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Quantification of neutrophil adhesion to skeletal muscle venules following ischemia-reperfusion
M Goldberg1, D Serafin, B Klitzman
1Plastic Surgery Research Laboratories, Duke University Medical Center, Durham, North Carolina 27710.
Abstract:
Ischemia-reperfusion is encountered in a wide variety of surgical situations. The damage resulting from ischemia-reperfusion may be due, in part, to the infiltration and activation of neutrophils into the reperfused tissue. The purpose of the study was to examine ischemia-reperfusion induced neutrophil activation in skeletal muscle. In control muscles, ischemia in the hamster right cremaster muscle was produced for 5 min after an initial 2 hr 55 min perfusion period. In ischemic muscles, ischemia was produced for 3 hr prior to reperfusion. After the clamps were removed, a video recording of the cremaster microvasculature was made using intravital fluorescence microscopy. Acridine orange was infused intravenously 10 min prior to video recording in order to selectively label and enhance the contrast of neutrophils. The number of neutrophils rolling along the endothelium of 40 to 60 microns-diameter venules in a 1-min period increased from 9.0 in control animal cremaster venules to 24.1 following ischemia-reperfusion (p less than .05; n = 11). The ischemia-reperfusion model developed in this study allows for the direct quantification of neutrophil adhesion in skeletal muscle and can be further used to assess pharmacologic minimization of neutrophil-mediated damage in skeletal muscle.
Insights
Ischemia-reperfusion injury in skeletal muscle leads to increased neutrophil activation. This study quantifies neutrophil rolling in hamster cremaster muscle following ischemia-reperfusion, providing a model for testing treatments.
Area of Science:
- Cardiovascular Biology
- Surgical Research
- Immunology
Background:
- Ischemia-reperfusion (I/R) injury is a significant complication in various surgical scenarios.
- Neutrophil infiltration and activation in reperfused tissues contribute to cellular damage.
- Understanding neutrophil behavior during I/R is crucial for developing therapeutic strategies.
Purpose of the Study:
- To investigate and quantify ischemia-reperfusion induced neutrophil activation in skeletal muscle.
- To establish a reliable model for assessing neutrophil adhesion in skeletal muscle microvasculature.
Main Methods:
- A hamster right cremaster muscle model was utilized to induce ischemia for 3 hours followed by reperfusion.
- Intravital fluorescence microscopy with acridine orange labeling was employed to visualize and quantify neutrophils.
- Neutrophil rolling along venule endothelium was measured in control and I/R subjected muscles.
Main Results:
- Neutrophil rolling significantly increased from 9.0 in control venules to 24.1 following ischemia-reperfusion (p < .05).
- The developed model allows for direct quantification of neutrophil adhesion in skeletal muscle.
- A marked increase in neutrophil adhesion was observed post-ischemia-reperfusion.
Conclusions:
- Ischemia-reperfusion significantly activates neutrophils in skeletal muscle microvasculature.
- This study presents a validated model for quantifying neutrophil adhesion in skeletal muscle.
- The model can be instrumental in evaluating pharmacological interventions aimed at reducing neutrophil-mediated damage.