Evaluation of the middle cerebral artery occlusion techniques in the rat by in-vitro 3-dimensional micro- and nano computed tomography.
BACKGROUND
Animal models of focal cerebral ischemia are widely used in stroke research. The purpose of our study was to evaluate and compare the cerebral macro- and microvascular architecture of rats in two different models of permanent middle cerebral artery occlusion using an innovative quantitative micro- and nano-CT imaging technique.


METHODS
4h of middle cerebral artery occlusion was performed in rats using the macrosphere method or the suture technique. After contrast perfusion, brains were isolated and scanned en-bloc using micro-CT (8 mum)3 or nano-CT at 500 nm3 voxel size to generate 3D images of the cerebral vasculature. The arterial vascular volume fraction and gray scale attenuation was determined and the significance of differences in measurements was tested with analysis of variance [ANOVA].


RESULTS
Micro-CT provided quantitative information on vascular morphology. Micro- and nano-CT proved to visualize and differentiate vascular occlusion territories performed in both models of cerebral ischemia. The suture technique leads to a remarkable decrease in the intravascular volume fraction of the middle cerebral artery perfusion territory. Blocking the medial cerebral artery with macrospheres, the vascular volume fraction of the involved hemisphere decreased significantly (p < 0.001), independently of the number of macrospheres, and was comparable to the suture method. We established gray scale measurements by which focal cerebral ischemia could be radiographically categorized (p < 0.001). Nano-CT imaging demonstrates collateral perfusion related to different occluded vessel territories after macrosphere perfusion.


CONCLUSIONS
Micro- and Nano-CT imaging is feasible for analysis and differentiation of different models of focal cerebral ischemia in rats.
Department of Radiology, Justus-Liebig University Giessen, Giessen, Germany. alexander.langheinrich@radiol.med.uni-giessen.de
Alexander C
AC
Langheinrich
Mesut
M
Yeniguen
Anne
A
Ostendorf
Simone
S
Marhoffer
Marian
M
Kampschulte
Georg
G
Bachmann
Erwin
E
Stolz
Tibo
T
Gerriets
D000818:Animals; D004195:Disease Models, Animal; D021621:Imaging, Three-Dimensional / Q000379:methods*; D020244:Infarction, Middle Cerebral Artery / Q000000981:diagnostic imaging / Q000473:pathology / Q000503:physiopathology; D008297:Male; D008833:Microcirculation / Q000502:physiology; D036103:Nanotechnology / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013536:Suture Techniques; D014057:Tomography, X-Ray Computed / Q000379:methods*