Ultrastructural changes in giant cell tumor of bone cultured cells exposed to quercetin.

Giant cell tumor of bone (GCTB) is a primary bone tumor that affects skeletally mature people and whose main treatment is surgical. Because there are few pharmacological alternatives for the treatment of this tumor to find other molecules or compounds that could be potential therapeutic agents is desirable. Quercetin is a flavonoid with described antitumoral effect in different types of cancer cell lines that could be a possible option in GCTB treatment. However, there is no literature about the effect of quercetin on GCTB. In the present paper, we reported the ultrastructural changes in GCTB cells exposed to quercetin and also determined the expression of RIP1K, Caspase 3 and Caspase 8 on the exposed cells. For this purpose, GCTB sample was obtained from one patient and cultured. Quercetin affected all the histological components of the GCTB. The ultrastructural changes consisted mainly in necroptosis, autophagocytosis and secondary necrosis. This is the first report about quercetin effects on giant cell tumor of bone cultured cells. Further studies in other models could be done to support the use of quercetin as a complementary treatment in giant cell tumor of bone.Abbreviations: Giant cell tumor of bone (GCTB); transmission electron microscopy (TEM); reverse transcription - polymerase chain reaction (RT-PCR); receptor interacting protein kinase 1 (RIP1K); Dulbecco's Modified Eagle's Medium (DMEM).

A novel model to culture cells from giant cell tumor of bone using three-dimensional (3D) polycaprolactone scaffold.

Two-dimensional (2D) culture of cells from giant cell tumor of bone (GCTB) is affected by loss of the multinucleated giant cells in subsequent passages. Therefore, there is limited time to study GCTB with all its histological components in 2D culture. Here, we explored the possibility of culturing GCTB cells on a polycaprolactone (PCL)-printed scaffold. We also evaluated the viability of the cultured cells and their adherence to the PCL scaffold at day 14 days using immunofluorescence analysis with calcein, vinculin, and phalloidin. Using the histological technique with hematoxylin and eosin staining, we observed all the histological components of GCTB in this 3D model. Immunohistochemical assays with cathepsin K, p63, and receptor activator of nuclear factor (NF)-κB ligand (RANKL) yielded positive results in this construct, which allowed us to confirm that the seeded cells maintained the expression of GCTB markers. Based on these findings, we concluded that the PCL scaffold is an efficient model to culture GCTB cells, and the cell viability and adherence to the scaffold can be preserved for up to 14 days. Moreover, this model can also be used in subsequent studies to assess in vitro cell-cell interactions and antineoplastic efficacy of certain agents to establish a treatment against GCTB.

Catecholamine levels and gene expression of their receptors in tissues of adults with osteosarcoma.

AIM: The purpose of this work was to identify and measure catecholamines, their metabolites, and the gene expression of catecholamine receptors in osteosarcoma tissue. MATERIALS AND METHODS: The levels of 3,4-dihydroxyphenylacetic acid, norepinephrine, serotonin, and 5-hydroxyindoleacetic acid in cancer tissue and in adjacent and non-oncological bone tissue were analysed by high-performance liquid chromatography, and the gene expression of catecholamine receptors and of dopamine ß-hydroxylase, monoaminoxidase, ki67, and Runx2 in the osteosarcoma tissue, tissue adjacent to the tumour, non-oncological bone, and human brain tissue was analysed by RT-PCR. RESULTS: We found significantly higher levels of 3,4-dihydroxyphenylacetic acid and norepinephrine in the cancer sample than in adjacent and non-oncological bone. We found that ß-adrenergic receptors and dopaminergic receptors, dopamine ß-hydroxylase, ki67, Runx2, and serotonergic receptor gene expression were significantly higher in tumour tissue than in adjacent and non-oncological bone. CONCLUSION: Catecholamines and their receptors could be potential molecular markers for osteosarcoma progression.

MicroRNA expression in relation with clinical evolution of osteosarcoma.

