Irene Sulzbacher

Investigation of apparent diffusion constant as an indicator of early degenerative disease in articular cartilage.

To investigate the apparent diffusion constant (ADC) as a prospective magnetic resonance imaging (MRI) marker of early degeneration in articular cartilage.

Expression of Platelet-Derived Growth Factor-α Receptor Is Associated With Tumor Progression in Clear Cell Renal Cell Carcinoma

Platelet-derived growth factors (PDGFs) exert their biologic function by binding to 3 different tyrosine kinase receptor isoforms. Especially the PDGF-alpha alpha receptor binds PDGF proteins with high specificity, which results in growth stimulation. The expression of the receptor and its ligand was studied in human renal cell carcinomas (RCCs) by immunohistochemical analysis, and the expression of PDGF-alpha alpha receptor and PDGF-AA was correlated with clinicopathologic parameters of patients with clear cell RCC (CCRCC). In CCRCC, the mean expression of PDGF-alpha alpha receptor and PDGF-AA was 38.8% (range, 0.0%-96.0%) and 18.4% (range, 0.0%-90.0%), respectively. PDGF-alpha alpha receptor expression was significantly higher in grade 3 and grade 4 tumors compared with grade 1 and grade 2 tumors (P = .027; Mann-Whitney test), and high receptor expression correlated with tumor progression in univariate analysis (P = .0253; log-rank test), while PDGF-AA expression had no prognostic influence on the outcome of patients with CCRCC. Therefore, immunohistochemical detection of high PDGF-alpha alpha receptor expression in CCRCC is associated with adverse outcomes. Furthermore, the PDGF receptor-factor interaction loop may be considered as a possible target for novel therapeutic strategies.

Expression of osteopontin and vascular endothelial growth factor in benign and malignant bone tumors.

Abstract. Osteopontin (OPN), one of the major non-collagenous proteins of bone matrix, is together with vascular endothelial growth factor (VEGF) a potent angiogenic protein. In this study we determined the expression of OPN in benign and malignant bone tumors and investigated the prognostic influence of OPN expression on the outcome of osteosarcoma patients. Fifty-seven osteosarcomas and 11 osteoblastomas as well as 5 bone specimens with remodeling sites were immunohistochemically analyzed for expression of OPN and VEGF. OPN was not detected in osteoblasts of remodeling sites. Osteoblastoma osteoblasts as well as osteoclastlike giant cells and osteosarcoma mononuclear cells showed variable staining. In osteosarcomas OPN and VEGF expression correlated with each other (r=0.390, P=0.003, Spearmans test). Although osteosarcoma patients with high VEGF expression showed a trend towards shorter overall survival (P=0.0841, log-rank test), OPN expression had no influence on patients overall or on disease-free survival. Our data indicate that expression of this protein might be upregulated in bone neoplasia. Although OPN expression correlates with VEGF expression in osteosarcomas, OPN expression does not provide predictive information about the outcome of osteosarcoma patients.

Platelet-derived growth factor-alpha receptor expression supports the growth of conventional chondrosarcoma and is associated with adverse outcome.

Bone cells are important targets of platelet-derived growth factors (PDGFs) because they stimulate proliferation of osteoblasts and chondrocytes. In this study we wanted to determine the expression of PDGF-AA and PDGF-&agr; receptor in conventional chondrosarcomas and to compare the results with those obtained from benign enchondromas and non-neoplastic cartilage tissue. Sixty-seven chondrosarcomas, 20 enchondromas, and 5 specimens of healthy cartilage as well as 7 specimens of hypertrophic callus cartilage were immunohistochemically analyzed for the expression of PDGF-AA and PDGF-&agr; receptor, respectively. Additionally, the proliferation activity was examined with the MIB-1 antibody. Clinical follow-up data were available from 53 patients. A significant overexpression of receptor and factor was found in chondrosarcomas as compared with enchondromas (PDGF-AA p = 0.013, PDGF-&agr; receptor p <0.001). MIB-1 values were significantly higher in chondrosarcomas (p <0.001). In healthy joint cartilage no staining was detectable, whereas reactive cartilage of callus formation showed high expression levels. PDGF-&agr; receptor expression was significantly higher in grade 3 chondrosarcomas compared with grade 2 (p = 0.022) and grade 1 tumors (p = 0.002). Survival analysis documented a significantly shorter overall survival for patients with high PDGF-&agr; receptor expression (p = 0.0172, log-rank test). Because PDGF-&agr; receptor expression positively correlates with the aggressiveness of chondrosarcoma, it may be considered as a possible target for novel therapeutic strategies.

