Perma hand silk

CLP procedure was performed (38 (link)) on mice that were anesthetized with ketamine/xylazine (University of Iowa, Office of Animal Resources). Briefly, the abdomen was shaved and disinfected with Betadine (Purdue Products), and a midline incision was made. The distal third of the cecum was ligated with Perma-Hand Silk (Ethicon), punctured once (“moderate”) or twice (“severe”) using a 25-gauge needle, and a small amount of fecal matter extruded. The cecum was returned to abdomen, the peritoneum was closed with 641G Perma-Hand Silk (Ethicon), and skin sealed using surgical Vetbond (3M). Following surgery, 1 mL PBS was administered s.c. to provide post-surgery fluid resuscitation. Bupivacaine (Hospira) was administered at the incision site, and flunixin meglumine (Phoenix) was administered for postoperative analgesia. This procedure created a septic state characterized by loss of appetite and body weight, ruffled hair, shivering, diarrhea, and/or periorbital exudates with 0–10% mortality rate, or 25–50% mortality for moderate and severe sepsis, respectively, similar to our previous reports (39 (link)). Sham mice underwent identical surgery excluding cecal ligation and puncture. Removal of punctured cecum as source control is not attempted here due to the short duration of the experiments - most mice were analyzed/sacrificed in the first 2–4 days post CLP surgery.

Mice were anesthetized with ketamine/xylazine (University of Iowa, Office of Animal Resources), the abdomen was shaved and disinfected with Betadine (Purdue Products), and a midline incision was made (28 (link)). The distal third of the cecum was ligated with Perma-Hand Silk (Ethicon), punctured once (for CLP₂₀) or twice (for CLP₅₀) using a 25-gauge needle, and a small amount of fecal matter extruded out of each puncture. The cecum was then returned to abdomen, the peritoneum was closed with 641G Perma-Hand Silk (Ethicon), and skin sealed using surgical Vetbond (3M). Following surgery, 1 mL PBS was administered s.c. to provide post-surgery fluid resuscitation. Bupivacaine (Hospira) was administered at the incision site, and flunixin meglumine (Phoenix) was administered for postoperative analgesia. Sham mice underwent identical surgery excluding cecal ligation and puncture. Clinical disease was scored according to the following system: Grooming 0-normal, 1-piloerection, 2-ruffled; Gait 0-normal, 1-ataxic, 2-none; Mobility 0-normal, 1-reduced, 2-immobile; Body Position 0-Full extension, 1-hunched, 2-moribund.

CLP procedure was performed on mice that were anesthetized with ketamine/xylazine (University of Iowa, Office of Animal Resources) as previously described (47 (link)). Briefly, the abdomen was shaved and disinfected with Betadine (povidone iodine; Purdue Products), and a midline incision was made. The distal third of the cecum was ligated with PERMA-HAND Silk (Ethicon) and one puncture was made with a 25-gauge needle, and a small amount of fecal matter extruded. The cecum was returned to abdomen, the peritoneum was closed with 641G PERMA-HAND Silk (Ethicon), and skin was sealed using surgical Vetbond (3M). Following surgery, 1 ml PBS was administered s.c. to provide post-surgery fluid resuscitation. Bupivacaine (Hospira) was administered at the incision site, and flunixin meglumine (Phoenix) was administered for postoperative analgesia. This procedure created a septic state characterized by loss of appetite and body weight, ruffled hair, shivering, diarrhea, and/or periorbital exudates with 0–10% mortality rate for moderate sepsis similar to previous reports (8 (link), 47 (link)). Sham mice underwent identical surgery excluding CLP.