The effects of hUCMSC-sEVs on osteogenic/odontogenic differentiation of hDPSCs and tube formation ability of HUVECs

doi:10.19439/j.sjos.2025.06.001

Abstract

Abstract: PURPOSE: To investigate the effects of small extracellular vesicles (sEVs) derived from human umbilical cord mesenchymal stem cells(hUCMSCs) on the migration and osteogenic/odontogenic differentiation ability of human dental pulp stem cells(hDPSCs), the migration and tube formation ability of human umbilical vein endothelial cells (HUVECs) and their possible mechanisms. METHODS: hUCMSCs, hDPSCs and HUVECs were cultured and identified, hUCMSC-sEVs and LPSpre-hUCMSC-sEVs were isolated and identified, HUVECs and hDPSCs were assigned to 4 kinds of treatments, including the negative control group(NC), the positive control group(PC), the hUCMSC-sEVs group and the LPSpre-hUCMSC-sEVs group. Cell migration ability was detected by Transwell and wound healing assays. Tube formation capacity of HUVECs was assessed by tube formation experiment. The osteogenic/odontogenic differentiation ability of hDPSCs was evaluated by alizarin red staining and RT-qPCR. High-throughput small RNA sequencing was used to define miRNA profiles in sEVs. RESULTS: hUCMSCs, hDPSCs, HUVECs, hUCMSC-sEVs and LPSpre-hUCMSC-sEVs were successfully isolated and identified. Compared with NC group, both LPSpre-hUCMSC-sEVs group and hUCMSC-sEVs group promoted migration of hDPSCs, migration and tube formation of HUVECs. There was no significant difference between LPSpre-hUCMSC-sEVs group and hUCMSC-sEVs group in promoting migration of hDPSCs (P>0.05). LPSpre-hUCMSC-sEVs group was stronger than hUCMSC-sEVs group in promoting migration and tube formation of HUVECs(P<0.05). The calcium salt deposition in LPSpre-hUCMSC-sEVs group and hUCMSC-sEVs group was higher than that in PC group, and the calcium salt deposition in LPSpre-hUCMSC-sEVs group was higher than that in hUCMSC-sEVs group(P<0.05). The mRNA expression levels of ALP, OSX, OCN and RUNX2 in PC group, hUCMSC-sEVs group and LPSpre-hUCMSC-sEVs group were higher than those in NC group(P<0.05), and hUCMSC-sEVs group and LPSpre-hUCMSC-sEVs group were higher than PC group(P<0.05). In addition, the expression levels of OCN and RUNX2 in LPSpre-hUCMSC-sEVs group were higher than those in hUCMSC-sEVs group(P<0.05), while there was no significant difference in ALP and OSX between LPSpre-hUCMSC-sEVs group and hUCMSC-sEVs group(P>0.05). The expression level of DSPP in PC group and hUCMSC-sEVs group was higher than that in NC group, but the difference was not statistically significant(P>0.05). The expression level of DSPP in LPSpre-hUCMSC-sEVs group was higher than that in PC group (P<0.05). The most highly expressed miRNAs including hsa-miR-21-5p, hsa-let-7a-5p, hsa-miR-100-5p, hsa-miR-26a-5p and hsa-miR-222-3p, and differentially expressed miRNAs including hsa-miR-199a-3p, hsa-miR-122-5p, hsa-miR-1246 and hsa-miR-615-3p were detected, which may be the key factors of LPSpre-hUCMSC-sEVs. CONCLUSIONS: Small extracellular vesicles derived from human umbilical cord mesenchymal stem cells can promote migration and osteogenic/odontogenic differentiation of hDPSCs, as well as migration and angiogenesis of HUVECs, and LPS can enhance these effects, which may be related to miRNAs which are the most abundantly and diffferentially expressed in LPSpre-hUCMSC-sEVs.

Key words: Lipopolysaccharide, Small extracellular vesicles, Human umbilical cord mesenchymal stem cells, Human dental pulp stem cells, Human umbilical vein endothelial cells, Osteogenic/odontogenic differentiation

CLC Number:

R781.05

Kang Jingyi, Wei Rixia, Deng Huidan, Li Quanjie, Wu Yu. The effects of hUCMSC-sEVs on osteogenic/odontogenic differentiation of hDPSCs and tube formation ability of HUVECs[J]. Shanghai Journal of Stomatology, 2025, 34(6): 561-570.

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