Biomedical subjects

Joerg-Christian Tonn

Publications and source records attributed to Joerg-Christian Tonn.

6 recordsLinked to original sources

PubMed2006

Effects of Raf-1 siRNA on human cerebral microvascular endothelial cells: a potential therapeutic strategy for inhibition of tumor angiogenesis.

The serine/threonine kinase Raf-1 is involved in the regulation of tumor cell survival, proliferation and metastasis formation, and has therefore emerged as a promising target for cancer therapy. In addition, Raf-1 activity mediates proliferation of endothelial cells thereby promoting angiogenesis and invasive growth of various tumors, including highly vascularized malignant glioblastoma. The aim of this study was to evaluate the effects of small inhibitory RNA (siRNA) directed against Raf-1 on viability, proliferation and motility in glioma cells and cerebral endothelial cells. Half-quantitative RT-PCR and Western blotting revealed efficient siRNA-mediated Raf-1 down regulation in glioma cells (U373, U251) and in human cerebral microvascular endothelial cells (HCMEC). Surprisingly, Raf-1 gene silencing failed to affect cell survival, proliferation or migration activity in the glioblastoma cell lines. In HCMEC, however, pronounced decrease of cell survival and significant inhibition of tube formation was achieved by Raf-1 siRNA compared to non-functional siRNA or vehicle controls. In conclusion, Raf-1 silencing appears as a potential therapeutic strategy to inhibit brain tumor angiogenesis and thereby outgrowth of highly vascularized glioblastoma multiforme, whereas direct cytotoxic effects of Raf-1 knockdown in tumor cells may vary.

Analysis of Variance↗

PubMed2006

Isolation and culture of microvascular endothelial cells from gliomas of different WHO grades.

Gliomas are the most common intrinsic brain tumors. The degree of vascularization corresponds to malignancy and is related to prognosis. In order to retrieve information about tumor behavior in situ, the use of primary tissue material for experiments is advantageous. With increasing evidence for the importance of microenvironment and vascularization in tumor biology, we concentrated on the isolation of endothelial cells (EC) from primary tumor material to investigate the role of endothelium within tumor tissue. We developed a method for isolation and purification of tumor-derived endothelial cells. EC were isolated and cultivated from normal brain using tissue digestion and Percoll density gradient centrifugation resulting in a <95% of EC culture. For isolation of EC from gliomas of different malignancy grades a combination of tissue digestion, Percoll gradient centrifugation and magnetic bead sorting by anti-CD31, -VE-Cadherin and -CD 105 was employed. This approach provided a purity of <98%. Cells were classified and characterized by testing expression of CD105, CD31, VE-Cadherin, vWF, UEA-1 and measuring DiI-Ac-LDL-uptake. To exclude contamination, staining and negative selection with anti-SMA, -GFAP, and -CD68 was performed. Tumors were histopathologically diagnosed according to WHO classification. We isolated EC from normal brain (NBEC, n = 11), low-grade gliomas WHO II (LGEC, n = 22), and high-grade gliomas WHO III & IV (HGEC, n = 11). There were no clear differences in EC morphology between the different tumor grades. However, a significantly higher proliferation rate of HGEC compared to LGEC was observed as well as distinctive antigen expression. Already in early passages isolated EC showed a rapid change in antigen expression indicating a phenotypic shift under culture conditions. We could establish a protocol for reliable and reproducible isolation and culture of EC from gliomas with different WHO grading. In first phenotypical and functional analyses, NBEC, LGEC and HGEC show remarkable differences. EC from all tumors could be grown in culture. However, passage related changes of EC phenotype demand very early passages to work with.

Brain Neoplasms↗

PubMed2006

Cavernous malformations of the brainstem: three-dimensional-constructive interference in steady-state magnetic resonance imaging for improvement of surgical approach and clinical results.

OBJECTIVE: The indications for resection of cavernous malformations (CMs) of the brainstem include neurological deficits, (recurrent) hemorrhage, and surgically accessible location. In particular, knowledge of the thickness of the parenchymal layer and of the CM's spatial relation to nuclei, tracts, cranial nerves, and vessels is critical for planning the surgical approach. We reviewed the operative treatment of 13 patients with 14 brainstem CMs, with special regard to refined three-dimensional (3D)-constructive interference in steady-state (CISS) magnetic resonance imaging (MRI). METHODS: Patients were evaluated neurologically and by conventional spin-echo/fast spin-echo and 3D-CISS MRI. Surgery was performed with the use of microsurgical techniques and neurophysiological monitoring. RESULTS: Eleven CMs were located in the pons/pontomedullary region; 10 of the 11 were operated on via the lateral suboccipital approach. Three CMs were located near the floor of the fourth ventricle and operated on via the median suboccipital approach, with total removal of all CMs. Results were excellent or good in 10 patients; one patient transiently required tracheostomy, and two patients developed new hemipareses/ataxia with subsequent improvement. Not only did 3D-CISS sequences allow improved judgment of the thickness of the parenchymal layer over the lesion compared with spin-echo/fast spin-echo MRI, but 3D-CISS imaging also proved particularly superior in demonstrating the spatial relation of the lesion to fairly "safe" entry zones (e.g., between the trigeminal nerve and the VIIth and VIIIth nerve groups) by displaying the cranial nerves and vessels within the cerebellopontine cistern more precisely. CONCLUSION: Surgical treatment of brainstem CMs is recommended in symptomatic patients. Especially in patients with lesions situated ventrolaterally, the 3D-CISS sequence seems to be a valuable method for identifying the CM's relation to safe entry zones, thereby facilitating the surgical approach.

