Overexpressed vascular endothelial growth factor in adipose derived stem cells attenuates fibroblasts and skin injuries by ultraviolet radiation.

Yueping Mao,Yina Wang,Yue Xia,Xiaoyuan Xie

doi:10.1042/bsr20190433

Abstract

Adipose-derived stem cells (ADSCs) and vascular endothelial growth factor (VEGF) contribute to the healing of wound. The purpose of the present study was to investigate the role of VEGF produced by ADSCs in the protection of fibroblasts and skin of mice from ultraviolet (UV) radiation. ADSCs and fibroblasts were extracted from adipose and skin on the abdomen of mice by enzyme digestion methods. ADSCs surface markers were detected using flow cytometry, and immunofluorescence was used to identify fibroblasts. The expression of VEGF in modified ADSCs with lentivirus was determined. Fibroblasts were injured by UV radiation and co-cultured with ADSCs carrying overexpressed VEGF or normal VEGF. Cell cycle was assessed by flow cytometry. Mice were treated with UV radiation dorsally and injected with ADSCs containing overexpressed VEGF or normal VEGF. mRNA and protein levels of cell senescence-related genes were measured by qPCR and western blot. It was found that ADSCs with overexpressed VEGF not only promoted the effect of ADSCs on down-regulating senescence-associated (SA)-β-Gal, p21 and matrix metalloproteinase (MMP)-1, the healing of wound injured by UV radiation and up-regulating collagen I expression in fibroblasts and wound, but also on inhibiting cell cycle arrest in fibroblasts injured by UV radiation and preventing the skin from photoaging caused by UV radiation. VEGF expression in ADSCs played a key role in protecting skin fibroblasts from ageing, which further allowed the skin to resist photoaging, thereby promoting the recovery of wound injured by UV radiation.

Highlights

Three main types of UV radiation are characterized in terms of its wavelength as follows: UVA, UVB, and UVC
To obtain and identify adipose-derived stem cell (ADSC), the cells extracted from abdominal subcutaneous adipose tissues were cultured for 48 h and observed under an inverted phase contrast microscope, and their immunophenotypings were identified by flow cytometry
The identification of immunophenotyping by flow cytometry exhibited that these cells expressed a significantly high level of CD29 (96.3%) and CD44 (95.4%), while the CD31 (4.27%) and CD45 (0.64%) were low expressed (Figure 1B), indicating that these cells we extracted from primary tissues and could be seen as ADSCs

Summary

Introduction

Three main types of UV radiation are characterized in terms of its wavelength as follows: UVA, UVB, and UVC. In 1990s, it has been proved that in the process of skin repair, fibroblasts were a class of important cells that were almost involved in the entire progress of wound healing [4,5]. The migration and proliferation of dermal fibroblasts, which are the key processes in wound repair, largely affect wound contraction, deposition of extracellular matrix, and reconstruction of tissues [4,5]. Adipose-derived stem cells (ADSCs) were one type of mesenchymal stem cells that can be collected and cultured from excess adipose tissue from human or mouse [8,9].

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