Extracellular vesicles from 3D cultured dermal papilla cells improve wound healing via Krüppel-like factor 4/vascular endothelial growth factor A -driven angiogenesis

Yunwei Wang; Kuo Shen; Yulin Sun; Peng Cao; Jia Zhang; Wanfu Zhang; Yang Liu; Hao Zhang; Yang Chen; Shaohui Li; Chaolei Xu; Chao Han; Yating Qiao; Qingyi Zhang; Bin Wang; Liang Luo; Yunshu Yang; Hao Guan

doi:10.1093/burnst/tkad034

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Extracellular vesicles from 3D cultured dermal papilla cells improve wound healing via Krüppel-like factor 4/vascular endothelial growth factor A -driven angiogenesis

更新时间：2024-02-04

- Extracellular vesicles from 3D cultured dermal papilla cells improve wound healing via Krüppel-like factor 4/vascular endothelial growth factor A -driven angiogenesis
- Extracellular vesicles from 3D cultured dermal papilla cells improve wound healing via Krüppel-like factor 4/vascular endothelial growth factor A -driven angiogenesis
- Burns & Trauma 2023年11卷第1期
- 作者机构：
  
  1. Department of Burns and Cutaneous Surgery
  2. Fourth Military Medical University
  3. Department of Plastic Surgery
  4. Tongji University School of Medicine
  5. Department of Burns and Plastic Surgery, General Hospital of Ningxia Medical University
  6. Department of Dermatology
  7. Department of hair diagnosis and treatment, Peking University Shougang Hospital
- 作者简介：
- 基金信息：
  
  This work was supported by the National Natural Science Foundation of China (No. 82272268) and the Industry-University-Research Innovation Fund of China (2021JH030).
- DOI：10.1093/burnst/tkad034
  中图分类号：
- 纸质出版日期：2023，
  
  网络出版日期：2023-10-30，
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Yunwei Wang, Kuo Shen, Yulin Sun, 等. Extracellular vesicles from 3D cultured dermal papilla cells improve wound healing via Krüppel-like factor 4/vascular endothelial growth factor A -driven angiogenesis[J]. Burns & Trauma, 2023,11(1).

Yunwei Wang, Kuo Shen, Yulin Sun, Peng Cao, Jia Zhang, Wanfu Zhang, Yang Liu, Hao Zhang, Yang Chen, Shaohui Li, Chaolei Xu, Chao Han, Yating Qiao, Qingyi Zhang, Bin Wang, Liang Luo, Yunshu Yang, Hao Guan, Extracellular vesicles from 3D cultured dermal papilla cells improve wound healing via Krüppel-like factor 4/vascular endothelial growth factor A -driven angiogenesis, Burns & Trauma, Volume 11, 2023, tkad034, https://doi.org/10.1093/burnst/tkad034
Yunwei Wang, Kuo Shen, Yulin Sun, 等. Extracellular vesicles from 3D cultured dermal papilla cells improve wound healing via Krüppel-like factor 4/vascular endothelial growth factor A -driven angiogenesis[J]. Burns & Trauma, 2023,11(1). DOI： 10.1093/burnst/tkad034.

Yunwei Wang, Kuo Shen, Yulin Sun, Peng Cao, Jia Zhang, Wanfu Zhang, Yang Liu, Hao Zhang, Yang Chen, Shaohui Li, Chaolei Xu, Chao Han, Yating Qiao, Qingyi Zhang, Bin Wang, Liang Luo, Yunshu Yang, Hao Guan, Extracellular vesicles from 3D cultured dermal papilla cells improve wound healing via Krüppel-like factor 4/vascular endothelial growth factor A -driven angiogenesis, Burns & Trauma, Volume 11, 2023, tkad034, https://doi.org/10.1093/burnst/tkad034 DOI：

摘要

Abstract

BackgroundNon-healing wounds are an intractable problem of major clinical relevance. Evidence has shown that dermal papilla cells (DPCs) may regulate the wound-healing process by secreting extracellular vesicles (EVs). However

low isolation efficiency and restricted cell viability hinder the applications of DPC-EVs in wound healing. In this study

we aimed to develop novel 3D-DPC spheroids (tdDPCs) based on self-feeder 3D culture and to evaluate the roles of tdDPC-EVs in stimulating angiogenesis and skin wound healing.MethodsTo address the current limitations of DPC-EVs

we previously developed a self-feeder 3D culture method to construct tdDPCs. DPCs and tdDPCs were identified using immunofluorescence staining and flow cytometry. Subsequently

we extracted EVs from the cells and compared the effects of DPC-EVs and tdDPC-EVs on human umbilical vein endothelial cells (HUVECs) in vitro using immunofluorescence staining

a scratch-wound assay and a Transwell assay. We simultaneously established a murine model of full-thickness skin injury and evaluated the effects of DPC-EVs and tdDPC-EVs on wound-healing efficiency in vivo using laser Doppler

as well as hematoxylin and eosin

Masson

CD31 and α-SMA staining. To elucidate the underlying mechanism

we conducted RNA sequencing (RNA-seq) of tdDPC-EV- and phosphate-buffered saline-treated HUVECs. To validate the RNA-seq data

we constructed knockdown and overexpression vectors of Krüppel-like factor 4 (KLF4). Western blotting

a scratch-wound assay

a Transwell assay and a tubule-formation test were performed to detect the protein expression

cell migration and lumen-formation ability of KLF4 and vascular endothelial growth factor A (VEGFA) in HUVECs incubated with tdDPC-EVs after KLF4 knockdown or overexpression. Dual-luciferase reporter gene assays were conducted to verify the activation effect of KLF4 on VEGFA.ResultsWe successfully cultured tdDPCs and extracted EVs from DPCs and tdDPCs. The tdDPC-EVs significantly promoted the proliferation

lumen formation and migration of HUVECs. Unlike DPC-EVs

tdDPC-EVs exhibited significant advantages in terms of promoting angiogenesis

accelerating wound healing and enhancing wound-healing efficiency both in vitro and in vivo. Bioinformatics analysis and further functional experiments verified that the tdDPC-EV-regulated KLF4/VEGFA axis is pivotal in accelerating wound healing.Conclusions3D cultivation can be utilized as an innovative optimization strategy to effectively develop DPC-derived EVs for the treatment of skin wounds. tdDPC-EVs significantly enhance wound healing via KLF4/VEGFA-driven angiogenesis.

关键词

Keywords

Extracellular vesiclesDermal papilla cells3D spheroidsWound healingAngiogenesis

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