Evaluation of predicted patellofemoral joint kinematics with a moving-axis joint model.

The main objectives of this study were to expand the moving-axis joint model concept to the patellofemoral joint and evaluate the patellar motion against experimental patellofemoral kinematics. The experimental data was obtained through 2D-to-3D bone reconstruction of EOS images and segmented MRI data utilizing an iterative closest point optimization technique. Six knee model variations were developed using the AnyBody Modeling System and subject-specific bone geometries. These models consisted of various combinations of tibiofemoral (hinge, moving-axis, and interpolated) and patellofemoral (hinge and moving-axis) joint types. The newly introduced interpolated tibiofemoral joint is calibrated from the five EOS quasi-static lunge positions. The patellofemoral axis of the hinge model was defined by performing surface fits to the patellofemoral contact area; and the moving-axis model was defined based upon the position of the patellofemoral joint at 0° and 90° tibiofemoral-flexion. In between these angles, the patellofemoral axis moved linearly as a function of tibiofemoral-flexion, while outside these angles, the axis remained fixed. When using a moving-axis tibiofemoral joint, a hinge patellofemoral joint offers (-5.12 ± 1.23 mm, 5.81 ± 0.97 mm, 14.98 ± 2.30°, -4.35 ± 1.95°) mean differences (compared to EOS) while a moving-axis patellofemoral model provides (-2.69 ± 1.04 mm, 1.13 ± 0.80 mm, 12.63 ± 2.03°, 1.74 ± 1.46°) in terms of lateral-shift, superior translation, patellofemoral-flexion, and patellar-rotation, respectively. Furthermore, the model predictive capabilities increased as a direct result of adding more calibrated positions to the tibiofemoral model (hinge-1, moving-axis-2, and interpolated-5). Overall, a novel subject-specific moving-axis patellofemoral model has been established; that produces realistic patellar motion and is computationally fast enough for clinical applications.

Development and validation of a subject-specific moving-axis tibiofemoral joint model using MRI and EOS imaging during a quasi-static lunge.

The aims of this study were to introduce and validate a novel computationally-efficient subject-specific tibiofemoral joint model. Subjects performed a quasi-static lunge while micro-dose radiation bi-planar X-rays (EOS Imaging, Paris, France) were captured at roughly 0°, 20°, 45°, 60°, and 90° of tibiofemoral flexion. Joint translations and rotations were extracted from this experimental data through 2D-to-3D bone reconstructions, using an iterative closest point optimization technique, and employed during model calibration and validation. Subject-specific moving-axis and hinge models for comparisons were constructed in the AnyBody Modeling System (AMS) from Magnetic Resonance Imaging (MRI)-extracted anatomical surfaces and compared against the experimental data. The tibiofemoral axis of the hinge model was defined between the epicondyles while the moving-axis model was defined based on two tibiofemoral flexion angles (0° and 90°) and the articulation modeled such that the tibiofemoral joint axis moved linearly between these two positions as a function of the tibiofemoral flexion. Outside this range, the joint axis was assumed to remain stationary. Overall, the secondary joint kinematics (ML: medial-lateral, AP: anterior-posterior, SI: superior-inferior, IE: internal-external, AA: adduction-abduction) were better approximated by the moving-axis model with mean differences and standard errors of (ML: -1.98 ±â€¯0.37 mm, AP: 6.50 ±â€¯0.82 mm, SI: 0.05 ±â€¯0.20 mm, IE: 0.59 ±â€¯0.36°, AA: 1.90 ±â€¯0.79°) and higher coefficients of determination (R2) for each clinical measure. While the hinge model achieved mean differences and standard errors of (ML: -0.84 ±â€¯0.45 mm, AP: 10.11 ±â€¯0.88 mm, SI: 0.66 ±â€¯0.62 mm, IE: -3.17 ±â€¯0.86°, AA: 11.60 ±â€¯1.51°).

Drug repurposing: sulfasalazine sensitizes gliomas to gamma knife radiosurgery by blocking cystine uptake through system Xc-, leading to glutathione depletion.

