3 0 silk suture

This study was carried out in accordance with the recommendations in the Animal (Scientific Procedures) Act 1986. The Home Office approved the study protocol (licence number 70/2716). 250–300 g Sprague-Dawley rats (n = 100) were sacrificed by CO₂ inhalation. The abdomen was sterilized with 70% ethanol and a U-shaped incision was performed to expose the abdominopelvic cavity. The abdominal inferior vena cava (IVC) and portal vein (PV) were identified and the PV was cannulated with a 24G cannula (BD, UK), which was secured in place with a 3–0 silk suture (Ethicon, UK). The abdominal IVC was ligated using silk sutures proximal to the right renal vein and the IVC was sectioned. The diaphragm was used as a holding point to release the whole liver from the supporting tissue. The whole procedure was carried out with special caution not to damage the Glisson’s capsule, which surrounds the organ.

Mice were killed by CO₂ inhalation. The abdomen was cleaned with 70% ethanol and a U-shaped incision was performed to expose the abdominopelvic cavity. The abdominal inferior vena cava (IVC) and portal vein (PV) were identified and the PV was cannulated with a 24G cannula (Becton Dickinson, Franklin Lakes, NJ), which was secured in place with a 3-0 silk suture (Ethicon, Somerville, NJ). The abdominal IVC was sectioned and 200 U of heparin injected into the portal vein. The diaphragm was used as a holding point to release the whole liver from the supporting tissue. The whole procedure was carried out with special caution not to damage Glisson’s capsule, which surrounds the organ.

A preoperative digital periapical radiograph was taken with cone indicator and reference marker, placed over the sensor. Conventional re-root canal treatment was performed, patients were recalled after 3 to 6 months for evaluation of periapical healing, those patients in which periapical healing was not observed were scheduled for periapical surgery. Local anesthesia 1:80,000 lidocaine with epinephrine was administered and a full-thickness mucoperiosteal flap was elevated. After identifying the periapical lesion site, a window was created by removing the cortical bone with a small round bur #2 (Mani, Japan) in a slow-speed handpiece. The periapical lesion was removed with the help of curettes (Hilbro, Japan). Apicoectomy and retrograde cavity preparation was performed with the help of ultrasonic tips (Pro ultra, DENTSPLY Maillefer, Switzerland) and the retrograde filling was done with MTA (Pro-root MTA, DENTSPLY Tulsa Dental Specialties, USA). The flap was then repositioned and sutured with a 3/0 silk suture (ETHICON, LLC).
Tissue from the periapical lesion was collected and divided immediately into two fragments: one fragment was transferred in RNA later buffer filled 1.5 mL sterilized centrifuge tubes and kept in − 80 °C until homogenized to extract RNA, the second fragment was used for histopathological analysis.

Whole stomachs were harvested from female and male Sprague Dawley rats, weighing approximately 200–250 g. After sacrifice, the abdominal wall was sterilized with 70% ethanol and a midline incision was performed to expose the abdominal cavity. Celiac artery was cannulated with a 27G cannula (Introcan Certo, B. Braun Medical AG, Germany) secured in place with a 3–0 silk suture (Ethicon, UK). Hepatic artery was ligated and portal vein sectioned. The stomach was harvested and barbed to luer-lock connectors (Cole Parmer, US) were used to cannulate cardias and pylorus.
Stomachs were decellularised using a detergent-enzymatic treatment (DET) as previously described. Both the vasculature and the organ lumen were perfused with a 1 ml/min flow rate using a Masterflex L/S variable speed roller pump (Masterflex, US). Each DET cycle was consisting of: 24 h perfusion with deionized water (18.2 MU/cm), 4 h with 4% sodium deoxycholate (Sigma, UK), 1 h with deionized water, 3 h 22,5 mg/l DNase-I in 0,9% sodium chloride (NaCl, Sigma, UK) and 1,11 g/l calcium chloride (CaCl₂, Sigma, UK). All organs after DET were preserved at 4 °C, in PBS (Gibco, UK) and antibiotics (Penicillin/streptomycin 1%) and irradiated to eliminate all contaminants.