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Biomedical subjects

A Myoui

Publications and source records attributed to A Myoui.

At least 19 recordsLinked to original sources

Comparison of the activities of multinucleated bone-resorbing giant cells derived from CD14-positive cells in the synovial fluids of rheumatoid arthritis and osteoarthritis patients.

OBJECTIVE: To investigate the morphology and function of multinucleated bone-resorbing giant cells derived from CD14-positive cells in the synovial fluids (SF) of patients with rheumatoid arthritis (RA) or osteoarthritis (OA). METHODS: CD14-positive cells were obtained by magnetic-activated cell sorting of primary cultures of mononuclear cells from the SF. Multinucleated bone-resorbing giant cells were induced from the CD14-positive cells in the presence or absence of cytokines. We examined various characteristics, including osteoclast markers, fusion index and bone-resorption activities of the multinucleated giant cells. RESULTS: Multinucleated giant cells were induced from the CD14-positive cells in the SF of the RA and OA patients by the addition of interleukin (IL)-3, IL-5 and IL-7, or granulocyte-macrophage colony-stimulating factor (GM-CSF), respectively. These multinucleated giant cells were positive for tartrate-resistant acid phosphatase (TRAP), carbonic anhydrase II, actin, vitronectin receptor and the calcitonin receptor. However, the average values for the number of nuclei, fusion index and bone-resorption functions of the SF cells from the RA patients were significantly higher than those derived from the OA patients. CONCLUSION: These results suggest that the induction and activities of multinucleated bone-resorbing giant cells may play a pivotal role in bone destruction, and that these processes may be enhanced significantly in RA patients.

Adult↗

Calcium hydroxyapatite ceramic implants in bone tumour surgery. A long-term follow-up study.

We reviewed the results of 51 patients with benign bone tumours treated by curettage and implantation of calcium hydroxyapatite ceramic (CHA). The mean follow-up was 11.4 years (10 to 15.5). Post-operative fractures occurred in two patients and three had local recurrences; three had slightly limited movement of the adjacent joint and one had mild osteoarthritis. There were no allergic or neoplastic complications. In all cases, radiographs showed that the CHA was well incorporated into the host bone. Statistical analysis showed that absorption of the implanted CHA was greater in males (odds ratio, 6.2; 95% CI, 1.6 to 23.7) and younger patients (odds ratio, 0.6 for increase in age of 10 years; 95% CI, 0.91 to 0.99). However, the implanted CHA was not completely absorbed in any patient. We conclude that CHA is a useful and safe bone substitute for the treatment of benign bone tumours.

Adolescent↗

Expression and distribution of transcripts for sonic hedgehog in the early phase of fracture repair.

Localization and expression of mRNAs for sonic hedgehog (Shh) at a fracture site in the early phase postfracture were investigated by in situ hybridization and reverse transcription and polymerase chain reaction (RT-PCR). A closed fracture was made in the midshaft of the right tibia of 5-week-old ICR mice, and fractured sites were harvested prefracture (day 0) and on days 2 and 12. In situ hybridization revealed that transcripts for Shh were not detected on day 0, but they were detected in proliferating callus-forming cells in the periosteum and the surrounding tissue, and in the medullary cavity prior to apparent new cartilage and bone formation. Gli 1 (a signaling mediator for Shh) and bone morphogenetic protein-4 transcripts were colocalized with those for Shh transcripts on day 2. The RT-PCR showed that Shh mRNA was detected in the PCR product from day 2, but not from days 0 and 12. These findings are the first description about the activation of Shh gene in the early postfracture reaction.

Animals↗

Localization of bone morphogenetic protein-2 in human osteoarthritic cartilage and osteophyte.

OBJECTIVES: To examine the localization of bone morphogenetic protein (BMP)-2 mRNA and protein in human osteoarthritic (OA) articular cartilage and osteophyte. DESIGN: Five normal, four growing and 14 OA human cartilage samples, graded histomorphologically by Mankin Score, were studied by in situ hybridization and immunohistochemistry for the expression of BMP-2. RESULTS: BMP-2 mRNA was present in chondrocytes in neonatal growing articular cartilage, but was scarcely present in normal adult articular cartilage. In OA articular cartilage, BMP-2 mRNA and protein were detected in both clustering and individual chondrocytes in moderately or severely damaged OA cartilage. In moderately damaged OA cartilage, BMP-2 mRNA was localized in both upper and middle zone chondrocytes, but was not detected in deep layer chondrocytes. In severely damaged OA cartilage, cellular localization of BMP-2 mRNA was extended to the deep zone. In the area of osteophyte formation, BMP-2 mRNA was intensely localized in fibroblastic mesenchymal cells, fibrochondrocytes, chondrocytes and osteoblasts in newly formed osteophytic tissue. The pattern of BMP-2/4 immunolocalization was associated with that of mRNA localization. CONCLUSIONS: BMP-2 mRNA and BMP-2/4 were detected in cells appearing in OA tissues. BMP-2 was localized in cells of degenerating cartilage as well as osteophytic tissue. Given the negative localization of BMP-2 in normal adult articular cartilage, BMP-2 might be involved in the regenerating and anabolic activities of OA cells, which respond to cartilage damage occurring in osteoarthritis.

