Carprofen

Following 12 weeks of High Fat Diet animals underwent Vertical Sleeve Gastrectomy or sham surgery, as previously described (11 (link), 12 (link)). Three days before bariatric or sham surgery, animals were exposed to liquid diet (20% dextrose) and remained on this diet for up to four days post operatively. Following this, mice were returned to high fat/high sucrose diet until euthanasia and tissues harvested ten weeks post bariatric surgery. Anaesthesia was induced and maintained with isoflurane (1.5-2%). A laparotomy incision was made, and the stomach was isolated outside the abdominal cavity. A simple continuous pattern of suture extending through the gastric wall and along both gastric walls was placed to ensure the main blood vessels were contained. Approximately 60% of the stomach was removed, leaving a tubular remnant. The edges of the stomach were inverted and closed by placing two serosae only sutures, using Lembert pattern (31 (link)). The initial full thickness suture was subsequently removed. Vicryl absorbable sutures (8.0) were used. Sham surgeries were performed by isolating the stomach and performing a 1 mm gastrotomy on the gastric wall of the fundus. All animals received a three-day course of analgesics Carprofen (Bayer, UK) and a five-day course of SC antibiotic injections (Enrofloxacin 10mg/kg). A survival of 80-90% was achieved.

Pancreatic islets were isolated by injecting collagenase-containing medium into the pancreatic duct, followed by separation from the exocrine tissue by gradient density and handpicking. The islets were cultured for 24 h in RPMI 1640 medium containing 11 mM glucose supplemented with 2 mM l-glutamine, 10% heat-inactivated foetal calf serum, 100 IU/mL penicillin, and 100 μg/mL streptomycin^{74 (link)}. For transplantation, 10–20 islets were aspirated with a 27-gauge blunt eye cannula (BeaverVisitec, UK) connected to a 100 μL Hamilton syringe (Hamilton) via 0.4-mm polyethylene tubing (Portex Limited). Prior to surgery, mice were anaesthetised with 2–4% isoflurane (Zoetis) and placed in a stereotactic frame to stabilise the head. The cornea was incised near the junction with the sclera, being careful not to damage the iris. Then, the blunt cannula, pre-loaded with islets, was inserted into the ACE and islets were expelled (average injection volume 20 µL for 10 islets). Carprofen (Bayer, UK) and eye ointment were administered post-surgery.

Three days before bariatric or sham surgery, animals were exposed to liquid diet (20% dextrose) and remained on this diet for up to 4 days post-operatively. Following this, mice were returned to high-fat/high-sucrose diet until euthanasia and tissues harvested 10 weeks post bariatric surgery. Anaesthesia was induced and maintained with isoflurane (1.5–2%). A laparotomy incision was made, and the stomach was isolated outside the abdominal cavity. A simple continuous pattern of suture extending through the gastric wall and along both gastric walls was placed to ensure the main blood vessels were contained. Approximately 60% of the stomach was removed, leaving a tubular remnant. The edges of the stomach were inverted and closed by placing two serosae only sutures, using Lembert pattern. The initial full thickness suture was subsequently removed. Sham surgeries were performed by isolating the stomach and performing a 1 mm gastrotomy on the gastric wall of the fundus. All animals received a 3-day course of analgesics Carprofen (Bayer, UK) and a 5-day course of SC antibiotic injections (enrofloxacin 10 mg/kg).