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PubMed2006

Infectivity enhancement for adenoviral transduction of canine osteosarcoma cells.

The full realization of conditionally replicative adenoviruses (CRAds) for cancer therapy has been hampered by the limited knowledge of CRAd function in vivo and particularly in an immunocompetent host. To address this issue, we previously proposed a canine adenovirus type 2 (CAV2)-based CRAd for clinical evaluation in canine patients with osteosarcoma (OS). In this study, we evaluated infectivity-enhancement strategies to establish the foundation for designing a potent CAV2 CRAd with effective transduction capacity in dog osteosarcoma cells. The results indicate that the native CAV2 fiber-knob can mediate increased binding, and consequently gene transfer, in both canine osteosarcoma immortalized and primary cell lines relative to previously reported Ad5 infectivity-enhancement strategies. Gene delivery was further enhanced by incorporating a polylysine polypeptide onto the carboxy terminus of the CAV2 knob. This vector demonstrated improved gene delivery in osteosarcoma xenograft tumors. These data provide the rationale for generation of infectivity-enhanced syngeneic CAV2 CRAds for clinical evaluation in a dog osteosarcoma model.

Adenoviridae↗
PubMed2002

Two novel tumor suppressor gene loci on chromosome 6q and 15q in human osteosarcoma identified through comparative study of allelic imbalances in mouse and man.

We have performed a comparative study of allelic imbalances in human and murine osteosarcomas to identify genetic changes critical for osteosarcomagenesis. Two adjacent but discrete loci on mouse chromosome 9 were found to show high levels of allelic imbalance in radiation-induced osteosarcomas arising in (BALB/cxCBA/CA) F1 hybrid mice. The syntenic human chromosomal regions were investigated in 42 sporadic human osteosarcomas. For the distal locus (OSS1) on mouse chromosome 9 the syntenic human locus was identified on chromosome 6q14 and showed allelic imbalance in 77% of the cases. Comparison between the human and mouse syntenic regions narrowed the locus down to a 4 Mbp fragment flanked by the marker genes ME1 and SCL35A1. For the proximal locus (OSS2) on mouse chromosome 9, a candidate human locus was mapped to chromosome 15q21 in a region showing allelic imbalance in 58% of human osteosarcomas. We have used a combination of synteny and microsatellite mapping to identify two potential osteosarcoma suppressor gene loci. This strategy represents a powerful tool for the identification of new genes important for the formation of human tumors.

Alleles↗
PubMed2003

Osteoblasts and MG-63 osteosarcoma cells behave differently when in contact with ProRoot MTA and White MTA.

AIM: To test the hypothesis that MG-63 osteosarcoma cells and primary osteoblasts react differently to ProRoot trade mark MTA (mineral trioxide aggregate) and White MTA by: (i) investigating the attachment of primary osteoblasts and MG-63 osteosarcoma cells to ProRoot trade mark MTA and White MTA; and (ii) comparing the osteogenic behaviour of both cell lines in contact with these endodontic materials. METHODOLOGY: Primary osteoblasts were harvested from foetal rat calvaria by sequential digestion and MG-63 osteosarcoma cells were purchased. Cells were exposed to ProRoot trade mark MTA and White MTA prepared according to the manufacturer's instructions. All samples and controls were prepared in quadruplicate. After 6, 9 and 13 days exposure to MTA, the cells were fixed and prepared for SEM examination. In addition, both the cell types were grown to confluence and exposed to beta-glycerophosphate and dexamethasone to assess mineralized nodule formation as a function of osteogenic behaviour. RESULTS: The number of cells on the surface of the culture dish and on top of the materials increased in all samples throughout the 3 time periods, except for White MTA where no primary osteoblasts were visible on top of the material by the end of 13 days. After exposing cells to differentiation medium nodules were observed in cultures of primary osteoblasts, but not of MG-63 osteosarcoma cells. CONCLUSIONS: Under the conditions of this study, whilst primary osteoblasts initially bound to White MTA, they did not survive on the surface by the end of 13 days. Primary osteoblasts formed mineralized nodules when exposed to differentiation medium, whilst MG-63 cells did not form nodules. As MG-63 cells do not behave osteogenically by forming mineralized nodules, and primary osteoblasts are more sensitive than MG-63 osteosarcoma cells to White MTA in cell culture, primary osteoblasts are more appropriate than MG-63 cells for testing endodontic materials in cell culture.