Aged↗

Frequent expression of smooth muscle markers in malignant fibrous histiocytoma of bone.

BACKGROUND/AIMS: Malignant fibrous histiocytoma (MFH) of bone, a relatively rare primary malignant bone tumour, is a distinct clinicopathological entity as opposed to MFH derived from soft tissue. Although the true histogenesis of this condition is still controversial, a considerable number of cases of MFH in soft tissue show positive immunohistochemical reactivity for muscle markers such as desmin, common muscle actin (HHF35), and alpha smooth muscle actin (SMA), suggesting that MFH cells are myofibroblastic in nature. METHODS: This study investigated immunoreactivity for several different muscle markers in 19 cases of MFH of bone together with reverse transcription polymerase chain reaction (RT-PCR) analysis on frozen tissue samples that were available in four cases, and compared the data with those found in 11 cases of osteosarcoma and 11 cases of soft tissue MFH treated over the same period. RESULTS: Immunohistochemistry revealed that MFH of bone showed relatively frequent expression of smooth muscle markers, including calponin (nine cases), alpha-SMA (nine cases), and SM22alpha (18 cases), and this was confirmed by RT-PCR analysis. However, only one, two, and three cases of MFH of bone showed positive staining for desmin, MyoD1, and HHF35, respectively. Similarly, 11 osteosarcoma cases were relatively frequently positive for alpha-SMA (five cases), calponin (four cases), and SM22alpha (seven cases), and less frequently positive for desmin (one case), MyoD1 (none), and HHF35 (none). In contrast, very few MFH of soft tissue cases (n = 11) showed positive reactivity for all of these muscle markers. It has recently been reported that human bone marrow stromal cells also express various kinds of smooth muscle markers including alpha-SMA and calponin. CONCLUSIONS: These results suggested that MFH of bone may derive from mesenchymal stromal cells in bone marrow and has a more myofibroblastic differentiation than soft tissue MFH.

Actins↗

Mechanisms of age-related decline in insulin-like growth factor-I dependent proteoglycan synthesis in rat intervertebral disc cells.

STUDY DESIGN: Age-related fluctuations in insulin-like growth factor-I dependent proteoglycan synthesis in rat intervertebral disc cells were investigated. OBJECTIVES: The purpose of this study was to determine whether synthetic responses to insulin-like growth factor-I decline with age and to explore the possibility that an age-related increase in the expression of insulin-like growth factor binding proteins suppresses matrix synthesis in intervertebral disc cells. SUMMARY AND BACKGROUND DATA: Several studies have reported that the responsiveness of chondrocytes to insulin-like growth factor-I decreases with age and furthermore that this phenomenon may be related to increased expression of insulin-like growth factor binding proteins by chondrocytes. MATERIALS AND METHODS: Nucleus pulposus tissue and cells were obtained from the coccygeal vertebrae of 8-week-old, 40-week-old, and 120-week-old rats. Age-related changes in the expression of insulin-like growth factor-I and its receptor were assessed together with insulin-like growth factor-I dependent proteoglycan synthesis by the cultured nucleus pulposus cells. Also, western blot analysis of insulin-like growth factor binding protein-1 was carried out, and further examination was performed of insulin-like growth factor-I signal transduction through tyrosine phosphorylation of insulin receptor substrate-1, which is a signal transducer of insulin-like growth factor-I. RESULTS: Semiquantitative reverse transcription polymerase chain reaction analysis indicated that the expression of insulin-like growth factor-I receptor in 120-week cells decreased clearly in comparison with the cells of younger animals. By contrast, insulin-like growth factor-I dependent proteoglycan synthesis decreased with age, and the sharpest decline of synthesis was found between 8-week and 40-week cells, although the level of insulin-like growth factor-I/insulin-like growth factor-I receptor gene expression was maintained in 40-week-old animals. Consistent with the results of proteoglycan synthesis, the expression of phosphorylated insulin receptor substrate-1 decreased with age. Thus, the expression of insulin-like growth factor binding protein-1 and proteoglycan synthesis was investigated by use of Long R3 insulin-like growth factor-I, which was not influenced by insulin-like growth factor binding proteins. Insulin-like growth factor binding protein-1 was strongly expressed in 40-week cells in comparison with the expression in 8-week cells. Furthermore, proteoglycan synthesis in 40-week cells supplemented with Long R3 insulin-like growth factor-I was upregulated in comparison with that in 40-week cells supplemented with insulin-like growth factor-I. CONCLUSION: The present findings indicate that the age-related decline in insulin-like growth factor-I dependent proteoglycan synthesis in nucleus pulposus is caused, at least in part, by the increase in insulin-like growth factor binding proteins at the early stages of aging, and further suggest that a loss of proteoglycan synthesis during the late stages of aging is caused by the downregulation of insulin-like growth factor-I receptor in addition to an increase in insulin-like growth factor binding proteins.