Expression of c-kit in human osteosarcoma and its relevance as a prognostic marker

Aims: To examine the prognostic relevance of c-kit expression in human osteosarcomas and to evaluate the mutation status in exon 9 and exon 11 of the c-kit gene. Methods: c-kit expression was examined in 100 human osteosarcomas by immunohistochemistry using paraffin embedded tumour tissues, and capillary sequencing of genomic DNA was performed to search for mutations in exons 9 and 11 of the c-kit gene. Results: 20 osteosarcomas showed c-kit expression ranging from 5% to 90% (mean 5.9%; SD 16.74%). Furthermore, DNA sequences of exon 9 and exon 11 of the c-kit gene were not altered in these tumours. Overall and disease free survival analysis did not reveal any differences between patients with osteosarcoma with c-kit expression and those with c-kit negative tumours. Conclusions: C-kit expression is not a prognostic marker in patients with osteosarcoma. The protein expression is not linked to mutations in exon 9 or exon 11 of the c-kit gene. Therefore, these exons may not function as targets for treatment modalities based on the suppression of c-kit tyrosine kinase activity.

Expression of platelet-derived growth factor-alpha receptor in human osteosarcoma is not a predictor of outcome

Aims: The aims of this study were to examine the prognostic relevance of platelet‐derived growth factor‐α receptor (PDGFRA) expression in human osteosarcomas and to evaluate the mutation status of exon 12 and exon 18 of the PDGFRA gene. Methods: PDGFRA expression was examined in 100 human osteosarcomas by immunohistochemistry using paraffin embedded tumour tissues, and capillary sequencing of genomic DNA was performed to search for mutations in exons 12 and 18 of the PDGFRA gene. Results: Ninety‐six osteosarcomas showed PDGFRA expression ranging from 4% to 90% (mean 40%, median 37.5%, SD 27.11%). Furthermore, DNA sequence of exon 12 and exon 18 of the PDGFRA gene were not altered in 40 tumours with high PDGFRA expression. Overall and disease‐free survival analysis did not reveal any differences between osteosarcoma patients with high PDGFRA expression and patients with low PDGFRA expression. Conclusions: The protein expression is not linked to mutations in exon 12 or exon 18 of PDGFRA gene. Therefore, treatment modalities based on the suppression of PDGFRA tyrosine kinase activity may need further investigation. PDGFRA expression is not a prognostic marker for osteosarcoma patients.

Periosteal osteoblastoma: a case report and a review of the literature.

Osteoblastomas located on the surface of cortical bone, so‐called periosteal (juxtacortical) osteoblastomas, are extremely rare. A 24‐year‐old man complained of pain and swelling in the left knee. The clinical and radiological investigation showed a tumor located in the posterior portion of the distal shaft of the femur. The radiological differential diagnosis included parosteal osteosarcoma, periosteal chondroma and periostitis ossificans. A frozen section was obtained and histology revealed an osteoblastoma with large epithelioid‐appearing osteoblasts consistent with an aggressive osteoblastoma. An en bloc resection of the tumor was performed and the definitive histology of the whole specimen revealed a typical osteoblastoma. The authors draw attention to the fact that periosteal osteoblastoma is a rare tumor that could be mistaken clinically and histologically for other and more common tumors at this location.

