✓ A primate model was used to determine whether oxyhemoglobin (OxyHb), methemoglobin (MetHb), or bilirubin is likely to be responsible for cerebral vasospasm following subarachnoid hemorrhage (SAH). Forty cynomolgus monkeys were randomly assigned to one of five groups. On Day 0, each animal underwent angiography followed by right craniectomy and placement of an Ommaya reservoir with its catheter adjacent to the right middle cerebral artery (MCA). The animals received intrathecal injections twice a day for 6 days of one of the following solutions: mock cerebrospinal fluid (CSF); OxyHb; MetHb; bilirubin; or supernatant fluid from an incubated mixture of autologous blood and mock CSF. On Day 7, angiography was repeated and the animals were killed. Comparison of angiograms obtained on Day 0 and Day 7 of the experiment showed significant vasospasm of the right MCA and the right anterior cerebral and internal carotid arteries in the animal groups that had received OxyHb or supernatant fluid. There was a smaller reduction in diameter of the same vessels in the bilirubin group (not statistically significant), while no effects were observed in the groups receiving MetHb or mock CSF. Electron microscopy of the right MCA's gave results consistent with the angiographic findings. One monkey in the OxyHb group developed a delayed-onset right MCA infarction. These data suggest that OxyHb is the cause of cerebral vasospasm following SAH.
R. Loch Macdonald, Bryce K. A. Weir, Tim D. Runzer, Michael G. A. Grace, J. Max Findlay, Kenichi Saito, David A. Cook, Bruce W. Mielke and Kenji Kanamaru
J. Max Findlay, R. Loch Macdonald, Bryce K. A. Weir and Michael G. A. Grace
✓ It is generally believed that surgery in the face of angiographic vasospasm is dangerous due to an increased incidence of postoperative cerebral ischemia. One theory is that arterial narrowing is exacerbated by surgical manipulation of vasospastic vessels during aneurysm dissection and clipping. This theory was tested in a primate model of cerebral vasospasm and the results reported.
Six monkeys underwent baseline cerebral angiography, followed by induction of subarachnoid hemorrhage (SAH) on both sides of the circle of Willis. An equal amount of fresh autologous blood clot was placed around each internal carotid, anterior cerebral, and middle cerebral artery. Six days later, angiography was repeated and the right craniectomy was reopened for clot evacuation and surgical manipulation of the right cerebral arteries, including placement of a temporary aneurysm clip on the right middle cerebral artery. The left cerebral arteries were not exposed or manipulated, and served as controls. Twenty-four hours later angiography was repeated, then the animals were killed. Equal and significant vasospasm (> 40% reduction in vessel caliber compared to baseline, p < 0.05) was seen in the middle cerebral arteries on both sides of the circle of Willis in all animals 6 and 7 days after SAH. There was no significant change in the severity of vasospasm on Day 7 compared with Day 6 in the right cerebral arteries. Increased risk of postoperative cerebral ischemia for surgery in the peak vasospasm period may be due to mechanisms other than increased arterial narrowing precipitated by surgical manipulation.
R. Loch Macdonald, Bryce K. A. Weir, James D. Young and Michael G. A. Grace
✓ It is unclear if vasospasm after subarachnoid hemorrhage (SAH) is predominantly due to smooth-muscle contraction, proliferative vasculopathy, or other changes within the arterial wall such as fibrosis or change in smooth-muscle phenotype. In this study, immunohistochemistry was used to examine changes in extracellular and cytoskeletal proteins in cerebral arteries after SAH that might support one of these mechanisms. Following baseline cerebral angiography, bilateral SAH was created in nine monkeys. Three animals each were killed 7, 14, or 28 days after SAH. Cerebral angiography was repeated on Day 7 in all animals and immediately prior to sacrifice in animals killed on Days 14 and 28. Both middle cerebral arteries and four control basilar arteries were examined using fluorescent antibody techniques with antisera to α-actin, myosin, fibronectin, fibrinogen, vimentin, desmin, laminin, and collagens (types I, III, IV, and V).
