A new embolus injection method to evaluate intracerebral hemorrhage in New Zealand white rabbits

Abstract

The rabbit large clot embolic stroke model has been used for over 23 years to study methods to manipulate hemorrhage and to test drugs and devices for safety, because the rabbit model is particularly sensitive to embolism-induced hemorrhage. This study refined the original embolization procedure using an automated, pump-assisted injection method to introduce large blood clots or macroscopic emboli into the middle cerebral artery (MCA) via an indwelling carotid artery catheter. The study shows that rapid injection of blood clots (3 ml/30 s) produced a model where there is a high hemorrhage incidence rate (79%) and a high stroke success rate (63%), compared to a low stroke success rate (19%) with no hemorrhages when clots were injected at a slow rate (3 ml/90 s). The rapid injection method, which produces a high hemorrhage rate, is particularly useful to study neuroprotective agents to attenuate embolism-induced hemorrhage. In addition, we show that manual injection of blood clots, which produces a lower baseline hemorrhage rate (41%) with a similar stroke success rate (65%), may allow investigators to study pharmacological agents to either up or down-regulate hemorrhage incidence. Lastly, we show that in the rabbit embolic stroke model, hemorrhages are adjacent to areas of 2,3,5-triphenyltetrazolium (TTC)-negative tissue, normally associated with infarcted or ischemic tissue. Thus, there is clear separation of ischemia and hemorrhage in the model, suggesting that therapeutics that are neuroprotective may also be useful to limit the evolution of ischemic damage associated with a hemorrhage, if not attenuate hemorrhage itself.