The Antioxidant Effect of Small Extracellular Vesicles Derived from Aloe vera Peels for Wound Healing
The Antioxidant Effect of Small Extracellular Vesicles Derived from Aloe vera Peels for Wound Healing
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DOI：
10.1007/s13770-021-00367-8
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发表时间：
2021-07-27
影响因子：
3.6
通讯作者：
Cho, Yong Woo
中科院分区：
文献类型：
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作者：
Kim, Min Kang;Choi, Young Chan;Cho, Yong Woo
背景：来自植物的细胞外囊泡（EVs）已成为美容和治疗应用的潜在候选者。在这项研究中，我们从芦荟皮（a - ev）中分离出ev，并研究了a - ev的抗氧化和伤口愈合潜力。方法：采用超离心和切向流过滤分离a - ev，采用透射电镜和纳米颗粒跟踪分析对其进行表征。采用WST-1检测和流式细胞术观察a - ev的细胞毒性和细胞摄取情况。采用超氧化物歧化酶（SOD）活性测定和细胞抗氧化活性（CAA）测定评价a - ev的抗氧化作用。采用人角质形成细胞（HaCaT）和成纤维细胞（HDF）体外划痕实验评估伤口愈合潜力。采用定量RT-PCR方法分析核因子-红细胞2相关因子2 （Nrf2）及其相关基因的表达。结果：a - ev呈圆形，直径在50 ~ 200 nm之间。a - ev在人皮肤细胞上表现出良好的细胞相容性，并通过网格蛋白、小泡介导的内吞作用和膜融合被内化到HaCaT细胞中。SOD活性和CAA测定显示，a - ev具有抗氧化活性，并以剂量依赖性方式降低h2o2处理的HaCaT细胞内ROS水平。划痕实验表明，A- ev增强了HaCaT和HDF的迁移能力。此外，a - ev显著上调h2o2处理的HaCaT细胞中Nrf2、HO-1、CAT和SOD基因的mRNA表达。我们的研究结果表明，a - ev可以通过Nrf2激活抗氧化防御机制和伤口愈合过程。结论：a - ev作为一种潜在的皮肤再生剂具有广阔的应用前景。
Background: Extracellular vesicles (EVs) derived from plants have emerged as potential candidates for cosmetic and therapeutic applications. In this study, we isolated EVs from Aloe vera peels (A-EVs) and investigated the antioxidant and wound healing potential of A-EVs. Methods: A-EVs were isolated by ultracentrifugation and tangential flow filtration and were characterized using transmission electron microscopy, nanoparticle tracking analysis. The cytotoxicity and cellular uptake of A-EVs were investigated by WST-1 assay and flow cytometry. The antioxidant effect of A-EVs was evaluated by superoxide dismutase (SOD) activity assay and cellular antioxidant activity (CAA) assay. The wound healing potential was assessed by in vitro scratch assay using human keratinocytes (HaCaT) and fibroblasts (HDF). The expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and their associated genes was analyzed by quantitative RT-PCR. Results: A-EVs displayed a round shape and had diameters from 50 to 200 nm. A-EVs showed good cytocompatibility on human skin cells and were internalized into HaCaT cells via clathrin-, caveolae-mediated endocytosis, and membrane fusion. The SOD activity and CAA assays exhibited that A-EVs had antioxidant activity and reduced intracellular ROS levels in H2O2-treated HaCaT cells in a dose-dependent manner. A scratch assay showed that A-EVs enhanced the migration ability of HaCaT and HDF. Moreover, A-EVs significantly upregulated the mRNA expression of Nrf2, HO-1, CAT, and SOD genes in H2O2-treated HaCaT cells. Our findings reveal that A-EVs could activate the antioxidant defense mechanisms and wound healing process via the Nrf2 activation. Conclusion: Overall results suggest that the A-EVs are promising as a potential agent for skin regeneration.