Glioblastomas (GBMs) are aggressive brain tumors that always recur after radiotherapy. Cystine, mainly provided by the system X(c)(-) antiporter, is a requirement for glioma cell synthesis of glutathione (GSH) which has a critical role in scavenging free radicals, for example, after radiotherapy. Thus, we hypothesized that the X(c)(-)-inhibitor sulfasalazine (SAS) could potentiate the efficacy of radiotherapy against gliomas. Here, we show that the catalytic subunit of system X(c)(-), xCT, was uniformly expressed in a panel of 30 human GBM biopsies. SAS treatment significantly reduced cystine uptake and GSH levels, whereas it significantly increased the levels of reactive oxygen species (ROS) in glioma cells in vitro. Furthermore, SAS and radiation synergistically increased DNA double-strand breaks and increased glioma cell death, whereas adding the antioxidant N-acetyl-L-cysteine (NAC) reversed cell death. Moreover, SAS and gamma knife radiosurgery (GKRS) synergistically prolonged survival in nude rats harboring human GBM xenografts, compared with controls or either treatment alone. In conclusion, SAS effectively blocks cystine uptake in glioma cells in vitro, leading to GSH depletion and increased ROS levels, DNA damage and cell death. Moreover, it potentiates the anti-tumor efficacy of GKRS in rats with human GBM xenografts, providing a survival benefit. Thus, SAS may have a role as a radiosensitizer to enhance the efficacy of current radiotherapies for glioma patients.

A novel brain metastases model developed in immunodeficient rats closely mimics the growth of metastatic brain tumours in patients.

AIMS: brain metastasis is a common cause of mortality in cancer patients, and associated with poor prognosis. Our objective was to develop a clinically relevant animal model by transplanting human biopsy spheroids derived from metastatic lesions into brains of immunodeficient rats. METHODS: nine different patient brain metastases from four different primary cancers were implanted into brains of immunodeficient rats. The xenografts were compared with patient tumours by magnetic resonance imaging, histochemistry, immunohistochemistry and DNA copy number analysis. RESULTS: after transplantation, tumour growth was achieved in seven out of nine human brain metastases. Spheroids derived from four of the metastases initiated in the rat brains were further serially transplanted into new animals and a 100% tumour take was observed during second passage. Three of the biopsies were implanted subcutaneously, where no tumour take was observed. The animal brain metastases exhibited similar radiological features as observed clinically. Histological comparisons between the primary tumours from the patients, the patient brain metastases and the derived xenografts showed striking similarities in histology and growth patterns. Also, immunohistochemistry showed a strong marker expression similarity between the patient tumours and the corresponding xenografts. DNA copy number analysis between the brain metastases, and the corresponding xenografts revealed strong similarities in gains and losses of chromosomal content. CONCLUSION: we have developed a representative in vivo model for studying the growth of human metastatic brain cancers. The model described represents an important tool to assess responses to new treatment modalities and for studying mechanisms behind metastatic growth in the central nervous system.

Gamma knife surgery of meningiomas involving the cavernous sinus: long-term follow-up of 100 patients.

OBJECTIVE: Resection of meningiomas involving the cavernous sinus often is incomplete and associated with considerable morbidity. As a result, an increasing number of patients with such tumors have been treated with gamma knife surgery (GKS). However, few studies have investigated the long-term outcome for this group of patients. METHODS: 100 patients (23 male/77 female) with meningiomas involving the cavernous sinus received GKS at the Department of Neurosurgery at Haukeland University Hospital, Bergen, Norway, between November 1988 and July 2006. They were followed for a mean of 82.0 (range, 0-243) months. Only 2 patients were lost to long-term follow-up. Sixty patients underwent craniotomy before radiosurgery, whereas radiosurgery was the primary treatment for 40 patients. RESULTS: Tumor growth control was achieved in 84.0% of patients. Twelve patients required re-treatment: craniotomy (7), radiosurgery (1), or both (4). Three out of 5 patients with repeated radiosurgery demonstrated secondary tumor growth control. Excluding atypical meningiomas, the growth control rate was 90.4%. The 1-, 5-, and 10-year actuarial tumor growth control rates are 98.9%, 94.2%, and 91.6%, respectively. Treatment failure was preceded by clinical symptoms in 14 of 15 patients. Most tumor growths appeared within 2.5 years. Only one third grew later (range, 6-20 yr). The complication rate was 6.0%: optic neuropathy (2), pituitary dysfunction (3), worsening of diplopia (1), and radiation edema (1). Mortality was 0. At last follow-up, 88.0% were able to live independent lives. CONCLUSION: GKS gives long-term growth control and has a low complication rate. Most tumor growths manifest within 3 years following treatment. However, some appear late, emphasizing the need for long-term follow-up.

