Safety

Safety for siRNAs

Blast analysis

We performed blast analysis on NCBI when designing the siRNA. The results (Fig.1) showed that our siRNA does not inhibit the expression of other genes besides the corresponding targets. Therefore, the safety was verified.

Safety for therapeutic sEVs

Validate that our therapeutic sEVs can inhibit the apoptosis of DPC induced by TP

We planned to see if our therapeutic sEVs (which load with 5αR-siRNA-1, Piezo1-siRNA-5, and mRNA-β-catenin) can inhibit apoptosis of DPC in the presence of androgen in a gesture to prove our therapeutic sEVs are safe. Therefore, we added testosterone propionate (TP) to the cell culture medium and co-cultured therapeutic sEVs with DPC for 30h. Then the apoptotic status of DPC was detected using Annexin V-FITC Apoptosis Detection Kit (from Beyoncé) by flow cytometry. As is shown in Fig.2, TP can induce apoptosis while apoptosis was significantly inhibited after the addition of sEVs. These findings indicated that our sEVs can be used as a treatment option and may have therapeutic implications for patients with AGA.