• Title/Summary/Keyword: Osteosarcoma cells

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ANALYSES OF THE EXPRESSION PROFILES OF GENES RESPONSIBLE FOR THE GROWTH OF OSTEOBLAST UPON VELVET ANTLERS TREATMENT (녹용이 조골세포(造骨細胞) 성장에 미치는 유전자 발현 profile 분석에 대한 연구)

  • Lee, Jong-Woo;Kim, Doeg-Kon
    • The Journal of Pediatrics of Korean Medicine
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    • v.16 no.1
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    • pp.39-74
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    • 2002
  • Recently, increased attention has been paid to the growth of the height of children and adolescents. To accelerate growth, velvet antlers are typically used in Oriental medicine. The present study investigated the effects of velvet antlers of velvet antlers on bone growth using the cell line of Human Osteosarcoma (Hos), derived from the bone-generating cells essential to bone growth. In order to give certain stress to this Hos, the medium contained 1% FBS was used for culturing for Hos cell instead of 10% in control. In this condition of which the proliferation had been significantly decreased, the ethanol extract of upper part of velvet antlers was added, As a result, the cells proliferation rate was significantly increased. Using Oligonucleotide DNA microarray, comparison and analyses were done to see what kind of specific genes would be differentially expressed. The result showed that as opposed to the control group, the stressed group indicated a decrease in the expressions of 6 kinds of genes such as, Id1, retinoid X receptor(RXRB) and 14-3-3 epsilon, etc. The velvet antler treated group, as opposed to the control group, showed a decreased in the expressions of 8 kinds of genes such as Id1, etc. and an increase in the expressions of 24 kinds of genes. The number of genes that showed differences in the velvet antler treated group compared with the stressed group was 7 the expression of 1 kind of gene was decreased, and the expressions of 6 kinds of genes were increased. Considering the mechanism by which velvet antlers affected the growth of osteoblast through reviewing the functions of these genes, the following results were attained. The constraint in the proliferation of Hos cells resulting from the medium contained 1% FBS seems to be caused by three important factors: 1) the decrease of the expression of 14-3-3 epsilon involved in the signal transduction and metabolism of growth, 2) the decrease of the expression of Id1 gene involved in the metabolism of bone formation, and 3) the decreased of expression of RXRB gene involved in the metabolism of retinoci acid. It is suggested that the improvement of the cell proliferating effects by velvet antler treatment, in stressed condition si mediated by increment of 6 genes particularly 14-3-3 epsilon, RXRB, and IGF2, with are the crucial factors for the cell growth and differentiation, metabolism of retinoic acid and osteoblast proliferation, respectively.

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Inhibitory Effects of Type IV Collagenase by Disulfiram (Disulfiram에 의한 type IV collagenase 저해효과)

  • Sin, Doo-Il;Park, Jae-Bok;Park, Kwan-Kyu;Cho, Chang-Ho;Oh, Hoon-Kyu;Choi, Chang-Hyuk;Cho, Hyun-Ji;Chang, Young-Chae
    • Journal of Life Science
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    • v.16 no.6
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    • pp.964-971
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    • 2006
  • The cancer cells, characterized by local invasion and distant metastasis, are very dependant on extracellular matrix. The expression of matrix metalloproteinases (MMPs) has been implicated in the invasion and metastasis of cancer cells. Among the human MMPs, matirx metalloproteinase-2 (MMP-2) and matrix metalloproteinse-9 (MMP-9) are key enzymes that degrade type IV collagen of the matrix. Here, we studied the effect of disulfiram, an anti-tumor compound, on the suppression of the tumor invasion and the activity of MMP-2, MMP-9 in human osteosarcoma cells (U2OS). Disulfiram had the type IV collagenase inhibitory activity, the effect of inhibition of gene and protein expression, and these inhibitions were responsible for blocking invasion through cell mediated and non-cell mediated pathways. In conclusion, disulfiram inhibited expression of MMP-2 and MMP-9, and regulated the invasion of U2OS, Caki-1 and Caski. These observations raise the possibility of clinical therapeutic applications for disulfiram used as a potential inhibitor of cancer invasion.

