Related Experiment Videos
A method for long duration anaesthesia for a new hindlimb ischaemia-reperfusion model in mice
A M Wiersema1, R Dirksen, W J Oyen
1Department of Surgery, St Radboud University Hospital, Nijmegen, The Netherlands.
Abstract:
To study the relationship between ischaemia-reperfusion and multiple organ dysfunction syndrome (MODS), a new anaesthesia method was required to be applied to C57BL/6 mice. These mice are also used in a well accepted, standardized model for MODS using intraperitoneally administered zymosan (zymosan induced general inflammation, ZIGI). The aim was to develop a new model for ischaemia-reperfusion with 6 h of anaesthesia. This and further specific requirements for the combination of ischaemia-reperfusion and the ZIGI method, made us select inhalational anaesthesia using isoflurane in oxygen. This study evaluates whether long-term anaesthesia confounds the results of ischaemia-reperfusion and the ZIGI model. In addition the benefits of using the analgesic buprenorphine were evaluated. Ischaemia was induced with a tourniquet around the hindlimb. Ischaemia and reperfusion were verified by imaging a radioactive tracer with a gamma-camera. It was established that anaesthesia with isoflurane in oxygen caused little perturbation of body temperature and respiratory rate. A survival rate of 89% without noteworthy influence on organs was obtained. Buprenorphine proved to provide adequate analgesia and had no influence on measured parameters. In our experimental setting, this model with long duration anaesthesia allowed us to induce ischaemia and reperfusion of the hindlimb without perturbation of measurements. It also allowed good exposure of the abdomen and facilitated combination with the ZIGI model.
Insights
A new long-duration anesthesia model using isoflurane in oxygen was developed for mice undergoing ischemia-reperfusion injury. This method effectively combined with a general inflammation model without significantly affecting outcomes or organ function.
Area of Science:
- * Physiology and Anesthesiology
- * Surgical Research and Inflammation Models
Background:
- * Investigating the link between ischemia-reperfusion and multiple organ dysfunction syndrome (MODS) requires precise animal models.
- * Existing models for MODS in C57BL/6 mice, like the zymosan-induced general inflammation (ZIGI) model, necessitate specific experimental conditions.
- * A need existed for a reliable anesthesia method supporting prolonged procedures and integration with inflammation models.
Purpose of the Study:
- * To develop and evaluate a novel, long-duration anesthesia model for inducing hindlimb ischemia-reperfusion in mice.
- * To assess the compatibility of this anesthesia method with the ZIGI model for studying MODS.
- * To determine the impact of prolonged isoflurane anesthesia on ischemia-reperfusion and ZIGI model outcomes.
Main Methods:
- * Development of a 6-hour anesthesia protocol using isoflurane in oxygen for C57BL/6 mice.
- * Induction of hindlimb ischemia using a tourniquet, followed by reperfusion.
- * Verification of ischemia-reperfusion using gamma-camera imaging of a radioactive tracer.
- * Evaluation of the analgesic effects and physiological impact of buprenorphine.
Main Results:
- * Isoflurane anesthesia maintained stable body temperature and respiratory rate with minimal perturbation.
- * The developed model achieved an 89% survival rate with no significant adverse effects on organs.
- * Buprenorphine provided effective analgesia without influencing measured physiological parameters.
- * The long-duration anesthesia facilitated hindlimb ischemia-reperfusion and abdominal access for combined ZIGI model application.
Conclusions:
- * Long-duration isoflurane anesthesia is a viable method for inducing hindlimb ischemia-reperfusion in mice.
- * This anesthesia protocol does not confound results from ischemia-reperfusion or ZIGI models.
- * The model supports the combined study of ischemia-reperfusion and general inflammation in MODS research.