Shedding Light on Osteosarcoma Cell Differentiation: Impact on Biomineralization and Mitochondria Morphology.

Osteosarcoma (OS) is the most common primary malignant bone tumor and its etiology has recently been associated with osteogenic differentiation dysfunctions. OS cells keep a capacity for uncontrolled proliferation showing a phenotype similar to undifferentiated osteoprogenitors with abnormal biomineralization. Within this context, both conventional and X-ray synchrotron-based techniques have been exploited to deeply characterize the genesis and evolution of mineral depositions in a human OS cell line (SaOS-2) exposed to an osteogenic cocktail for 4 and 10 days. A partial restoration of the physiological biomineralization, culminating with the formation of hydroxyapatite, was observed at 10 days after treatment together with a mitochondria-driven mechanism for calcium transportation within the cell. Interestingly, during differentiation, mitochondria showed a change in morphology from elongated to rounded, indicating a metabolic reprogramming of OS cells possibly linked to an increase in glycolysis contribution to energy metabolism. These findings add a dowel to the genesis of OS giving new insights on the development of therapeutic strategies able to restore the physiological mineralization in OS cells.

Mass Spectrometric-Based Proteomics for Biomarker Discovery in Osteosarcoma: Current Status and Future Direction.

Due to a lack of novel therapies and biomarkers, the clinical outcomes of osteosarcoma patients have not significantly improved for decades. The advancement of mass spectrometry (MS), peptide quantification, and downstream pathway analysis enables the investigation of protein profiles across a wide range of input materials, from cell culture to long-term archived clinical specimens. This can provide insight into osteosarcoma biology and identify candidate biomarkers for diagnosis, prognosis, and stratification of chemotherapy response. In this review, we provide an overview of proteomics studies of osteosarcoma, indicate potential biomarkers that might be promising therapeutic targets, and discuss the challenges and opportunities of mass spectrometric-based proteomics in future osteosarcoma research.

Cytological diagnosis of osteosarcoma with emphasis on diagnostic pitfalls.

INTRODUCTION: Fine needle aspiration is a well-established technique for evaluating primary and secondary bony lesions. With use in selected cases, it achieves a diagnostic yield comparable to biopsies. METHODS: Cases of osteosarcoma (OS) with available histological follow-up were retrieved over a 10-year period. Detailed morphological evaluation was done, with special emphasis on pitfalls in the diagnosis of OS on cytology and the various variants of OS. RESULTS: Of the 41 cases with available follow-up histology, 56% were correctly diagnosed as OS on cytology. The most common false-negative cytological diagnosis of OS, in 17% cases, was giant cell tumour. The possible explanations for this included low cellularity, minimal atypia, absence of typical osteoid, misinterpretation of metachromatic osteoid material as fibro-collagenous material and non-availability of radiology at time of aspiration. CONCLUSION: A triple-phase evaluation including clinical evaluation, appropriate radiological correlation and cytology/histopathology, is important to clinch an accurate diagnosis.

In vitro and in vivo studies of antiosteosarcoma activities of formononetin.

Osteogenic sarcoma (OGS) is a primary bone cancer, characterized by aggressive neoplasm from mesenchymal oncogenesis. However, the clinical therapeutic regimen against OGS is limited. Therefore, potential medication warrants to be further developed. Our previous study indicates that formononetin (FN) exerts effective pharmacological activity against OGS. This study aimed to further decipher the molecular mechanism behind this benefit. Patients with OGS were recruited for clinical data assay and immunoassay. Human OGS cell line (U2OS) and tumor-bearing nude mice were subjected to a battery of biochemical analyses and immunoassays for integrative evaluation of FN-exerted anti-OGS effects. In human data, OGS samples showed increased expressions of ERα, p-PI3KCATyr317 , and p-AKT Ser473 proteins, followed by notably upregulated miR-375 content in comparison with that in OGS-free. In addition, FN-treated U2OS cells showed inhibited cell proliferation, elevated lactic dehydrogenase production and lowered endogenous miR-375 level in cells. Further, reduced immunopositive cells of Ki-67, p-PI3KCA Tyr317 , and p-AKT Ser473 were observed by the treatments of FN, while the intracellular Bax- and Apaf-1-positive cells were increased dose-dependently. Beneficially, FN-treated tumor-bearing mice exhibited reduced tumor mass and intercellular miR-375 expression. Meanwhile, immuno-labeled cells and proteins of Bax, Caspase-3, and Apaf-1 in FN-treated mice were increased dose-dependently, whereas ERα, p-PI3KCA Tyr317 , and p-AKT Ser473 positive cells and proteins were downregulated, respectively. Collectively, our current results elucidate that FN exerts effective therapeutic benefits against OGS, and the pharmacological mechanism may be related to promoting cell apoptosis by inactivating intracellular miR-375/ERα-PI3K/AKT cascaded pathway.