EXPRESSION OF VS38 IN OSTEOBLASTS AND STROMA CELLS OF BONE TUMORS

VS38 is a mouse monoclonal antibody which recognizes an intracytoplasmic antigen of 64 kilodaltons present in normal and neoplastic cells. It was reported to be of potential value in identifying myeloma or plasmacytoma in bone marrow or other tissues. During diagnostic analysis of bone marrow biopsies we noticed a consistent staining of osteoblasts with VS38. This led us to investigate the immunoreactivity in a range of bone lesions. 58 lesions were examined in the current study, including benign and malignant tumors as well as tumor like lesions and processes with reactive bone formation. The Streptavidin-peroxidase complex technique was applied on paraffin embedded sections, with 3-amino-9-ethylcarbazole serving as chromogen. All osteoblasts of reactive origin and part of the osteoblasts of benign neoplasms showed positive immunostaining. The antibody stained stroma cells in 81.25% (13/16) of the cases of benign osteogenic tumors and in 82.35% (14/17) of the osteosarcomas. Additionally, VS38 labelling was observed in bone tumors of fibrohistiogenic origin such as nonossifying fibroma and giant cell tumor, the histogenesis of which is still being debated. On the whole, it can be concluded that antibody VS38 lacks specificity as a plasma cell marker. It shows, however, a striking affinity to osteoblasts and in part also to stroma cells of osteogenic and fibrohistiogenic bone tumors.

Bcl‑2 correlates with localization but not outcome in human osteosarcoma

bcl-2 is a member of the bcl-2 family that inhibits apoptosis, plays a crucial role in cell viability and is expressed in various types of tumors. With respect to inconsistent results in previous studies, the aim of the present study was to generate a clear hypothesis with regards to the value of bcl-2 expression as a predictive or prognostic factor in human osteosarcoma. The expression of bcl-2 was examined immunohistochemically in 49 patients with high-grade osteosarcoma and the results were correlated with localization, histological response to chemotherapy, survival and the occurrence of metastases. In patients with osteosarcoma, 21/49 cases (43%) were positive for bcl-2 expression and the remaining cases were negative. A significantly higher expression of bcl-2 was observed in central tumors located in the pelvis (83 vs. 37% positive; P<0.05). The bcl-2 expression status revealed no statistically significant correlation with response to chemotherapy, with 57% of patients with bcl-2-positive tumors showing a good response and 43% showing a poor response. No significant difference was observed when comparing survival or occurrence in bcl-2-positive and -negative tumors. In conclusion, the results of the present study indicate that, despite higher bcl-2 expression in central osteosarcoma, the expression in high-grade osteosarcoma is not a reliable prognostic or predictive marker.

High-resolution 3 T MR microscopy imaging of arterial walls.

PurposeTo achieve a high spatial resolution in MR imaging that allows for clear visualization of anatomy and even histology and documentation of plaque morphology in in vitro samples from patients with advanced atherosclerosis. A further objective of our study was to evaluate whether T2-weighted high-resolution MR imaging can provide accurate classification of atherosclerotic plaque according to a modified American Heart Association classification.MethodsT2-weighted images of arteries were obtained in 13 in vitro specimens using a 3 T MR unit (Medspec 300 Avance/Bruker, Ettlingen, Germany) combined with a dedicated MR microscopy system. Measurement parameters were: T2-weighted sequences with TR 3.5 sec, TE 15–120 msec; field of view (FOV) 1.4 × 1.4; NEX 8; matrix 192; and slice thickness 600 μm. MR measurements were compared with corresponding histologic sections.ResultsWe achieved excellent spatial and contrast resolution in all specimens. We found high agreement between MR images and histology with regard to the morphology and extent of intimal proliferations in all but 2 specimens. We could differentiate fibrous caps and calcifications from lipid plaque components based on differences in signal intensity in order to differentiate hard and soft atheromatous plaques. Hard plaques with predominantly intimal calcifications were found in 7 specimens, and soft plaques with a cholesterol/lipid content in 5 cases. In all specimens, hemorrhage or thrombus formation, and fibrotic and hyalinized tissue could be detected on both MR imaging and histopathology.ConclusionHigh-resolution, high-field MR imaging of arterial walls demonstrates the morphologic features, volume, and extent of intimal proliferations with high spatial and contrast resolution in in vitro specimens and can differentiate hard and soft plaques.