Aluminum Compounds↗
PubMedDate not supplied

Accuracy of radiography, nuclear scintigraphy, and histopathology for determining the proximal extent of distal radius osteosarcoma in dogs.

OBJECTIVE: To compare the accuracy of radiography, nuclear scintigraphy, and histopathology for determining the proximal extent of distal radius osteosarcoma in dogs. STUDY DESIGN: Retrospective clinical study. SAMPLE POPULATION: Twenty client-owned dogs taken to Colorado State University with osteosarcoma of the distal radius. METHODS: Medical records of 20 dogs with confirmed osteosarcoma that underwent a limb-salvage procedure were reviewed. Measurements were performed directly from the lateral view of each radius, from both the scintigram and the radiograph, to determine the length of the radius and the distance from the proximal extent of the tumor to the distal radiographic or scintographic extent of the radius. The ratio of distal radial involvement to total radius length was calculated. A similar ratio was also determined using the macroslide, which included the entire portion of bone that was excised during the limb-salvage procedure. All 3 methods of measurement were compared. RESULTS: Nuclear scintigraphy significantly overestimated tumor length when compared with macroslide specimen measurements. Radiography also overestimated tumor length, but these results were not significantly different from macroslide specimen measurements. CONCLUSIONS: Both radiography and nuclear scintigraphy overestimate the extent of distal radius osteosarcoma in dogs when compared with histopathologic macroslides of the same lesions. Nuclear scintigraphy overestimated tumor extent to a larger degree than did radiography. CLINICAL RELEVANCE: Although radiography is a more accurate method of measurement of the extent of distal radius osteosarcoma, because nuclear scintigraphy overestimates tumor length to a greater degree, scintigraphy may provide a larger margin of safety for determining the site of proximal osteotomy during a limb-salvage procedure. However, caution should be taken when utilizing scintigraphy, because this method may overestimate length of radius involved to such an extent as to cause the surgeon to believe that a patient is not a suitable limb-salvage candidate.

Animals↗
PubMedDate not supplied

[The importance of conventional radiographs in the diagnosis of osteosarcoma].

We report about a 12-year-old boy with a history of recurrent patella luxation. Due to persistent pain in the distal left femur a MRI examination was performed in another hospital which suggested a malignant bone tumor. Without validation of the MRI findings by conventional radiographs bone biopsy was performed. Histopathological examination yielded the diagnosis of a chondroblastic osteosarcoma. Before initiating polychemotherapy, plain radiographs for the first time, a nuclear imaging study and an additional MRI examination were performed in our hospital. The results of these studies made the diagnosis of an osteosarcoma unlikely. In particular, plain radiographs did not show any osseous lesion which was characteristic of an osteosarcoma. To establish a definite diagnosis biopsy was repeated with resection of the bone area which showed suspicious changes in MRI studies. An osteosarcoma was ruled out by histopathological examination. The pathologic changes detected in MRI were rated as bone bruise on plain radiographs and seemed to be of traumatic origin. Our case report emphasises the importance of conventional radiographs in establishing the diagnosis of an osteoarcoma respectively bone tumors and tumor-like lesions in general. They still remain the mainstay in diagnosing bone forming tumors. MRI imaging studies may show changes which mimick solid lesions but in deed can be of traumatic origin. Without informing the pathologist about the exact origin of the specimen, histopathological examination may lead to the misdiagnosis of a chondroblastic osteosarcoma if specimen, like in this case report, represents epiphyseal tissue showing cartilaginous areas with reactive bone formation.

Biopsy↗
PubMedDate not supplied

[Osteosarcoma and Ewing's sarcoma--The most frequent malignant bone tumors in children--therapy and outcome].