Angiography showed that vasospasm was most severe on Day 7, present but resolving on Day 14, and completely resolved by Day 28. Microscopic study of arterial sections and blinded review of microphotographs of arterial sections by five independent observers did not reveal changes in intensity of density of staining for collagens, desmin, myosin, laminin, or α-actin in the tunica media of tunica adventitia. Fibronectin immunoreactivity increased 14 days after SAH. Seven days after SAH, occasional areas of tunica media showed immunoreactivity to fibrinogen. On Day 28, intimal thickening was observed in four of six middle cerebral arteries and this tissue demonstrated immunoreactivity to α-actin, myosin, vimentin, desmin, fibronectin, laminin, and each type of collagen. No significant increases in the number of intimal cells showing immunoreactivity to α-actin were seen and no significant changes in the hydroxyproline content of cerebral arteries developed at any time after SAH. These results suggest that rigidity and lumen narrowing of vasospasm are not due to increased arterial collagen, although other proteins in the arterial wall or an alteration in cross-linking of existing proteins could produce these changes. There is no indication that smooth-muscle contractile proteins change during vasospasm or that increases in the number of α-actin-containing myointimal cells contribute to vasospasm. The occurrence of intimal thickening and increased tunica media fibronectin after vasospasm suggests that vasospasm damages smooth muscle, possibly as a result of intense prolonged smooth-muscle contraction.
R. Loch Macdonald, M. Christopher Wallace and John R. W. Kestle
✓ The postoperative angiograms in 66 patients who underwent craniotomy for clipping of 78 cerebral aneurysms were reviewed. Indications for urgent postoperative angiography included neurological deficit or repeat subarachnoid hemorrhage. Routine postoperative angiograms were carried out in the remaining patients. Postoperative angiograms were reviewed to determine the incidence of unexpected findings such as unclipped aneurysms, residual aneurysms, and unforeseen major vessel occlusions. Logistic regression analysis was used to test if the following were factors that predicted an unexpected finding on postoperative angiography: aneurysm site or size; the intraoperative impression that residual aneurysm was left or a major vessel was occluded; intraoperative aneurysm rupture; opening or needle aspiration of the aneurysm after clipping; or development of a new neurological deficit after surgery. Kappa values were calculated to assess the agreement between some of these clinical factors and unexpected angiographic findings.
Unexpected residual aneurysms were seen in three (4%) of the 78 occlusions. In addition, three aneurysms were completely unclipped (4%); these three patients were returned to the operating room and had their aneurysms successfully obliterated. There were nine unexpected major vessel occlusions (12%); six of these resulted in disabling stroke and two patients died. Of six major arteries considered to be occluded intraoperatively and shown to be occluded by postoperative angiography, two were associated with cerebral infarction. Logistic regression analysis showed that a new postoperative neurological deficit predicted an unforeseen vessel occlusion on postoperative angiography. Factors could not be identified that predicted unexpected residual aneurysm or unclipped aneurysm.
The inability to predict accurately the presence of residual or unclipped aneurysm suggests that all patients should undergo postoperative angiography. Since a new postoperative neurological deficit is one factor predicting unexpected arterial occlusion, intraoperative angiography may be necessary to help reduce the incidence of stroke after aneurysm surgery. With study of more patients or of factors not examined in this series, it may be possible to select cases more accurately for intraoperative or postoperative angiography.
Bozena A. M. Vollrath, Bryce K. A. Weir, R. Loch Macdonald and David A. Cook
✓ An investigation was undertaken of the mechanism by which oxyhemoglobin and its analog methemoglobin might cause cerebrovascular spasm. The effect of these compounds on the levels of intracellular inositol triphosphate and calcium in cultured primate cerebrovascular smooth-muscle cells and the contractile action of oxyhemoglobin on isolated rings of primate cerebral arteries were also examined. Oxyhemoglobin, but not methemoglobin, produces a transient but highly significant increase in the intracellular levels of inositol triphosphate. Intracellular calcium levels in these cells are increased by thrombin, aluminum tetrafluoride, and oxyhemoglobin, and the sustained elevation in intracellular calcium is prevented by ethyleneglycol tetra-acetic acid and the phospholipase C inhibitor neomycin. Removal of the oxyhemoglobin after as long as 48 hours' incubation with this compound allowed cells to rapidly reduce their intracellular calcium levels to near normal. Oxyhemoglobin produced contractions of isolated rings of both normal and spastic cerebral arteries, although the response of spastic vessels was significantly smaller. This effect was inhibited by neomycin. The addition of neomycin relaxed arteries that were contracted with oxyhemoglobin, 5-hydroxytryptamine, or potassium chloride. It is thus likely that activation of phospholipase C is a critical step in the development of vasospasm, but the transient nature of the response to inositol triphosphate suggests that the sustained contraction may arise from other phospholipase C-dependent mechanisms.
