A technical description of the brain tumor window model: an in vivo model for the evaluation of intraoperative contrast agents.

The evaluation of candidate optical contrast agents for brain tumor delineation in ex vivo models may not accurately predict their activity in vivo. This study describes an in vivo model system designed to assess optical contrast agents for brain tumor delineation. The brain tumor window (BTW) model was created by performing biparietal craniectomies on 8-week-old Sprague-Dawley rats, injecting 9L glioma cells into the cortex and bonding a cover slip to the cranial defect with cyanoacrylate glue. Tumor growth was followed serially and occurred in an exponential fashion. Once tumors on the cortical surface achieved a 1mm radius, intravenous contrast agents were injected while the appearance of the cortical surface was recorded. Computerized image analysis was used to quantitatively evaluate visible differences between tumor and normal brain. Tumor margins became readily apparent following contrast administration in the BTW model. Based on red component intensity, tumor delineation improved fourfold at 50 min post-contrast administration in the BTW model (P<0.002). In summary, window placement overlying an implanted glioma is technically possible and well tolerated in the rat. The BTW model is a valid system for assessing the in vivo activity of optical contrast agents.

The brain tumor window model: a combined cranial window and implanted glioma model for evaluating intraoperative contrast agents.

OBJECTIVE: Optical contrast agents for brain tumor delineation have been previously evaluated in ex vivo specimens from animals with implanted gliomas and may not reflect the true visual parameters encountered during surgery. This study describes a novel model system designed to evaluate optical contrast agents for tumor delineation in vivo. METHODS: Biparietal craniectomies were performed on 8-week-old Sprague-Dawley rats. 9L glioma cells were injected intraparenchymally. A cover slip was bonded to the cranial defect with cyanoacrylate glue. When the tumor radius reached 1 mm, Coomassie Blue was administered intravenously while the appearance of the cortical surface was recorded. Computerized image analysis of the red/green/blue color components was used to quantify visible differences between tumor and nonneoplastic tissue and to compare delineation in the brain tumor window (BTW) model with the conventional 9L glioma model. RESULTS: The tumor margin in the BTW model was poorly defined before contrast administration but readily apparent after contrast administration. Based on red component intensity, tumor delineation improved 4-fold at 50 minutes after contrast administration in the BTW model (P < .002). The conventional 9L glioma model overestimated the degree of delineation compared with the BTW model at the same dose of Coomassie Blue (P < .03). CONCLUSION: Window placement overlying an implanted glioma is technically possible and well tolerated in the rat. The BTW model is a valid system for evaluating optical contrast agents designed to delineate brain tumor margins. To our knowledge, we have described the first in vivo model system for evaluating optical contrast agents for tumor delineation.

Effect of electrical stimulation of the cervical spinal cord on blood flow following subarachnoid hemorrhage.

OBJECT: Cervical spinal cord stimulation (SCS) increases global cerebral blood flow (CBF) and ameliorates cerebral ischemia according to a number of experimental models as well as some anecdotal reports in humans. Nonetheless, such stimulation has not been systematically applied for use in cerebral vasospasm. In the present study the authors examined the effect of cervical SCS on cerebral vasoconstriction in a double-hemorrhage model in rats. METHODS: Subarachnoid hemorrhage (SAH) was induced with 2 blood injections through an indwelling catheter in the cisterna magna. Spinal cord stimulation was applied 90 minutes after induction of the second SAH (Day 0) or on Day 5 post-SAH. Measurements of the basilar artery (BA) diameter and cross-sectional area and regional CBF (using laser Doppler flowmetry and (14)C-radiolabeled N-isopropyl-p-iodoamphetamine hydrochloride) were obtained and compared between SAH and sham-operated control rats that did not receive SCS. RESULTS: At Day 0 after SAH, there were slight nonsignificant decreases in BA diameter and cross-sectional area (89 +/- 3% and 81 +/- 4%, respectively, of that in controls) in no-SCS rats. At this time point, BA diameter and crosssectional area were slightly increased (116 +/- 6% and 132 +/- 9%, respectively, compared with controls, p < 0.001) in SCS-treated rats. On Day 5 after SAH, no-SCS rats had marked decreases in BA diameter and cross-sectional area (64 +/- 3% and 39 +/- 4%, respectively, compared with controls, p < 0.001) and corrugation of the vessel wall. These changes were reversed in rats that had received SCS (diameter, 110 +/- 9% of controls; area, 106 +/- 4% of controls; p < 0.001). Subarachnoid hemorrhage reduced CBF at Days 0 and 5 post-SAH, and SCS increased flows at both time points, particularly in regions supplied by the middle cerebral artery. CONCLUSIONS: Data in this study showed that SCS can reverse BA constriction and improve global CBF in this SAH model. Spinal cord stimulation may represent a useful adjunct in the treatment of vasospasm.