Aging↗

Ossification of the ligamentum flavum associated with osteoblastoma: a report of three cases.

We report three cases of spinal osteoblastoma with ossification of the ligamentum flavum (OLF) adjacent to the tumor. The patients in this report, all young adults, had no symptoms except for back pain. Computed tomography (CT) demonstrated a typical radiolucent nidus in the spinal pedicle/lamina with a dense sclerotic rim. In addition, ectopic bone formation at the insertion point of the ligamentum flavum adjacent to the tumor was clearly illustrated. Magnetic resonance imaging (MRI) revealed the tumor and surrounding inflammatory responses, but OLF was not detected clearly. Histological examination revealed endochondral ossification of the ligamentum flavum that is quite unusual for normal young adults. Immunohistochemical assays in one case demonstrated that bone morphogenetic protein (BMP)-2/4 was expressed in the osteoblastic tumor cells. This case raises the possibility that BMPs secreted from the tumor cells triggered ectopic ossification in the spinal ligament.

Adult↗

Immunohistochemical detection of parathyroid hormone-related peptide, Indian hedgehog, and patched in the process of endochondral ossification in the human.

Parathyroid hormone-related peptide (PTHrP), Indian hedgehog (Ihh), and patched (Ptc; a receptor for Ihh) were immunolocalized in tissue undergoing endochondral ossification in the human. PTHrP, Ihh, and Ptc were immunolocalized in prehypertrophic and hypertrophic chondrocytes in mature cartilage matrix. PTHrP and Ptc were immunostained in proliferating chondrocytes and perichondrial cells, whereas Ihh was not. PTHrP, Ihh, and Ptc showed positive immunostaining in osteoblasts in the bone-forming area. In the bone resorption site, PTHrP was immunolocalized in osteoclasts, whereas Ihh and Ptc were not. The present findings indicated that PTHrP, Ihh, and Ptc were associated with the process of endochondral ossification, and suggested the possible involvement of Ihh and PTHrP signaling in the regulation of proliferation and hypertrophy of chondrocytes in human chondrogenesis.

Bone Neoplasms↗

Localization and expression of osteopontin in the rotator cuff tendons in patients with calcifying tendinitis.

Calcifying tendinitis of rotator cuff tendons is a common and painful condition caused by ectopic calcification in humans. To examine the involvement of osteopontin (OPN), a potent regulator of calcium deposition on connective tissues, localization and expression of OPN protein and messenger (m)RNA were investigated in human tissue samples of calcified rotator cuff tendons. Immunohistochemistry demonstrated that OPN was localized in cells surrounding the calcified area. OPN was localized in two distinct cell types, i.e., fibroblast-like cells negative for CD68 and tartrate-resistant acid phosphatase (TRAP) and multinucleated macrophages positive for CD68 and TRAP. In situ hybridization revealed that the mRNA expression of OPN in these cells coincided with the immunohistochemistry results, and these results were supported by reverse transcriptase polymerase chain reaction analysis using human OPN-specific oligonucleotides. Cells located away from the calcified area did not express OPN. The present findings indicate the involvement of OPN in the process of calcification of rotator cuff tendons and suggest that OPN plays a role in such painful disorders through the actions of at least two cell types.

Acid Phosphatase↗

Involvement of BMP-2 signaling in a cartilage cap in osteochondroma.

This study describes the distributions of bone morphogenetic protein (BMP)-2 as well as mRNAs for BMP receptor type IB (BMPRIB). collagen types II (Col II) and III (Col III) in a growing "cartilage cap" of osteochondroma. In situ hybridization and immunohistochemical study were performed using histological sections obtained during surgery. BMP-2 was detected in mesenchymal cells in the outer fibrous layer and chondrocytes in the inner cartilaginous matrix, positive for Col III and Col II, respectively. BMPRIB mRNA was distributed in chondrocytes. This is the first study to provide observational evidence of the involvement of BMP-2 signaling in the pathogenesis of cartilage cap of osteochondroma. and suggests the role of BMP-2 in the growth of cartilage cap in osteochondroma.