Treatment of vestibular schwannomas. Why, when and how?

Sporadic vestibular schwannoma (VS) causes unilateral hearing loss, tinnitus, vertigo and unsteadiness. In many cases, the tumour size may remain unchanged for many years following diagnosis, which is typically made by MRI. In the majority of cases the tumour is small, leaving the clinician and patient with the options of either serial scanning or active treatment by gamma knife radiosurgery (GKR) or microneurosurgery. Despite the vast number of published treatment reports, comparative studies are few, and evidence is no better than class III (May, 2006). The predominant clinical endpoints of VS treatment include tumour control, facial nerve function and hearing preservation. Less focus has been put on symptom relief and health-related quality of life (QOL). It is uncertain if treating a small tumour leaves the patient with a better chance of obtaining relief from future hearing loss, vertigo or tinnitus than by observing it without treatment. Recent data indicate that QOL is reduced in untreated VS patients, and may differ between patients who have been operated and patients treated with GKR. In the present paper we review the natural course and complaints of untreated VS patients, and the treatment alternatives and results. Furthermore, we review the literature concerning quality of life in patients with VS. Finally, we present our experience with a management strategy applied to more than 300 cases since 2001.

In vivo secretory potential and the effect of combination therapy with octreotide and cabergoline in patients with clinically non-functioning pituitary adenomas.

The secretory capacity, in vivo, of clinically non-functioning pituitary adenomas may possibly predict tumour volume reduction during intensive medical therapy. Ten patients (mean (range) 53 years (26-73)) with clinically non-functioning macroadenomas, > or = 10 mm were studied. The secretory capacity of the adenomas was examined using basal, NaCl and TRH-stimulated LH, FSH and alpha-subunit levels. The effect on tumour volume of 6 months' therapy with the combination of a somatostatin analogue, octreotide 200 microg x 3/day and a dopamine-D2-agonist, cabergoline 0.5 mg x 1/day was studied. The basal LH, FSH and alpha-subunit levels were determined before and during 6 months' therapy with octreotide and cabergoline, and MR scans were used to evaluate tumour volume before and during this period of therapy. Octopus-perimetry was used to examine the visual fields. A reduction in tumour volume (mean +/- SEM (range); 30% +/- 4% (18-46%)) during 6 months of combination therapy with octreotide and cabergoline was recorded only in patients with in vivo secretory potential. Tumour volume was not reduced in four patients: in three of these patients it remained unchanged while in one patient it was observed to have increased (by 14%). Of the six patients with pretherapy secretory capacity, one displayed a very high basal level of alpha-subunit (74 microg/l) despite unmeasurable levels of LH and TSH, and an FSH-level of 1 IU/l. The other five patients presented paradoxical LH, FSH and/or alpha-subunit responses to TRH. A reduction in basal levels of LH, FSH and/or alpha-subunit was observed in all six patients, and the maximum reduction of at least one of the hormonal levels was 66% +/- 7% (50-98%). The basal levels of LH, FSH and alpha-subunit in the 10 patients were (mean +/- SEM (range)), 3.0 IU/l +/- 1.0 (0.0-7.4), 12.7 IU/l +/- 5.0 (0.0-39.0) and 9.0 IU/l +/- 7.0 (0.2-74.0). During six months of therapy with octreotide and cabergoline, the basal levels of LH, FSH and alpha-subunit were reduced by > or = 50% in seven patients - including the six patients with in vivo secretion prior to therapy. No new visual field defects were detected during therapy and no deterioration of existing visual field defects was recorded. The medical therapy was well tolerated. The in vivo basal and TRH-stimulated secretory capacity of LH, FSH and alpha-subunit predicted tumour reduction following intensive medical therapy in all of our patients with non-functioning pituitary adenomas.

Staging uterine cervical carcinoma with low-field MR imaging.

