Experimental Subarachnoid Haemorrhage Models in Rats

Journal of Neuroscience Methods, 2000

The rat endovascular filament model has been utilized to study subarachnoid hemorrhage (SAH). Because the severity of the hemorrhage with this model has proven difficult to modulate, we attempted to vary the hemorrhage by modifying filament size, and compared this model to the blood injection method with regards to acute physiological responses and hemorrhage size. SAH was achieved using either a 3-0 or 4-0 filament, or by injecting 0.3 cc of autologous blood into the cisterna magna. Peak ICP elevations were lowest in the 4-0 filament group. CBF decreased acutely and rose from its nadir in all three models with the injection model demonstrating the earliest recovery. In the injection group, mean arterial blood pressure rose acutely and remained elevated, whereas in the 3-0 group, MABP rose transiently and in the 4-0 group it did not rise significantly. Histologically, there was less subarachnoid blood in the 4-0 group vs. the injection or 3-0 groups and a different distribution of blood in the two experimental models. Varying filament size provides a method to modulate the severity of SAH in the filament model. In addition, the rat endovascular filament and blood injection models produce different distribution of blood and physiological responses.

2002

rebral artery vasospasm is a major cause of death and disability in patients experiencing subarachnoid hemorrhage (SAH). Currently, little is known regarding the impact of SAH on small diameter (100-200 m) cerebral arteries, which play an important role in the autoregulation of cerebral blood flow. With the use of a rabbit SAH model and in vitro video microscopy, cerebral artery diameter was measured in response to elevations in intravascular pressure. Cerebral arteries from SAH animals constricted more (ϳtwofold) to pressure within the physiological range of 60-100 mmHg compared with control or sham-operated animals. Pressure-induced constriction (myogenic tone) was also enhanced in arteries from control animals organ cultured in the presence of oxyhemoglobin, an effect independent of the vascular endothelium or nitric oxide synthesis. Finally, arteries from both control and SAH animals dilated as intravascular pressure was elevated above 140 mmHg. This study provides evidence for a role of oxyhemoglobin in impaired autoregulation (i.e., enhanced myogenic tone) in small diameter cerebral arteries during SAH. Furthermore, therapeutic strategies that improve clinical outcome in SAH patients (e.g., supraphysiological intravascular pressure) are effective in dilating small diameter cerebral arteries isolated from SAH animals.

Journal of Neuroscience Methods, 2008

Although the pathophysiology of post-angioplasty restenosis has been extensively studied in extracranial arteries using transluminal vascular injury model in rodents, it is still not well known in the intracranial arteries, which have quite different structures from extracranial arteries. Here, we examined whether 1-min placement of modified intraluminal suture could induce an injury in the internal carotid artery (ICA) in rats and observed temporal profile of histological change after the injury. HE staining showed that the injured intracranial ICA was dilated, while the media was markedly thinned at 1 day after injury. The internal elastic lamina was not observed, and the media contained few cells. At 1 week after injury, a thin layer of neointimal hyperplasia was observed on the luminal side of the internal elastic lamina. Neointimal hyperplasia developed until at least 4 weeks after injury. Morphometric analysis demonstrated that the healing process of the injury was related to arterial remodeling. Immunohistochemical staining for ␣-smooth muscle actin and electron microscopic analysis showed that the neointima was composed of smooth muscle cells. Re-endothelialization was observed from 1 to 4 weeks after injury by immunohistochemical staining for von Willebrand's factor and electron microscopic analysis. Vascular endothelial growth factor was expressed in neointima on days 7 and 14. Interestingly, superoxide anion was not increased in injured arteries on day 3, when the infiltration of macrophages was intensive, but increased on day 7, when infiltrating macrophages almost disappeared. These findings might shed new light on pathophysiology of post-angioplasty restenosis in intracranial arteries.

Acta Neurochirurgica, 1992

Haemodynamic instability is of great importance in clinical management of patients with subarachnoid haemorrhage (SAH). The significance of angiographicaUy demonstrable vasospasm for disturbances of cerebral blood flow (CBF) and cerebral autoregulation has not yet been clarified.

