Endothelial cells and endothelin-1 promote the odontogenic differentiation of dental pulp stem cells

It has been established that dental pulp stem cells (DPSCs) serve an important role in the restoration and regeneration of dental tissues. DPSCs are present in blood vessels and also exist in the vessel microenvironment in vivo and have a close association with endothelial cells (ECs). The present study aimed to evaluate the influence of ECs and their secretory product endothelin-1 (ET-1) on the differentiation of DPSCs. In the present study, cells were divided into four groups: i) a DPSC-only control group; ii) a DPSC with ET-1 administration group; iii) a DPSC and human umbilical vein endothelial cell (HUVEC) direct co-culture group; and iv) a DPSC and HUVEC indirect co-culture group using a Transwell system. Reverse transcription-quantitative polymerase chain reaction was used to detect the expression of the odontoblastic differentiation-associated genes, including dentin sialoprotein (DSP) and dentin matrix acidic phosphoprotein 1 (DMP-1) at days 4, 7, 14 and 21. Alizarin Red S staining, immunofluorescence and western blot analyses were also conducted to assess the differentiation of the DPSCs in each group. The highest expression levels of odontoblastic differentiation-associated genes were observed on day 7 and in the two co-culture groups were increased compared with the DPSC-only and DPSC + ET-1 culture groups at all four time points. However, expression levels in the DPSC + ET-1 group were not downregulated as notably as in the co-culture groups on days 14 and 21. The Transwell group exhibited the greatest ability for odontoblastic differentiation compared with the other groups according to staining with Alizarin Red S, immunofluorescence and western blot analysis results. According to the results of the present study, the culture solution with HUVECs affected the differentiation of DPSCs. In addition, ET-1 may promote the odontoblastic differentiation of DPSCs.