BACKGROUND: Osteosarcoma and Ewing's sarcoma are the most frequent malignant bone tumors in children and young adults with relatively poor overall survival rates. METHODS: Between January 1980 and December 1994, 175 children with osteosarcoma and 64 children with Ewing's sarcoma were treated at the author's institution. 22 children had synchronous metastases, 19 patients had a pathologic fracture. Both groups were treated systemically with chemotherapy regimens (COSS and CESS). Local therapy was amputation or tumor resection and endoprosthetic replacement or biological reconstruction with wide or radical resection margins. In case of Ewing's sarcoma in 35 patients postoperative radiation therapy was done. RESULTS: Five-year overall survival rate for osteosarcoma and Ewing's sarcoma patients is about 63 %, ten-year survival rate for osteosarcoma patients is 60.2 %, for Ewing's sarcoma patients 54.5 %. Prognostic factors significantly influencing overall survival rates are tumor response to chemotherapy (p values = 0.0056 and 0.013, respectively), surgical treatment with adequate resection margins (p value = 0.0001 for osteosarcoma patients) and development of postoperative metastases (p value = 0.0001 for both groups). CONCLUSION: For both groups of malignant bone tumors systemic chemotherapy as well as adequate surgical therapy are necessary to reduce the rates of local recurrences and to achieve better survival rates.

Adolescent↗
PubMed1995

Expression of P-glycoprotein in high-grade osteosarcomas in relation to clinical outcome.

BACKGROUND: Increased levels of P-glycoprotein occur in some osteosarcomas. In this study we determined the relation between P-glycoprotein status and outcome in patients with high-grade osteosarcoma. METHODS: P-glycoprotein status was determined immunohistochemically in specimens of osteosarcoma of the extremities (stage II) from 92 patients who were treated with surgery and chemotherapy. The P-glycoprotein status was analyzed in relation to the length of event-free survival. RESULTS: The presence of increased levels of P-glycoprotein in the osteosarcoma was significantly associated with a decreased probability of remaining event-free after diagnosis (P = 0.002). In a multivariate analysis, P-glycoprotein status (P = 0.001) and the extent of tumor necrosis after preoperative chemotherapy (P = 0.04) were independent predictors of clinical outcome. The risk of adverse events was increased substantially (rate ratio, 3.37; 95 percent confidence interval, 1.60 to 7.10) among patients with increased levels of P-glycoprotein in tumor cells, as compared with patients who did not have increased levels of P-glycoprotein in tumor cells. CONCLUSIONS: In patients with high-grade osteosarcoma treated with surgery and chemotherapy, the presence of increased levels of P-glycoprotein in tumor cells is associated with a significantly increased risk of adverse events and is independent of the extent of necrosis after preoperative chemotherapy.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2003

Cutaneous metastasis of osteosarcoma.

Cutaneous metastases of solid tumors have been reported with a frequency that ranges from 0.7% to 4.4% with recent studies reporting 9% and 10%. Osteosarcomas seldom metastasize to the skin. The most frequent metastatic sites are to the lungs, bones, and kidneys. We report 2 cases of osteosarcoma that metastasized to the skin. A 75-year-old patient with known osteosarcoma of the tibia developed cutaneous nodules overlying the site of the primary tumor. A 46-year-old woman with a history of osteosarcoma of the knee developed a nodule on her scalp. Histologic examination of both lesions revealed metastatic osteosarcoma. This is a rare event that, to our knowledge, has previously only been reported 6 times.

Aged↗
PubMedDate not supplied

Osteosarcoma, seasonality, and environmental factors in Wisconsin, 1979-1989.

Proxy exposure measures and readily available data from the Wisconsin Cancer Reporting System were used to contrast 167 osteosarcoma cases with 989 frequency-matched cancer referents reported during 1979-1989. Differences in potential exposure to water-borne radiation and fluoridated drinking water, population size for the listed place of residence, and seasonality were assessed. An association was found between osteosarcoma and residence in a population of less than 9,000 (odds ratio = 1.6, 95% confidence interval = 1.1-2.4). In addition, an association between month of birth (May through July versus other months of birth) and osteosarcoma among individuals who were less than 25 y of age (odds ratio = 1.9, 95% confidence interval = 1.1-3.4). Overall, no association was found between potential exposure to fluoridated drinking water and osteosarcoma (odds ratio = 1.0, 95% confidence interval = 0.6-1.5). The association between osteosarcoma and water-borne radiation was weak and was not significant statistically (odds ratio = 1.5, 95% confidence interval = 0.8-2.8).

