脐静脉内皮细胞外泌体和内皮祖细胞外泌体对hDPSCs增殖及迁移能力的比较研究

doi:10.13591/j.cnki.kqyx.2022.11.004

Abstract

Abstract: Objective Human umbilical vein endothelial cells-derived exosome (HUVECs-exo) and endothelial progenitor cells-derived exosome (EPCs-exo) were co-cultured with hDPSCs respectively, to study and compare their effects on proliferation and migration of hDPSCs, in order to provide reference for the application of exosomes in pulp regeneration. Methods Exosomes derived from human umbilical vein endothelial cells and endothelial progenitor cells were isolated by ultracentrifugation in vitro. The morphologies of both exosomes were observed by transmission electron microscopy. Meanwhile, the size of exosomes was measured by NTA. Western blot was used to analyze the exosome surface markers of exosomes including CD9, CD63 and TSG101. Both exosomes were applied to human dental pulp stem cells according to the concentration gradient of 5, 10 and 20 μg/mL, respectively. The proliferation of dental pulp stem cells was detected by CCK-8 assay and migration ability of dental pulp stem cells was detected by cell scratch wound healing assay and Transwell assay. Results Under transmission electron microscope, both exosomes exhibited a discoid morphology, with particle size ranging from 30 nm to 150 nm. Western blot indicated the expression of exosome surface markers including CD9, CD63 and TSG101 in both exosomes. Compared with the control group, two kinds of exosomes with different concentrations didn't promote the proliferation of human dental pulp stem cells (P>0.05). The cell scratch wound healing test and Transwell test showed that both exosomes could promote the migration of human dental pulp stem cells compared with the control group, and the difference was statistically significant (P<0.05), in which the concentration of 10 μg/mL exosomes had the most significant effect (P<0.01). There was no significant difference in the effect of two exosomes (P>0.05). Conclusion Exosomes from the two kinds of cells did not significantly promote the proliferation of human dental pulp stem cells, but could improve the migration of human dental pulp stem cells, especially at the concentration of 10 μg/mL.

Key words: umbilical vein endothelial cells, endothelial progenitor cells, human dental pulp stem cells, exosome, dental pulp regeneration

CLC Number:

R349.5

ZHANG Xue, XU Zhaoying, JIANG Pengfei, PAN Shuang. Comparative study on the proliferation and migration of umbilical vein endothelial cells-derived exosome and endothelial progenitor cells-derived exosome on human dental pulp stem cells[J]. Stomatology, 2022, 42(11): 979-983.

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Comparative study on the proliferation and migration of umbilical vein endothelial cells-derived exosome and endothelial progenitor cells-derived exosome on human dental pulp stem cells

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