PURPOSE: To assess the validity of low-field MR in staging cervical cancer compared to clinical staging. MATERIAL AND METHODS: A total of 95 women entered the study over a 3-year period. MR examinations with a 0.1 T resistive magnet using a body coil and clinical staging according to the FIGO recommendations (1988) were performed within 2 weeks from clinical diagnosis. T1- and T2-weighted sequences were obtained in transversal and sagittal acquisitions, and an additional T1 before and after contrast (randomisation to 0.1 or 0.3 mmol/kg b.w. gadodiamide). Treatment decisions on surgery or radiation therapy were made solely on the clinical staging. RESULTS: Sixty-one patients were found to be eligible for surgery. In 5 women, the pathological results revealed a more advanced stage of the disease than assessed by clinical staging. MR correctly staged 4 of the 5 but otherwise tended to overstate the disease. Contrast enhancement significantly reduced this trend (p<0.05) regardless of the contrast medium dose used. Divided into two groups, an operable (less than stage 2b) and an inoperable group (more than stage 2a), the clinical staging correctly classified 57 patients (accuracy 92%) compared to 52 patients with MR using contrast enhancement (accuracy 84%). The specificity was no higher than 31%, whereas the reproducibility of the MR assessment was fairly good with kappa values around 0.65 for both intra- and inter-observer variations. CONCLUSION: In the present set-up, clinical assessment was superior to low-field MR in staging cervical cancer. When using contrast enhancement, the staging accuracies of low-field MR were comparable to the ones reported for techniques with higher tesla values, whereas the specificity and reproducibility errors were lower. The method, therefore, needs to be optimised.

Acoustic neuroma--treatment modalities. Surgery, gamma-knife or observation?

We present our results of 211 patients with acoustic neuroma over a period of 10 years, 1988-97. We operated on 100 and 111 had Gamma-knife (GK) treatment (69 were available to follow-up). The results are excellent for surgery on small and intracanalicular tumours. In tumours of the same size, surgery and GK treatment give comparable, but somewhat different, results. In the GK group of 54 primary treated patients, 3 patients had to be operated on and another 4 developed hydrocephalus. A group of 35 acoustic tumours was observed for more than 3 years. Nineteen did grow (54%). Hearing was unchanged in 23%. We performed surgery in 11 patients and gave 2 patients GK treatment because of tumour growth of > 2 mm in diameter a year. We conclude that either treatment is effective for small and medium-sized acoustic neuromas. Hearing preservation was best in the GK-treated group (80%), compared to only 12.5% in the group operated via the suboccipital route. Larger tumours and most medium-sized tumours should be operated, as should smaller tumours with persistent symptoms of vertigo and pain. Medical contraindications to surgery or reluctance to undergo surgery make GK treatment a good alternative. Treatment of residual tumours with the GK could also be a solution to a difficult problem.

[Atypical endometrial hyperplasia. Prognosis and course]. / Atypisk hyperplasi i endometriet. Prognose og forløb.

The treatment of patients with the diagnosis atypical endometrial hyperplasia has been disputed during the last decades. The aim of the study was to evaluate the treatment of these patients and analyse the progression rate to invasive carcinoma of the endometrium. Fifty-seven patients with atypical hyperplasia were examined and treated from 1976 through 1991. The medical records were examined retrospectively and the pathology slides were revised by one pathologist in accordance with the 1975 WHO recommendations. Thirty-one (54%) patients were on oestrogen treatment as monotherapy at the time of diagnosis. Forty-two patients had a hysterectomy performed within five months, and five patients had a hysterectomy performed 10 to 61 months after diagnosis. A total of 18 out of 57 patients (31.6%) had or developed endometrial carcinoma all with myometrial invasion: 14 stage I with < or = 50% myometrial invasion, three stage I with > 50% myometrial invasion, and one stage IV. There was no significant difference in age, body mass index, parity or hormone replacement treatment between the group with endometrial carcinoma and the group without endometrial carcinoma. We conclude that unopposed oestrogen treatment and nulliparity are the main risk factors for atypical hyperplasia and that hysterectomy is the appropriate treatment for patients with atypical hyperplasia of the endometrium.

Infection of rat brain cell aggregates with neurovirulent and nonneurovirulent strains of herpes simplex virus type 1.

