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经内镜十二指肠和直肠肌肉活检采样:一种在猪模型中检测肌间神经元的生存研究。

Endoscopic muscle biopsy sampling of the duodenum and rectum: a pilot survival study in a porcine model to detect myenteric neurons.

机构信息

Developmental Endoscopy Unit, Division of Gastroenterology and Hepatology, Mayo Clinic College of Medicine, Rochester, Minnesota, USA.

Enteric NeuroScience Program, Division of Gastroenterology and Hepatology, Mayo Clinic College of Medicine, Rochester, Minnesota, USA.

出版信息

Gastrointest Endosc. 2018 Feb;87(2):600-606. doi: 10.1016/j.gie.2017.07.023. Epub 2017 Jul 20.

Abstract

BACKGROUND AND AIMS

Small bowel and colorectal muscle biopsy sampling requires a surgical approach. Advancing our understanding of the pathophysiology of motility disorders, such as functional bowel disorders, intestinal pseudo-obstruction, and slow-transit constipation, is hindered by our inability to noninvasively obtain muscularis propria (MP) for evaluation of multiple cell types, including myenteric neurons. The aims of this study were to determine (1) technical feasibility, reproducibility, and safety of performing duodenal endoscopic muscle biopsy sampling (dEMB) and rectal endoscopic muscle biopsy sampling (rEMB) using a clip-assist technique and (2) the presence of myenteric neurons in tissue samples.

METHODS

Five 40-kg pigs were studied. Each animal underwent a dEMB and rEMB procedure. dEMB was performed using a single resection clip-assist technique. An over-the-scope clip was advanced to the duodenum. Tissue was suctioned into the cap and the clip deployed. The pseudopolyp of the duodenal wall created was then resected using snare electrocautery. rEMB was performed using a double resection clip-assist technique. EMR was initially performed to uncover the underlying MP using a band ligation technique. An over-the-scope clip was then advanced to the exposed MP. The MP was retracted and suctioned into the cap and the clip deployed. The pseudopolyp of the MP was resected using snare electrocautery. An antibody to protein gene product 9.5 was used to determine the presence of myenteric neurons in the samples. Animals were kept alive for 2 weeks, at which time an upper endoscopy and necropsy were performed.

RESULTS

dEMB and rEMB were successfully performed in all animals with no procedural adverse events using this "no hole" (close then cut) approach. Mean procedure times for dEMB and rEMB were 23.7 ± 2.5 minutes and 13.25 ± 2.8 minutes, respectively. Mean length of resected full-thickness duodenal wall was 13.25 ± 4.3 mm and rectal MP was 12.5 ± 1.7 mm. Hematoxylin and eosin stain and antibody to protein gene product 9.5 confirmed the presence of MP with inner circular, outer longitudinal, and intermuscular layers, including myenteric neurons, in all samples. Clinical course was uneventful in all animals. Repeat upper endoscopy at 2 weeks showed well-healed dEMB sites. Necropsy in all animals showed no perforation, fluid collection, or abscess at the dEMB and rEMB sites.

CONCLUSIONS

Based on this preclinical study, dEMB and rEMB appear to be technically feasible, reproducible, and safe. Sufficient MP tissue was obtained to identify myenteric neurons. These promising results are a step toward successful and safe implementation of these techniques into clinical practice for tissue diagnosis of muscle-based pathologies.

摘要

背景与目的

小肠和结直肠肌肉活检采样需要手术方法。我们对运动障碍的病理生理学的理解,例如功能性肠病、假性肠梗阻和慢传输性便秘,受到限制,因为我们无法非侵入性地获得肌肉层(MP)来评估多种细胞类型,包括肌间神经元。本研究的目的是确定:(1)使用夹辅助技术进行十二指肠内镜下肌肉活检采样(dEMB)和直肠内镜下肌肉活检采样(rEMB)的技术可行性、可重复性和安全性;(2)组织样本中存在肌间神经元。

方法

对 5 只 40 公斤重的猪进行了研究。每只动物都接受了 dEMB 和 rEMB 手术。dEMB 使用单切夹辅助技术进行。将过镜夹推进到十二指肠。组织被吸入帽中并展开夹子。然后使用圈套电烙术切除十二指肠壁的假息肉。rEMB 使用双切夹辅助技术进行。首先使用结扎带技术进行 EMR,以暴露出下方的 MP。然后将过镜夹推进到暴露的 MP。将 MP 缩回并吸入帽中并展开夹子。使用圈套电烙术切除 MP 假息肉。使用针对蛋白基因产物 9.5 的抗体来确定样本中是否存在肌间神经元。动物存活 2 周,此时进行上消化道内镜检查和尸检。

结果

所有动物均成功完成了 dEMB 和 rEMB,没有使用这种“无孔”(先封闭再切割)方法的手术不良事件。dEMB 和 rEMB 的平均手术时间分别为 23.7±2.5 分钟和 13.25±2.8 分钟。十二指肠全层切除的平均长度为 13.25±4.3mm,直肠 MP 为 12.5±1.7mm。苏木精-伊红染色和针对蛋白基因产物 9.5 的抗体证实,所有样本均存在具有内环形、外纵形和肌间层的 MP,包括肌间神经元。所有动物的临床病程均无异常。2 周时的重复上消化道内镜检查显示 dEMB 部位愈合良好。所有动物的尸检均未在 dEMB 和 rEMB 部位发现穿孔、积液或脓肿。

结论

基于这项临床前研究,dEMB 和 rEMB 似乎在技术上是可行的、可重复的和安全的。获得了足够的 MP 组织来识别肌间神经元。这些有希望的结果是成功和安全地将这些技术应用于肌肉相关疾病的组织诊断的临床实践的重要一步。

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