Chapter 7 The aim of this study is to investigate the role of IR injury in PFL in a murine model that mimics the DIEP flap. PFL caused by insufficient intrinsic vascular supply, either arterial or venous, and the effect of IR injury on PFL were investigated. Methods Experiments and euthanasia of animals were performed in accordance with the institutional animal care guidelines and had the approval of the Maastricht University Medical Center Animal Care Committee. Animals were kept in separate housing for the duration of the study in order to prevent allocannibalism of (partial) necrotic flap tissue. Eight‐ to 12‐week‐old Swiss Webster mice were used from the Charles River Laboratories. All mice were anesthetized with ketamine (62.5 mg/kg) and medetomidine (400 g/kg) intraperitoneally. After the disappearance of righting reflex, 1.5 mL of saline was admitted subcutaneously for fluid balance during the procedure. Anesthesia was maintained with ketamine (17.5 mg/kg/h) and medetomidine (112 g/kg/h) subcutaneously. Visagel was used for cornea protection. Thereafter, the pertinent surgical site was depilated and disinfected with topical antibacterial preparation (70% alcohol). Core body temperature was controlled and maintained with the use of an automated heating pad during the procedure. Temgesic was subcutaneously administered on the dorsum during the first three days for postoperative pain relief (0.1 mg/kg body weight), and the animals were checked for normal behavior and habitual status. The mice were separated into three experimental groups (Table 7.1). In the nonischemia group, the flap was raised without ischemia. In the subsequent groups, IR injury I and IR injury II, there was an increasing ischemia time of 30 and 60 minutes, respectively. In order to maintain comparability, anesthesia duration was standardized for all study groups (90 minutes). Table 7.1 Study groups. 1 Nonischemic (n7) Flap model, distal ischemia model 2 IR injury* I (n7) Distal ischemia compared with group 1 and 30 min** IR injury 3 IR injury* II (n7) Distal ischemia compared with group 1 and 60 min** IR injury * ** IR injury = ischemia‐reperfusion injury, min = minutes. The DIEP/TRAM flap mouse model based on the unilateral deep epigastric artery has been described before.15,16 The abdominal skin is marked with a template creating a skin paddle measuring 2 cm in the craniocaudal direction and 3 cm in a transverse direction (Figure 7.1A). 102
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