The development of an improved preclinical mouse model of intracerebral hemorrhage using double infusion of autologous whole blood

Abstract

The present study was conducted in mice to validate a double blood infusion model of intracerebral hemorrhage (ICH) that does not use anticoagulant. We investigated the effect of intrastriatal infusion of blood on hematoma volume, neurologic function, brain edema and swelling, and markers of neuroinflammation and oxidative DNA damage. Anesthetized C57BL/6 adult male mice were infused in the left striatum with 4 µl of blood over 20 min at 0.2 µl/min; the needle was left in place for 7 min, and the remaining 6 µl of blood was then infused over 30 min. The injection needle was slowly withdrawn 20 min after the second injection. Sham-operated control mice received only needle insertion. The hematoma produced in this model was primarily restricted to the striatum, and the mice demonstrated severe neurologic deficits that appeared within 60 min and remained evident at 72 h. Brain water content and swelling were significantly increased and were associated with a marked increase in ICH-induced neutrophil infiltration, microglial/macrophage and astrocyte activation, cytochrome c release, and oxidative DNA damage. Other groups have mixed the anticoagulant heparin with the infused blood, an agent that could affect in vivo clot formation. We believe that this double blood infusion model that does not use anticoagulant improves upon the procedure and provides an easy and reproducible alternative for inducing ICH in mice; it should be useful for studying the pathophysiology of ICH and for testing potential pharmaceutical and surgical interventions.

Cerebroprotection by the neuronal PGE2 receptor EP2 after intracerebral hemorrhage in middle-aged mice

“…We also used a combination of morphologic criteria to define microglia and macrophages as either resting or activated. 31 …”

A Critical Appraisal of Experimental Intracerebral Hemorrhage Research

“…For now, the most widely used rodent models of ICH for neuroprotection, rehabilitation, and stem cell studies are the direct infusion of whole blood into the brain (Bullock et al, 1984) and the injection of bacterial collagenase (Rosenberg et al, 1990), an enzyme that damages the basal lamina (extracellular matrix) thereby causing bleeding. In both cases, some investigators also add heparin (Belayev et al, 2003;Del Bigio et al, 1996), but this is not typical and is a potential confounding factor (Wang et al, 2008b), albeit with some clinical relevance. Common also are simplified models that involve injecting blood components, such as thrombin and iron, into the brain (Andaluz et al, 2002).…”

Neuroinflammation Extends Brain Tissue at Risk to Vital Peri-Infarct Tissue: A Double Tracer [11C]PK11195- and [18F]FDG-PET Study

“…These multifaceted responses follow specific temporospatial patterns (Hallenbeck et al, 1986;Schroeter et al, 1994;Schroeter et al, 1999;Mabuchi et al, 2000;Schroeter et al, 2001;Wang et al, 2008) and are termed 'neuroinflammation.' Although responses of inflammatory cells are only part of the complex process of neuroinflammation, in this study, we use them as a surrogate marker for the localization of neuroinflammatory processes overall.…”