Effects and Mechanism of Plasma-Activated Medium on Angiogenesis of Vascular Endothelial Cells

Yue Hao,Na Liu,Xingmin Shi,Guanjun Zhang,Xiangni Wang,Guimin Xu,Yixin Cui

doi:10.3390/app11209603

Abstract

After cell culture medium is treated with low temperature plasma (LTP), the liquid is rich in reactive oxygen and nitrogen species (RONS), and becomes plasma-activated medium (PAM). PAM, as the supplier of RONS, can affect the angiogenesis of cells. The purpose of this study is to investigate the effects and related mechanism of PAM on human umbilical vein endothelial cells (HUVECs). Cell viability and cell cycle were evaluated after HUVECs were treated with PAM for 24 h. Changes in cell angiogenesis, migration and adhesion, secretion of cytokines such as VEGF and bFGF, expression of VEGFR-2 and phosphorylation of the key proteins in the MEK/ERK signaling pathway, concentrations of H2O2 and NO2− in PAM and in cells were also investigated. The results showed that PAM obtained by LTP treatment had dual effects on the angiogenesis of HUVECs: PAM obtained by short-term LTP treatment promoted the angiogenesis of HUVECs, while PAM obtained by long-term LTP treatment inhibited the angiogenesis of HUVECs. The mechanism may be that PAM treatment changes the content of RONS, affects the VEGF-VEGFR-2 signaling pathway, and ultimately affects the angiogenesis of HUVECs.

Highlights

After the cell culture medium is treated with low temperature plasma (LTP) jet [1] for a period of time, the ions, electrons, neutral particles, ultraviolet rays, reactive oxygen and nitrogen species (RONS) and other substances react with the culture medium [2] and produce plasma-activated medium (PAM)
The results of MTT and cell cycle experiments in this study showed that PAM produced by LTP treatment for 15–30 s could promote the proliferation and cell viability of human umbilical vein endothelial cells (HUVECs) after 24 h culture, and the proportion of cells in S phase increased
PAM produced by short-term LTP treatment can promote the proliferation, angioPAM produced by short-term LTP treatment can promote the proliferation, angiogenesis, migration and adhesion of HUVECs, while PAM produced by long-term LTP

Summary

Introduction

After the cell culture medium is treated with low temperature plasma (LTP) jet (atmospheric pressure plasma jet, APPJ) [1] for a period of time, the ions, electrons, neutral particles (atoms, gas molecules, etc.), ultraviolet rays, RONS and other substances react with the culture medium [2] and produce plasma-activated medium (PAM). Related studies have shown that short-term PAM treatment can promote the proliferation of fibroblasts [3], induce osteoblast differentiation [4], inhibit over 20 kinds of cancer cells cultured in vitro [5], including melanoma cells [6], prostate cancer cells [7], glioma cells [8], lymphoma cells [9], and liver cancer cells [10]. Studies have shown that under the same treatment mode and time, when LTP simultaneously treats normal cells and tumor cells, it inhibits the activity of tumor cells, but significantly reduces the invasion, metastasis and adhesion of tumor cells, and has no obvious damage to normal cells [12,13]. What’s more, PAM can be kept stable for several days under low temperature (−80 ◦ C), dark and sealed storage, and the concentration of active ingredients remains unchanged [14]

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Conclusion