Characterization of an improved double hemorrhage rat model for the study of delayed cerebral vasospasm.

Rats have been shown in several studies to develop delayed cerebral vasospasm after subarachnoid hemorrhage (SAH). However, the rat model has not yet been fully validated, and there are number of methodological issues. In this study, we present an improved double hemorrhage rat model for studies of delayed cerebral vasospasm. Double hemorrhage was induced at an interval of 24h using a PE-10 catheter introduced into the cisterna magna through a parieto-occipital burr hole. Blood volumes consisted of 0.3/0.2 ml (Group 1) or 0.2/0.1 ml (Group 2). Regional cerebral blood flow (CBF) was measured with (14)C-IMP, and compared to a sham operated control group. Additionally, the diameter and cross-sectional area of basilar artery (BA) was measured microscopically at four different levels. Mortality in Group 1 was 40%, compared to 1.5% in Group 2 and further experiments focused on the latter. Immediately following the second hemorrhage, the vessel diameter and cross-sectional area of BA were not significantly changed, but by day 5 these had decreased to 64+/-3% and 39+/-4% of control (p<0.001), respectively. CBF changes were global, affecting all cerebral areas to a similar degree. Immediately following second SAH induction, CBF diminished to 62-73% of control (p<0.05). By day 1, CBF had slightly increased above normal (101-120%), but there was then a second progressive reduction in flow in all areas examined at day 5 reaching 70-85% of baseline (p<0.05). CBF then normalized by day 7 and 9. In this study, we present a new technique for creation of double SAH leading to delayed cerebral vasospasm in rats.

Mechanisms of spinal cord stimulation in ischemia.

OBJECT: The goal of this study was to assess the duration of neuroprotection after SCS. Nearly 40 years after the first description of spinal cord stimulation (SCS), the mechanisms underlying its physiological effects remain unclear. It is known that SCS affects activity in the nervous system on a broad scale. Local neurohumoral changes within the dorsal horn of the spinal cord have been described, as have changes in cortical activation in a number of brain regions. Spinal cord stimulation has even been found to have profound effects on sympathetic vascular tone, a discovery that has led to its use in ameliorating blood flow in the limbs, heart, and brain. METHODS: In an effort to delineate the limits of neuroprotection offered by SCS, the authors have studied its use in an experimental model of permanent middle cerebral artery (MCA) occlusion in rats. The investigators applied SCS in a delayed fashion 3, 6, or 9 hours after MCA occlusion. The results are reported and mechanisms underlying the physiological effects of SCS are reviewed, with particular attention being paid to the effect of SCS on cerebral blood flow in the setting of cerebral ischemia. CONCLUSIONS: The authors found that SCS applied as late as 6 hours postischemia significantly reduces stroke volumes, whereas SCS applied 9 hours after ischemia fails to reduce stroke injury.

Evidence for a central pathway in the cerebrovascular effects of spinal cord stimulation.

OBJECTIVE: Cervical spinal cord stimulation (SCS) augments cerebral blood flow (CBF) in a number of animal models. The mechanisms underlying the cerebrovascular effects of SCS are not yet well delineated. In this study, we analyzed two alternative pathways in CBF alterations induced by SCS in rats, one involving direct modulation of sympathetic outflow and the other through central vasomotor influence. METHODS: Resection of the superior cervical ganglion (SCG), SCS alone, or SCS after SCG removal was performed in adult male Sprague-Dawley rats. CBF was measured with (14)C-inosine monophosphate radiotracer studies. In another set of experiments, SCS was performed after spinalization at the cervicomedullary junction or after laminectomy alone. RESULTS: Baseline CBF in the SCG removal group was 71 +/- 8 ml/100 g/min, similar to controls. SCS alone significantly increased blood flow to 100 +/- 10 ml/100 g/min (P < 0.05). Animals that underwent SCS after SCG removal demonstrated a similar robust augmentation in CBF. SCS-induced changes in CBF were completely attenuated by spinalization. CONCLUSION: The profound effects of spinal cord transection on SCS-induced CBF augmentation, together with the lack of effect of surgical sympathectomy, suggest that the mechanisms underlying the effects of SCS involve central influences rather than cervical sympathetic outflow. These findings suggest a possible role for brainstem vasomotor centers in the cerebrovascular effects of SCS.