Osteosarcoma is the most common malignant bone tumor. Early diagnosis remains a major challenge, mainly because of the lack of specific biomarkers. We performed miRNAs expression analysis through qPCR in affected and paired healthy bone derived from osteosarcoma patients. Hierarchical clustering using the top ten miRNAs with differential expression showed two main clusters. One integrated by patients with the presence of metastasis or relapse and the other without these complications. Further pathway enrichment analysis reduced to four main miRNAs, hsa-miR-486-3p, hsa-miR-355-5p, hsa-miR-34a-5p and hsa-miR-1228-3p. Afterwards, we compared patients with and without metastasis, the function enrichment analysis along with review of relevant literature, showed that hsa-miR-93-5p and hsa-miR-28-5p were associated with metastasis development. Our results support the relevance of miRNAs in the pathogenesis of osteosarcoma and contribute with evidence regarding the potential role of miRNAs as potential biomarkers. More studies are needed to define the most informative miRNAs in osteosarcoma patients.

Circulating miR-215-5p and miR-642a-5p as potential biomarker for diagnosis of osteosarcoma in Mexican population.

Osteosarcoma is the most common malignant bone neoplasia affecting individuals in the second decade of life. The survival rate has not been improved during the last 25 years, in part because of the lack of specific markers. The microRNAs have been identified as important regulators of gene expression, experimental evidence suggests these molecules as key players in cancer development and progression. To identify miRNAs differentially expressed in serum from patients with osteosarcoma compared to healthy donors in Mexican population. Fifteen osteosarcoma patients and fifteen age and sex matched healthy individuals were recruited. Two pools of total RNA extracted from serum per study group were prepared and the miRNA expression profiles were analyzed through TaqMan Low Density Arrays. Validation was carried out through RT-qPCR using individual TaqMan assays for those miRNAs differentially expressed. Fifteen miRNAs were differentially expressed in osteosarcoma patients compared to healthy controls. Overexpression of miR-215-5p and miR-642a-5p was confirmed by validation through RT-qPCR. The expression analysis of miRNAs from serum in osteosarcoma patients revealed differential expression of miR-215-5p and miR-642a-5p. Both microRNAs are potential markers for osteosarcoma diagnosis.

Dedifferentiated giant-cell tumor of bone with an undifferentiated round cell mesenchymal component.

The dedifferentiated giant-cell tumor of the bone is a very rare variant of the giant-cell tumor (GCT). We report the clinical, radiographic and histological findings of a dedifferentiated GCT in which the dedifferentiated component consisted of small round cells. We also comment on previously reported cases of dedifferentiated GCT, discuss the clinical implications of this dual histology, and analyze the information published about the coexistence of similar genetic abnormalities in GCT and small round cell tumors of the bone.

Diagnosis of a dedifferentiated chondrosarcoma of the pelvis by fine needle aspiration. A case report.

BACKGROUND: Dedifferentiated chondrosarcomas account for approximately 10% of all chondrosarcomas. There are few reports that describe the cytologic findings in dedifferentiated chondrosarcomas. In all these cases it was only present in the smears as a high grade sarcoma component. CASE: A 58-year-old man with a bone lesion in the right acetabular zone was admitted to the hospital. The clinical diagnosis was chondrosarcoma. A fine needle aspiration biopsy was performed. The smears of the sample had different characteristics in composition. Some slides had mostly abundant chondroid matrix with low to moderate cellularity. Other slides were cellular and contained mainly pleomorphic, spindled-shaped cells. The typical appearance of dedifferentiated chondrosarcoma was present in some slides that showed focal zones with chondroid matrix adjacent to or surrounded by pleomorphic spindled cells. The composition of the surgical specimen (slightly greater cartilaginous component than the sarcomatous one) allowed us to make a straightforward diagnosis of dedifferentiated chondrosarcoma. CONCLUSION: This is the first report that describes the presence of both components, high and low grade, by fine needle aspiration biopsy in dedifferentiated chondrosarcoma.

[Frequency of bone neoplasms in children]. / Frecuencia de neoplasias óseas en niños.

INTRODUCTION: Pediatric age is the group mainly affected by bone tumors. The objectives of this work were: (1) To determine which are the bone tumors and pseudotumoral lesions more frequent in the group of age from 0 to 14 years; (2) To determine if there are differences in the frequency of the tumors and pseudotumoral lesions between different subgroups and (3) To determine if there is any difference in the frequency of the different tumors according to gender. METHODS: We conducted a cross-sectional study based on the Pathology files of the National Institute of Rehabilitation. Inclusion criteria were: all cases between January 2000 and December 2006 diagnosed by hystopathological means as bone tumor or pseudotumoral lesion, with an age of presentation between 0 to 14 years were included. RESULTS: Frequency of bone tumors and pseudotumoral lesions in this series were: osteocondroma (44%), osteosarcoma (15.2%), fibrous dysplasia (10.4%) and encondroma (8.8%). We observed differences in the bone tumors or pseudotumor-like lesions frequency between the 3 subgroups of age studied.

