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 Vol. 11, Issue 1, (2023)
- 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：
DOI：10.1093/burnst/tkad034
CLC：Published：2023，
Published Online：30 October 2023，
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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, 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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