〔Abstract〕 Objective To develop an extracellular vesicles(EVs) co-loading system by synergistically deliver tumor necrosis factor related apoptosis inducing ligand(TRAIL)/verteporfin(VPF) combination to induce apoptosis and inhibit proliferation of OSCC cells. Methods TRAIL/VPF co-loaded EVs(MSCT-EVs/VPF) was purified and collected though ultracentrifugation and dialysis. The expression of CD63, CD9 and TRAIL was detected by BCA to confirm the origin of EVs. High performance liquid chromatography(HPLC) was used to detect the drug loading of VPF and draw the release curve in vitro. Cytotoxicity and half inhibitory concentration(IC50) were detected by MTT assay. The apoptosis rate was detected by flow cytometry. Finally, Western blot was used to detect the effects of MSCT-EVs/VPF on the expression of apoptosis related proteins and Yap in SCC25 cells. Results MSCT-EVs/VPF particles were round and well dispersed with a diameter of about 100 nm. The drug loading of the nano system was about(15.43 ± 0.44)%, 57.8% of VPF was released in 10 h and 82.5% in 45 h; MSCT-EVs and VPF could inhibit the growth of SCC25 tumor cells in a dose-dependent manner, showing good synergistic effect in the ratio of 10:1-5:1(CI<1, wt%). At the ratio of 100:5～100:15(Mass ratio of MSCT-EVs to VPF, wt%), the IC50of MSCT-EVs/VPF was significantly lower than that of free MSCT-EVs + free VPF group(P<0.05), and showed a more effective inhibition. The high inhibitory effect of MSCT-EVs/VPF on squamous cell carcinoma cells was partly due to the regulation of Caspase-3, Bax, BCL-2, mTOR, p-mTOR and YAP. Conclusion EVs delivery of a fixed proportion of TRAIL/VPF shows high inhibitory effect on oral squamous cell carcinoma cells, which provides a new idea for the treatment of multidrug-resistant tumors.