Experimental transplantation gliomas in the adult cat brain

Experimental brain tumours were produced in adult cats by stereotactic xenotransplantation of the rat glioma clone F98. Regional ATP, glucose and lactate were measured after 2-4 weeks on coronal cryostat sections by substrate-induced bioluminescence, potassium content was imaged by the histochemical sodium cobaltinitrite method, and regional pH by incubating cryostat sections with the fluorescent pH-indicator umbelliferone. The regional biochemical alterations were correlated with magnetic resonance imaging and tissue water content. Biochemical changes were heterogeneous in tumours but exhibited a rather uniform pattern in peritumoural oedema. ATP was consistently reduced, glucose and lactate were increased and pH was more alkaline than in normal white matter. The decrease of ATP matched the increase of water, indicating that ATP decline represents fractional dilution in the oedematous tissue rather than break-down of energy metabolism. The increased lactate levels, therefore, may originate from the tumour and not from a metabolic disturbance in the peritumoural oedematous tissue. The implications of this interpretation for the pathogenesis of peritumoural oedema are discussed.     

A study on peritumoural brain oedema around meningiomas by CT and MRI scanning

Summary There is a great variability in the amount of peritumoural brain oedema accompanying meningiomas. In a previous study it was found that the degree of brain oedema in the white matter around meningiomas correlated with disruption of the layers (especially the cerebral cortex), which separate the tumour from the white matter, as well as with the size and histological subtype of the tumour.In the present study comprising 9 meningiomas, the volume of oedema was calculated by integration of the cross-sectional oedematous areas on serial MRI slices. The volume of oedema was zero in 3 cases and ranged from 11 to 176.4 ml in the other 6 cases. The MRI-scans also showed disruption of the cortex in all cases, ranging from slight to severe. T₁ and T₂ measurements were made at the level of maximum extension, using a mixed sequence at a field strength of 1.5 T. From the T₂ values tissue water content in % was calculated using the equations: WC=39.36/(R₂ + 37.2) for cortex, and WC=29.63/(R₂ + 27.8) for white matter. These had been obtained by correlating water content with relaxation rates, measured in vitro on human brain autopsy specimens which were subjected to hydration with distilled water or dehydration by hyperosmolar solutions. Mean water content amounted to 82.53% for normal cortex, 74.72% for normal white matter, and 84.59% for oedematous white matter around the tumour.On the assumption that the spread of contrast agent marks the advancement of the front of oedema produced by the tumour, CT-studies were made before, and at 1 1/2, 3 and 6h after contrast infusion. The increase in diameter of the contrast-stained area on the CT-scan allows calculation of the excess of oedema production per unit tumour volume. Of 6 tumours with oedema (mean peritumoural water content of 91% and mean volume of oedema of 69.2 ml) the production excess at the steady-state was 0.18–1.08 ml/h/cm³ tumour volume, whereas 3 tumours without associated oedema had a production excess of 0.03–0.12 ml/h/cm³. Moreover, penetration of the cortex seems to constitute a separate factor determinig the spread of oedema.     

Factors affecting the extension of peritumoural brain oedema. A CT-study

Summary In human brain tumours the extension of peritumoural brain oedema may vary considerably. 37 brain tumours of various pathology and 2 abscesses were examined to identify the factors and mechanisms responsible for the oedema spreading. Peritumoural oedema profiles were determined towards the white matter and ventricle by measuring the CT-numbers of consecutive tissue blocks of 3.0–3.6 mm from the tumour to the normal white matter or the ventricle. It was found that neither the size of the tumour nor the histology has a close relationship to the amount of peritumoural oedema. The distance of oedema spreading rather is determined by the amount of fluid accumulation in the white matter immediately bordering the tumour. This relationship corresponds to a semilogarithmic function and represents the relation between the tumour-adjacent accumulation of extracellular fluid volume and the distance of extracellular fluid movement. The analysis of this relation leads to the suggestion that pressure gradients and bulk flow are involved in the development of human peritumoural oedema.     