OSteosarcoma of Ethmoid Sinus-A Rare Entity.

Osteogenic sarcoma of the Craniofacial region are uncommon.They have been mainly reported in the mandible and the maxilla, Primary osteosarcoma arising from ethmoid sinus is an extremely rare condition. We report a case of 34 year old male, presented with gradually increasing swelling over the right medial canthus with history of double vision for one year. Clinically we thought it was a mucous retention cyst in the ethmoid area. After doing Contrast enhanced CT, we proceeded with transnasal endoscopic removal of the cyst. Histopathological examination of the specimen surprisingly turned out to be a low grade Osteogenic sarcoma. Patient was treated with adjuvant chemotherapy.

Extra-Skeletal Osteosarcoma of the Prostate After Treated Prostatic Acinar Adenocarcinoma: A Case Report and Review of the Literature.

Extra-skeletal osteosarcoma is a rare form of malignant soft tissue sarcoma. Its occurrence in the prostate gland is particularly uncommon. In this case report, we present a patient diagnosed with osteosarcoma arising within the prostatic gland. A 58-year-old man was initially diagnosed with Gleason 8 prostate acinar adenocarcinoma following a transurethral resection (TUR) of the prostate. This diagnosis was accompanied by locoregional involvement and multiple bone metastases. The patient underwent a treatment regimen including complete androgen blockade, chemotherapy, greenlight laser prostate vaporization, and palliative radiotherapy. After treatment, he achieved a complete biochemical response, and his bone metastases remained stable. However, at 16 months post-diagnosis, clinical follow-up by means of radiological examinations revealed an increase in the size of the prostatic lesion, along with additional infiltration of the tumor into the rectum and bladder walls. Remarkably, a mesenchymal tumor proliferation with intratumor calcifications was observed. A subsequent TUR biopsy of the prostate showed a malignant tumor spindle and ovoid cell proliferation with high-grade nuclear atypia, necrosis, and islets of osteoid formation, leading to a final diagnosis of high-grade prostatic extra-skeletal osteosarcoma. Despite undergoing chemotherapy, the patient's condition progressed with the development of pulmonary and liver metastases, culminating in his demise.

The whole transcriptome analysis using FFPE and fresh tissue samples identifies the molecular fingerprint of osteosarcoma.

Osteosarcoma is a form of bone cancer that predominantly impacts osteoblasts, the cells responsible for creating fresh bone tissue. Typical indications include bone pain, inflammation, sensitivity, mobility constraints, and fractures. Utilising imaging techniques such as X-rays, MRI scans, and CT scans can provide insights into the size and location of the tumour. Additionally, a biopsy is employed to confirm the diagnosis. Analysing genes with distinct expression patterns unique to osteosarcoma can be valuable for early detection and the development of effective treatment approaches. In this research, we comprehensively examined the entire transcriptome and pinpointed genes with altered expression profiles specific to osteosarcoma. The study mainly aimed to identify the molecular fingerprint of osteosarcoma. In this study, we processed 90 FFPE samples from PathWest with an almost equal number of osteosarcoma and healthy tissues. RNA was extracted from Paraffin-embedded tissue; RNA was sequenced, the sequencing data was analysed, and gene expression was compared to the healthy samples of the same patients. Differentially expressed genes in osteosarcoma-derived samples were identified, and the functions of those genes were explored. This result was combined with our previous studies based on FFPE and fresh samples to perform a meta-analysis. We identified 1,500 identical differentially expressed genes in PathWest osteosarcoma samples compared to normal tissue samples of the same patients. Meta-analysis with combined fresh tissue samples identified 530 differentially expressed genes. IFITM5, MMP13, PANX3, and MAGEA6 were some of the most overexpressed genes in osteosarcoma samples, while SLC4A1, HBA1, HBB, AQP7 genes were some of the top downregulated genes. Through the meta-analysis, 530 differentially expressed genes were identified to be identical among FFPE (105 FFPE samples) and 36 fresh bone samples. Deconvolution analysis with single-cell RNAseq data confirmed the presence of specific cell clusters in FFPE samples. We propose these 530 DEGs as a molecular fingerprint of osteosarcoma.

