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基于菠萝蛋白酶的酶促清创对皮肤细胞的影响。

Effect of Bromelain-Based Enzymatic Debridement on Skin Cells.

作者信息

Schulz Alexandra, Fuchs Paul Christian, Oplaender Christian, Valdez Leandra Börner, Schiefer Jennifer Lynn

机构信息

Department of Plastic Surgery, Hand Surgery, Burn Center, University of Witten/Herdecke, Cologne-Merheim Medical Center (CMMC), Germany.

Cell and Molecular Laboratory, Department of Trauma and Hand Surgery, Heinrich Heine University, Düsseldorf, Germany.

出版信息

J Burn Care Res. 2018 Jun 13;39(4):527-535. doi: 10.1093/jbcr/irx011.

Abstract

Several reports have concluded that enzymatic debridement based on Bromelain (NX) is selective and efficient. Although clinical trials showed that viable tissue is not damaged at the macroscopic level, the effect on the cellular level is largely unknown. The current study is meant to close this gap by evaluating whether NX has an effect on vital cells of the human dermis on a cellular level. In an experimental in vitro study design, the effect of NX on human keratinocytes, fibroblasts, and macrophages was analyzed. Enzymatic treatment was performed for 4 hours by using either cell culture medium or phosphate-buffered saline as diluting agent for NX. Cell viability and relative cell number in relation to untreated control cells were determined using a resazurin-based assay. In addition, the development of enzyme activity during clinical treatment was analyzed: wound fluid collected from a burn wound at different points of debridement was applied on collagen-elastin disks to prove enzymatic digestion activity. Both keratinocytes and fibroblasts were damaged by NX even at low concentrations. Both cell types showed improved survival when a medium was used for dissolving NX. Macrophages appeared to resist NX treatment more efficiently than the other cell types. In the clinical trial, NX activity in the wound fluid decreased clearly following 4 hours of enzymatic debridement. NX induces toxicity of vital skin cells in vitro. However, macrophages appear to be more resistant against NX treatment in vitro. The inflammatory responses of vital cells in the burn wound itself are likely to inhibit NX activity. The effect of this inflammatory process on NX activity will have to be investigated in future studies.

摘要

多项报告得出结论,基于菠萝蛋白酶(NX)的酶促清创具有选择性且高效。尽管临床试验表明在宏观层面上存活组织未受损伤,但在细胞层面的影响在很大程度上尚不清楚。当前的研究旨在通过评估NX在细胞层面上是否对人真皮的重要细胞产生影响来填补这一空白。在一项体外实验研究设计中,分析了NX对人角质形成细胞、成纤维细胞和巨噬细胞的影响。使用细胞培养基或磷酸盐缓冲盐水作为NX的稀释剂进行4小时的酶促处理。使用基于刃天青的检测方法测定与未处理的对照细胞相关的细胞活力和相对细胞数量。此外,分析了临床治疗期间酶活性的变化:将在清创不同时间点从烧伤创面收集的创面渗出液应用于胶原 - 弹性蛋白圆盘以证明酶消化活性。即使在低浓度下,NX也会损伤角质形成细胞和成纤维细胞。当使用培养基溶解NX时,这两种细胞类型的存活率均有所提高。巨噬细胞似乎比其他细胞类型更能有效抵抗NX处理。在临床试验中,酶促清创4小时后,创面渗出液中的NX活性明显下降。NX在体外可诱导重要皮肤细胞产生毒性。然而,巨噬细胞在体外似乎对NX处理更具抗性。烧伤创面本身重要细胞的炎症反应可能会抑制NX活性。这一炎症过程对NX活性的影响将有待未来的研究进行探究。

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