Corticosteroid therapy of experimental tumour oedema

Summary Experimental brain tumours were produced in cats by stereotactic implantation of 4 million suspended cells of a rat glioma clone into the internal capsule. Three weeks after implantation a spherical tumour developed with a diameter of up to 10 mm which was surrounded by vasogenic white matter oedema. In untreated animals water content in the peritumoural white matter increased from 69.1 ± 0.9 to 80.0 ± 0.8 ml/100 g w. w., and regional blood flow reciprocally decreased from 32.2. ± 5.6 to 18.9 ± 0.05 ml/100 g/min. A single injection of a crystalline suspension of 10 mg/kg dexamethasone given intramuscularly one week before the animals were killed, led to a significant amelioration of brain oedema. Peritumoural white matter water content decreased to 73.0 ± 0.5 ml/100 g w. w. and blood flow rose to 35.7 ± 2.8 ml/100 g/min. These changes were accompanied by parallel shifts of electrolyte content but they did not correlate with EEG activity, as assessed by Fourier frequency analysis. Corticosteroids did not prevent extravasation of peroxidase or Evans blue across the tumour vessels. The beneficial effect, therefore, is attributed to either an acceleration of resorption or an inhibition of the spread of oedema from the tumour into the peritumoural brain tissue.     

Formation and propagation of brain oedema fluid around human brain metastases. A CT study

Summary Computerized tomography (CT) was used to examine the timecourse of the propagation of extravasated contrast medium from small brain metastases into the peritumoural oedematous white matter, following infusion of 200 ml of meglumine amidtrizoate for 3 hours. Four patients with a metastatic brain tumour were examined. CT scans at identical levels were taken 1.5, 3, 6, 9, and 12 hours after start of contrast infusion. Following 4–7 days of dexamethasone treatment (8–12mg/day i.v.) the examination was repeated. A contrast-enhanced area was observed surrounding the clearly delineated tumours, expanding gradually in a circular fashion into the peritumoural white matter oedema. The expanding circular enhancement was measured planimetrically on the various scans. From these values, the increase in radius/hr respectively in volume/hr was calculated, assuming a spherical geometry. This enabled a determination of the rate of oedema fluid formation and of the speed of oedema fluid propagation. The formation rate of oedema fluid amounted to 0.5–3.2ml/hour and the speed of oedema fluid spreading to 1.9 mm/hour. Following treatment with dexamethasone the formation rate of oedema fluid is reduced by 30–50%. The important clinical implications of these new findings are discussed.     

Examination of the metabolism of oedematous brain tissue

Glucose and gas metabolism as well as water, sodium, and potassium content of normal and oedematous white matter slices of rabbit brain have been determined. A cutting technique for the preparation of the slices is described. Although the increase of the relative water content in oedematous white matter is much more pronounced than in oedema of the grey matter, we observed only one significant change: the pyruvate production in vitro is about 20% higher in slices prepared from oedematous brain.     

Experimental transplantation gliomas in the adult cat brain

Summary In adult cats experimental brain tumours were produced by stereotactical xenotransplantation of the rat glioma clone F 98 into the internal capsule of the left hemisphere. Two to four weeks after transplantation tumours and peritumoural oedema were investigated by magnetic resonance imaging (MRI), electrophysiological recording and analysis of tissue content of water, electrolytes and extravasated serum proteins.Spherical tumours with a diameter of about 10 mm developed at the injection site and were surrounded by massive white matter oedema. Water content in peritumoural white matter increased from 2.63 ± 0.17 to 3.65 ± 0.19 ml/g d.w. (means ± SD), sodium from 187±11 to 351±55 eq/g d.w. and calcium from 7.4±1.1 to 13.3 ± 1.3 ± 1.3 eq/g d.w. Potassium and magnesium did not change. Oedema development was associated with the extravasation of 18.0 ± 16.8mg/g d.w. albumin and 15.8 ± 12.2 mg/g d.w. immunoglobulin. The calculated electrolyte content of oedema fluid approximated that of plasma but the serum protein content was about 40% lower. The ratio of low (albumin) to high (immunoglobulin) molecular weight proteins was the same in blood and oedema fluid. It is, therefore, concluded that peritumoural oedema consist of two components,a whole plasma extravasate and a protein-free ultra-filtrate.Peritumoural oedema could be clearly detected by MRI but differentiation between tumour and oedema was only possible after contrast enhancement with gadolinium-DTPA. The ratios of the intensities of the MR signal correlated linearly with the water content within white matter. MRI, in consequence, allows quantification of oedema provided a reference area with normal water content is present.     