[Bone tumor frequency in adults and elderly]. / Frecuencia de neoplasias óseas en adultos maduros y adultos mayores.

INTRODUCTION: We consider of great importance knowing the types of bone tumors and tumor-like lesions in adults and the elderly, which are increasing in proportion in our country. OBJECTIVES: (1) To analyze the frequency of bone tumors and tumor-like lesions in adults and elderly; (2) to determine any difference between these two groups; (3) and to analyze the most frequent sites of involvement. MATERIAL AND METHODS: We conducted a cross-sectional descriptive study that consisted in a retrolective review of all cases classified as a bone tumor or a tumorlike condition from year 2000-2006. We included all patients that presented with a bone tumor or tumor-like condition above 40 years-old. RESULTS: The most frequent diagnoses in patients between 40 and 60 years-old were plasmocytoma/myeloma (19.4%); giant cell tumor (14.1%) and metastases (12.3%). In patients older than 60 years, prevalence of bone tumor and tumor-like lesion was respectively: metastases (37.2%); plasmocytoma/myeloma (11.8%) and malignant fibrous histiocytoma (6.7%). Several differences were observed between these two groups.

[Prevalence of bone neoplasms in adolescents and young adults]. / Frecuencia de neoplasias óseas en adolescentes y adultos jóvenes.

INTRODUCTION: The objectives of this work were: (1) To determine the prevalence of bone neoplasms and pseudoneoplasms lesions in a group of adolescents (15 to 19 years), (2) To determine the more frequent bone neoplasms and pseudoneoplasms lesions in a group of young adults (29 to 39 years), (3) To determine if there are differences in the frequency of bone neoplasms and pseudoneoplasms lesions between the group of adolescents and the group of young adults and (4) To determine if there is any difference according to gender. MATERIAL AND METHODS: We conducted a descriptive, cross-sectional study that consisted in the review of the Pathology Department files in the National Institute of Rehabilitation. We included all patients with a histopathologic diagnosis of bone neoplasms or pseudoneoplasms lesion from January 2000 to December 2006 within the age of presentation from 15 to 19 or 20 to 39 years. RESULTS: The five more frequent bone neoplasms and pseudoneoplasms lesions in adolescents were osteochondroma, osteosarcoma, chondroblastoma, non-ossifying fibroma and aneurysmal bone cyst. In young adults the five more frequent were: giant cell neoplasm, osteosarcoma, osteochondroma, fibrous dysplasia and enchondroma.

[Partial internal hemipelvectomy for a secondary giant pelvic chondrosarcoma. A case report]. / Hemipelvectomía parcial interna por condrosarcoma secundario gigante de pelvis. Reporte de caso.

The purpose of this paper is to present the orthopedic-oncologic and functional outcomes of internal partial hemipelvectomy for a secondary giant pelvic chondrosarcoma in a patient with multiple hereditary osteochondromatosis.

[Results of the experimental repair of osteochondral lesions in a pig model using tissue engineering]. / Resultados en la reparación experimental de lesiones osteocondrales en un modelo porcino mediante ingeniería de tejidos.

OBJECTIVE: To repair experimental osteochondral knee lesions in pigs using tissue engineering. MATERIAL AND METHODS: Eight 40-kg pigs underwent surgery. Cartilage and periosteal biopsies of their control knee were taken. Cartilage and periosteal cells were independently isolated, cultured and seeded in biodegradable PGA and PLA polymers that were fixed on the bottom of an osteochondral defect in the pig's experimental knee, with bioabsorbable Mitek implants. Four months later the pigs were sacrificed and the knees were analyzed with nuclear magnetic resonance imaging (NMRI), macroscopic assessment, histology, electron microscopy (EM), scanning electron microscopy (SEM) and SEM element analysis. RESULTS: All the defects were filled with cartilage-like tissue according to the NMRI evaluation and the visual examination. Hyaline-like cartilage was obtained in 3 defects and fibrocartilage in 5. The EM showed chondrocytes in the repair tissue. The SEM showed appropriate integration to the bone and the surrounding tissue. SEM element analysis showed sulphurized matrix attached to the bone with calcium and phosphates as predominant elements. DISCUSSION: Tissue engineering enabled the production of tissues similar to normal ones. The polymer fixation system was effective.

