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Stem cells have attracted great interest in the development of tissue engineering. However, the self-regeneration and multi-differentiation capabilities of stem cells are easily impaired during cell transplantation. Recent studies have demonstrated that <i>Sapindus mukorossi</i> (<i>S. mukorossi</i>) seed oil has various positive biological effects. However, it is not yet clear whether <i>S. mukorossi</i> seed oil can increase the growth and differentiation of dental pulp mesenchymal stem cells (DPSCs). The aim of this study is to investigate the effects of <i>S. mukorossi</i> seed oil on the proliferation and differentiation of DPSCs. DPSCs with and without <i>S. mukorossi</i> seed oil, respectively, were evaluated and compared. The viabilities of the cells were assessed by MTT tests. The osteogenetic and odontogenetic capacities of the DPSCs were tested using Alizarin red S staining and alkaline phosphatase (ALP) activity assays. In addition, real-time PCR was performed to examine the gene expression of ALP, BMP-2 and DMP-1. Finally, extracellular matrix vesicle secretion was detected via scanning electron microscopy. No significant difference was observed in the viabilities of the DPSCs with and without <i>S. mukorossi</i> seed oil, respectively. However, under osteogenic and odontogenic induction, <i>S. mukorossi</i> seed oil increased the secretion of mineralized nodules and the ALP activity of the DPSCs (<i>p</i> < 0.05). The ALP gene expression of the differentiation-induced DPSCs was also enhanced. Finally, a greater secretion of extracellular matrix vesicles was detected in the DPSCs following odontogenic induction complemented with <i>S. mukorossi</i> seed oil. Overall, the present results show that <i>S. mukorossi</i> seed oil promotes the osteogenic/odontogenic differentiation and matrix vesicle secretion of DPSCs.