A study of electrical impedance of oedematous cerebral tissue during operations

Summary In 11 patients, most of whom had cerebral tumours, electrical impedance was recorded during the introduction of an impedance electrode during operation. After removal of the lesion impedance was measured in the tissue adjacent to the walls of the wound cavity. The readings were correlated with water and electrolyte content and with histological findings in the specimens taken from the same places. Electrical impedance appeared to be lowered in regions invaded by tumour and also in the surrounding oedematous areas of white matter. Unless primarily affected, cerebral cortex usually did not show oedema.The increase of water in the oedematous area was accompanied by a proportional increase of tissue Na⁺ and a decrease of tissue K⁺. The method of impedance measurement does not allow the delineation of a tumour from the surrounding oedema.     

In vivo estimation of water content in cerebral white matter of brain tumour patients and normal individuals: Towards a quantitative brain oedema definition

Summary The building stones in MR imaging, namely the relaxation times (RT) correlate in certain circumstances well with water content of cerebral white matter (WM). It is, therefore, possible to estimate regional in vivo water content using RT imaging with an accuracy within ±2–4%. The purpose of the present prospective investigation was to study the RT's of cerebral WM and thereby estimate normal in vivo water content. Furthermore, the method was used to study whether water content was different in the apparently normal cerebral hemisphere of brain tumour patients compared to normal individuals. RT data of the non-tumour bearing hemisphere in 42 brain tumour patients were compared to 35 normal individuals. Results: 1) Water content was significantly higher in the posterior WM as compared to frontal WM in both normal individuals and apparently normal WM in brain tumour patients and this appears to be a true physiological difference. 2) Water content of normal WM correlated positively with age. 3) There was no significant difference between RT data from the normal hemisphere in histologically different types of brain tumours. 4) Water content was significantly higher in the apparently normal WM of brain tumour patients compared to normal individuals. 5) Based on the present results, a definite lower threshold of white matter oedema could be established as 75% water content, corresponding to a T1 of 0.850 seconds.     

Resorption of peritumoural oedema in cerebral gliomas during dexamethasone treatment evaluated by NMR relaxation time imaging

Summary Peritumoural brain oedema is a prominent feature of malignant brain tumours. Glucocorticoids diminish the neurological symptoms and signs caused by the oedema and reduce the abnormally high cerebral water content. The exact mechanisms of action of the glucocorticoids are unknown.The present study investigates the influence of dexamethasone on NMR relaxation time T₁ in peritumoural oedema in 13 patients with gliomas. It is shown that NMR T₁ images can be used as a potent monitor of brain oedema, and that dexamethasone significantly reduces mean T₁ after 1, 3, and 7 days of treatment by 2%, 6%, and 13% respectively.Using an image histogram analysis technique the term superoedema was defined as the 50% of the total oedema area with the highest t₁, corresponding to the highest water content. It is shown, that with this technique the treatment effect of steroids on superoedema was a reduction of 13%, 33%, and 57% after 1, 3, and 7 days of treatment respectively. The mean change after 24 hours of treatment was statistically significant (p < 0.01).The method can be used in all situations where the anti-oedematous effect of a given treatment is to be monitored.     

