PTHrP and Bcl-2: essential regulatory molecules in chondrocyte differentiation and chondrogenic tumors.

Human chondrosarcomas (CS) are a frequent form of malignant bone tumors. The accurate distinction between benign solitary enchondroma and conventional CS of bones is a major diagnostic goal. Although the histological characteristics of chondrogenic tumors and the grading of CS (G1 to G3) have been defined by several authors, immunohistochemical markers for the different entities and grades are still missing and the mechanisms of tumorigenesis remain poorly understood. In addition to the emerging evidence that parathyroid hormone-related peptide (PTHrP) plays a critical role in endochondral bone formation we have recently reported that Bcl-2 lies downstream of PTHrP in the regulation of chondrocyte differentiation. To further characterize chondrogenic tumors and to determine whether PTHrP and the regulation of Bcl-2-expression is of relevance to tumorigenesis, we analyzed the expression of both PTHrP and Bcl-2 on a series of 23 cases of solitary enchondroma (9 cases) and primary CS (14 cases) using light and confocal microscopy. While all 9 enchondromas exhibited a detectable level of PTHrP-expression only, 2 showed low levels of immunoreactivity for Bcl-2. In sharp contrast, strong coexpression of Bcl-2 and PTHrP was found in 11 (composed of 3 CS G3, 7 CS G2, and one dedifferentiated CS) out of 14 CS, while the expression level of these proteins was below the detection limit in two CS G1 and one dedifferentiated CS. To verify this data 3 cases each of enchondroma, CS G2, and CS G3 respectively, were subjected to quantitative confocal analysis, after double labeling for PTHrP and Bcl-2. The results showed a significant increase in the expression of both PTHrP and Bcl-2, in malignant CS versus the benign enchondromas. Most interestingly, the levels of expression of both PTHrP and Bcl-2 correlated with the degree of malignancy of the chondrogenic tumors. These results therefore suggest that both PTHrP and Bcl-2 play a role in the tumorigenesis of chondrogenic tumors and further indicate that both proteins may participate in the same pathway regulating chondrocyte differentiation.

P-glycoprotein expression in high grade central osteosarcoma and normal bone cells. An immunohistochemical study.

One important mechanism by which multidrug resistance is mediated is the mdr1 gene product, P-glycoprotein (Pgp). Even though chemotherapy, in the treatment of high grade central osteosarcoma (hgc-OS), has led to dramatic improvements in survival rate, a certain percentage of patients still show only a poor response to chemotherapy. To further characterize a potential connection between Pgp and chemotherapy as well as the role of Pgp in tumorigenesis of osteosarcoma, we analyzed Pgp-expression in hcg-OS. Immunohistochemistry was performed on 68 hgc-OS samples from 58 patients using the monoclonal antibody JSB-1; in addition, Pgp-expression in normal bone cells was studied in 5 human epiphyseal growth plates. 70.5% of all cases stained positive for P-glycoprotein, while 29.5% of the cases were negative. Cases investigated after chemotherapy showed a higher incidence (82.9%) of positive P-glycoprotein immunostaining than cases prior to chemotherapy (64.4%). The Pgp-expression of 34 biopsies was compared with chemotherapy, as determined at the surgical specimen. In these cases, however, no correlation could be established between P-glycoprotein expression of the biopsy and the later response to chemotherapy. 48.4% of the cases with biopsies, initially positive for Pgp, showed a good response in the surgical specimen, while only 27.2% of Pgp-positive biopsies were later classified as non-responders. In the normally growing skeleton, positive immunostaining was detected in the area of mineralization of epiphyseal growth plates. Osteoclasts, hypertrophic chondrocytes, and cuboidal osteoblasts showed Pgp-expression, while there was a lack of Pgp in the majority of osteocytes and chondrocytes in the resting and proliferating zone. These data therefore suggest that P-glycoprotein expression in hgc-OS resembles, at least in part, the phenotype of active bone cells. These results may explain why P-glycoprotein, by using immunohistochemistry, in biopsies of osteosarcomas is insufficient to predict the response to chemotherapy.

The thickness of human vertebral cortical bone and its changes in aging and osteoporosis: a histomorphometric analysis of the complete spinal column from thirty-seven autopsy specimens.

The object of this study was to analyze the cortical thickness (Ct.Th) of the ventral and dorsal shell of the vertebral bodies throughout the human spine in aging and in osteoporosis. Therefore, the complete front column of the spine of 26 autopsy cases (aged 17-90, mean 42 years) without diseases affecting the skeleton and of 11 cases (aged 58-92, mean 77 years) with proven osteoporosis were removed. A sagittal segment prepared through the center of all vertebral bodies was undecalcified, embedded in plastic, ground to a 1 mm thick block, and stained using a modification of the von Kossa method. The analysis included the measurement of the mean cortical thickness of both the ventral and dorsal shell, respectively (from the third cervical to the fifth lumbar vertebral body). The qualitative investigation of the structure of the cortical ring completed the analysis. The presented data revealed a biphasic curve for both the ventral and dorsal shell, skeletally intact with high values of the cortical thickness in the cervical spine (285 microm), and a decrease in the thoracic (244 microm) and an increase in the lumbar spine (290 microm). The mean thickness of the ventral shell is in general greater than the thickness of the dorsal shell in both skeletally normal and osteoporotic cases. The cortical thickness of the spine showed no gender-specific differences (p = NS). There was a slight decrease of the cortical thickness with aging; however, this decrease and the correlation of cortical thickness to age was only significant below vertebral body T8 (r = 0.225-0.574; p(r) < 0.05-0.005). Most interestingly, however, osteoporosis presents itself with a highly significant loss of cortical thickness throughout the whole spine. This decrease of cortical thickness was more marked in the dorsal shell (p < 0.05) than in the ventral shell (ventral from C3 to T6 [p < 0.05] below T6 [p = NS]). We therefore conclude that in osteoporosis the loss of spinal bone mass is not only a loss of trabecular structure but also a loss of cortical thickness. Furthermore, these results may explain the development of regions of least resistance within the spine in aging and the clustering of osteoporotic fractures in the lower thoracic and lumbar spine.

Frequent reduction or loss of DCC gene expression in human osteosarcoma.

The 'deleted in colon carcinoma' (DCC) gene has been considered a candidate tumour-suppressor gene that encodes for a transmembrane protein with strong structural similarity to members of the superfamily of neural cell adhesion molecules. It has been mapped to the chromosomal region 18q21.1 and it is implicated in cellular differentiation and developmental processes. In human osteosarcoma allelic loss frequently occurs on the long arm of chromosome 18, suggesting a possible involvement of the DCC gene in the pathogenesis of this tumour entity. In the present study the mRNA and protein expression and rearrangements at the DNA level of the DCC gene were addressed in 25 osteosarcomas and several tumour cell lines, including osteosarcoma- and colon carcinoma-derived cell lines. Using an reverse transcriptase polymerase chain reach in (RT-PCR)-based approach DCC expression was found to be lost or substantially reduced in 14 of 19 high-grade osteosarcomas, in three of six lower grade osteosarcomas and most of the tumour cell lines, in contrast to normally differentiated osteoblasts. Immunohistochemical studies on DCC protein expression of 14 selected tumours correlated well with the RT-PCR-based results. In view of the putative tumour-suppressor characteristics of the DCC gene its loss or reduction of expression could be a specific event in the development or progression of many high-grade osteosarcomas.

[Comparative DNA cytometric and cytogenetic ploidy determination of chondrosarcoma and osteosarcoma]. / Vergleichende DNA-zytometrische und zytogenetische Ploidiebestimmungen an Chondrosarkomen und Osteosarkomen.

10 chondrosarcomas and 10 osteosarcomas were examined using cytogenetics and DNA-image-cytometry. Cytogenetically 6 of 10 chondrosarcomas and 4 of 10 osteosarcomas showed hyperdiploid tumorcells. By DNA-cytometry in 8 of 10 chondrosarcomas and 9 of 10 osteosarcomas hyperdiploid tumorcells resp. hyperdiploid stemlines were detected. This discrepancy reflects an in-vitro-selection depending on the different entities. In 7 aneuploid clones of chondrosarcomas the chromosomal ploidy was calculated using the relative length of the chromosomes and compared with the DNA-ploidy of the native tumor. There was a close relation between both parameters of nuclear DNA-content. The interpretation of cytogenetic results is improved using a combination of karyotypic and DNA-cytometric examination. This is particularly important for the search for relations between numeric chromosomal aberrations and morphological parameters (grading).

Heterogeneity of the skeleton: comparison of the trabecular microarchitecture of the spine, the iliac crest, the femur, and the calcaneus.

The objective of this study was to elucidate the structure of cancellous bone and its age-related changes at different skeletal sites. Therefore, the lumbar spine, iliac crest, femur, and calcaneus of 12 age- and sex-matched skeletal healthy autopsy cases (6 females, 6 males, aged 28-84 years, mean 54 years) were removed. The following analysis includes an evaluation of the trabecular bone volume (BV/TV, %) and the trabecular interconnection (TBPf, mm-1) as well as a qualitative investigation of the structure of trabecular bone. BV/TV shows the following mean values: lumbar spine, 8.3% (+/- 0.8%); iliac crest, 11.5% (+/- 1.6%); intertrochanteric, 10.2% (+/- 1.2%); femoral neck, 15.8% (+/- 1.6%); and calcaneus, 15.4% (+/- 2.0%). There are significant differences between the BV/TV of the femoral neck and that of the lumbar spine as well as between that of the calcaneus and the lumbar spine (p < 0.01). However, a positive correlation can be seen between the bone mass of the spine and that of all other investigated sites (r = 0.67 to r = 0.80; pr < 0.1). The trabecular interconnection of the lumbar spine (2.7 mm-1, SEM +/- 0.2 mm-1) and the femoral neck (0.3 mm-1, SEM +/- 0.3 mm-1) differs significantly. Only these two sites show a significant positive correlation of TBPf (r = 0.60; pr < 0.1). Age-dependent alteration of the spine and the femoral neck in bone mass and bone structure is nearly the same. The trabecular microarchitecture of the iliac crest varies systematically. A region 4-10 cm behind and 1-3 cm below the anterior superior iliac spine turns out to be the most suitable biopsy site because of its closest relation to the lumbar bone mass. However, drawing information about the trabecular interconnection within the lumbar spine by measurement of the iliac crest at any site seems to be impossible. The horizontal specimens reveal a vertical running tubular spongiosa pattern that is arranged in concentric rings starting from the dorsal shell like a honeycomb. The comparison of TBPf in horizontal and vertical planes and its age-related changes indicates the age-related bone loss to be predominantly a loss of horizontal trabeculae. Thus, the presented data provide further information about the skeletal distribution and heterogeneity of the trabecular microarchitecture.

Architecture and distribution of cancellous bone yield vertebral fracture clues. A histomorphometric analysis of the complete spinal column from 40 autopsy specimens.

The objective of this study was to analyze the structure of cancellous bone and its significance for vertebral fractures. Therefore, the complete spinal column from 40 autopsy cases (18 without diseases affecting the skeleton and 12 osteoporotic) was removed and sectioned in the sagittal plane to a thickness of 1 mm. A surface-stained block grinding technique allowed combined two- and three-dimensional histomorphometric analysis, which included an evaluation of the trabecular bone volume (BV/TV, in %) and the trabecular interconnection (TBPf, in mm). In addition, qualitative investigation of the structure of trabecular bone was done. The distribution of trabecular bone volume within the spinal column of a normal skeleton shows a curve, with the highest values in the cervical spine and a decline in the thoracic and lumbar spine. Osteoporosis presents itself with a pathologically diminished trabecular bone volume, whereas the distribution within the spine is comparable to that of the controls. Osteoporotic patients show an apparently reduced trabecular interconnection. It is important that the measured values for TBPf are not only in general higher, but also more widely dispersed. The age-related decrease of trabecular bone mass is due to the transformation from plates to rods. This is quantitatively indicated by the close correlation of BV/TV and TBPf (P < 0.001, r = 0.85). The bone loss in osteoporosis is a loss of structure and a loss of whole trabeculae, which is caused by perforations. It involves a gradual change from normal bone. However, the polyostic heterogeneity in osteoporosis is immense. These structural differences demonstrate the development of regions of least resistance within the spine, serving as an explanation of osteoporotic fractures. Due to the polyostotic heterogeneity it is impossible to define a threshold mineral content for crash fractures by diagnostic measurements at any reference site.

[Morphologic characteristics of chondroblastoma. A retrospective study of 56 cases of the Hamburg bone tumor register]. / Morphologische Charakteristika des Chondroblastoms. Eine retrospektive Untersuchung an 56 Fällen des Hamburger Knochentumorregisters.

Representing only about 1% of all primary bone tumors, chondroblastoma constitutes a very rare bone tumor entity. 56 cases of chondroblastoma, that had been collected by the Hamburg Bone Tumor Registry from 1972 to 1995, were examined histologically together with the radiological and clinical findings. In addition immunohistochemistry with antibodies against S 100, PGM1, LCA and the proliferationmarker MIB 1 was performed. The mean age was 20.4 years and male patients being the majority with a gender ratio of 2.7:1. Predominant localisation was the epiphyses of the long bones, although almost 40% of the tumors were located at untypical sites. Usually a well-circumscribed lysis could be seen on plain X-Ray examination, however partial cortical destruction could be observed in one third of the cases. Histologically characteristic was a polygonal cell component with a weblike chonroid matrix, sometimes with a plane-like appearance. 5 cases showed a distinct nuclear polymorphism making a distinction from osteosarcoma difficult. Using immunohistochemistry all tumors except for one showed positive reaction for S 100 protein. Although the histogenesis of chondroblastoma is not completely understood, morphological findings as well as the observed reactivity with the S 100 protein indicate the chondroid origin. No reactivity for PGM 1 (CD 68) or LCA could be detected. All chondroblastoma showed a low rate of proliferation, thereby being distinguishable from high malignant bone tumors. In general chondroblastoma show a benign biological behavior. Different behavior was observed in 2 cases. One relapse located in the pelvis revealed local aggressive growth while in another case in the humerus a malignant transformation had taken place.

[P-glycoprotein expression in osteosarcoma]. / P-Glykoproteinexpression beim Osteosarkom.

One of the mechanisms by which multidrug resistance is mediated, is the mdr1 gene product, P-glycooprotein. Immunohistochemistry was performed for 63 osteosarcomas of 54 patients to investigate P-glycoprotein expression using the monoclonal antibody JSB-1. Most of the patients were children or adolescents who had received treatment under the framework of the Cooperative Osteosarcoma Study Group. In addition P-glycoprotein expression was assayed in five growth plates. Of all cases 68.5% stained positive for P-glycoprotein. Cases that had received chemotherapy showed a higher incidence (80.9%) of positive P-glycoprotein immunostaining than cases that had not received chemotherapy (66.6%). No relation could be established between P-glycoprotein expression and the response to chemotherapy, since the majority of P-glycoprotein positive biopsies showed a good response in the surgical specimen after chemotherapy. Furthermore, 42.9% of P-glycoprotein negative biopsies were classified as non-responders in the later surgical specimen. In addition to P-glycoprotein expression in osteosarcomas positive immunostaining was also detected in osteoblasts, osteocytes, osteoclasts as well as in some chondroblasts. The results indicate that P-glycoprotein expression in osteosarcomas also exists prior to chemotherapy and resembles the phenotype of normal bone tissue. However, the determination of P-glycoprotein by using immunohistochemistry in biopsies of osteosarcomas cannot predict the response to chemotherapy.

[Cell proliferation in bone tumors. Immunohistologic study of Ki-67 protein expression]. / Zellproliferation bei Knochentumoren.

Bone tumors represent a group of tumors of various dignity. In spite of this single tumor entities may display strong morphological resemblance to each other which can in turn result in profound difficulties in differential diagnosis. The biological behaviour of a tumor is mainly determined by its rate of proliferation. In this study the rate of proliferation of 64 bone tumors (30 high-grade central osteosarcomas, 6 low-grade osteosarcomas, 8 giant cell tumors, 8 aneurysmatic bone cysts, 5 osteoidosteomas/osteoblastomas, 7 fibrous dysplasias and 5 cases of a myositis ossificans) were analysed. Immunohistochemistry was performed on paraffin-embedded tissue sections using the MIB-1 monoclonal antibody. MIB-1 recognizes the proliferation-associated Ki-67 protein which is expressed during the active phases of the cell cycle but cannot be detected in senescent cells. Among high-grade central osteosarcomas a significantly higher rate of proliferation (average value 30%) was found in comparison with low-grade osteosarcomas and other benign intraosseous bone tumors. This approach proved to be very useful in the distinction between high-grade and low-grade osteosarcomas as well as bone-forming intraosseous tumors. However distinguishing low-grade osteosarcomas from benign bone tumors by determining only the rate of proliferation was not possible, although interestingly, the proliferative rate of myositis ossificans, a purely reactive lesion, was in the range of the values determined for high-grade osteosarcoma.

[Solitary bone cysts. Morphologic variation, site, incidence and differential diagnosis]. / Solitäre Knochenzysten. Morphologische Variationsbreite, Lokalisation, Häufigkeit und Differentialdiagnose.

Analysis of 402 solitary bone cysts demonstrates the wide morphological variation of this cystic lesion with regard to histology and radiology. Aside from metaphyseal location in femur (33%) and humerus (23%), solitary bone cysts are also often located in calcaneus (11%), tibia (11%) and pelvis (10%). Most patients are in the second decade of life. Differentiation between this benign lesion and malignant bone tumors is very important in daily clinical routine. The diagnosis cannot be based solely on radiological findings because of the variation of solitary bone cysts and the special forms, such as calcifying solitary bone cyst. Therefore, exact histological diagnosis is of particular importance.

[Spongiosa structure and polyostotic heterogeneity in osteoporosis. Mechanism of bone transformation, morphology, clinical significance]. / Spongiosastruktur und polyostotische Heterogenität bei Osteoporose.

Osteoporosis is the most frequent generalized bone disease. The clinical expression of postmenopausal osteoporosis is characterized by spontaneous fractures of the vertebral bodies which affect mainly the trabecular bone structure. Thus the morphological bone transformation in osteoporosis is of clinical relevance. The object of this study was to analyze quantitatively and qualitatively the distribution of three-dimensional structure of cancellous bone in the human spine in osteoporosis. Therefore the complete anterior column of the spine and bone biopsies of the iliac crest of 11 autopsy cases with proven osteoporosis and 26 autopsy cases without primary or secondary bone disease were removed. A 1-mm-thick prepared sagittal section through the center of all vertebral bodies was embedded undecalcified in plastic and stained on the surface using a modification of the von Kossa method. This technique allowed combined two- and three-dimensional measurements simultaneously; these included evaluation of trabecular bone volume, trabecular interconnection, trabecular thickness, and trabecular number. The quantitative spine deformity index and qualitative analysis of the trabecular bone structure completed the investigation. The bone loss in osteoporosis is a loss of structure and the loss of whole trabeculae caused by perforations. The age-related decrease of trabecular bone mass is due to the transformation from plates to rods. Patient with osteoporosis show pathologically diminished trabecular bone volume and apparently reduced trabecular interconnection, while trabecular thickness and trabecular number show age-dependent change. The polyostotic heterogeneity in osteoporosis is immense. Neighboring vertebral bodies show differences of up to 100% in bone volume and bone structure. Due to this fact it is impossible to define a threshold for osteoporotic fractures. At the moment the transformation and loss of trabecular bone structure in osteoporosis is assumed to be irreversible; therefore, early prophylaxis is necessary to prevent clinical manifestations of these changes of bone.

[Periosteal osteosarcoma. Histologic characteristics, preparation technique, growth pattern and differential diagnosis]. / Das periostale Osteosarkom. Histologische Charakteristika, Präparationstechnik, Wachstumsmuster und Differentialdiagnose.

Periosteal osteosarcoma is a distinct bone tumor entity with characteristic morphological features within the group of juxtacortical osteosarcoma. Periosteal osteosarcoma is predominantly located in the long tubular bones, especially in the tibia and femur and is situated on the outer circumference of the tumor-bearing bone (saucerization phenomenon). In contrast to parosteal osteosarcoma, periosteal osteosarcoma is less differentiated and is believed to have a worse prognosis. In this work the histological features are described with predominantly chondroblastic differentiation of 14 cases with periosteal osteosarcoma. A horizontal preparation technique of periosteal osteosarcoma specimens allows comparison with computed tomography and is the optimal method to detect an invasion of the medullary cavity. Further studies are necessary to clarify if neoadjuvant chemotherapy could improve the prognosis of certain patients.

Structural heterogeneity within the axis: the main cause in the etiology of dens fractures. A histomorphometric analysis of 37 normal and osteoporotic autopsy cases.

Fractures of the odontoid process are potentially serious injuries; Type II and III fractures, as described by Anderson and D'Alonzo, are seen in the emergency room especially in young adolescents and individuals over 60 years of age. The etiology of these fractures is still controversial. Malunion and nonunion in both types of fractures are presumed to be due to insufficient external or internal fixation, but this theory has not been fully explained. To examine these issues, the authors expanded their prior studies of the anatomy of the axis. For histomorphometric analysis of cancellous and cortical bone, the axis was removed in 37 autopsies (26 normal and 11 osteoporotic cases) and sectioned in the sagittal plane to a thickness of 1 mm using a surface-stained block-grinding technique. The base of the dens is the region of least resistance for fractures because of its reduced trabecular bone volume, a poorer trabecular interconnection, and a cortical thickness one-third that of the axis. In all cases, trabeculae were disconnected from the trabecular lattice, and in 30%, microcallus formations were demonstrated in the base of the dens. A special filigree type of trabeculae in the base of the dens is often seen in patients with osteoporosis; microarchitectural differences of cancellous bone between the base of the dens and the other regions of the axis are also markedly increased. The authors infer from the data that the bone structure of the axis is responsible for the location, distribution, and frequency of fractures of the odontoid process in normal healthy bone and this frequency is greatly increased in individuals with osteoporosis. The deficiency of bone mass within the base also suggests a new explanation for the occurrence of nonunions, even after treatment of fractures of the base of the dens.

Intervertebral variation in trabecular microarchitecture throughout the normal spine in relation to age.

The vertebral bodies of the complete spine (C-3-L-5) were removed in 26 autopsy cases without evidence for primary or secondary bone disease (13 males aged 19-79 years and 13 females aged 17-90 years). A sagittal segment through the center of all vertebral bodies was embedded undecalcified in hydroxyethylmethacrylate and processed to so-called surface stained block grindings. Histomorphometric analysis of the complete segment was performed using a computer-assisted image analysis system (IBAS 2000). The structural parameters investigated were bone volume (BV/TV) and trabecular interconnection quantificated by trabecular bone pattern factor (TBPf). A close correlation of BV/TV and TBPf was found in all vertebral bodies irrespective of vertebral region (r = 0.8, p < 0.001). This indicates that the age-related decrease of trabecular bone mass is primarily the consequence of the transformation from plates to rods and the loss of whole trabecular structures. This basic principle is valid throughout the complete spine. However, the systematic analysis of vertebral trabecular bone from C-3 to L-5 revealed a significant intervertebral variation of trabecular microarchitecture. The density of trabecular structure of cervical vertebrae is much higher than that of thoracic and lumbar vertebrae (p < 0.001). The extent of age-related loss of trabecular bone mass and structure showed a decrease within the spine from the caudal to the cranial region (p < 0.05). The loss of bone volume in individuals between the ages of 30 and 80 years in the lumbar spine was 53%, whereas in the thoracic spine the decrease was 41%, and in the cervical spine only 24%.(ABSTRACT TRUNCATED AT 250 WORDS)

Calcifying solitary bone cyst: morphological aspects and differential diagnosis of sclerotic bone tumours.

Fourteen solitary bone cysts (SBC) with large areas of calcification (7 in the femur, 4 in the humerus, and 1 each in the pelvis, the tibia and the scapula) and 402 SBC from the Hamburg Bone Tumour Registry were reviewed in a retrospective study. The analysis was done with emphasis on the clinical, radiological and histological appearances. SBC are well known lesions, but calcifying SBC (CSBC) or extensive extragnathic cement-like bone productions are rare. The clinical and radiological differential diagnosis includes fibrous dysplasia, chondroma, low-grade chondrosarcoma and osteosarcoma. Bits of this cement-like matrix are detectable within the wall of approximately 70% (278 of 402) of SBC from the registry. CSBC are changed SBC. The intraoperative confirmation of the diagnosis on a frozen section by the bone pathologist leads to curettage which is currently the most common therapy in this benign lesion.

Polyostotic heterogeneity of the spine in osteoporosis. Quantitative analysis and three-dimensional morphology.

It was the aim of this study to record quantitatively and qualitatively the distribution of the three-dimensional microarchitecture throughout the human spine in osteoporosis. Bone biopsies of the iliac crest and the complete spine of 26 autopsy cases without skeletal disease and 11 female patients with proven osteoporosis were removed. Grindings of all vertebrae by a technique which we developed allowed two- and three-dimensional measurements simultaneously. The analysis included an evaluation of trabecular bone volume, trabecular interconnection, and trabecular thickness, as well as a qualitative investigation of the structure of cancellous bone. The bone loss in osteoporosis is a loss of structure. The relative loss of the trabecular microarchitecture is greater in the iliac crest than in the lumbar spine. It is a gradual change from normal bone to osteoporosis. Transformation from plates to rods and the loss of whole trabeculae are caused by perforations. The polyostotic heterogeneity in osteoporosis is remarkable. Adjacent vertebrae may show differences of up to 100% in bone structure and bone volume. This explains the difficulties in early diagnosis of osteoporosis. Due to the polyostotic heterogeneity it is impossible to define a threshold mineral content for osteoporotic fractures.

[Apoptosis: bcl-2 as a key protein for programmed cell death]. / Apoptose: Bcl-2 ein Schlüsselprotein für den programmierten Zelltod.

The realization that malignant tumors may grow not only due to proliferation but also because cells don't die when they should provides absolute new directions in basic and clinical tumor research. Thus, apoptosis, the programmed cell death is very hot now in the scientific community. In the center of interest are the regulation mechanisms of apoptosis and its significance in different tumor entities. Recently, numerous studies on apoptosis during embryogenesis, different stages of tissue development and in epithelial and hormone dependent tumors have been published. The data suggested an important role of bcl-2 in regulation of apoptosis. In this view the demonstration of bcl-2 oncogene expression in osteosarcoma, a mesenchymal tumor, is of potential great interest.

[Osteosarcoma--apoptosis and proliferation. Study of bcl-2 expression]. / Osteosarkom--Apoptose und Proliferation. Untersuchung zur bcl-2-Expression.

The relationship between the growth of tumors and the expression of the protooncogene Bcl-2 could be shown in epithelial tumors. A bcl-2 expression leads to a prolonged cell survival due to an inhibition of apoptosis. The potential meaning of bcl-2 expression in mesenchymal tumors remains still unknown. The fact, that the heterogenous group of osteosarcoma is not sufficiently characterized at present, suggested to investigate the bcl-2 expression in osteosarcoma. Thus, immunohistochemistry was used to analyze 47 specimens of different osteosarcomas of 36 patients. Sixteen cases (46%) showed a strong expression of bcl-2 and 13 cases (35%) were moderately positive for bcl-2. Seven cases (19%) were negative for bcl-2. The heterogenous, negative up to strong expression of bcl-2 yield clues, that the Bcl-2 controlled regulation of programmed cell death could be an important factor of cellular kinetics. Additionally the cellular proliferation rate was determined with the monoclonal antibody MIB 1, directed against the Ki-67 epitope. The data of bcl-2 expression and cellular proliferation rate lead to a classification correlating with the histological classification. To verify the importance of apoptosis in the genesis of mesenchymal tumors and whether Bcl-2 may play an important role as a predictive factor for the prognosis of osteosarcoma, further investigations will be needed.