Adolescent↗
PubMed2001

Induction of osteosarcoma and acute myeloid leukaemia in CBA/H mice by the alpha-emitting nuclides, uranium-233, plutonium-239 and amercium-241.

PURPOSE: To compare tumour induction in CBA/H mice, principally osteosarcoma and acute myeloid leukaemia, resulting from exposure to the alpha-emitting nuclides, uranium-233, plutonium-239 and americium-241, and to relate differences between the three nuclides to the pattern of dose delivery within tissues. MATERIALS AND METHODS: Each nuclide was administered intraperitoneally in citrate solution to three groups of adult male CBA/H mice at levels of activity which gave estimated life-time average skeletal doses of about 0.25-0.3 Gy, 0.5-1 Gy and 1-2 Gy. Animals were carefully monitored and sacrificed as soon as they showed signs of ill health; tumours were identified by standard histopathological techniques. RESULTS: Statistical modelling by Cox regression showed that, considering all three nuclides together, there was a highly significant increase in risk of death from osteosarcoma or myeloid leukaemia with increasing dose rate. For osteosarcoma, the effect was significantly greater for 239Pu than 241Am, while separate analysis for 233U showed no significant increase with increasing dose rate. For example, the increase in relative risk of death from osteosarcoma for an increase in life-time average dose rate to bone of 1 mGyd(-1) was 4.2 (2.7-6.5) for 239Pu, 2.3 (1.4-3.4) for 241Am and 1.1 (0.4-3.1) for 233U. For myeloid leukaemia, there was no significant difference between 239Pu and 241Am in the effect of dose rate. The increase in relative risk from myeloid leukaemia for an increase in average dose rate of 1 mGyd(-1) was 1.8 (1.1-2.8) for 239Pu, 2.0 (1.4-2.9) for 241Am and 1.5 (0.8-2.7) for 233U. Significant increases in renal and hepatic carcinomas were also recorded in animals exposed to 233U and 241Am, respectively. Studies of the distribution of the nuclides within the skeleton, published separately, have shown differences in their retention in individual bones and within bone. The proportions of decays occurring near to endosteal bone surfaces and throughout bone marrow were in the order: 239Pu> 241Am>233U. CONCLUSIONS: For osteosarcoma, the relative effectiveness of the nuclides in terms of average bone dose, in the order 239Pu>241Am>233U, is consistent with the proportion of dose delivered near to endosteal surfaces. For myeloid leukaemia, the greater effectiveness of 239Pu and 241Am than 233U is consistent with their accumulation in marrow.

Americium↗
PubMed1975

Viruses in osteosarcomas induced by 226Ra. A study of the induction of bone tumours in mice.

Three groups (15 in each) of 3-month-old mice were injected intraperitoneally with 0-5 muCi 226Ra. They were male and female C3H/H and female CBA/H. The groups were matched by similar controls that were not injected. Osteosarcomas were found in 14/15 of the female C3H/H mice--animals known to carry mammary-tumour viruses in milk--compared with 6/15 in their male counterparts. In CBA/H female mice, which have a very low incidence of both mammary tumours and leukaemia, osteo-sarcomas developed in 7/15 of the animals. The CBA/H mice lived about 5-8 months longer and revealed their osteosarcomas at a correspondingly later time than C3H/H mice. Virus particles were observed in each of the three mammary tumours and in 15 out of the 17 osteosarcomas examined by electron microscopy. No definitive statement can be made from this experiment whether or not osteosarcoma was caused by a virus; but the results suggest that viruses may be a contributory factor in the development of osteosarcoma in these animals.

Animals↗
PubMed1992

A23187-induced translocation of 5-lipoxygenase in osteosarcoma cells.