Spatial and quantitative distribution of human peritumoural brain oedema in computerized tomography

Computerized tomography (CT) was used to study the pathways of oedema spreading in man. Based on the assumption that local changes in CT numbers in oedematous white matter closely correspond to changes in tissue water content, CT numbers of consecutive tissue blocks of 3.0-3.6 mm were examined in the main directions of oedema spreading: towards the deep white matter, towards the cortex and towards the ventricle. Tumours with oedema grade II and III showed a reduction of CT number of 10 +/- 1.8. The corresponding increase in water content of about 10-12% seems to be an upper limit of fluid accumulation in the white matter. From this oedema centre, water content very slowly and gradually decreased along the oedema projection into the deep white matter. In contrast, if oedema reached the cortex of adjacent gyri, the decline in water content was very sharp. A similar observation was made in the external capsule where oedema sharply declined at the border to the adjacent grey matter, putamen and claustrum. Oedema projection towards the ventricle showed a nearly uniform magnitude from the centre to the ventricular lining, suggesting a certain resistance by a limited capacity of transependymal drainage of oedema fluid. It is assumed that the spatial distribution and extension of oedema around a brain tumour is determined by a system of differential resistance to fluid movement in the following order: grey matter--ventricular lining--white matter.     

Angiogenesis and Brain Oedema in Intracranial Meningiomas: Influence of Vascular Endothelial Growth Factor

Summary The correlation between angiographic neovascularization, peritumoural brain oedema (PTBOe) and the expression of vascular endothelial growth factor (VEGF) , was analysed in 30 patients with intracranial meningiomas. Pre-operative angiograms were examined for the existence of either an exclusively dural tumour blush or an additionally pial tumour supply from cerebral arteries. Furthermore the presence of macroscopic tumour-neovascularization and dysplastic changes of tumour-draining cerebral veins was evaluated. VEGF expression was investigated on histological tissue samples, using immunohistochemical techniques. VEGF immunohistochemistry and neuroradiological evaluations were performed in double blind fashion. Tumour volume and the amount of oedema were calculated by computerized tomography (CT) or magnetic resonance imaging (MRI). The oedema-tumour volume ratio was defined as oedema index (OeI). Compared to VEGF-negative meningiomas, tumours with striking VEGF staining revealed a significant higher mean oedema index (OeI=4,2 vs. OeI=1,5; p<0.018), and a higher oedema incidence (91,7% vs. 44,4%; p<0.046). Equally, meningiomas with additionally tumour supply from cerebral arteries were associated with a significant higher mean OeI (OeI=4.1 vs. OeI=1.2; p<0.01) and oedema incidence (94,7% vs. 20,0%; p<0,0023) than meningiomas with exclusively tumour supply from dural arteries. All meningiomas with striking VEGF-expression were associated with vascular tumour supply from cerebral arteries, but VEGF-negative tumours only in 50% (p<0.029). These data suggest a link between VEGF-expression, arterial tumour supply and peritumoural brain oedema. The development of tumour supply from cerebral arteries may be important for formation of meningioma-related oedema. Therefore, VEGF may represent a potent mediator in the evolution of this type of vascularization in meningiomas.     

Rapid clinical deterioration in a patient with multi-focal glioma despite corticosteroid therapy: a quantitative MRI study

A patient with high-grade multi-focal glioma deteriorated rapidly despite high dose corticosteroid therapy (dexamethasone: 16 mg/day). MRI was used to measure diffusion tensor parameters and longitudinal relaxation time (T1) values of peritumoural oedematous brain before and after commencing steroid treatment. Forty-eight hours after steroid treatment there was no evidence of brain oedema reduction. Specifically, regions of oedematous brain showed a significant increase in mean diffusivity () with a significant decrease in diffusion anisotropy (p < 0.05), but without any change in T1 values. These quantitative MRI data were mirrored by the rapid deterioration seen when assessing the patient clinically. This case shows that quantitative MRI can not only measure steroid treatment response but also failure in patients with malignant gliomas.     

Immunofluoroscopic investigation of extravasation of serum proteins in human brain tumours and adjacent structures

Summary Forty-seven neurosurgical human specimens taken from malignant gliomas and peritumoural brain structures were investigated for extravasation of serum proteins. Serum proteins were visualized microscopically in paraffin-embedded material using a double layer immunofluorescence technique. Proteins accumulated in the tumour and in the adjacent peritumoural white matter, and were mainly located extracellularly. Intracellular uptake was observed in some but not all tumour cells, in reactive astrocytes and, occasionally, in oligodendrocytes and neurons. Diffuse infiltration of products was present in necrotic, cystic and haemorrhagic regions. The distribution of extravasated proteins corresponded precisely to that previously observed in transplanted tumours in cats (Hossmannet al. 1979), suggesting that the pathophysiology of human tumour oedema is similar to that of the experimental material. Since all patients were operated without corticosteroid therapy, the present results can be used as a reference for forthcoming studies on the effect of corticosteroids on peritumorous oedema.     