Rat brain cell aggregates represent a three-dimensional tissue culture system of brain tissue in the form of small, multicellular spheroids. In the present work, we have infected these "minibrains" with neurovirulent, nonneurovirulent, and nonreplicating strains of HSV-1. The neurovirulent strains 17(+) and KOS(M) spread rapidly through the aggregates, while the nonreplicating ICP4 deletion mutant KD6 infected cells only at the periphery of the aggregates. Spread and replication of the nonneurovirulent strains RE6 and tk-7, and to some extent also of R13/1, were restricted. The interaction between different strains of HSV-1 and the rat brain cell aggregates is thus comparable to that seen in the brain, suggesting that the aggregates represent a useful tool for studying HSV-1 infection of brain tissue in vitro.

Effects of DFMO on glioma cell proliferation, migration and invasion in vitro.

The polyamine inhibitor DL-alpha-difluoromethylornithine (DFMO) is a specific irreversible inhibitor of ornithine decarboxylase which is a rate-limiting enzyme in the polyamine bio-synthesis pathway. The present study describes the effects of DFMO on glioma cell proliferation, migration and invasion using multicellular spheroids from three glioma cell lines (GaMg, U-251 Mg and U-87 Mg). 10 mM DFMO reduced cell migration in the three cell lines by about 30-50%. 1 mM putrescine, added together with DFMO inhibited the DFMO effect. A stronger effect was observed in the growth assay where 10 mM DFMO reduced the spheroid growth, for all cell lines, by 90%. This effect was also reversed by adding 1 mM of putrescine. In vitro tumor cell invasion experiments indicated after 3 days of confrontation, an extensive invasion also after 10 mM DFMO treatment. The brain aggregate volumes were reduced to about the same extent as in the absence of drug, suggesting essentially no effects of DFMO on the invasive process. It is concluded that the tumor spheroids retained their ability to invade normal brain tissue even after DFMO exposure. However, DFMO inhibited spheroid growth and cell migration which supports the notion that cell growth, migration and invasion are biological properties that are not necessarily related to each other.

[Patients with early stages of endometrial cancer should be spared adjuvant radiotherapy. Danish Endometrial Cancer Group]. / Patienter med tidlige stadier af endometriecancer bør skånes for adjuverende strålebehandling.

In an attempt to create uniform nationwide guidelines for the management of all stages of endometrial carcinoma, and to limit the use of adjuvant radiation therapy in stage I disease to high-risk patients only, a protocol was developed by the Danish Endometrial Cancer group (DEMCA). From September 1986 through August 1988, 1214 women in Denmark with newly diagnosed carcinoma of the endometrium have been treated according to this protocol. This figure represents all endometrial carcinomas diagnosed in Denmark during this two-year period. The primary treatment was total abdominal hysterectomy and bilateral salpingo-oophorectomy, no preoperative radiation therapy was delivered. In 1039 cases no macroscopic residual tumour and/or microscopic tumor tissue in the resection margins was found following surgery. Based on surgery and histopathology, these patients were classified as: P-stage I low risk (n = 641), P-stage I high risk (n = 235), P-stage II (n = 105) and P-stage III, Group 1 (n = 58). No postoperative radiation therapy was given to P-I low risk cases. P-I high risk, P-II, and P-III (Group 1) cases received external radiation therapy. Recurrence rate at 68-92 months follow-up was 45/641 (7%) in P-I low risk, 36/235 (15%) in P-I high risk, 30/105 (29%) in P-II, and 27/58 (47%) in P-III (Group 1) cases. Fifteen of 17 vaginal recurrences in P-I low risk cases were salvaged (mean observation time 61 months). In this population-based investigation it has been shown that P-stage low-risk patients are adequately treated by total abdominal hysterectomy and bilateral salpingo-oophorectomy, and that no pre- or postoperative radiation therapy is necessary.

[Cerebral metastases treated with stereotaxic gamma radiation. 6-year experience with the "gamma knife" at the Haukeland hospital]. / Hjernemetastaser behandlet med stereotaktisk gammabestråling. Seks års erfaring med bruk av "gammakniven" ved Haukeland Sykehus.