In a previous study, osteosarcoma cells expressing both 5-lipoxygenase (5-LO) and 5 lipoxygenase-activating protein (FLAP) synthesized leukotrienes upon A23187 stimulation (Dixon, R. A. F., R. E. Diehl, E. Opas, E. Rands, P. J. Vickers, J. F. Evans, J. W. Gillard, and D. K. Miller. 1990. Nature (Lond.). 343:282-284). Osteosarcoma cells expressing 5-LO but not expressing FLAP were unable to synthesize leukotrienes. Thus, it was determined that FLAP was required for the cellular synthesis of leukotrienes. To examine the role of FLAP in A23187-induced translocation of 5-LO to a membrane fraction, we have studied the A23187-stimulated translocation of 5-LO in osteosarcoma cells expressing both 5-LO and FLAP, and in osteosarcoma cells expressing 5-LO only. We demonstrate that in cells expressing both 5-LO and FLAP, 5-LO translocates to membranes in response to A23187 stimulation. This 5-LO translocation is inhibited when cells are stimulated in the presence of MK-886. In osteosarcoma cells expressing 5-LO but not expressing FLAP, 5-LO is able to associate with membranes following A23187 stimulation. In contrast to the cells containing both 5-LO and FLAP, MK-886 is unable to prevent 5-LO membrane association in cells transfected with 5-LO alone. Therefore, we have demonstrated that in this cell system, 5-LO membrane association and activation can be separated into at least two distinct steps: (1) calcium-dependent movement of 5-LO to membranes without product formation, which can occur in the absence of FLAP (membrane association), and (2) activation of 5-LO with product formation, which is FLAP dependent and inhibited by MK-886 (enzyme activation).

5-Lipoxygenase-Activating Proteins↗
PubMed1986

Production of a monoclonal antibody against a human osteosarcoma xenograft.

Monoclonal antibodies (MAbs) against a human sarcoma xenograft carried in BALB/c nu/nu mice were produced by immunizing BALB/c mice with tumor cells and fusing their spleens with the SP2/O-Ag 14 mouse myeloma cell line. Hybridoma supernatants were screened using cryostat tissue sections and an immunoperoxidase reaction for ability to stain osteosarcoma xenograft tumor cells but not tonsil lymphocytes. Of 73 supernatants tested, 19 reacted with both osteosarcoma tumor cells and lymphocytes, while three reacted only with osteosarcoma. One of the latter hybridomas was cloned by limiting dilution to establish a line producing an IgG1 MAb (OS-1). By immunoperoxidase, this MAb stained tumor cells in surgical biopsies of primary (6 of 7) and metastatic (1) osteosarcomas and showed limited cross-reactivity with other tumors. It also cross-reacted with some basement membranes, endothelium and muscular media of blood vessels, and smooth muscle, but not with parenchymal cells of various normal tissues. This MAb may prove useful for the immunohistochemical confirmation of a diagnosis of osteosarcoma in surgical pathology.

Animals↗
PubMed2006

Osteosarcoma: evaluation of information on the internet.

The information about osteosarcoma on the existing websites was examined. The purpose of this study was to evaluate the quality of Internet information available to patients on the topic of osteosarcoma. The secondary purpose was to rank the identified websites with respect to the caliber of relevant information provided by the websites sponsors' identity. A great many of people in the world "surf" the World Wide Web, searching for medical information. The information on the Internet varies dramatically in terms of content and quality for lack of uniformed standard with respect to the medical publications on the Internet. Five search engines searched the search terms "osteosarcoma." The first 25 links displayed by each engine were evaluated for a theoretical total of 125 websites. According to the information context, sponsor identity we evaluated each website. An information quality score of 0 to 26 was generated for each site. A score of 20 or greater was thought to be "high-quality" information website. Eighty-nine unique websites were identified, among which only 8 (9%) scored 20 or more on the information quality score; 32 scored 10 or less. The overall mean information quality score was 11.5. The websites with highest mean scores were academic organization and affiliated hospitals of university. The quality of Internet information on osteosarcoma is variable. Less than 10% of relevant websites were of high information value. The rank list of high quality websites from our information quality score should provide useful information of osteosarcoma.