Jaw Fibro-Osseous Lesions: Use of a Predictive Index in Grading Probable Malignant Changes and a Review of Cases.

Statement of the Problem: Fibro-osseous lesions (FLs), may rarely exhibit malignant features likewise undergo malignant transformation. Awareness of these features can assist in screening for potentially malignant cases and identifying low-grade central osteogenic sarcoma (LGCOS) that may mimic FLs. Purpose: The objective of this study was to determine the usability of an index in predicting malignant changes in jaw FLs. Materials and Method: This was a retrospective study where hematoxylin and eosin (H&E) slides and archival records of fibrous dysplasia (FD) and ossifying fibroma (OF) cases were reviewed. The sections were assessed for permeation of marrow spaces, stromal growth pattern, cytologic atypia, mitotic activity, and pattern of bone growth, which are parameters for diagnosing LGCOS. The predictive histologic index of malignancy (PHIM) was determined by a sum of the scores and graded as 0=nil, 1=low, 2 & 3=moderate, and 4 & 5=high. Data were presented using descriptive analysis. Results: Ninety-three cases of FLs met the inclusion criteria, consisting of 40(43%) cases of FD and 53(57%) cases of OF. The peak age of presentation for FD and OF was 2nd and 3rd decade. There was a female preponderance of 1:1.6. The maxilla was the most common site affected by FD, while the mandible was most commonly affected by OF. For FD cases, the PHIM was moderate in 10(25%) cases and low in 21(52.5%) cases. Similarly, for OF cases, 30(56.6%) cases had low grade PHIM while 10(17%) cases had moderate grade PHIM. Conclusion: The PHIM depicted low to moderate malignancy grade in some of the cases studied. Follow up studies would be necessary to assess the PHIM.

Role of bone scintigraphy (bone scan) in skeletal osteosarcoma: A retrospective audit and review from tertiary oncology centre.

Introduction: Bone scan is a investigation which uses radionuclide phosphonate compound for whole skeletal survey. In this current study we have done the analysis of the role of bone scan in skeletal osteosarcoma at tertiary oncology care centre. Material & methods: This is a retrospective study conducted in a tertiary oncology centre from January 2022 to February 2023. A total of 92 patients with skeletal OGS were included in our study undergone 99 mTCcMDP whole body bone scan. 99 mTc MDP was prepared freshly every morning and dose for each patient were calculated as per EANM guidelines. Images were acquired 2-3 h of post injection. All images were acquired at GE infinia dual head machine with peak setting at 140Kev and LEAP collimator. Suspicious lesions on planer bone scan were correlated with SPECT fused with CT. All the bone scans were reviewed retrospectively by two independent nuclear medicine physicians. Results: In this study group, 86 patients with biopsy proven skeletal OGS underwent 99 mTCcMDP bone scan of which 63 were males and 23 were females (2.7:1) with age of study group ranging from 7years to 48years. Patients referred for bone scan were retrospectively categorized in two groups, first group patients (52) were referred for initial staging of disease and second group of patients (34) were referred for follow-up or re-staging of the disease. Total 09 patients showed distant skeletal metastases on bone scan, out of which 05 were in initial staging group and 04 in follow up group. Conclusion: Osteosarcoma has propensity to metastasize to many sites in the body however most common site being lung followed by skeletal, nodal and rarely soft tissue metastasis. Bone scan enjoys a optimal sensitivity in case of osteosarcoma to detect skeletal metastasis but have low specificity. However being a cost effective and faster investigation makes it a wise investigation of choice in case of osteosarcoma for skeletal metastasis evaluation.