[Epidemiology of bone and soft-part tumors of the foot and knee]. / Epidemiología de tumores óseos y partes blandas del pie y tobillo.

DESIGN: Report of cases. OBJECTIVE: Review of the cases in a period of 10 years with bone and soft tumors in foot and ankle, to knowing epidemilogical, clinic and patologic anatomy parameters to describe the behavior. MATERIALS AND METHODS: Review of 166 cases from 1991 to 2000 and been analyze with descriptive statistic, association measurment for inside stratum, with odds ratio, hipótesis test with chi square for qualitative date and t to Student for quantitative date. RESULTS: 166 patients within 2 years to 78 years old, 81 with bone tumors and 79 with soft tumors, mostly benign, the most frequent was in the soft tissue ganglion and oseal benign exostosis in bone, 6 different malignant tumors, the principal affected zone were the toes, as a difference to literature, the most affected age group was 10 to 20 years followed to the 30 to 40 years old, we report 42 different patological diagnostics results to soft tissue and osseous tissue. CONCLUSION: clinical features is not a useful parameter to differentiate between malign or benign tumors and does not allow to establish the biological behavior, we propose the diagnostic algorithm that includes the intentional clinical probe, comparative X-ray in three projections with soft technique, in suspicion to malignant lesion may require CT scan, MRI, osseous scan and finally biopsy which will improve the final outcome.

[Use of fibrin glue in combination with autologous bone graft as bone enhancer in posterolateral spinal fusion. An experimental study in New Zealand rabbits]. / Liofilizado de fibrina combinado con injerto óseo autólogo en artrodesis posterolateral. Estudio en conejos de Nueva Zelanda.

BACKGROUND: Currently there are different strategies to increase the fusion rate in spine surgery in the presence of autologous bone graft. The use of fibrin glue has multiple applications in surgery, but there is controversy about the use of fibrin glue as a bone enhancer. METHODS: The purpose of the study was to determinate the effectiveness of fibrin glue as a bone enhancer in posterolateral arthrodesis in New Zealand rabbits. Posterolateral arthrodesis was done in ten New Zealand rabbits at the level of L5-L6 using autologous bone graft in the right side (control side) and autologous bone graft plus fibrin glue in the left side (study side). The rabbits were harvested at 8 weeks, obtaining the lumbar spine for radiological, manual palpation and light microscopic analysis. RESULTS: Solid arthrodesis was obtained in 100% of the controls and in only 60% of the study animals. There were no differences among methods for determination of solid arthrodesis whether by radiological, manual palpation or light microscopic analysis. In 40% of non-unions, only in one (10%) was fibrocartilage obtained. In the remaining 30%, only inflammatory cells were obtained in the gap between the transverse process. CONCLUSIONS: Fibrin glue does not have a positive effect in the success of solid fusion in posterolateral arthrodesis in rabbits. The use of fibrin glue significantly decreased the rate of solid fusion; therefore, we do not recommend its use as a bone enhancer.

Accuracy of intraoperative consultation for bone tumors: experience in an orthopedic hospital.

BACKGROUND: Intraoperative consultation is an integral part of surgical pathology. However, there are few reports concerning intraoperative pathology consultation exclusively for bone tumors. METHOD: Our hospital is an orthopedic institution that specializes in the diagnosis and treatment of bone tumors, and we retrospectively reviewed the intraoperative consultations for bone tumors on our service from January 2002 to February 2006. The objectives of this work were to: (1) evaluate the clinical reasons for intraoperative consultation on bone tumors; (2) identify discordant diagnoses between the intraoperative consultation diagnosis and the definitive diagnosis; (3) determine in how many cases it was possible to establish a specific diagnosis; and (4) analyze the accuracy of the intraoperative consultation diagnosis on the bone tumors. RESULTS: The two most common reasons for an intraoperative consultation were to rule out malignancy and to determine the adequacy of the resection margins. The third most common reason was to establish a diagnosis. In the latter cases, it was possible to establish a specific diagnosis in 86.3% of the cases; and we had only one discordant case. CONCLUSIONS: Intraoperative consultation on bone tumors has problems and limitations. They are due mainly to the characteristics of the material that comprise the specimen from this type of lesion. It is important that similar assays be carried out at medical institutions that specialized in bone tumors to determine the effectiveness rates of intraoperative consultation in this field throughout the world. This is the first series that has analyzed the accuracy of intraoperative consultation on bone tumors with respect to the clinical reasons for the consultation.