During the last 6 years we have treated 32 patients with 45 metastases to the brain in the Gamma Knife unit. 21 of these were treated exclusively with the Gamma Knife. The remaining 11 patients received radiosurgery for recurrent disease after surgery and whole-brain irradiation (six patients), new metastases after whole-brain irradiation alone (three patients) or for local regrowth after surgery (two patients). The range of tumour volume was 0.1-43.3 cm3 (median 2.4 cm3). Marginal tumour dose was 5-30 Gy (median and mean: 25 Gy) to the 30-70% isodose-volume line according to tumour volume and localization. 19 patients died during the period of follow-up. Only three patients died from their intracranial metastases. Thus, local growth control was achieved in 29 patients. 16 patients died from extracranial manifestations. The average survival time for the patients who died during the observation period was 11 (1-37 months), and the survival time for patients still alive was 10-75 (median 14, average 29) months. Mean observation period for all patients was 17 (1-75) months. Brain metastases are physically and biologically ideal lesions to treat with radiosurgery. Stereotactic radiosurgery applied to radiographically small and distinct metastases is safe, non-invasive and highly effective. The treatment requires only a short stay in hospital, and is much less inconvenient to the patient than open surgery or whole-brain irradiation. Radiosurgery can be used on lesions inaccessible to open neurosurgery or resistant to classical fractionated radiotherapy. Gamma Knife treatment has become our first choice for patients with less than four intracranial metastases with diameters less than 3-3.5 cm.

Migratory patterns of lac-z transfected human glioma cells in the rat brain.

Malignant brain tumors are characterized by extensive tumor-cell infiltration into the normal brain tissue. The present work describes the migratory behavior of human glioma cells transplanted into the adult rat brain with the aim of exploiting the extent of active cell migration and passive cell displacement within the central nervous system. To detect every transplanted tumor cell, a stably bacterial beta-galactosidase (lac-z) transfected human glioma cell line was used. To distinguish between an active cell migration process and passive cell displacement, rat brains were also implanted with inert fluorescent polystyrene microspheres and the distribution of tumor cells and microspheres was studied 1 hr and 3 days after implantation. One hour after implantation the tumor cells were strictly localized at the implantation site. However, 3 days after implantation, both tumor cells and microspheres showed an extensive distribution within the brain. Confirming earlier neuropathological and experimental studies, it is shown that the lac-z-transfected glioma cells had the capacity to move within the Virchow-Robin and subarachnoid spaces. However, since fluorescent microspheres were also found in these areas, this spread of tumor cells may be primarily mediated by the extensive cerebrospinal fluid flow that exists within the brain. Three days after implantation, the glioma cells also showed an active migration over the corpus callosum. In comparison, the fluorescent microspheres showed only limited spread along the callosal body. It is concluded that the bacterial lac-z gene can be stably transfected into human glioma cells and, since every tumor cell can be visualized within the brain, this model provides a tool for studying the mechanisms behind tumor-cell invasion of the brain.

Glioma cell invasion visualized by scanning confocal laser microscopy in an in vitro co-culture system.

Confrontation cultures between glioma spheroids and brain cell aggregates are well established in glioma research, and the model reflects several similarities to the in vivo brain tumour invasive process. The lipid-binding fluorescent carbocyanine dyes DiO (3,3'-dioctadecyloxacarbocyanine perchlorate) and DiI (1,1'-dioctadecyl-3,3,3,'3,'-tetramethylinocarbocyanine perchlorate) are widely used in cell biology as tracers for studying cell movement. Mature brain cell aggregates grown from fetal rat brain cells, and spheroids initiated from two glioma cell lines (GaMg and D-54Mg) were stained with DiO and DiI, respectively. Penetration of DiI and DiO into the tumour spheroids and brain aggregates was studied by confocal laser scanning microscopy (CLSM). After 48 h of dye exposures, the tracers had almost completely penetrated the tumour spheroids and brain aggregates. Light-microscopic sections of the specimens indicated that the dye incorporation had little effect on cellular morphology. Cell migration from DiI stained D-54Mg and GaMg spheroids was similar to that observed from unstained spheroids. Growth was also unaffected after 48 h of DiI exposure. Gioma cell invasion was assessed by CLSM using co-cultures of DiI -stained spheroids and DiO-stained brain cell aggregates. Optical sections revealed a gradual decrease in remaining brain volume, indicating a progressive invasive process. Single tumour cells were identified deep within the brain aggregates. In addition normal brain cells were also identified in the tumour spheroids. It is concluded that vital staining can be used to identify both normal cells and tumour cells during tumour cell invasion in vitro. The method may provide the possibility for studying the kinetics of single normal and tumour cell movement in individual tumour/brain co-cultures.