Long term survival in adult osteosarcoma patients treated with a two-drug regimen: Final results of the OSAD93 phase II study of the FSG-GETO.

RATIONALE: We report a phase II trial (OSAD93) testing CDDP with ifosfamide (IFO), without doxorubicin in neoadjuvant phase, in adult osteosarcoma with a 25 years follow-up. PATIENTS AND METHODS: This is a multicentric phase II study of neoadjuvant chemotherapy with IFO and CDDP in localized high-grade osteosarcoma of patients. Patients received 4 pre-operative courses of IFO 9 g/m2 and CDDP 100 mg/m2 on day 4 (SHOC regimen), followed by local treatment. Doxorubicin was added post-operatively (HOCA regimen) in patients with > 10 % residual tumor cells. A Good Histological Response (GHR), ie ≤ 10 % residual tumor cells in > 30 % of patients, was the primary objective. Disease-free survival (DFS), overall survival (OS) and toxicity were secondary objectives. RESULTS: From Jan 1994 to Jun 1998, 60 patients were included. Median age was 27 (range: 16-63). Primary tumor sites were limbs (76 %), trunk, head or neck (24 %). After neoadjuvant SHOC, grade 3-4 and febrile neutropenia, thrombopenia, and re-hospitalization occurred in 58 %, 17 %, 17 % and 22 % of SHOC courses and in 76 %, 28 %, 47 %, 47 % of HOCA courses, respectively. GHR was obtained in 16/60 (27.5 %) patients. With a median follow-up of 322 months, the DFS and OS were 51.8 % and 64.4 % at 5 years. At 10 years, DFS and OS were 49.9 % and 64.4 %. At 25 years, DFS and OS were 47.8 % and 55.9 %. No long-term cardiac toxicity was observed. Three patients developed a second malignancy (one fatal) after 300 months. CONCLUSION: Though the primary endpoint of OSAD93 was not met, this pre-operative doxorubicin-free regimen led to excellent long-term survival with limited toxicity in localized osteosarcoma.

Serum Osteopontin as a Potential Marker for Metastasis and Prognosis in Primary Osteogenic Sarcoma: A Systematic Review.

Osteosarcoma (OS), a primary malignant bone tumor, poses significant challenges in diagnosis and prognosis. It is a painful medical burden, and treating it is still a difficult issue. Osteopontin (OPN), a multifunctional extracellular matrix protein, has emerged as a promising biomarker in this context. This systematic review explores the role of OPN as a diagnostic and prognostic marker in OS, highlighting its potential in enhancing early detection, monitoring disease progression, and predicting patient outcomes. Various studies have demonstrated elevated levels of OPN in OS patients, correlating with tumor aggressiveness, metastatic potential, and poor prognosis. In addition, OPN's involvement in tumor microenvironment regulation and metastatic processes underscores its clinical relevance as a biomarker. For this systematic review, comprehensive literature searches were conducted in the PubMed databases for research published between the database's establishment and November 11, 2022. Out of the nine studies that were available for analysis, a higher level of OPN in primary osteogenic sarcoma patients indicates a poorer prognosis and higher incidence of metastasis. OS has not shown commensurable progress with concerns to treatment approches and survical outcomes. However, the discovery of a biological marker that can predict metastasis and severity will be a groundbreaking development for advancements in OS diagnosis and treatment. Therefore, understanding the intricate interplay between OPN and OS pathogenesis holds promise for improving patient management and developing targeted therapeutic strategies.

Beyond the bones: Extraskeletal osteosarcoma of the thigh.

Extraskeletal osteosarcoma (ESOS) is a rare malignant mesenchymal soft tissue tumor that usually arises in the lower extremities. It is typically a high-grade malignancy that represents only around 1%-2% of all soft tissue sarcomas and 2%-4% of all osteosarcomas. In this report, we describe a case of a 67-year-old female who presented with a 4-day history of a painless lump in her posterior right thigh. Workup utilizing different imaging modalities yielded a diagnosis of ESOS. The radiologic features of ESOS, as well as the current treatment paradigm and prognosis of this rare malignancy, will be discussed based on a review of the literature.

