Schreiber Dominik, Jost Viktor, Bischof Michael, Seebach Kristina, Lammers Wim Jep, Douglas Rees, Schäfer Karl-Herbert
Dominik Schreiber, Viktor Jost, Michael Bischof, Kristina Seebach, Karl-Herbert Schäfer, Enteric Nervous System Group, University of Applied Sciences Kaiserslautern, Zweibrücken 66482, Germany.
World J Gastroenterol. 2014 Dec 28;20(48):18216-27. doi: 10.3748/wjg.v20.i48.18216.
To evaluate and characterize motility patterns from small intestinal gut segments depending on different perfusion media and pressures.
Experiments were carried out in a custom designed perfusion chamber system to validate and standardise the perfusion technique used. The perfusion chamber was built with a transparent front wall allowing for optical motility recordings and a custom made fastener to hold the intestinal segments. Experiments with different perfusion and storage media combined with different luminal pressures were carried out to evaluate the effects on rat small intestine motility. Software tools which enable the visualization and characterization of intestinal motility in response to different stimuli were used to evaluate the videotaped experiments. The data collected was presented in so called heatmaps thus providing a concise overview of form and strength of contractility patterns. Furthermore, the effect of different storage media on tissue quality was evaluated. Haematoxylin-Eosin stainings were used to compare tissue quality depending on storage and perfusion mode.
Intestinal motility is characterized by different repetitive motility patterns, depending on the actual situation of the gut. Different motility patterns could be recorded and characterized depending on the perfusion pressure and media used. We were able to describe at least three different repetitive patterns of intestinal motility in vitro. Patterns with an oral, anal and oro-anal propagation direction could be recorded. Each type of pattern finalized its movement with or without a subsequent distension of the wavefront. Motility patterns could clearly be distinguished in heatmap diagrams. Furthermore undirected motility could be observed. The quantity of the different patterns varies and is highly dependent on the perfusion medium used. Tissue preservation varies depending on the perfusion medium utilized, therefore media with a simple composition as Tyrode solution can only be recommended for short time experiments. The more complex media, MEM-HEPES medium and especially AQIX(®) RS-I tissue preservation reagent preserved the tissue much better during perfusion.
Perfusion media have to be carefully chosen considering type and duration of the experiments. If excellent tissue quality is required, complex media are favorable. Perfusion pressure is also of great importance due to the fact that a minimum amount of luminal pressure seems to be necessary to trigger intestinal contractions.
根据不同的灌注介质和压力,评估和描述小肠肠段的运动模式。
在定制设计的灌注室系统中进行实验,以验证和标准化所使用的灌注技术。灌注室的前壁为透明材质,便于进行光学运动记录,还配备了定制的固定装置来固定肠段。进行了不同灌注和储存介质与不同腔内压力相结合的实验,以评估对大鼠小肠运动的影响。使用能够可视化和描述肠道运动对不同刺激反应的软件工具来评估录像实验。收集的数据以所谓的热图形式呈现,从而提供收缩模式的形式和强度的简要概述。此外,评估了不同储存介质对组织质量的影响。使用苏木精 - 伊红染色来比较取决于储存和灌注模式的组织质量。
肠道运动具有不同的重复运动模式,这取决于肠道的实际情况。根据所使用的灌注压力和介质,可以记录和描述不同的运动模式。我们能够在体外描述至少三种不同的肠道重复运动模式。可以记录具有向口、向肛和口 - 肛传播方向的模式。每种模式在波前随后扩张或不扩张的情况下完成其运动。运动模式在热图中可以清晰区分。此外,还可以观察到无定向运动。不同模式的数量各不相同,并且高度依赖于所使用的灌注介质。组织保存情况因所使用的灌注介质而异,因此像台氏液这样成分简单的介质仅推荐用于短期实验。更复杂的介质,如MEM - HEPES培养基,尤其是AQIX(®) RS - I组织保存试剂,在灌注过程中能更好地保存组织。
必须根据实验类型和持续时间仔细选择灌注介质。如果需要优异的组织质量,复杂介质更有利。由于似乎需要最小量的腔内压力来触发肠道收缩,灌注压力也非常重要。