Moorldi2 hir

8-weeks-old male immunodeficient nude mice (NU/J) (Jackson Laboratory, Strain #002019) were used of this assay. Briefly, the dorsal skin of the mice was depilated 24 hours before the procedure. The mice were anesthetized by continuous 1–3% isoflurane administration throughout the duration of the procedure. The back skin was cleaned and sterilized with betaine and 70% ethanol. A sterile 6 mm biopsy punch was used to create a wound on the back skin of each animal. A 0.5 mm thick silicon sheet was placed around the wound, and it was secured with eight interrupted sutures of non-absorbable 6–0 Nylon (Henry Schein, cat: 1016053). After surgery, the mice were treated with EFF or control EVs at a final concentration of 1.8 ×10⁸ EVs/mL in saline solution, and control mouse were treated with saline solution. Finally, a single dose of buprenorphine was administered subcutaneously to control pain. The animals were housed individually after surgery. Skin perfusion was assessed at 10 days after EV treatment, immediately before euthanasia using a high-resolution laser doppler imager (MOORLDI2-HIR, Moor Instruments). All animal procedures performed in this study were approved by the Institutional Animal Care and Use Committee at The Ohio State University (protocol # 2016A00000074-R2).