表皮干细胞来源的外泌体通过促进人皮肤成纤维细胞的增殖和迁移来改善伤口愈合。
Epidermal stem cell-derived exosomes improve wound healing by promoting the proliferation and migration of human skin fibroblasts.
作者信息
Kang Deni, Wang Xiaoxiang, Chen Wentao, Mao Lujia, Zhang Weiqiang, Shi Yan, Xie Julin, Yang Ronghua
机构信息
Department of Burn and Plastic Surgery, Guangzhou First People's Hospital, Guangzhou Medical University, 1 Panfu Road, Yuexiu District, Guangzhou City, Guangdong Province, 510180, China.
Department of Burns, The First Affiliated Hospital of Sun Yat-Sen University, 58 Zhongshan Second Road, Yuexiu District, Guangzhou City, Guangdong Province, 510062, China.
出版信息
Burns Trauma. 2024 Dec 16;12:tkae047. doi: 10.1093/burnst/tkae047. eCollection 2024.
BACKGROUND
Epidermal stem cells (ESCs) are primarily located in the basal layer of the epidermis and play a crucial role in wound healing. ESCs-derived exosomes (ESCs-Exo) are emerging as promising candidates for skin regeneration and wound healing. However, the underlying mechanisms remain unclear. This study aims to investigate the role and mechanisms of ESCs-Exo in promoting the proliferation, migration, and collagen synthesis of human skin fibroblasts (HSFBs).
METHODS
This study generated, isolated, and characterized ESC-Exos. The effects of ESCs-Exo on the proliferation of human skin fibroblasts (HSFBs) were detected via Cell Counting Kit-8 (CCK8), 5-Ethynyl-2'-deoxyuridine (EdU), and Proliferating Cell Nuclear Antigen (PCNA) and Marker of Proliferation Ki-67 (MKI67) gene expression methods. The effect of ESCs-Exo on the migration of HSFBs was detected via a transwell assay and a scratch test. The concentrations of collagen secreted by the HSFBs and the mRNAs of the two kinds of collagen expressed by the HSFBs were analyzed. We also analyzed the phosphorylation of Protein Kinase N1 (PKN1) and the expression of cyclins via western blotting. Finally, the effect of ESCs-Exo on wound healing was verified by animal experiments, and the key genes and signaling pathways of ESCs-Exo were excavated by transcriptomic analysis.
RESULTS
Western blotting revealed that the exosomes of ESCs highly expressed established markers such as Alix, CD63, and CD9. ESC-Exos significantly promoted HSFB proliferation and migration in a dose-dependent manner, as well as HSFB collagen synthesis, and effectively increased the ratio of collagen III/I. In addition, bioinformatics analysis showed that the expression of key gene C-X-C motif chemokine ligand 9 was lower in the ESCs-Exo group, which may promote wound healing by regulating PKN1-cyclin and tumor necrosis factor signaling pathways. Animal experiments demonstrated that ESCs-Exo could reduce inflammation and accelerate wound healing.
CONCLUSIONS
In this study, we found that ESCs-Exo may improve wound healing by promoting the proliferation and migration of HSFBs.
背景
表皮干细胞（ESCs）主要位于表皮基底层，在伤口愈合中起关键作用。表皮干细胞来源的外泌体（ESCs-Exo）正成为皮肤再生和伤口愈合的有前景的候选物。然而，其潜在机制仍不清楚。本研究旨在探讨ESCs-Exo在促进人皮肤成纤维细胞（HSFBs）增殖、迁移和胶原合成中的作用及机制。
方法
本研究制备、分离并鉴定了ESCs-Exos。通过细胞计数试剂盒-8（CCK8）、5-乙炔基-2'-脱氧尿苷（EdU）、增殖细胞核抗原（PCNA）和增殖标志物Ki-67（MKI67）基因表达方法检测ESCs-Exo对人皮肤成纤维细胞（HSFBs）增殖的影响。通过Transwell实验和划痕实验检测ESCs-Exo对HSFBs迁移的影响。分析HSFBs分泌的胶原蛋白浓度以及HSFBs表达的两种胶原蛋白的mRNA。我们还通过蛋白质印迹分析了蛋白激酶N1（PKN1）的磷酸化和细胞周期蛋白的表达。最后，通过动物实验验证ESCs-Exo对伤口愈合的影响，并通过转录组分析挖掘ESCs-Exo的关键基因和信号通路。
结果
蛋白质印迹显示ESCs的外泌体高表达已确定的标志物，如Alix、CD63和CD9。ESCs-Exos以剂量依赖性方式显著促进HSFBs增殖和迁移，以及HSFBs胶原合成，并有效增加III型与I型胶原蛋白的比例。此外，生物信息学分析表明，ESCs-Exo组中关键基因C-X-C基序趋化因子配体9的表达较低，其可能通过调节PKN1-细胞周期蛋白和肿瘤坏死因子信号通路促进伤口愈合。动物实验表明ESCs-Exo可减轻炎症并加速伤口愈合。
结论
在本研究中，我们发现ESCs-Exo可能通过促进HSFBs的增殖和迁移来改善伤口愈合。
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