A Novel Laser-Doppler Flowmetry Assisted Murine Model of Acute Hindlimb Ischemia-Reperfusion for Free Flap Research

Tolga Taha Sönmez, Othman Al-Sawaf, Gerald Brandacher, Isabella Kanzler, Nancy Tuchscheerer, Mersedeh Tohidnezhad, Anastasios Kanatas, Matthias Knobe, Athanassios Fragoulis, René Tolba, David Mitchell, Thomas Pufe, Christoph Jan Wruck, Frank Hölzle, Elisa Anamaria Liehn

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Article numbere66498
JournalPloS one
Volume8
Issue number6
DOIs
StatePublished - Jun 20 2013

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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    Sönmez, T. T., Al-Sawaf, O., Brandacher, G., Kanzler, I., Tuchscheerer, N., Tohidnezhad, M., Kanatas, A., Knobe, M., Fragoulis, A., Tolba, R., Mitchell, D., Pufe, T., Wruck, C. J., Hölzle, F., & Liehn, E. A. (2013). A Novel Laser-Doppler Flowmetry Assisted Murine Model of Acute Hindlimb Ischemia-Reperfusion for Free Flap Research. PloS one, 8(6), [e66498]. https://doi.org/10.1371/journal.pone.0066498