Effects of Scutellariae Radix on Neuronal Apoptosis of Spinal Cord Contusion Injury in Rats (황금(黃芩)이 척수압박손상 흰쥐의 척수신경세포 자연사에 미치는 영향)

  • Bahn, Hyo-Jung;Jo, Jong-Jin;Kim, Bum-Hoi;Park, Seong-Ha;Shin, Jung-Won;Kim, Seong-Joon;Sohn, Nak-Won
    • Journal of Korean Medicine Rehabilitation
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    • v.21 no.4
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    • pp.13-22
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    • 2011
  • Objectives : This Study was performed to evaluate the effects of Scutellariae Radix(SR) water-extract on the tissue and neuronal apoptosis of the spinal cord injury(SCI). Methods: SCI was induced by mechanical contusion following laminectomy of 10th thoracic vertebra in Sprague-Dawley rats. SR was orally given once a day for 7 days after SCI. Neuronal apoptosis was examined with terminal deoxynucleotidyl transferase-mediated dUTPnick-end labeling(TUNEL) assay. Bax (Bcl-2-asociated X protein), Bcl-2(B-cell blastoma 2), c-Fos(FBJ osteosarcoma oncogene) expressions were examined using immuno-histochemistry. Individual TUNEL and immuno-labeled cells expressing Bax, Bcl-2 and c-Fos were counted on the same level in peri-damaged region and in ventral horn. Results: 1. SR significantly reduced number of TUNEL labeled apoptotic cells induced by the spinal cord contusion injury. 2. SR significantly reduced Bax positive cells expression on the motor neuron in the ventral horn induced by the spinal cord contusion injury. 3. SR strengthened Bcl-2 expression on the motor neuron in the ventral horn induced by the spinal cord contusion injury. 4. SR reduced c-Fos expression on the motor neuron in the ventral horn induced by the spinal cord contusion injury. Conclusions : These results suggest that SR plays an inhibitory role against neuronal apoptosis and has significant effects for locomotor disfunction induced by SCI.

Bizarre Parosteal Osteochondromatous Proliferation of the Femur (Nora's Lesion) - A Case Report - (대퇴골에 발생한 Bizarre Parosteal Osteochondromatous Proliferation of the Femur (Nora 병변) - 증례 보고 -)

  • Shin, Bong-Kyung;Cho, Hyun-Deuk;Yum, Bum-Woo;Choi, Jong-Sang;Kim, Chul-Hwan
    • The Journal of the Korean bone and joint tumor society
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    • v.5 no.3
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    • pp.178-182
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    • 1999
  • We report a case of bizarre parosteal osteochondromatous proliferation of the right femur in an 18-year-old man. Roentgenograms showed a calcific mass attached to the underlying cortex with a broad base. Histologically, the lesion showed hypercellular cartilaginous tissue with maturation into trabecular bone, which contained spindle cells and lymphocytes in the intertrabecular spaces. Bizarre parosteal osteochondromatous proliferations are a form of heterotopic ossification and should not be mistaken for osteosarcoma or chondrosarcoma.

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The Analysis of the Cytokine Expression in Musculoskeletal Tumors (근골격계 종양에서 사이토카인 발현의 분석)

  • Lee, Joon-Han;Kwak, Eun-Seok;Shon, Oog-Jin;Kim, Hee-Sun;Shin, Duk-Seop
    • Journal of Yeungnam Medical Science
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    • v.20 no.2
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    • pp.187-196
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    • 2003
  • The cytokines are the hormone-like proteins, which are produced in the mononuclear cells. They have many roles, such as immune mediators, cell differentiations, angiogenesis. The chemokines have chemotactic effects which control the host immune response. There were few reports about the cytokines associated with musculoskeletal tumors. From late 1980s, the cytokine studies of bone tumors such as osteosarcoma were started, but most studies for benign and malignant musculoskeletal tumors were left to be explored. To evaluate the characteristics of the cytokines in variable musculoskeletal tumors, tissues were obtained from the seven patients who visited the Yeungnam University hospital from February to July 2000. They were lipoma (1 case), parosteal osteoma (1 case), enchondroma (2 cases), pigmented villonodular synovitis (1 case), ganglion (1 case), and metastaic squamous cell carcinoma (1 case). The gene experession of the cytokines were analyzed by RNase protection assay (RPA) and reverse transcription-polymerase chain reaction (RT-PCR). The lipoma and parosteal osteoma expressed MIP-$1{\beta}$, and IP-10 genes. The two enchondromas showed different results, one expressed all of MIP-$1{\alpha}$, MIP-$1{\beta}$ and IP-10 genes but the other expressed none of above. The pigmented villonodular synovitis strongly expressed MIP-$1{\alpha}$ and IP-10 when compared with the other cases. The ganglion did not express all of the chemokines mentioned above. And the metastatic squamous cell carcinoma expressed all of the chemokines and especially IP-10 was highly expressed. Even though this study has only a few cases, these results provide a basis for the cytokine mediating network study in musculoskeletal tumors.