Bone Morphogenetic Protein 2↗

Detection of SYT-SSX fusion gene in peripheral blood from a patient with synovial sarcoma.

This report describes a case involving a 22-year-old pregnant woman with synovial sarcoma in the thigh. The patient recognized an elastic hard mass accompanied by a dull pain in the anteromedial portion of the right thigh. Magnetic resonance imaging delineated a deep soft-tissue mass measuring 9 x 7 x 6 cm. Histologic diagnosis of poorly differentiated synovial sarcoma was made based the results of an open biopsy. In this patient, the SYT-SSX fusion gene transcript was detected by nested polymerase chain reaction (PCR) in the peripheral blood collected before biopsy. Two months after wide local resection of the tumor, multiple lung metastases developed. This is the first reported case in which tumor cells were detected by nested PCR in the peripheral blood of a patient with synovial sarcoma. These findings suggest that circulating tumor cells should be monitored because they may serve as a prognostic indicator for synovial sarcoma.

Adult↗

SYT-SSX fusion proteins in synovial sarcomas: detection and characterization with new antibodies.

To identify and characterize the SYT-SSX fusion proteins in synovial sarcomas, we developed two polyclonal antibodies against the N-terminal part and for the C-terminal part of the SYT-SSX2 protein. Specificity was demonstrated on COS-7 cells transfected with two subtypes of SYT-SSX fusion genes, SYT-SSX1 and SYT-SSX2. Both antibodies recognized a single protein of 61 kDa in an immunoprecipitation of the transfected COS-7 cell lysates. These antibodies also detected the native protein of 61 kDa in the lysate of a human synovial sarcoma cell line (HS-SY41) with immunoprecipitation, and in extracts of human synovial sarcomas with western blot analysis. An immunohistochemical study, using human synovial sarcoma tissues, demonstrated that the SYT-SSX fusion proteins localized in the nucleus of the tumor cells. These antibodies provide a useful method for studying the expression of the SYT-SSX fusion proteins.

Antibodies, Neoplasm↗

Immunohistochemical detection of cathepsin D, K, and L in the process of endochondral ossification in the human.

Cathepsins D, K, and L were immunolocalized in tissue undergoing endochondral ossification in the human. Cathepsins D, K, and L were localized in osteoclasts and chondroclasts attached to bone matrix and cartilage matrix, respectively. Cathepsins D and L were immunostained in chondrocytes. Immunolocalization of cathepsin D was limited to hypertrophic chondrocytes adjacent to the osteochondral junction. In contrast, cathepsin L was immunolocalized in both proliferating and hypertrophic chondrocytes. In the bone marrow space, cathepsins D, K, and L were localized in multinucleated cells. Cathepsin D was diffusely detected in mononuclear bone marrow cells which were negative for cathepsins K and L. The present findings indicated that cathepsins K, D, and L were associated with the process of endochondral ossification in the human, and suggested that these cathepsins share roles in bone and cartilage turnover in the human.

Bone Marrow↗

Involvement of multinucleated giant cells synthesizing cathepsin K in calcified tendinitis of the rotator cuff tendons.

OBJECTIVES: Calcified tendinitis of the shoulder joint is a common painful condition. Resorption of the calcium deposits is one of the key events in the pathogenesis of this disease. The aim of this study was to examine whether the multinucleated giant cells that appear in this condition have osteoclast phenotypes. METHODS: Immunohistochemical and RNA in situ hybridization analysis of cathepsin K, a marker for osteoclasts, was performed in human surgical samples. RESULTS: The multinucleated cells located near the calcium deposits were positive for cathepsin K protein and mRNA. Reverse transcription-polymerase chain reaction using human cathepsin K-specific oligonucleotide primers confirmed that synthesis of cathepsin K mRNA occurs in the tissues of calcified rotator cuffs. CONCLUSION: The multinucleated giant cells which appear in the resorption area of calcium deposits in calcified tendinitis have the osteoclast phenotype.

Arthrography↗

Intraoperative extracorporeal autogenous irradiated bone grafts in tumor surgery.

