title: A novel laser-Doppler flowmetry assisted murine model of acute hindlimb ischemia-reperfusion for free flap research.
abstract: Suitable and reproducible experimental models of translational research in reconstructive surgery that allow in-vivo investigation of diverse molecular and cellular mechanisms are still limited. To this end we created a novel murine model of acute hindlimb ischemia-reperfusion to mimic a microsurgical free flap procedure. Thirty-six C57BL6 mice (n = 6/group) were assigned to one control and five experimental groups (subject to 6, 12, 96, 120 hours and 14 days of reperfusion, respectively) following 4 hours of complete hindlimb ischemia. Ischemia and reperfusion were monitored using Laser-Doppler Flowmetry. Hindlimb tissue components (skin and muscle) were investigated using histopathology, quantitative immunohistochemistry and immunofluorescence. Despite massive initial tissue damage induced by ischemia-reperfusion injury, the structure of the skin component was restored after 96 hours. During the same time, muscle cells were replaced by young myotubes. In addition, initial neuromuscular dysfunction, edema and swelling resolved by day 4. After two weeks, no functional or neuromuscular deficits were detectable. Furthermore, upregulation of VEGF and tissue infiltration with CD34-positive stem cells led to new capillary formation, which peaked with significantly higher values after two weeks. These data indicate that our model is suitable to investigate cellular and molecular tissue alterations from ischemia-reperfusion such as occur during free flap procedures.
authors.affiliation: Department of Oral and Maxillofacial Surgery, Medical Faculty, RWTH Aachen University, Aachen, Germany. ttahas13@gmail.com
authors.forename: Tolga Taha
authors.initials: TT
authors.lastname: Sönmez
authors.forename: Othman
authors.initials: O
authors.lastname: Al-Sawaf
authors.forename: Gerald
authors.initials: G
authors.lastname: Brandacher
authors.forename: Isabella
authors.initials: I
authors.lastname: Kanzler
authors.forename: Nancy
authors.initials: N
authors.lastname: Tuchscheerer
authors.forename: Mersedeh
authors.initials: M
authors.lastname: Tohidnezhad
authors.forename: Anastasios
authors.initials: A
authors.lastname: Kanatas
authors.forename: Matthias
authors.initials: M
authors.lastname: Knobe
authors.forename: Athanassios
authors.initials: A
authors.lastname: Fragoulis
authors.forename: René
authors.initials: R
authors.lastname: Tolba
authors.forename: David
authors.initials: D
authors.lastname: Mitchell
authors.forename: Thomas
authors.initials: T
authors.lastname: Pufe
authors.forename: Christoph Jan
authors.initials: CJ
authors.lastname: Wruck
authors.forename: Frank
authors.initials: F
authors.lastname: Hölzle
authors.forename: Elisa Anamaria
authors.initials: EA
authors.lastname: Liehn
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D058752:Free Tissue Flaps / Q000098:blood supply*; D006614:Hindlimb / Q000098:blood supply*; D017078:Laser-Doppler Flowmetry; D051379:Mice; D008810:Mice, Inbred C57BL; D055806:Microvessels / Q000378:metabolism / Q000503:physiopathology; D018482:Muscle, Skeletal / Q000098:blood supply / Q000473:pathology / Q000503:physiopathology; D018919:Neovascularization, Physiologic; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D012038:Regeneration; D015427:Reperfusion Injury / Q000503:physiopathology*; D012867:Skin / Q000098:blood supply / Q000473:pathology / Q000503:physiopathology
chemical_list: D019408:Platelet Endothelial Cell Adhesion Molecule-1