Epidermal stem cell-derived exosomes improve wound healing by promoting the proliferation and migration of human skin fibroblasts

Deni Kang; Xiaoxiang Wang; Wentao Chen; Lujia Mao; Weiqiang Zhang; Yan Shi; Julin Xie; Ronghua Yang

doi:10.1093/burnst/tkae047

Epidermal stem cell-derived exosomes improve wound healing by promoting the proliferation and migration of human skin fibroblasts

Burns Trauma. 2024 Dec 16:12:tkae047. doi: 10.1093/burnst/tkae047. eCollection 2024.

Authors

Deni Kang¹, Xiaoxiang Wang², Wentao Chen³, Lujia Mao⁴, Weiqiang Zhang⁵, Yan Shi⁶, Julin Xie², Ronghua Yang¹

Affiliations

¹ 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.
² Department of Burns, The First Affiliated Hospital of Sun Yat-Sen University, 58 Zhongshan Second Road, Yuexiu District, Guangzhou City, Guangdong Province, 510062, China.
³ Department of Medical cosmetology, Foshan Second People's Hospital, 78 Weiguo Road, Chancheng District, Foshan City, Guangdong Province, 528000, China.
⁴ Department of Burn and Plastic Surgery, Guangzhou First People's Hospital, South China University of Technology, 1 Panfu Road, Yuexiu District, Guangzhou City, Guangdong Province, 510180, China.
⁵ The First Clinical Medical College, Guangdong Medical University, 2 Wenming East Road, Xiashan District, Zhanjiang City, Guangdong Province, 524002, China.
⁶ Department of Plastic, Medical Center of Burn Plastic and Wound Repair, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, No. 17 Yongwai Zheng Street, Donghu District, Nanchang City, Jiangxi Province, 330006, China.

Abstract

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.

Keywords: Epidermal stem cells; Exosomes; Human skin fibroblasts; Migration; Proliferation; Skin regeneration; Wound healing.