A novel laser-Doppler flowmetry assisted murine model of acute hindlimb ischemia-reperfusion for free flap research

PLoS One. 2013 Jun 20;8(6):e66498. doi: 10.1371/journal.pone.0066498. Print 2013.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Disease Models, Animal
  • Free Tissue Flaps / blood supply*
  • Hindlimb / blood supply*
  • Laser-Doppler Flowmetry
  • Mice
  • Mice, Inbred C57BL
  • Microvessels / metabolism
  • Microvessels / physiopathology
  • Muscle, Skeletal / blood supply
  • Muscle, Skeletal / pathology
  • Muscle, Skeletal / physiopathology
  • Neovascularization, Physiologic
  • Platelet Endothelial Cell Adhesion Molecule-1 / metabolism
  • Regeneration
  • Reperfusion Injury / physiopathology*
  • Skin / blood supply
  • Skin / pathology
  • Skin / physiopathology

Substances

  • Platelet Endothelial Cell Adhesion Molecule-1

Grants and funding

This study was supported by a grant from the START program (129/10) and by a grant from the Interdisciplinary Centre for Clinical Research (IZKF) within the Faculty of Medicine of RWTH Aachen University (T9-3; T9-5). The funders had nor role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.