Adipose mesenchymal stem cell-derived exosomes stimulated by hydrogen peroxide enhanced skin flap recovery in ischemia-reperfusion injury
Adipose mesenchymal stem cell-derived exosomes stimulated by hydrogen peroxide enhanced skin flap recovery in ischemia-reperfusion injury
复制标题
过氧化氢刺激的脂肪间充质干细胞来源的外泌体增强了缺血再灌注损伤中皮瓣的恢复
DOI：
10.1016/j.bbrc.2018.04.065
复制
发表时间：
2018-06-02
影响因子：
3.1
通讯作者：
Han, Yan
中科院分区：
文献类型：
--
作者：
Bai, Yun;Han, Yu-di;Han, Yan
背景：间充质干细胞(MSC)来源的外切体已被认为是治疗缺血性疾病或损伤的新候选方法，并可能成为细胞治疗的替代疗法。本研究旨在探讨脂肪间充质干细胞(ADSC)来源的外切体能否在缺血再灌注(I/R)损伤时对皮瓣起到保护作用，并诱导新生血管形成。方法：采用结扎左上腹浅血管(SIEA)后血液灌流的方法，观察外切体在体内移植皮瓣I/R损伤中的作用。将正常ADSC来源的Exosome(ADSC-exos)和经H_2O_2预处理的ADSC来源的Exosome(H_2O_2-ADSC-exos)注入皮瓣内。应用激光多普勒血流灌注成像技术(LDPI)检测组织瓣的血液灌流单位(BPU)，免疫组织化学(IHC)检测血管内皮细胞CD31标记的微血管密度。苏木精-伊红染色(H&E)和TdT介导的生物素化dUTP缺口末端标记(TUNEL)技术检测皮瓣炎性细胞浸润和细胞凋亡情况。结果：体内实验显示，与I/R组相比，Exosome组术后第5天的皮瓣存活率和毛细血管密度显著增加，炎症反应和细胞凋亡明显减少(P<；0.05)。此外，与正常外切体相比，双氧水-ADSC-exos有更好的结果(P<；0.05)。ADSC-exos能显著促进人脐静脉内皮细胞(HUVEC)的增殖(P<；0.05)，但不同微环境来源的Exosome之间无统计学差异(P>；0.05)。HUVEC与H_2O_2-ADSC-exos共培养后，细胞迁移率增加，形成的索状结构明显多于ADSC-exos组和对照组(P<；0.05)。结论：ADSC-exOS能提高I/R损伤后皮瓣存活，促进新生血管形成，减轻炎症反应和细胞凋亡。使用特定的微环境进行体外干细胞培养，如含有低浓度的过氧化氢，将促进定制外切体的开发，用于皮瓣移植的无细胞治疗应用。(C)2018 Elsevier Inc.保留所有权利。
Background: Mesenchymal stem cell (MSC)-derived exosomes have been recognized as new candidates for the treatment of ischemic disease or injury and may be an alternative treatment for cell therapy. This aim of the study was to evaluate whether exosomes derived from adipose mesenchymal stem cell (ADSC) can protect the skin flap during ischemia-reperfusion (I/R) injury and induce neovascularization.Methods: To investigate the effects of exosomes in the I/R injury of flap transplantation in vivo, flaps were subjected to 6 h of ischemia by ligating the left superficial inferior epigastric vessels (SIEA) followed by blood perfusion. Exosomes derived from normal ADSC (ADSC-exos) and exosomes derived from ADSC preconditioned with H2O2 (H2O2-ADSC-exos) were injected into the flaps. Then, the blood perfusion unit (BPU) of the flaps was measured by Laser Doppler Perfusion Imaging (LDPI) and microvessel density was determined by the endothelial with cell marker CD31 with Immunohistochemistry (IHC) staining. Inflammatory cell infiltration of the skin flap and apoptosis were detected by hematoxylin & eosin staining (H&E) and the TdT-mediated biotinylated dUTP nick end-labeling (TUNEL) technique.Results: In vivo, exosomes significantly increased flap survival and capillary density compared to I/R on postoperative day 5, and decreased the inflammatory reaction and apoptosis in the skin flap (P < 0.05). Furthermore, H2O2-ADSC-exos had better outcomes compared to normal exosomes (P < 0.05). ADSC-exos could significantly increase human umbilical vein endothelial cell (HUVEC) proliferation (P < 0.05), but no statistic difference was found in exosomes derived from different microenvironments (P > 0.05). HUVEC co-cultured with H2O2-ADSC-exos increased the migration ratio and generated more cord-like structures compared to ADSC-exos and the control group (P < 0.05).Conclusion: ADSC-exos can enhance skin flap survival, promote neovascularization and alleviate the inflammation reaction and apoptosis in the skin flap after I/R injury. The use of a specific microenvironment for in vitro stem cell culture, such as one containing a low concentration of H2O2, will facilitate the development of customized exosomes for cell-free therapeutic applications in skin flap transplantation. (C) 2018 Elsevier Inc. All rights reserved.