IL-6 KO mice develop experimental amoebic liver infection with eosinophilia.

Interleukin 6 (IL-6) is a multifunctional cytokine that regulates various aspects of the immune response, such as acute phase reaction and hematopoiesis, and is an important signal that coordinates activities of liver cells, macrophages, and lymphocytes. Amoebic liver lesions have been studied, usually in hamsters, due to the problem of abscess development in mice. We report here the development of an experimental amoebic liver abscess (ALA) model in mice deficient in IL-6. Axenically grown amoebae were injected directly into the livers of C57BL/6 wild type (WT) and IL-6 KO -/- mice; the abscesses produced were counted and the inflammatory process was examined on 5, 10, and 20 days postinfection. Our results showed that IL-6 KO -/- mice develop ALA, in contrast to the WT strain, which usually do not have signs of abscess or infection. Histological analysis of the abscesses showed extended inflammatory response, mainly mediated by eosinophils, which strongly infiltrate the abscess in IL-6 K -/- mice. The present results suggest that in mice, IL-6 could play a role in the resistance against ALA.

Fine needle aspiration cytology of low grade fibromyxoid sarcoma. Report of a case with histologic correlation.

BACKGROUND: Low grade fibromyxoid sarcoma, first described in 1987, is a rare sarcoma characterized by a bland and deceptively benign histologic appearance but with aggressive behavior. CASE: A 51-year-old female presented with a history of a recurrent and slowly growing mass in the left foot. Fine needle aspiration biopsy showed an abundant myxoid background with occasional thick bands of collagen. Tumor cells present in the myxoid background were spindle shaped, with focally mild or a light degree of nuclear enlargement, hyperchromasia and pleomorphism CONCLUSION: Low grade fibromyxoid sarcoma has particular cytologic features. Besides a careful cytologic evaluation of all the components, clinical and radiographic correlation is necessary to make the correct diagnosis.

[Obesity and steatohepatitis. Histologic aspects]. / Obesidad y esteatohepatitis. Aspectos histológicos.

The traditional concept of adipose tissue as a passive reservoir for energy storage is no longer valid because it has been demonstrated that adipose tissue is a complex, essential, and highly active metabolic and endocrine organ that not only responds to afferent signals from traditional hormone systems and the central nervous system (CNS), but also expresses and secretes factors with important endocrine functions. These factors include leptin and other cytokines. Adipose tissue is also a major site for metabolism of sex steroids and glucocorticoids. The important endocrine function of adipose tissue is emphasized by adverse metabolic consequences of both adipose tissue excess and deficiency. Adipose tissue excess, particularly in visceral compartment, is associated with insulin resistance, hyperglycemia, dyslipidemia, hypertension, and prothrombotic and proinflammatory states. Liver is one of the principal targets of lipid-associated damage by mechanisms that involve apoptosis activation by source of tumoral necrosis factor-alpha and caspase activation and liberation of oxygen-reactive species by oxidative stress and enzymatic chains such as P450, CYP2E1, and CYP3A4, resulting in a continuum involving non alcohol-related fatty liver, non-alcoholic steatohepatitis with or without fibrosis, and liver cirrhosis. This work presents an overview of endocrine functions of adipose tissue and its influence on mechanisms of liver damage.

Efecto de la metilprednisolona en la amibiasis hepática experimental / Effect of methylprednisolone in experimental amebic liver abscess

Para examinar si la extensión de la necrosis tisular en la amibiasis hepática experimental en hamsters es proporcional a la magnitud de la respuesta inflamatoria inicial se comparó la evolución morfológica de las lesiones en presencia y en ausencia de dosis altas de metilprednisolona a las 10 hrs, 24 hrs, 72 hrs y 7 días. Se observó en los animales con metilprednisolona una respuesta inflamatoria inicial menor que en los controles y una extensión de las lesiones menor a los 7 días de evolución. La metilprednisolona no altera ni la actividad proteolítica ni la eritrofagocitosis de Entamoeba histolytica in vitro.