First case of medullary osteogenic sarcoma of the pelvis: 12-year follow-up of reconstruction with hemipelvis allograft after resection.

A 45-year-old male patient with low-grade central osteosarcoma (LGCO) in the periacetabular region underwent wide resection, fresh frozen hemipelvis allograft reconstruction, and total hip prosthesis. To the best of our knowledge, this case is the first example of low-grade osteogenic sarcoma in flat bones. Aseptic loosening of the acetabular cup was observed 44 months after the operation, and it was revised with a constrained acetabular cup. Recurrent subluxation due to constraint ring failure and cup malposition was observed at 89 months after the revision surgery. Revision operation was performed with cage + non-constrained cup. Twelve years after the first operation, he is in exceptionally functional and disease-free condition. He can walk unlimited distances without pain. Radiographs show complete union at the junction of the allograft and disease-free bone.

Limb Salvage Surgery in Osteosarcoma of the Femur Using Liquid-Nitrogen-Frozen Free Femur Autograft in a Developing Country.

Introduction: Limb salvage is a technically demanding and capital intensive procedure. In low-income countries, amputation followed by prosthesis use is widely used for the treatment of limb malignancies. This is associated with considerable morbidity and frequent non-adherence to prosthetic use. Therefore, we are reporting a case of osteosarcoma of the femur treated by the use of pre-operative chemotherapy, followed by frozen free femur autograft and intramedullary nailing, with excellent radiologic and functional results at 1 year of follow-up, in a developing country. Case Report: A 16-year-old female presented with the left thigh swelling and pain for 3 months. X-ray of the left femur showed an osteosclerotic tumor in the medial aspect of the middle third of the left femur, diagnosed as osteogenic sarcoma on biopsy. Pre-operative chemotherapy was given. En bloc resection of the tumor was done. The cleaned femur shaft was frozen in liquid nitrogen, reimplanted, and stabilized with an intramedullary nail. One year after surgery, the femur osteotomy sites are fully united and the patient is fully ambulant, without evidence of metastasis or recurrence. Conclusion: This case describes the successful use of a free frozen autograft of the femur in a 16-year-old girl with osteosarcoma of the left femur, done in a developing country, Kenya. It demonstrates the feasibility of limb salvage using this technique in low-income countries.

Osteosarcoma of the Distal Femur Presenting as Giant Cell Tumor: A Case Report.

The most frequent benign bone tumor, known as a giant cell tumor (GCT), typically develops in the second and third decades of life. GCTs of the bone that have already been diagnosed and have already undergone treatment with denosumab therapy, curettage or excision, or radiotherapy frequently develop malignant transformation. A very uncommon occurrence involves a GCT of the bone that has always been malignant. Here, we describe the case of a 25-year-old man with a large cell tumor of the distal femur discovered after six months of symptom onset. The MRI suggested an aneurysmal bone cyst or subsequent modifications of an aneurysmal bone cyst in a GCT. A biopsy was performed, and the results pointed to a benign GCT of the bone. There were no pleomorphic or hyperchromatic lesions, unusual mitoses, or cellular atypia. The patient was treated with tumor removal and internal fixation using plate osteosynthesis two weeks later. The samples were sent for histopathology. The report was suggestive of osteogenic sarcoma or malignant transformation of the giant cell. This could happen due to the possibility of a biopsy sample being taken from an area not representative of the tumor site, which is not uncommon since the osteosarcoma also contains areas of conventional GCT. Thus, osteosarcoma usually mimics conventional GCT of the bone.

[Synchronic multicentric osteosarcoma. A case in a 10 year old boy and review of the literature]. / Osteosarcoma multicéntrico sincrónico. Un caso en niño de 10 años y revisión de la literatura.