Humans↗
PubMed2006

CD99 acts as an oncosuppressor in osteosarcoma.

CD99 was recently reported to be under control of the osteoblast-specific transcription factor Cbfa1 (RUNX2) in osteoblasts, suggesting a role in the phato-physiology of these cells. No extensive information is available on the role(s) of this molecule in malignant phenotype, and osteosarcoma, in particular, has never been studied. We report that in 11 different cell lines and 17 clinical samples CD99 expression is either undetectable or very low. Being expressed in the normal counterpart, we tested the hypothesis that CD99 down-regulation may have a role in osteosarcoma development and progression. CD99-forced expression in two osteosarcoma cell lines significantly reduced resistance to anoikis, inhibited growth in anchorage independence as well as cell migration, and led to abrogation of tumorigenic and metastatic ability. Therefore, the molecule acts as a potent suppressor of malignancy in osteosarcoma. CD99 gene transfection induces caveolin-1 up-regulation and the two molecules were found to colocalize on the cell surface. Treatment with antisense oligonucleotides to caveolin-1 abrogates the effects of CD99 on migration. The findings point to an antioncogenic role for CD99 in osteosarcoma, likely through the regulation of caveolin-1 and inhibition of c-Src kinase activity.

12E7 Antigen↗
PubMed1999

Osteosarcoma metastatic to the kidneys without lung involvement.

Symptomatic renal metastasis is very rare in osteosarcoma. A few reported cases had pulmonary metastases before renal involvement. We present a case of metastatic renal osteosarcoma without pulmonary involvement. The patient presented with hematuria 8 years after initial treatment of osteosarcoma of the left distal femur. Computed tomography revealed a bilateral calcified renal mass. The patient died of disease 4 months after detection of the metastases. At autopsy, metastatic osteosarcoma was discovered in the bilateral kidney, both renal veins, inferior vena cava and right atrium. There was no pulmonary involvement. We emphasize the need for renal evaluation for the follow-up of patients with osteosarcoma.

Adult↗
PubMed1979

Some prognostic and epidemiologic factors in canine osteosarcoma.

Naturally occurring skeletal osteosarcomas in a series of 144 untreated dogs were found especially to involve the ends of the long bones of the forelimbs and affected predominantly older male dogs of giant and large breeds. Most tumors were large and partially necrotic and had extended into soft tissues. Of 12 host and tumor characteristics tested in the first part of the study, tumor diameter and volume were significantly associated with the presence of pulmonary metastases at autopsy. The second part of the study revealed that extension of the tumor into the soft tissues and localization of the tumor in the hind legs were associated with a poor prognosis, whereas the fibrosarcomatous type of tumor was associated, as in man, with a favorable prognosis. An association between the 12 characteristics tested was found in 11 of 78 combinations at the 5% level and in 5 combinations at the 1% level. Affected giant dogs were generally younger than affected small and medium-sized dogs. Especially in giant dogs, the osteosarcomas involved the long bones and were of relatively large diameters. The sarcomas in female dogs were larger in volume than those in males. Pure osteoblastic osteosarcomas were generally smaller than combined (chondroblastic and fibroblastic) osteosarcomas. Peritumorous lymphocytes and plasma cells were present in 50% of the dogs, especially in small and young dogs. When compared with a reference population, great Danes, rottweilers, German shepherds, and boxers were found to be overrepresented in the osteosarcoma group.

Animals↗
PubMed1984

High-grade surface osteosarcomas.

Nine patients (seven males and two females) had high-grade, ordinary-type osteosarcomas that were juxtacortically located. The ages of these patients ranged from 9 to 62 years at the time of diagnosis. The roentgenographic appearance of these tumors occasionally is similar to that of periosteal osteosarcomas. Histologically, these tumors were indistinguishable from ordinary high-grade intramedullary osteosarcomas. None of the nine tumors had histologic features of parosteal or periosteal osteosarcomas. All nine patients were treated by ablative surgery. Seven patients died of tumor. Because of its poorer prognosis, this tumor should be differentiated from other juxtacortical osteosarcomas.

Adolescent↗