Animals were anesthetized with 3% isoflurane and maintained under anesthesia with 2% isoflurane and oxygen. Analgesia was provided by subcutaneous buprenorphine simbadol (0.1 mg/kg) injection pre-operatively and 24 h after surgery. All surgical procedures were performed using sterile microsurgical technique with an operating microscope (Superlux 40, Carl Zeiss).
To create the CP-TIB nerve graft, a freshly axotomized common peroneal (CP) nerve branch was grafted to the acutely or chronically denervated tibial (TIB). To mimic a delayed graft, the TIB nerve branch was transected 2–5 mm distal to the sciatic trifurcation and sutured to the quadriceps muscle. After 8 weeks of delay7 (link), the proximal (CP) and distal (TIB) stumps were aligned and sutured into either end of a 5 mm silicone conduit (Tuzic, Siliclear tubing, 4.6 mm OD, 1.98 mm ID) with 10/0 ethilon sutures to produce the CP-TIB graft with an interposed silicone conduit for cell and protein isolation. Immediate repair was achieved similarly but with immediate CP-TIB cross-suture (graft).
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