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A comparative analysis of basic characteristics of several deproteinized bovine bone substitutes (수종의 탈단백 우골 이식재의 특성 비교 분석)

  • Yeo, Shin-Il;Park, Sung-Hwan;Noh, Woo-Chang;Park, Jin-Woo;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.39 no.2
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    • pp.149-156
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    • 2009
  • Purpose: Deproteinized bovine bone substitutes are commonly used in dental regenerative surgery for treatment of alveolar defects. In this study, three different bovine bone minerals - OCS-B (NIBEC, Seoul, Korea), Bio-Oss (Geistlich - Pharma, Switzerland), Osteograft/N - 300 (OGN, Dentsply Friadent Ceramed. TN, USA) - were investigated to analyze the basic characteristics of commercially available bone substitutes. Methods: Their physicochemical properties were evaluated by scanning electron microscopy, energy dispersive X-ray spectrometer (EDS), surface area analysis, and Kjeldahl protein analysis. Cell proliferation and alkaline phosphatase (ALP) activity of human osteosarcoma cells on different bovine bone minerals were evaluated. Results: Three kinds of bone substitutes displayed different surface properties. Ca/P ratio of OCS - B shown to be lower than other two bovine bone minerals in EDS analysis. Bio-Oss had wider surface area and lower amount of residual protein than OCS - B and OGN. In addition Bio - Oss was proved to have lower cell proliferation and ALP activity due to lots of residual micro particles, compared with OCS - B and OGN. Conclusions: Based on the results of this study, three bovine bone minerals that produced by similar methods appear to have different property and characteristics. It is suggested that detailed studies and quality management is needed in operations for dental use and its biological effects on new bone formation.

Fabrication of Nanofiber-Combined 3D Scaffolds using Dual-Head Deposition Technology (듀얼헤드 적층 기술을 이용한 나노섬유로 결합된 3D 인공지지체 제작)

  • Sa, Min-Woo;Lee, Chang-Hee;Kim, Jong Young
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.17 no.1
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    • pp.108-115
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    • 2018
  • In bone tissue engineering, polycaprolactone (PCL) is one of the most widely used biomaterials to manufacture scaffolds as a synthetic polymer with biodegradability and biocompatibility. The polymer deposition system (PDS) with four axis heads, which can dispense bio-polymers, has been used in scaffold fabrication for tissue engineering applications. A dual-head deposition technology of PDS is an effective technique to fabricate 3D scaffolds. The electrospinning technology has been widely used to fabricate porous and highly interconnected polymer fibers. Thus, PDS can fabricate nanofiber-combined hybrid scaffolds using fused deposition modeling (FDM) and electrospinning methods. This study aims to fabricate nanofiber-combined scaffolds with uniform nanofibers using PDS. The PCL nanofibers were fabricated and evaluated according to the fabrication process parameters. PCL nanofibers were successfully fabricated when the applied voltage, tip-to-collector distance, flow rate, and solution concentration were 5 kV, 1 cm, 0.1 ml/h, and 8 wt%, respectively. The cell proliferation was evaluated according to the electrospinning time. Scanning electron microscopy was used to acquire images of the cross-sectioned hybrid scaffolds. The cell proliferation test of the PCL and nanofiber-combined hybrid scaffolds was performed using a CCK-8 assay according to the electrospinning time. The result of in-vitro cell proliferation using osteosarcoma MG-63 cells shows that the hybrid scaffold has good potential for bone regeneration.

MMP-1 and TIMP-1 production in MG-63 cells stimulated with Prevotella nigrescens Lipopolysaccharide (Prevotella nigrescens lipopolysaccharides로 자극된 MG63 세포에서 분비되는 기질금속단백질 MMP-1과 TIMP-1의 수준에 관한 연구)

  • Yang Won-Kyung;Kim Mi-Ri;Shon Won-Jun;Lee In-Bog;Cho Byeong-Hoon;Um Chung-Moon;Son Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.29 no.5
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    • pp.470-478
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    • 2004
  • The purpose of this study is to monitor the secretion of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) produced by human osteosarcoma cell line (MG63) stimulated with Prevotella nigrescens lipopolysaccharides (LPS). and to compare the level of secretion before and after the treatment of calcium hydroxide on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. MG63 cells were stimulated by the LPS (0, 1, $10{\;}\mu\textrm{g}/ml$) or LPS($10{\;}\mu\textrm{g}/ml$) pretreated with 12.5 mg/ml of $Ca(OH)_2$ for 3 days. Total RNA was isolated from the cell. and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 and TIMP-1. The results were as follows. 1. MMP-1 mRNA expression at 48 hr was highly increased by stimulation with P. nigrescens LPS. The increase was dose-dependent. 2. When stimulated with ($1{\;}\mu\textrm{g}/ml$ of LPS. TIMP-1 mRNA expression was highly increased at 24 hr and 48 hr. However. TIMP-1 expression was suppressed at higher concentration ($10{\;}\mu\textrm{g}/ml$). 3. When P. nigrescens LPS was pretreated with $Ca(OH)_2$. MMP-1 and TIMP-1 gene expression was downregulated. The results of this study suggest that transcriptional regulation of MMP-1 and TIMP-1 by P. nigrescens LPS could be one of the important mechanisms in bone resorption of periapical inflammation. The result of calcium hydroxide on MMP-1 and TIMP-1 gene expression suppression shows that calcium hydroxide detoxified bacterial LPS and thus should be used the medication of choice for intracanal dressings in root canal infected with black-pigmented bacteria.