Effect of Hyperosmotic Solutions on Human Brain Tumour Vasculature

Summary  Reversible opening of the blood-brain barrier (BBB) has been used to increase delivery of chemotherapeutic agents into brain tumours, but it is complicated and requires general anaesthesia. Without affecting the normal BBB, and avoiding the complications of BBB modification by hyperosmotic solution, we tried an adequate minimal BBB disruption in brain tumours. Although the effect of BBB disruption on normal brain has been described, there are no reports of the effect of an impaired BBB on microcirculation. In this study, four patients underwent surgical resection of a glioblastoma multiforme (GM; n=1), astrocytoma (n=2), or metastatic brain tumour (n=1). Epicerebral microcirculation was observed in the operative field. Serial fluorescein microangiograms of the tumour and peritumoural area were obtained before and after BBB disruption was introduced intra-operatively by retrograde infusion of mannitol introducing a catheter via the temporal superficial artery back to the carotid bifurcation. On the initial microangiogram, staining by the fluorescein dye was observed in the GM and metastatic tumour but not in the astrocytoma; no extravasation of fluorescein dye was observed in the peritumoural areas. After BBB disruption, fluorescein perfusion increased and extravasation of fluorescein dye from the venules was observed in the GM and the metastatic tumour and in the peritumoural area of both lesions; BBB disruption started from venules in the peritumoural area without affecting the normal brain. However, such effects were not observed in the astrocytomas after BBB disruption nor in normal brain tissue in any patient. It appears that the integrity of the BBB is less stable in the peritumoural area of GM and metastatic brain tumours than it is in astrocytomas or normal brain. Osmotic BBB disruption may offer a method for achieving global delivery of therapeutic agents to brain tumours and peritumoural areas.     

Peritumoural brain oedema in intracranial meningiomas: Influence of tumour size, location and histology

Summary Peritumoural brain oedema was examined retrospectively in 175 patients with 179 intracranial meningiomas. The influence of tumour size, location and histology were investigated.Tumour volume and localization, and the presence of peritumoural brain oedema (PTBOe) were determined by computed tomography (CT). The oedema-tumour volume ratio was defined as Oedema Index (Oel). All patients underwent microsurgical removal of the tumour. Surgically resected meningiomas were classified histopathologically based on criteria of the new World Health Organization (WHO) classification. A close relationship was found between the tumour size and the incidence of peritumoural oedema: with increasing size of the tumour the incidence of oedema also rises, the oedema index, however decreases. Frontobasal and temporobasal meningiomas showed a significant increase in the oedema incidence and the mean oedema index. If major parts of the surface of meningiomas were adjacent to subarachnoid cisterns only a slight tendency for the development of oedema was observed. WHO-III-meningiomas showed a significantly higher oedema incidence (61.1% vs. 94.4%; p<0.004) and mean oedema index (Oel=2.7 vs. 3.7; p<0.0009) than WHO-I-meningiomas. Brain tissue was affected in 59 cases. 19 meningiomas with infiltration into adjacent brain parenchyma revealed a statistically significant increase in oedema incidence (94.7% vs. 51.7%; p<0.0003) and mean oedema index (Oel=3.9 vs. Oel=2.2; p<0.0001) when compared to tumours without any brain tissue involvement in the histopathological specimens. Tumours with large volume, fronto-temporo-basal location and anaplastic histology were not only associated with the highest incidence of oedema formation but also presented with an overproportionate infiltrative growth. Thus, a disruption of the arachnoid or a true brain infiltration may be an essential factor for the development of a PTBOe.     

