 倍）可以可靠地计数神经元。先天性巨结肠样本在近端肠神经节细胞密度上差异 8 倍，并且切除的肠道中 ENS 结构具有不同的形态。
组织透明化和 3 维成像比组织切片提供了更可靠的 ENS 结构信息。ENS 结构在儿童时期发生变化。3 维 ENS 解剖结构可能为包括先天性巨结肠症在内的人类肠道运动障碍提供新的见解。