Adolescent↗

PubMed2006

Technical description, phantom accuracy, and clinical feasibility for fiducial-free frameless real-time image-guided spinal radiosurgery.

OBJECT: The authors describe the technical application of the Xsight Spine Tracking System, data pertaining to accuracy obtained during phantom testing, and the initial clinical feasibility of using this fiducial-free alignment system with the CyberKnife in spinal radiosurgery. METHODS: The Xsight integrates with the CyberKnife radiosurgery system to eliminate the need for implantation of radiographic markers or fiducials prior to spinal radiosurgery. It locates and tracks spinal lesions relative to spinal osseous landmarks. The authors performed 10 end-to-end tests of accuracy using an anthropomorphic head and cervical spine phantom. Xsight was also used in the treatment of 50 spinal lesions in 42 patients. Dose planning was based on 1.5-mm-thick computed tomography slices in which an inverse treatment planning technique was used. All lesions could be treated using the fiducial-free tracking procedure. Phantom tests produced an overall mean targeting error of 0.52 +/- 0.22 mm. The setup time for patient alignment averaged 6 minutes (range 2-45 minutes). The treatment doses varied from 12 to 25 Gy to the median prescription isodose of 65% (40 to 70%). The tumor volume ranged between 1.3 and 152.8 cm3. The mean spinal cord volume receiving greater than 8 Gy was 0.69 +/- 0.35 cm3. No short-term adverse events were noted during the 1- to 7-month follow-up period. Axial and radicular pain was relieved in 14 of 15 patients treated for pain. CONCLUSIONS: Fiducial-free tracking is a feasible, accurate, and reliable tool for radiosurgery of the entire spine. By eliminating the need for fiducial implantation, the Xsight system offers patients noninvasive radiosurgical intervention for intra- and paraspinal tumors.

Adolescent↗

PubMed2005

Reproducibility of activation in four motor paradigms. An fMRI study.

Most functional imaging data are collected in single session experiments; little is known about the reproducibility or test-retest reliability of the activation patterns found in these experiments. In our study, 15 healthy volunteers performed four simple motor-paradigms ("Hand", "Foot", "Mouth" and "Tongue") for functional magnetic resonance imaging (fMRI) in 3 sessions on different days. Reproducibility of activations in four anatomical regions (pre- and postcentral gyri, paracentral lobule and the supplementary motor area) was measured in terms of voxels active in all sessions (common voxels) relative to voxels active in single sessions, giving reliability coefficients from 0 to 1. Two significance levels were used to identify active voxels. Reproducibility of activations was highest for foot and hand movements in the primary motorsensory areas; reliability coefficients were in the range of 0.62 to 0.78. Activations for mouth movements showed a very poor reproducibility. Application of the more stringent statistical threshold always led to a reduction of reproducible voxels. Reliability of fMRI data is not only a theoretical issue, but is of special practical importance in clinical settings such as integration of fMRI into neuronavigation for neurosurgical planning. Much care has to be taken if only single session data are available for interpretation.

Adult↗

PubMed2003

Influence of VEGF-R2 inhibition on MMP secretion and motility of microvascular human cerebral endothelial cells (HCEC).

Neovascularization and invasion are key features of malignant gliomas. Matrix metalloproteinases (MMPs) are supposed to play a major role mediating these processes. To analyze the expression patterns of MMPs in microvascular human cerebral endothelial cells (HCEC), we isolated endothelial cells from normal human brain microvessels. Characterization of cellular origin was performed by immunostaining, using the endothelial cell markers Ulex europaeus Agglutinin-1, von-Willebrand-Factor and Glucose-transporter-1. Contamination by other cell types was tracked by immunohistochemistry for GFAP (astrocytes), ASM (pericytes) and CD68 (macrophages). Secretion of MMPs was evaluated by ELISA and zymography. To determine whether HCEC show any difference in MMP expression compared to endothelial cells of other origin we analyzed human umbilical vein endothelial cells (HUVEC). HCEC show a decrease of MMP-3 and MMP-2 protein when treated with SU5416, a VEGF-R2 (KDR/flk-1) inhibitor, whereas MMP expression remained unchanged in HUVEC. To determine whether these findings show any effect in the motility of these cells we used a three-dimensional co-culture assay of avascular glioblastoma spheroids with primary HCEC spheroids. Untreated controls showed invasion of both cell populations into each other whereas treatment of the co-cultures with SU5416 resulted in complete inhibition of endothelial cell invasion hence indicating that flk-1 related motility of endothelial cells is critically involved in this process and can be studied with this assay. The results of different effects of anti-angiogenic treatment on proteolytic properties of two endothelial cell populations suggest that neovascularization of human brain tumors in vitro is dependent on the surrounding endothelial cell type and should therefore be studied with organ-specific human microvascular cerebral endothelial cells.

Angiogenesis Inhibitors↗