Antioxidant Effect of Hederagenin 3-O-b-D-Glucopyranosyl(1→3)-a-L-Rhamnopyranosyl(1→2)-a-L-Arabinopyranoside (HDL) Isolated from Root Bark of Ulmus davidiana (유근피로부터 분리한 hederagenin 3-O-b-D-glucopyranosyl(1→3)-a-L-rhamnopyranosyl(1→2)-a-L-arabinopyranoside (HDL)의 항산화 효과)

  • Bong, Jin-Gu;Park, Yoon-Yub
    • Journal of Life Science
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    • v.20 no.2
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    • pp.281-291
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    • 2010
  • We investigated the antioxidant effects of hederagenin 3-O-b-D-glucopyranosyl($1{\rightarrow}3$)-a-L-rhamnopyranosyl($1{\rightarrow}2$)-a-L-arabinopyranoside (HDL) isolated from root bark of Ulmus davidiana on the activity of enzymes related to reactive oxygen species (ROS) in human osteosarcoma U2OS cells. Cobalt chloride ($CoCl_2$), a transition metal, was used as an inducer of oxidative stress, generating hydrogen peroxide ($H_2O_2$) via increasing xanthine oxidase (XO) activity. The increased levels of $H_2O_2$, XO, ferritin, and ferritin iron by $CoCl_2$ were diminished effectively by co-treatment with HDL in U2OS cells. Furthermore, decreased levels of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) by $CoCl_2$ were highly increased by co-treatment with HDL in U2OS cells; however, the levels of glutathione peroxidase (GPx) did not change. The increased contents of TBARS related to lipid peroxidation were significantly reduced by HDL in U2OS cells. The concentration of GSH changed in a pattern that went against regulated TBARS by $CoCl_2$ and HDL. We examined the expression of p53, $p21^{CIP1/WAF1}$, and $p27^{KIP1}$ proteins related to oxidative stress and cell cycle regulation. As a result, the expression of $p27^{KIP1}$ modulated by $CoCl_2$ was not changed by HDL. However, the expression of p53 and $p21^{CIP1/WAF}$ increased by $CoCl_2$ was reduced by HDL in U2OS cells. Together with alteration of p53 and $p21^{CIP1/WAF1}$ proteins, the accumulated cells at G1 phase by $CoCl_2$ was decreased by HDL in U2OS cells. Our data suggests that HDL inhibits $CoCl_2$-generated ROS in U2OS cells, providing potentially new antioxidant compounds that are isolated from natural products.

Fine Needle Aspiration Cytology in Bone and Soft Tissue Tumor (골 및 연부 조직 종양에서의 미세 침 흡입 세포 검사)

  • Lee, Soo-Yong;Jeon, Dae-Geun;Lee, Jong-Seok;Kim, Sug-Jun;Cho, Kyung-Ja;Oh, Hyung-Ho;Park, Min-Hyo;Park, Hyun-Soo;Jang, Jin-Dae
    • The Journal of the Korean bone and joint tumor society
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    • v.2 no.1
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    • pp.33-37
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    • 1996
  • From January 1994 to August 1995 we performed 270 cases of fine needle aspiration cytology for bone and soft tissue lesions. Among them 137 cases were diagnosed histologically. Sixty-seven cases were benign and 70 cases were malignant. We analysed these on the aspect of sensitivity, specifity, positive predictive value, negative predictive value of the cytologic examination. Malignant cases consisted of bone tumor 29 cases; soft tissue tumor 25 cases; and metastatic tumor 16 cases. The sensitivity and specifity of the aspiration cytology for malignant tumor were 67.8% and 97%, respectively. The positive and negative predictive value of aspiration cytology were 97.4% and 72.7%, respectively. Among them 40 cases were primary bony lesions; 14 benign lesions, 26 malignamt bone tumors. Sensitivity of aspiration cytology for these primary bony lesion was 90%(18/20) and its specifity was 100%(13/13). Three cases showed atypical cells in cytology but with biopsy we could find them as one benign and two malignancies. Two cases of manignancy showed negative result in cytology; one was parosteal sarcoma of proximal femur and the other was osteosarcoma of proximal tibia which was biopsied already at other hospital. Although the overall sensitivity and specifity for malignant bone and soft tissue tumors were relatively low and not so predictable, the fine needle aspiration cytology for primary bony lesion was useful in the differential diagnosis of benign or malignancy.

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