• Title/Summary/Keyword: Osteoblast-like cells

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EFFECTS OF SEVERAL CYTOKINES ON THE FUNCTIONS OF FETAL RAT OSTEOBLAST-LIKE CELLS IN VITRO

  • Han, Hee-Sung;Kim, Jung-Keun;Chang, Young-IL
    • The korean journal of orthodontics
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    • v.25 no.6 s.53
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    • pp.689-696
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    • 1995
  • Effects of several cytokines($IL-1{\beta},\;TNF_{\alpha},\;and\;IFN_{\gamma}$) have been examined on fetal rat osteoblast-like cells. To investigate whether cytokines play direct causal roles in production of lysosomal enzyme, fetal rat osteoblast-like cells were treated with $IL-1{\beta},\;TNF_{\alpha},\;and\;IFN_{\gamma}$, respectively or combined. And acid phosphatase was determined by biochemical method. Alkaline phosphatase was assayed to determine the effects of $IL-1{\beta},\;TNF_{\alpha},\;and\;IFN_{\gamma}$ on the expression of this enzyme. And also experiment of calcified nodule formation was performed to assess the effects of cytokines on the bone-forming activity of osteoblast-like cells in vitro. Acid phosphatase activity was significantly increased by the addition of $IL-1{\beta}\;and\;TNF_{\alpha}$, whereas decreased by $IFN_{\gamma}$. However, no significant change:: in alkaline phosphatase activity was observed when the osteoblast-like cells were treated with $IL-1{\beta}\;and\;TNF_{\alpha}$. Interestingly, $IFN_{\gamma}$ showed stimulatory effect on alkaline phosphatase activity. The number of calcified nodules was decreased by treatment of cultures with 1 ng/ml $IL-1{\beta},\;20\;ng/ml\;TNF_{\alpha}$, and 500 u/ml $IFN_{\gamma}$ continuously for 21 days, while considerable number of calcified nodules were formed in control group of osteoblast-like cell in culture for 21 days. These results seem to suggest that cytokines may play crucial roles in bone remodeling through the direct action on the osteoblast-like cell.

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CELLULAR ATTACHMENT AND GENE EXPRESSION OF OSTEOBLAST-LIKE CELLS ON ZIRCONIA CERAMIC SURFACES

  • Pae, Ah-Ran;Lee, