Fine needle aspiration cytology of bone tumors

OBJECTIVE: To study the role of fine needle aspiration cytology (FNAC) in the diagnosis of bone tumors and its impact on therapeutic decisions. STUDY DESIGN: A group of 122 cases of bone tumor were evaluated by FNAC. Detailed diagnoses were compared with the available histology. RESULTS: Diagnostic accuracy of FNAC was 90.5% in this study. FNAC could differentiate between various round cell tumors such as Ewing's sarcoma and myeloma, among various giant cell-rich lesions of bone and between the benign and malignant chondroid bone tumors. Some uncommon variants were also correctly diagnosed. In metastatic bone tumors, the source of primary malignancy could not be indicated in the majority (52.9%) because of the poorly differentiated morphology. Osteoid or osteoid-like material was demonstrable in 63.6% cases of osteogenic sarcoma. A case of chondroblastic osteogenic sarcoma that was reported as chondrosarcoma was the only diagnostic error in the study. FNAC obviated the need of open biopsy in 63.8% patients, and therapeutic decisions were made according to the cytologic diagnoses. CONCLUSION: FNAC plays an important role in the early diagnosis of bone tumors by its accuracy, ease of use and rapidity and is helpful in making the therapeutic decisions.     

Use of carminomycin on children and adolescents with osteogenic sarcoma]

The therapeutic effect of carminomycin was studied in clinic at different treatment schemes with respect to 14 children and juvenile patients with osteogenic sarcoma. Pronounced local effect evident from disappearance of the pain and in some cases decrease of the metastatic tumor were noted in the patients with metastases of the osteogenic sarcoma to the bones or relapses of the primary tumor. Subjective improvement and objective effect were observed respectively in 90 and 53 per cent of the patients with metastases into the lungs and pronounced lung symptomatology.     

Demonstration of virus particles in Moloney murine sarcoma virus-induced periosteal bone in mice

Balb/c mice were inoculated intramuscularly with Moloney murine sarcoma virus in one of the hind legs. This led to the rapid development of a regressive sarcoma and also to the proliferation and osteogenic differentiation of cells in the adjacent periosteum. Examination of the tissues by transmission electron microscopy revealed the presence of type A and C virus particles within the sarcoma cells as well as within the cells of the newly formed bone. Extracellular type C virus particles were formed by budding from the cell surface and by release from disintegrating cells. No virus particles were found in the bone or the surrounding soft tissues of the contralateral, noninfected leg. These observations suggest that viral infection of periosteal cells are at least partly responsible for the osteogenic response associated with the virus-induced sarcoma. Production of growth factors by the sarcoma cells could also contribute to this process.     

High-dose methotrexate/leucovorin adjuvant chemotherapy of osteogenic sarcoma: biochemical effects in DNA-synthesis of bone marrow cells (author's transl)]

DNA metabolism in bone marrow cells was measured under high-dose Methotrexate/Leucovorin adjuvant chemotherapy of a patient with primary amputation of his right leg because of osteogenic sarcoma. The biochemical data showed that there was no rescue effect of Leucovorin after 200 mg/kg Methotrexate. Corresponding to this "biochemical failure" of the rescue effect the patient died from the complications of a long and very severe bone marrow suppression. To improve the safety of this therapeutic regimen the intravenous injection and in some cases a higher dose of Leucovorin is recommended.     

Mass Spectrometric Identification of Ancient Proteins as Potential Molecular Biomarkers for a 2000-Year-Old Osteogenic Sarcoma

Osteosarcoma is the most common primary malignant tumor of bone usually occurring in young adolescent and children. This disease has a poor prognosis, because of the metastases in the period of tumor progression, which are usually developed previous to the clinical diagnosis. In this paper, a 2000-year-old ancient bone remain with osteogenic sarcoma was analyzed searching for tumor biomarkers which are closely related to this disease. After a specific extraction SDS-PAGE gel electrophoresis followed by tryptic digestion was performed. After the digestion the samples were measured using MALDI TOF/TOF MS. Healthy bone samples from same archaeological site were used as control samples. Our results show that in the pathological skeletal remain several well known tumor biomarkers are detected such as annexin A10, BCL-2-like protein, calgizzarin, rho GTPase-activating protein 7, HSP beta-6 protein, transferrin and vimentin compared to the control samples. The identified protein biomarkers can be useful in the discovery of malignant bone lesions such as osteosarcoma in the very early stage of the disease from paleoanthropological remains.     

Human osteogenic sarcoma cells exhibit enhanced protein phosphorylation

Protein phosphorylation was compared in normal human cells and human osteogenic sarcoma cells. The phosphorylation of endogenous cellular protein substrates was measured by two independent methods, incubation of homogenized cells with [γ-³²P]ATP or labeling of intact cells with Na₂H³²PO₄. Phosphorylated proteins were identified by SDS-polyacrylamide gel electrophoresis and autoradiography. The stained protein bands of all four osteosarcoma cell lines were nearly identical to those of the normal cells. However, each of the osteosarcoma cell lines showed autoradiographic evidence of enhanced phosphorylation in many different protein bands which was neither cyclic AMP-dependent nor a function of cellular growth rate or density. When normal and tumor cell homogenates were mixed prior to incubation with [γ-³²P]ATP, the resulting phosphoprotein patterns resembled those obtained with the tumor cells alone. In addition, a surgically derived osteogenic sarcoma was cultured and an established line obtained; another portion of the fresh tumor was immediately homogenized and used in a phosphorylation assay. The same enhanced phosphorylation pattern was obtained with the homogenized fresh tumor as with the cell line established from it. These results suggest thathuman osteogenic sarcoma cells are able to perform a significantly increased amount of phosphorylation of endegenous cellular protein substrates when compared to normal human cells.     

Paleopathological study on a case of osteosarcoma

This report concerns a probable case of osteosarcoma found in a precontact Hawaiian skeleton from the east coast of Oahu Island, Hawaii. A young adult female showed a tumorous bone proliferation with a coarse, corallike appearance at the distal metaphyseal area of the left femur. In gross observation, a profusion of coalescing bone was extended to the surrounding space and also invaded the marrow space. X-ray films revealed spotted and ringed shadows in the shaft and a "sunburst appearance" in the lesion. Histological examination of the tumor bone fragment showed a great deal of primitive bone tissue formation without any systemic Haversian structure. The diagnosis of osteogenic osteosarcoma is much more compatible than other primary malignant bone tumors such as Ewing's sarcoma, fibrosarcoma, and chondrosarcoma or osteoplastic metastatic carcinoma of the bone when the location and morphology of the tumor are considered along with the age of the decedent.     

Immunospecificity of non-histone chromosomal proteins in 89Sr-induced osteogenic sarcoma (mouse).

Antibodies against non-histone chromosomal proteins for 89Sr-induced osteogenic sarcoma (mouse) were prepared by immunization of rabbits. The immunoreactivity of this antigen was then compared with those of non-histone chromosomal proteins from Ehrlich ascites tumor, normal mouse liver, and calf thymus by the method of quantitative microcomplement fixation. The non-histone chromosomal proteins of 98Sr-induced osteogenic sarcoma, fractionated by hydroxylapatite chromatography, exhibited significant affinity for the antibodies. Similar proteins from Ehrlich ascites tumor, normal mouse liver, or calf thymus were virtually inactive, indicating the tissue-specificity of 89Sr-induced osteogenic sarcoma proteins.     

A comparative evaluation of the clinical x-ray picture of uremic osteodystrophy before and after parathyroidectomy]

The authors present the results of clinical, x-ray, and biochemical studies carried out in 51 patients with uremic osteodystrophy, treated with hemodialysis, before and after parathyroidectomy. The patients were divided into 4 groups with various patterns of x-ray symptoms. Patients with x-ray signs of fibrous osteodystrophy made up group 1, the second group consisted of patients with a combination of fibrous osteodystrophy and osteomalacia with secondary hyperparathyrosis predominance; the third group, like the second one, included patients with the mixed form of uremic osteodystrophy, but with the predominance of the osteomalacic syndrome; Group 4 patients had no x-ray signs of bone changes, and the diagnosis of uremic osteodystrophy was confirmed by clinical laboratory evidence. Analysis of the clinical and x-ray data before and after parathyroidectomy has brought the authors to a conclusion that such an intervention was effective only in cases with manifest clinical and x-ray symptoms of fibrous osteodystrophy. In Group 2 patients with the mixed form of uremic osteodystrophy and less manifest osteomalacia as against fibrous osteodystrophy, subtotal or partial parathyroidectomy is advisable only in cases when conservative therapy is of no avail and fibrous dystrophy is progressing. Surgical treatment is contraindicated to patients in whom x-ray signs of osteomalacia predominate over fibrous osteodystrophy in the total picture of uremic osteodystrophy; it may result in a rapid progress of osteomalacia.     

Parathyroid hormone (PTH) and PTH-like protein (PLP) stimulate interleukin-6 production by osteogenic cells: a possible role of interleukin-6 in osteoclastogenesis

Osteogenic cells mediate PTH-stimulated osteoclastic bone resorption by a yet unidentified mechanism. We show that primairy rat osteoblast-like cells and the clonal osteogenic sarcoma cell line UMR-106 produce interleukin-6 (IL-6) and that bPTH(1-84) and synthetic hPLP(1-34) stimulate this production dose-dependently. With both peptides a close relation between IL-6 and cyclic-AMP production was found, though for PTH concentrations higher than 2.10(-8) M a clear dissociation was observed. Significant IL-6 activity was also detected in media of cultures of 17-day-old fetal mouse radii and metacarpals which was clearly stimulated by PTH. The source of IL-6 in these bone explants seems to be the osteogenic (cartilage) cells. Treatment of bone explants with IL-6 induced osteoclastic resorption which, however, depended on the bone resorption system used. This bone resorbing action of IL-6 is exerted probably through an effect on the formation of osteoclasts (osteoclastogenesis) rather than on the activation of already existing mature osteoclasts. We suggest that IL-6 produced by osteogenic cells may be a mediator in PTH-stimulated osteoclastic bone resorption.     

Osteogenic sarcoma of the mandible: a plea for radical initial surgery

A total of 24 cases of osteogenic sarcoma of the mandible have been reported to the SEER Program of the National Cancer Institute from 1973 to 1983. Out of 14 patients whose disease was staged as regional, only 9 were eligible for 5-year survival figures and only 2 are alive, for a 23 percent 5-year cure. For those patients staged as localized, only four of six eligible for 5-year cure survived, for a 66 percent 5-year survival. Considering all patients reported to SEER who were eligible for 5-year cure, the rate is 40 percent. We believe this reflects the practice of localized resection of these tumors. At Charity Hospital of Louisiana at New Orleans there have been 10 cases of osteogenic sarcoma of the mandible since 1948. There have been no 5-year survivors for those operated on by other services, usually by local resection. On the Tulane Plastic Surgery Service, a total of six cases of osteogenic sarcoma of the mandible were treated by radical surgery with 100 percent survival from 11 to 22 years.     

Genetics of rare mesenchymal tumors: Implications for targeted treatment in DFSP,ASPS, CCS,GCTB and PEComa

Soft tissue and bone sarcomas comprise a heterogeneous group of mesenchymal tumors that include roughly 130 distinct diagnostic entities. Many of them are exceptionally rare, with only few cases diagnosed worldwide each year. Development of novel targeted treatment in this group of tumors is of special importance since many sarcoma subtypes are resistant to conventional chemotherapy and the effective therapeutic options are limited. In this review we aim to discuss the molecular implications for targeted therapy in selected rare soft tissue and bone sarcoma subtypes, including dermatofibrosarcoma protuberans (DFSP), alveolar soft part sarcoma (ASPS), clear cell sarcoma (CCS), giant cell tumor of bone (GCTB) and perivascular epithelioid cell neoplasms (PEComas).This article is part of a Directed Issue entitled: Rare cancers.     

Roentgenologic characteristics of small bronchoalveolar cancer of the lung under conditions of image processing using linear filtration (roentgenomorphologic comparison)

X-ray and morphological signs were compared in 50 patients with histologically verified bronchioloalveolar cancer (BAC). Particular attention was drawn to a study of an x-ray picture after computer processing of a tomographic image using a special technique. The most characteristic x-ray signs of BAC were subpleural localization, the presence of a fragmented scar in a tumor, and ill defined outlines. The peculiarities of an x-ray picture of small-size BAC were determined by the morphological structure of a tumor, a degree of its differentiation that could be detected in most cases only after image processing by the linear filtration technique.     

Histogenesis of Ewing's sarcoma. An evaluation of intermediate filaments and endothelial cell markers

Four cases of Ewing's sarcoma, three in bone and one from an extraskeletal site, were studied immunohistologically using monospecific antibodies against intermediate filament proteins of keratin, vimentin, desmin and neurofilament types. All cases were also evaluated for the presence of Factor VIII-related antigen (FVIIIR:Ag) and for the binding of Ulex europaeus I lectin (UEA I), both of which are endothelial markers. In all cases the tumor cells contained vimentin but not keratin, desmin or neurofilaments. The tumor cells could not be decorated with either anti-FVIIIR:Ag or UEA I, whereas the vascular endothelium was positive for both markers. The vimentin-positivity indicates a mesenchymal derivation of Ewing's sarcoma, while the lack of endothelial markers argues against the proposed endothelial origin of this tumor.     

Arsenic trioxide promotes senescence and regulates the balance of adipogenic and osteogenic differentiation in human mesenchymal stem cells

Arsenic trioxide (ATO) as an anti-tumor drug could induce differentiation and apoptosis in tumor cells. Mesenchymal stem cells (MSCs) play important roles in the hematogenesis of bone marrow. Many reports have shown that the disorder of MSC adipogenic and osteogenic differentiation occurs in some diseases. However, reports about the effects of ATO on MSCs are limited. In this study, we found that 1 μM ATO promoted MSC senescence mainly through p21, although it had no effect on apoptosis at this dose. Furthermore, ATO promoted adipogenic differentiation, but inhibited osteogenic differentiation in MSCs. Our study also showed that CCAAT/enhancer-binding protein alpha C/EBPα and peroxisome proliferator-activated receptor gamma PPARγ might be involved in the regulation of adipogenic and osteogenic differentiation induced by ATO. Our results indicated that ATO may exert an anti-tumor effect by influencing bone marrow micro-environment. Moreover, it may regulate the adipogenic and osteogenic differentiation of MSCs.     

Immunotherapy of osteogenic sarcoma with transfer factor

Summary Fifteen patients with osteogenic sarcoma were treated with transfer factor derived from leukocytes of their household contacts. Eight of the fifteen patients were tumor-bearing, and transfer factor therapy was correlated with increased cell-mediated immunity in peripheral blood lymphocytes and with lymphocytic infiltrates into the tumors. There was no marked increased in survival time as compared with historical controls, but this therapy did not accelerate the disease, and there were no untoward side effects.Seven of the fifteen patients were disease-free when transfer therapy was initiated shortly after surgical removal of the primary tumor (five patients) or solitary pulmonary metastases (two patients). These patients received transfer factor injections every 2 weeks for 1–2 years. Six of the seven patients are disease-free 62–82 months after surgery. Compared with probabilities of 5-year survival computed from historical controls, this is significant at P<0.008. Abbreviations used in this paper are: CI, cytotoxicity index; GCT, giant cell tumor of bone; HHC, household contacts; HHC_os, household contacts of patients with osteogenic sarcoma; MIC, mononuclear inflammatory cell; TFCI, transfer factor cytotoxicity index; TSTF, tumor-specific transfer factor.     

Systemically administered rhBMP-2 promotes MSC activity and reverses bone and cartilage loss in osteopenic mice

Osteoporosis is a disease manifested in drastic bone loss resulting in osteopenia and high risk for fractures. This disease is generally divided into two subtypes. The first, post-menopausal (type I) osteoporosis, is primarily related to estrogen deficiency. The second, senile (type II) osteoporosis, is mostly related to aging. Decreased bone formation, as well as increased bone resorption and turnover, are thought to play roles in the pathophysiology of both types of osteoporosis. In this study, we demonstrate in murine models for both type I (estrogen deficiency) and type II (senile) osteopenia/osteoporosis that reduced bone formation is related to a decrease in adult mesenchymal stem cell (AMSC) number, osteogenic activity, and proliferation. Decreased proliferation is coupled with increased apoptosis in AMSC cultures obtained from osteopenic mice. Recombinant human bone morphogenetic protein (rhBMP-2) is a highly osteoinductive protein, promoting osteogenic differentiation of AMSCs. Systemic intra-peritoneal (i.p.) injections of rhBMP-2 into osteopenic mice were able to reverse this phenotype in the bones of these animals. Moreover, this change in bone mass was coupled to an increase in AMSCs numbers, osteogenic activity, and proliferation as well as a decrease in apoptosis. Bone formation activity was increased as well. However, the magnitude of this response to rhBMP-2 varied among different stains of mice. In old osteopenic BALB/c male mice (type II osteoporosis model), rhBMP-2 systemic treatment also restored both articular and epiphyseal cartilage width to the levels seen in young mice. In summary, our study shows that AMSCs are a good target for systemically active anabolic compounds like rhBMP-2.     

The development relationship between osteocytes and osteoclasts: a study using the quail-chick nuclear marker in endochondral ossification.

Interspecific grafts of limb buds and femurs on the chorioallantoic membrane of 5-day-old hosts and into the somatopleure of 3-day-old hosts were carried out between quail and chick embryos. Due to their different nuclear features, the cells of the two species can be identified in the chimeric bones resulting from the endochondral ossification which occurs in the explanted tissues. By following the cell lineage in the bone and marrow we were able to show that the hemopoietic and the osteogenic (comprising osteoblasts, osteocytes, and chondrocytes) cell lines have different embryological origins. The osteogenic line is derived from the limb bud mesenchyme, while the hemopoietic cells are brought into the bone marrow via the circulation. In the fixed cells of the marrow two categories have to be distinguished: the reticular cells originating from the bone rudiment and the endothelial cells which invade the cartilage and are of hematogenous origin. The osteoclasts belong to the hemopoietic cell line and are not derived from any cell type of the osteogenic line.     

Silencing of HMGA1 expression by RNA interference suppresses growth of osteogenic sarcoma

The expression of high mobility group protein A1 (HMGA1) protein has been closely related to various malignant and prognostic degrees of tumor. To investigate the influence of down-regulating HMGA1 on the tumor and the mechanism underlying antitumor of HMGA1, we transfected the HMGA1 shRNA vector into the osteogenic sarcoma MG-63 cell and observed the changes of cell proliferation, invasion abilities, and the tumor growth. HMGA1 gene expression could be efficiently inhibited, and cell proliferation, migration, invasion, and matrix metalloprotease level were also decreased. BALB/C nude mice injected with the MG-63 cells transfected HMGA1 shRNA showed the significant lower tumor weight, tumor volume, and longer tumor-forming time compared with the control group. Our results suggest that knockdown of HMGA1 could inhibit growth and metastasis potentials of MG-63 cells, which may be a therapeutic target protein for osteogenic sarcoma and may be of biological importance.