Extracellular space and electrolyte distribution in cortex and white matter of dog brain in cold induced oedema

Summary 24 hours after a circumscribed cold injury of the cortex dog brains were perfused from the lateral ventricle and the frontal subarachnoidal space to the cisterna magna with an artificial CSF containing trace amounts of³⁵S-labelled thiosulphate. Simultaneously the extracellular tracer was administered intravenously. Extracellular fluid volume was estimated and found to be increased from 10 to 15% in the oedematous cortex and from 10 to 27% in the oedematous white matter. The actual size of ECS in oedematous white matter, however, must be larger as indicated by the relative alterations of thiosulphate distribution, tissue water, sodium and chloride. Apparently a small part of the fluid accumulation affects the cellular compartment in oedematous white matter. It may be concluded from the close spatial correspondence of the spreading of I¹³¹ albumin and Evans blue, the increase in water and sodium content, and the enlargement of the TSS that the dilated extracellular channels are filled with a plasma like oedema fluid, derived from blood.This investigation was supported by a grant from the Deutsche Forschungsgemeinschaft.     

Emerging molecular mechanisms of brain tumour oedema

A common property of brain tumours is their ability to cause oedema in the surrounding brain. Oedema forms as a result of a leaky blood-tumour barrier and persists when the brain fails to clear the excess fluid. It is a significant source of morbidity and mortality. The principal anatomical component of the blood-brain barrier is the endothelial tight junction which opens in glioma microvessels. Multiple tight junction proteins have recently been identified, such as occludin, claudin, ZO-1, ZO-2 and ZO-3. We propose a model to explain tight junction opening in gliomas based on vascular endothelial growth factor secretion and loss of tight junction inducing factor production by tumour cells. The level of expression of the water channel aquaporin-4 in peritumoural astrocytes may determine the rate of oedema fluid clearance. The identification of the molecular mechanisms of brain tumour oedema may allow the design of novel anti-oedema medications.     

Cystic meningioma

Summary Meningiomas are usually solid tumours. Cystic changes in meningiomas are rare. These cysts may occur extratumoural, peritumoural or intratumoural. Diagnostic difficulties arise in cases of cystic meningiomas. Nine cases of cystic meningiomas were operated on in the past 6 years at the department of neurosurgery, University of Alexandra. The mean age of patients was 46.2 years. Male to female ratio was 1/2. Less oedema was observed in extratumoural cysts, and more oedema in intratumoural cysts. The fluid contained was dark brown or dirty yellow in intratumoural cysts (type C), xanthochromic in peritumoural cysts surrounded by tumour tissue (Type B), or clear fluid in extratumoural cysts (Type A). Based on these two observations it is proposed that the cyst fluid and peritumoural oedema may represent variable degrees of degeneration or secretion by tumour cells. Pre-operatively diagnostic criteria are presented.     

Treatment of experimental brain oedema following sudden decompression,surgical wound,and cold lesion with vasoprotective drugs and the proteinase inhibitor “Trasylol”

The study was performed on 81 cats with three models of experimental brain oedema: sudden decompression, surgical wound, and cold injury. During the experiments blood pressure, central venous pressure, and intracranial pressure were recorded. The blood-brain-barrier was tested with Evans blue solution. The gray and white matter tissue was sampled at the end of the experiment, and the water content and sodium and potassium concentrations were determined. The animals with the same experimental model were divided into three groups: untreated, treated with the vasoprotective agents, and treated with the protease inhibitor Trasylol. In the sudden decompression model after balloon deflation, white matter haemorrhages and oedema development were found in gray matter and basal nuclei. In animals treated with the vasoprotective drugs, haemorrhages were not observed, and oedematous changes were less pronounced. The Trasylol effect on oedema development was not significant in this model. In the surgical wound model, oedematous changes were observed after 24 hours following the lesion. Oedema occurred in the white matter, as in the animals with cold lesions. In both models--surgical wound and cold lesion--the beneficial effect of Trasylol was shown, while the effect of Aescorin was less evident. The results obtained seemed to testify to the usefulness of both Trasylol and vasoprotective drugs in the prevention and treatment of brain oedema in neurosurgical patients.