The osteosarcoma is a malignant neoplasm originating in bone-producing cells whose usual presentation is predominantly monostotic in long bones. The polyostotic expression is rare, variable and controversial, predominantly high-grade cellular lesions in children; low-grade lesions that affect long bones in adults and others that show a dominant lesion in long bones from which metastases arise. We present the case of a ten-year-old patient with painful tumors in metaphasis and diaphysis of the femur, tibia, fibula, clavicle, humerus, as well as in pelvis, vertebrae and base of the skull of synchronous appearance with blindness. Diagnosis was made through X-ray images, CT scan, magnetic resonance imaging (MRI), and incisional biopsy of the femur. Evidenced of absence of neoplastic manifestation in lung imaging, at the time of pathological diagnosis. Histological findings revealed a high-grade osteosarcoma.

Osteogenic Sarcoma Presenting with Skip, Lymph Nodal, Pulmonary, Pleural Metastases and Malignant Effusion: An Unusual Appearance on Bone Scan.

Osteogenic sarcoma is an aggressive malignant bone tumor arising from primitive mesenchymal bone-forming cells. Ossification is the characteristic feature of the metastases in osteosarcoma which aids their detection on 99mTc-MDP bone scintigraphy. Although lung is the most common site of metastasis in osteogenic sarcoma, pleural involvement with effusion and skip metastases are rare. Herein, we report a case of osteogenic sarcoma of distal femur with diffuse calcified pleural thickening leading to malignant pleural effusion, calcified lung, lymph node, and bone metastases illustrated on bone scan.

Sinonasal Osteosarcoma: Report of 14 New Cases and Systematic Review of the Literature.

Objective The objective of this study is to describe the clinical presentation, tumor characteristics, natural history, and treatment patterns of sinonasal osteosarcoma. Methods Fourteen patients who had been treated for osteosarcoma of the nasal cavity and paranasal sinuses at a tertiary care center were reviewed. In addition, a systematic review of the literature for osteosarcoma of the sinonasal cavity was performed. Results In a systematic review, including 14 patients from the authors' institution, 53 total studies including 88 patients were assessed. Median follow-up was 18 months (interquartile range: 8-39 months). The most common presenting symptoms were facial mass or swelling (34%), and nasal obstruction (30%). The most common paranasal sinus involved by tumor was the maxillary sinus (64%), followed by the ethmoid sinuses (52%). The orbit (33%), dura (13%) and infratemporal fossa (10%) were the most common sites of local invasion. The majority of patients underwent surgery followed by adjuvant therapy (52.4%). Increasing age was associated with decreased overall survival rate (unit risk ratio [95% confidence interval (CI)] = 1.02 [1.003-1.043]; p = 0.0216) and T4 disease was associated with decreased disease-specific survival rate (hazard ratio [HR] = 2.87; p = 0.0495). The 2- and 5-year overall survival rates were 68 and 40%, respectively, while 2- and 5-year disease-specific survival rates were 71% and 44%, respectively. Conclusion Sinonasal osteosarcomas are uncommon tumors and can pose a significant therapeutic challenge. Increasing age and T4 disease are associated with worse prognosis. This disease usually warrants consultation by a multidisciplinary team and consideration of multimodality therapy.

Unusual Presentation of Widely Metastatic Extraskeletal Osteosarcoma: Case Report.

Extraskeletal osteosarcoma is a highly aggressive malignant osteoid forming mesenchymal neoplasm arising from soft tissues which accounts for 1% of all soft tissue sarcomas. We report the case of a 46-year-old female with no significant past medical history presenting to an emergency department with a right lateral thigh mass following minor trauma. She was eventually found to have high grade extraskeletal osteosarcoma with rapid progression of disease resulting in patient demise. Differentiation of these lesions from alternative processes relies on specific imaging and pathologic features. Differential diagnoses include both benign and malignant etiologies such as myositis ossificans, soft tissue hemangiomas, and other malignant soft tissue neoplasms such as epithelial and synovial sarcoma.

Dural Metastasis in Spinal Canal Diagnosed on F-18 Fluorodeoxyglucose Positron Emission Tomography Contrast-Enhanced Computed Tomography Scan in Treated Case of Osteogenic Sarcoma of the Left Femur.

We describe a rare case of osteogenic sarcoma of the right femur; who 2 years after the treatment of primary site developed dural metastasis. Surveillance imaging for unusual pattern of metastasis may lead to earlier detection and treatment decision-making, which may improve survival and quality of life.