R. Loch Macdonald, M. Christopher Wallace and Terry J. Coyne
✓ Intracranial aneurysm surgery performed between 4 and 12 days after subarachnoid hemorrhage (SAH) has been associated with an increased risk of delayed cerebral ischemia and poor outcome compared to surgery performed before or after this time. To investigate whether this increased risk is due to aggravation of vasospasm, the angiograms obtained before and after surgery in 56 patients operated on at various times after aneurysmal SAH were studied. Vasospasm was quantitated by measuring the diameters of intracranial arteries and expressed as the ratio of the diameters of the intracranial arteries to the diameter of the extracranial internal carotid artery. Aggressive surgical clot removal was not performed at surgery. To correct for differences in prognostic factors for vasospasm between patients operated on at different times after SAH, multiple regression analysis was performed using the arterial diameter ratio during vasospasm as the dependent variable and the prognostic factors for vasospasm, including the time of surgery, as independent variables. Equations predicting the severity of vasospasm could be generated using the clinical grade on admission, patient age, and preoperative arterial diameter ratio.
The time of surgery had no effect on vasospasm. Cerebral infarction due to vasospasm developed in five (15%) of 34 patients operated on within 3 days after SAH and in four (20%) of 20 operated on between 4 and 12 days after SAH (p = 0.66). A good outcome for these two groups was achieved in 88% and 85%, respectively (p = 1.00). These results suggest that the timing of surgery does not affect the development of vasospasm. Any increased risk of cerebral ischemia associated with surgery performed between 4 and 12 days after SAH is due to factors other than aggravation of vasospasm.
R. Loch Macdonald, M. Christopher Wallace, Walter J. Montanera and Jennifer A. Glen
✓ To define the pathological effects of angioplasty on vasospastic arteries, 36 rabbits underwent angiography and induction of vasospasm by placement of blood-filled (vasospasm groups) or empty (control group) silastic sheaths around the cervical carotid arteries. Two (Day 2) or 7 days (Day 7) later, angiography was repeated and one carotid artery in each animal was dilated by balloon angioplasty. The rabbits were sacrificed 1 day, 7 days, or 3 to 4 weeks after angioplasty. Significant vasospasm developed after placement of silastic sheaths with blood (mean reductions in diameter 39% ± 6% at Day 2 and 48% ± 5% at Day 7). Arterial narrowing was less apparent in the control groups at Day 2 (24% ± 7%). Angioplasty performed on Day 2 significantly increased arterial diameters of vasospastic arteries (50% ± 7%; p < 0.05) but not those of control arteries (10% ± 6%, p > 0.05). Angioplasty performed on Day 7 increased the arterial diameters by a similar degree (47% ± 13%, not significant). Arteries remained dilated after angioplasty, although there was significant vasospasm 7 days after angioplasty when angioplasty was performed on Day 2. Blinded, semiquantitative histopathological study of the arteries showed that 3 to 4 weeks after angioplasty, there was significant endothelial proliferation and a trend for thinning of the tunica media. There were no significant changes in control arteries subjected to angioplasty. Angioplasty was not associated with significant arterial fibrosis as measured by hydroxyproline content (analysis of variance). The increase in endothelial proliferation and decrease in the thickness of the tunica media suggest that, in the rabbit model, angioplasty damages endothelial and smooth-muscle cells. This may be the basis for the observation that vasospastic arteries do not reconstrict after angioplasty.