2023, Vol. 27 ›› Issue (1): 54-58

Human periodontal ligament stem cells-derived exosomes interfere with the proliferation and differentiation of MC3T3-E1 cells

Lan Qian1, Gu Yangcong2, Xiao Xin1, Bi Xueting1, Li Na3   

Abstract: BACKGROUND: Human periodontal ligament stem cells had a strong osteogenic ability, but the effect of human periodontal ligament stem cells-derived exosomes on proliferation and osteogenic differentiation of MC3T3-E1 is not clear.
OBJECTIVE: To explore the effect of human periodontal ligament stem cells-derived exosomes on the proliferation and differentiation of MC3T3-E1 cells.
METHODS: Human periodontal ligament stem cells were isolated and cultured by enzyme digestion. Human periodontal ligament stem cells-derived exosomes were extracted by ultracentrifugation, and identified by transmission electron microscopy, nanoparticle tracking analysis and western blot assay. Effect of different mass concentrations of human periodontal ligament stem cells-derived exosomes on proliferation of MC3T3-E1 cells was detected by CCK-8 assay. The effect of 100 mg/L human periodontal ligament stem cells-derived exosomes on osteogenic mineralization of MC3T3-E1 cells was observed by alizarin red staining. Western blot assay was used to detect the phosphorylation levels of MEK and ERK in MC3T3-E1 cells before and after the intervention of 100 mg/L human periodontal ligament stem cells-derived exosomes.  
RESULTS AND CONCLUSION: (1) Transmission electron microscopy showed that exosomes were vesicles formed by lipid bilayer. Nanoparticle tracking analysis showed that exosomes were 50-120 nm in diameter and concentrated at 79.86 nm. Western blot assay showed that the exosomes contained expression levels of CD81, CD63 and TSG101. (2) Compared with the control group, human periodontal ligament stem cells-derived exosomes could promote the proliferation of MC3T3-E1 cells in a dose-dependent manner. (3) Compared with the control group, MC3T3-E1 cells were able to form many calcium nodules in the human periodontal ligament stem cell-derived exosome group. Compared with the control group, protein expression of p-MEK and p-ERK increased in MC3T3-E1 cells of the human periodontal ligament stem cell-derived exosome group. (4) It is indicated that human periodontal ligament stem cells-derived exosomes can promote the proliferation and osteogenic differentiation of MC3T3-E1 cells, which may be associated with the activation of MEK/ERK signaling pathway.
Key words: periodontal ligament stem cell, exosome, osteoblast, MC3T3-E1 cell, proliferation, differentiation