There are several procedures for reconstruction of bony defects after resection of malignant musculoskeletal tumors. The clinical results of intraoperative extracorporeal autogenous irradiated bone grafts in 20 patients with musculoskeletal tumors are discussed. The authors' method of treatment consists of: (1) wide en bloc resection of the tumor with involved bone; (2) curettage of the tumor from the resected bone; (3) extracorporeal irradiation with 50 Gy as a bolus single dose to the isolated bone; and (4) reimplantation of the irradiated bone into the host with fixation devices. Twelve bone sarcomas and eight soft tissue sarcomas with bone involvement were treated surgically with this reconstruction method after wide resection of the tumors. The irradiated bone was used as an intercalary graft in seven cases, as an osteoarticular graft in 11 cases, and as a hemicortical graft in two cases. The theoretical advantages of this method are certain sterilization of tumor cells with radiation, easy availability and good adaptation of size and shape, no risk of disease transmission, preservation of bone stock and ligamentous tissue, and no immunologic reaction. Radiologically, bony union occurred in 23 of 29 (79%) osteotomy sites. The overall radiographic evaluation rating was 74% and the functional rating was 73% according to the International Society of Limb Salvage rating system. Nonunion (20%) and infection (15%) were the two major complications. Preservation of the tendon insertions and ligamentous structures of the irradiated bone seemed to restore excellent joint function. No local recurrence was detected from the irradiated bones during the mean followup of 45 months. These results indicate intraoperative extracorporeal irradiated bone graft can be a widely applicable method for reconstruction in tumor surgery.

Adolescent↗

Mutation of the p53 gene in postradiation sarcoma.

Radiotherapy is known to cause secondary malignancies in the radiation field; postradiation sarcomas (PRS) are one example of such malignancies. Little is known about the genetic changes, including p53 gene alterations, that are thought to play a role in the tumorigenesis of human PRS. In the present study, p53 gene mutations were analyzed on paraffin-embedded specimens from 24 patients with PRS (4 men and 20 women) by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) followed by direct sequencing. The primary tumors of these patients were uterine cervical cancers in 14, breast cancers in 3, malignant lymphomas in 2, and others in 5. Total radiation doses ranged from 36 to 300 Gy (median, 60 Gy). The latent period between completion of radiation therapy and development of PRS ranged from 3 to 34 years (median, 10 years). Malignant fibrous histiocytoma was the most common PRS, accounting for 12 cases. PCR-SSCP revealed the aberrant mobility shifts of bands in 24 cases: 21 shifts in exon 5, 18 in exon 7, and 12 in exon 8. Direct sequencing of the SSCP product revealed a total of 58 mutations in 21 (88%) of 24 cases: 4 cases had a single mutation, 5 had 2 mutations, 5 had 3 mutations, 6 had 4 mutations, and 1 had 5 mutations. Although 31% of the mutations did not change an amino acid, every tumor had at least one mutation that did, which may have provided the selection pressure for expansion. The frequency of p53 gene mutation in sporadic soft tissue sarcomas was 20%. These findings highlighted the extraordinarily high frequency of p53 gene mutations in PRS. G:C to A:T transition at dipyrimidine sites was found in 14 (58%) of 24 cases. Collectively, these findings indicate that radiation is causative for soft tissue sarcomas via p53 gene mutations.

Adolescent↗

Neoadjuvant chemotherapy for pediatric osteosarcoma patients.

BACKGROUND: Since the first trial for chemotherapy in children with osteosarcoma in 1977, the survival rate has gradually improved. Currently, more than 60% of all patients are cured, mainly because of the introduction of intensive chemotherapy using doxorubicin, high dose methotrexate, and cisplatin. The increased survival rates have promoted efforts to improve the quality of survival through the use of limb salvage surgery rather than amputation. Improvements in chemotherapeutic efficacy should result in a more favorable outcome and better function of the affected limb. The current study evaluated factors that influence chemotherapy so that a higher survival rate could be obtained. METHODS: Three chemotherapy regimens comprised of doxorubicin, high dose methotrexate, cisplatin, and ifosfamide were retrospectively analyzed in 67 pediatric osteosarcoma patients. Twenty-three patients were treated with chemotherapy comprised of high dose methotrexate and doxorubicin (OOS-A regimen), 25 were treated with OOS-A with the addition of cisplatin (OOS-B), and 19 were treated with OOS-B with the addition of ifosfamide (OOS-C). RESULTS: The OOS-A regimen was poorest in terms of survival (40.6%) compared with the OOS-B (67.5%) and OOS-C regimens (72.5%) (P < 0.001). There was no significant difference in survival between the OOS-B and OOS-C regimens. In the OOS-B regimen, patients who received a higher relative dose intensity showed a better prognosis. CONCLUSIONS: These findings show that doxorubicin, high dose methotrexate, and cisplatin are the most potent drugs and suggest that it is more important to maintain the dose intensity of the regimen to improve survival rather than add a new drug.

Adolescent↗