Sympathetic mechanisms in cerebral blood flow alterations induced by spinal cord stimulation.

OBJECT: Cervical spinal cord stimulation (SCS) has been found to augment cerebral blood flow (CBF) in a number of animal models, although the mechanisms underlying the cerebrovascular effects of SCS are poorly described. In this study, the authors examined the role of sympathetic tone in CBF alterations induced by SCS in rats. METHODS: Spinal cord stimulation was performed at three intervals while CBF was monitored with laser Doppler flowmetry (LDF). Either hexamethonium (5, 10, or 20 mg/kg), prazosin (0.25, 0.5, or 1 mg/kg), idazoxan (0.5, 1, or 2 mg/kg), propranolol (1, 2, or 4 mg/kg), or vehicle was administered intravenously before the second stimulation. Changes in LDF values due to SCS were recorded as the percentage of change from baseline values and were analyzed. In vehicle-treated animals, SCS increased LDF values by 60.5 +/- 1.8% over baseline, whereas both high-dose hexamethonium and prazosin completely abolished the SCS-induced increases in LDF values. On the other hand, LDF values increased by 50.9 +/- 4% and 61.4 +/- 4% after SCS in the presence of idazoxan or propranolol, respectively. Administration of sympathetic nervous system blockers resulted in a variable degree of systemic hypotension as well. Nevertheless, induced hypotension without sympathetic blockade had only a minimal effect on SCS-induced augmentation of LDF values (48 +/- 1.4% over baseline). CONCLUSIONS: Sympathetic tone plays a major role in SCS-induced increases in CBF. This effect seems to be mediated primarily by alpha1-adrenergic receptors. Systemic hypotension alone cannot explain the effects of sympathetic blockade on the SCS response. Clinical use of SCS in the treatment of cerebral ischemia should take alpha1-adrenergic receptor sympathetic tone into account.

Spinal cord stimulation reducing infarct volume in a model of focal cerebral ischemia in rats.

OBJECT: The authors previously showed that spinal cord stimulation (SCS) increases cerebral blood flow in rats, indicating that this technique may be useful in the treatment of focal cerebral ischemia. In the present study, the neuroprotective potential of SCS in the setting of middle cerebral artery occlusion (MCAO) was investigated. METHODS: The authors induced permanent, focal cerebral ischemia by using either suture-induced occlusion or direct division of the MCA in Sprague-Dawley rats. Electrical stimulation of the cervical spinal cord was performed during cerebral ischemia. Cerebral blood flow was assessed using both laser Doppler flowmetry (LDF) and quantitative radiotracer analysis. Stroke volumes were analyzed after 6 hours of ischemia. Spinal cord stimulation resulted in a 52.7 +/- 13.3% increase in LDF values (nine animals). Following MCAO, LDF values decreased by 64.1 +/- 3.6% from baseline values (10 animals). Spinal cord stimulation subsequently increased LDF values to 30.9 +/- 13.5% below original baseline values. These findings were corroborated using radiotracer studies. Spinal cord stimulation in the setting of transcranial MCAO significantly reduced stroke volumes as well (from 203 +/- 33 mm3 [control] to 32 +/- 8 mm3 [MCAO plus SCS], seven animals in each group, p < 0.001). Similarly, after suture-induced MCAO, SCS reduced stroke volumes (from 307 +/- 29 mm3 [control] to 78 +/- 22 mm3 [MCAO plus SCS], 10 animals in each group, p < 0.001). CONCLUSIONS: A strategy of performing SCS for the prevention of critical ischemia is feasible and may have the potential for the treatment and prevention of stroke.