人脐静脉内皮细胞来源外泌体对人牙髓干细胞生物学行为的影响

Effects of human umbilical vein endothelial cells-derived exosomes on the biological behaviors of human dental pulp stem cells

目的评估人脐静脉内皮细胞来源的外泌体(human umbilical vein endothelial cells-derived exosomes,HUVECs-exo)对人牙髓干细胞(human dental pulp stem cells,hDPSCs)生物学行为的影响。方法通过超高速离心法提取HUVECs-exo,分别用0、5、10、20μg/mL的HUVECs-exo处理hDPSCs,通过CCK-8实验、Transwell实验和划痕实验检测细胞的增殖和迁移能力;通过碱性磷酸酶(alkaline phosphatase,ALP)染色、茜素红染色和Western blot研究HUVECs-exo对hDPSCs成骨/成牙分化的影响。结果CCK-8实验结果显示HUVECs-exo对hDPSCs的增殖无显著影响,Transwell和划痕实验结果显示5、10μg/mL的HUVECs-exo显著地促进了hDPSCs的迁移(P<0.05),ALP染色和茜素红染色结果显示其对hDPSCs的矿化无显著促进作用,Western blot结果显示HUVECs-exo未上调Runt相关转录因子2(runt-related transcription factor 2,RUNX2)和骨桥蛋白(osteopontin,OPN)的表达水平。结论HUVECs-exo对hDPSCs的增殖和成骨/成牙分化无显著影响,但能显著促进hDPSCs的迁移。

Objective To evaluate the effect of human umbilical vein endothelial cells-derived exosomes(HUVECs-exo)on the biological behaviors of human dental pulp stem cells(hDPSCs).Methods Ultracentrifuge was used to extract the HUVECs-exo.hDPSCs were treated with 0,5,10,20μg/mL HUVECs-exo and the proliferation and migration of hDPSCs were detected by CCK-8 assay,Transwell and scratch experiments;The effects on the osteogenic/odontogenic differentiation of hDPSCs was researched by alkaline phosphatase(ALP)staining,alizarin red staining and Western blot.Results The results of CCK-8 showed that the proliferation of hDPSCs was not significantly affected.The results of Transwell and scratch experiments showed that 5μg/mL and 10μg/mL HUVECs-exo significantly promoted hDPSCs migration(P<0.05).The results of ALP staining and alizarin red staining showed that the mineralization of hDPSCs was not significantly affected and Western blot analysis showed that the expression levels of runt-related transcription factor 2(RUNX2)and osteopontin(OPN)were not significantly up-regulated.Conclusion HUVECs-exo has no significant effect on hDPSCs cell proliferation and osteogenic/odontogenic differentiation,but could significantly promote migration.