Leptomeningeal tissue: a barrier against brain tumor cell invasion.

BACKGROUND: Primary brain tumors are characterized by an extensive infiltrative growth into the surrounding brain tissue. This process is confined to the central nervous system, and tumor cell metastasis to other organs is rare. However, other tumors of non-neural origin may frequently metastasize to the central nervous system. PURPOSE: The purpose of the present study was to examine the invasive behavior of different glioma cells into tissues of neural (brain aggregates) as well as non-neural origin (leptomeningeal tissue). Using the same target tissues, the invasive characteristics of two neural metastatic tumors (one malignant melanoma and one small-cell lung carcinoma) were also studied. This direct comparison of the invasive behavior between tumors of neural and non-neural origin provides valuable information regarding the mechanisms of glioma cell dissemination in the central nervous system. METHODS: The in vitro invasive behavior of human tumors of the central nervous system into human leptomeningeal tissue as well as into normal rat brain tissue was studied. For this purpose, a co-culture system consisting of tumor biopsy specimens, human leptomeningeal cell aggregates, and brain cell aggregates was established. Three glioblastomas, one oligodendroglioma, one meningioma, one small-cell lung carcinoma, and one malignant melanoma were studied. RESULTS: In co-cultures of gliomas and leptomeningeal cell aggregates, a well-defined border between the two tissues was observed. The brain cell aggregates, in contrast, were consistently invaded by the glioma cells. The brain metastases showed a different invasion pattern. The metastatic cells invaded and progressively destroyed leptomeningeal cell aggregates, whereas they did not invade the brain cell aggregates. Upon confrontation of the leptomeningeal tissue with the meningioma, a fusion of the two tissues was observed. Immunostaining of the leptomeningeal tissue showed a strong expression of the basement membrane components fibronectin, collagen type IV, and laminin with no expression of glial fibrillary acidic protein, neuron-specific enolase, or S-100 protein. CONCLUSIONS: The present study indicates that there may be important biologic differences between the invasive behavior of gliomas and non-neuroepithelial tumors. Our co-culture experiments suggest that leptomeningeal cells and associated acellular components may constitute a barrier against glioma cell invasion. However, this barrier may not be functional for metastatic tumors to the brain. The presence of glioma cells within the leptomeninges should not necessarily be taken as evidence of aggressive growth or as an indicator of malignancy.

Three-dimensional growth of glial cell lines affects growth factor and growth factor receptor mRNA levels.

We have studied, at the mRNA level, the influence of various defined growth conditions on the expression of TGF-alpha, PDGF-BB, EGF-R, PDGF-R alpha, and PDGF-R beta in five different glioma cell lines (D-37MG, D-54MG, D-263MG, GaMG, and U-251MG). RNA isolated from logarithmically growing, confluent monolayer cells or multicellular spheroids was analyzed. Northern blot experiments show that with a few exceptions, specific mRNA steady-state levels were considerably higher in cells grown in a three-dimensional organization relative to cells in the logarithmic growth phase.

Transfection of glioma cells with the neural-cell adhesion molecule NCAM: effect on glioma-cell invasion and growth in vivo.

The tumor growth and the invasive capacity of a rat glioma cell line (BT4Cn) were studied after transfection with the human transmembrane 140-kDa isoform of the neural-cell adhesion molecule, NCAM. After s.c. injection, the NCAM-transfected cells showed a slower growth rate than the parent cell line (BT4Cn). Upon intracerebral implantation with BT4Cn cells and different clones of NCAM-transfected cells, all animals developed neurological symptoms within 13-16 days. However, the tumors showed different growth characteristics. The NCAM-transfected BT4Cn cells were localized in the region of the injection site, with a sharply demarcated border between the tumor and brain tissue. In contrast, the parental cell line showed single-cell infiltration and more pronounced destruction of normal brain tissue. Using a 51Cr-release assay, spleen cells from rats transplanted with BT4Cn tumor cells generally showed a lower cytotoxic response than the spleen cells from rats transplanted with the transfected variants of BT4Cn cells, indicating that the transfection procedure in itself mediated an activation of the immune system. The present data suggest that NCAM may influence the malignant behavior of rat glioma cells in vivo.