Stroke, 1988

Cerebral blood flow was measured by a [ l4 C]butanol indicator fractionation technique in rats subjected to subarachnoid hemorrhage, in control rats, and in rats given injections of buffered saline into the subarachnoid space (sham hemorrhage). Cerebral blood flow was significantly decreased in both the subarachnoid hemorrhage and sham hemorrhage rats 3 hours after injection. However, blood flow returned to control levels by 24 hours, and measurement for 14 days after subarachnoid hemorrhage failed to show any delayed decrease in cerebral blood flow. Electron microscopic studies of basilar arteries from rats subjected to subarachnoid hemorrhage 72 hours before killing failed to show any of the morphologic changes that have been associated with vasospasm in humans or in higher animal models. Our studies indicate that the rat model of subarachnoid hemorrhage has limited applicability to the study of subarachnoid hemorrhage following ruptured cerebral aneurysms in humans. However, although rats are not a perfect model of this clinical condition, some pathophysiologic changes similar to those observed in human subarachnoid hemorrhage have been demonstrated in this model and deserve further investigation.

Neuroradiology, 2007

Exact quantification of vasospasm by angiography is known to be difficult especially in small vessels. The purpose of the study was to develop a new method for computerized analysis of small arteries and to demonstrate feasibility on cerebral angiographies of rats acquired on a clinical angiography unit. A new software tool analysing grey values and subtracting background noise was validated on a vessel model. It was tested in practice in animals with subarachnoid haemorrhage (SAH). A total of 28 rats were divided into four groups: SAH untreated, SAH treated with local calcium antagonist, SAH treated with placebo, and sham-operated. The diameters of segments of the internal carotid, caudal cerebral, middle cerebral, rostral cerebral and the stapedial arteries were measured and compared to direct measurements of the diameters on magnified images. There was a direct correlation between the cross-sectional area of vessels measured in a phantom and the measurements acquired using the new image analysis method. The spread of repeated measurements with the new software was small compared to the spread of direct measurements of vessel diameters on magnified images. Application of the measurement tool to experimental SAH in rats showed a statistically significant reduction of vasospasm in the SAH groups treated with nimodipine-releasing pellets in comparison to all the other groups combined. The presented computerized method for analysis of small intracranial vessels is a new method allowing precise relative measurements. Nimodipine-releasing subarachnoidal pellets reduce vasospasm, but further testing with larger numbers is necessary. The tool can be applied to human angiography without modification and offers the promise of substantial progress in the diagnosis of vasospasm after SAH.

Academic Emergency Medicine, 2002

Patients with essential hypertension (EH) have higher mortality rates from hemorrhage. How the complex physiologic changes seen in EH affect the response to uncontrolled hemorrhage has yet to be adequately described. Objective: To test the null hypothesis that there would be no difference in the hemorrhage volumes and hemodynamic responses to uncontrolled hemorrhage between hypertensive rats (SHRs) and normotensive rats (WKYs). Methods: Twenty-four adult rats (12 WKYs and 12 SHRs) were anesthetized with althesin via the intraperitoneal route. The femoral artery was cannulated by cutdown for mean arterial pressure (MAP) measurement and blood gas sampling. Twelve rats (6 WKYs and 6 SHRs) underwent uncontrolled hemorrhage by 50% tail amputation. Twelve rats (6 WKYs and 6 SHRs) served as non-hemorrhage controls. The MAP, base excess (BE), and cumulative blood loss were measured pre-hemorrhage and then every 15 minutes post-hemorrhage for 90 minutes. Data were reported as mean Ϯ standard error of the mean. Comparisons between control and uncontrolled hemorrhage groups were analyzed by analysis of variance (ANOVA) with repeated-measures post-hoc testing by Bonferroni. Statistical significance was defined by an alpha = 0.05. Results: Mortality rates were significantly higher (p < 0.05) for the SHRs (100%) as compared with the WKYs (33%). Changes in time-averaged MAP post-hemorrhage were significantly greater (p < 0.001) in the SHR group (88 Ϯ 10 mm Hg) as compared with the WKY group (48 Ϯ 4 mm Hg). Hemorrhage volume was significantly lower (p = 0.02) in the SHR group (3.7 Ϯ 0.5 mL) as compared with the WKY group (6.1 Ϯ 0.7 mL). Conclusions: Hypertensive rats had a higher mortality rate than normotensives from a comparable vascular injury with lower hemorrhage volumes.

Surgical Neurology International, 2011

Background: Double-injection models of subarachnoid hemorrhage (SAH) in rats are the most effective in producing vasospasm, delayed neurological deficits and infarctions. However, they require two large surgeries to expose the femoral artery and the atlanto-occipital membrane. We have developed a minimally-invasive modification that prevents confounding effects of surgical procedures, leakage of blood from the subarachnoid space and minimizes risk of infection. Methods: Rats are anesthetized and the ventral tail artery is exposed through a small (5 mm), midline incision, 0.2 mL of blood is taken from the artery and gentle pressure is applied for hemostasis. The rat is flipped prone, and with the head flexed to 90 degrees in a stereotactic frame, a 27G angiocath is advanced in a vertical trajectory, level with the external auditory canals. Upon puncturing the atlanto-occipital membrane, the needle is slowly advanced and observed for cerebrospinal fluid (CSF). A syringe withdraws 0.1 mL of CSF and the blood is injected into the subarachnoid space. The procedure is repeated 24 hours later by reopening the tail incision. At 8 days, the rats are euthanized and their brains harvested, sectioned, and incubated with triphenyltetrazolium chloride (TTC). Results: Rats develop neurological deficits consistent with vasospasm and infarction as previously described in double-injection models. Cortical and deep infarctions were demonstrated by TTC staining and on histopathology. Conclusions: A minimally invasive, double-injection rat model of SAH and vasospasm is feasible and produces neurological deficits and infarction. This model can be used to study neuroprotective treatments for vasospasm and delayed neurological deficits following SAH, reducing the confounding effects of surgical interventions.

Acta Neurochirurgica, 1991

Although the pathophysiology of chronic cerebral vasospasm following subarachnoid haemorrhage (SAH) is still unclear, it is certain that the amount of subarachnoid blood is predictive of the severity of cerebral vasospasm. Accordingly, massive subarachnoid haemorrhage (> 0.5 ml) was induced in adult rats via direct injection into the cisterna magna. Compared to other previously published models of experimental SAH in rats a much larger amount of blood was injected. The basilar artery was exposed 72 hours post subarachnoid haemorrhage and photographed under controlled conditions. The diameter of the artery was assessed by an image analyzer. A 50% reduction in diameter was found in 25 rats subjected to SAH as compared to 9 control rats and 4 rats with intracisternal saline injection. We conclude that when massive subarachnoid haemorrhage is induced, and direct measurements of the basilar artery are made, the rat can be used as a reliable model for investigation of SAH induced arterial vasospasm.

Turkish Neurosurgery, 2020

To compare the histological and angiographic measurements of the basilar artery in an experimental rabbit subarachnoid hemorrhage model. The basilar artery was measured using both histological and angiographic methods in experimental subarachnoid hemorrhage (SAH) and vasospasm rabbit models. New Zealand white rabbits were randomly categorized into two groups: control and SAH groups. The SAH group rabbits were operated on to create an experimental SAH. Both groups were examined angiographic and histological methods. On comparing the two methods, angiographic and histopathological measurements of the basilar artery were similar in the control group. However, in the SAH group, the difference between the angiographic and histopathological measurement methods was significant. Histopathological measurements of the basilar artery were lower than angiographic measurements, and the difference was statistically significant. In the angiographic method, although there was a marked decrease in basilar artery measurements in the SAH group, the differences between the groups was not statistically significant. However, in the histopathological method, measurement differences between the control and SAH groups were statistically significant. CONCLUSION: Histopathological measurements were shown to be more sensitive than angiographic methods in demonstrating cerebral vasopasm in experimental SAH rabbit models.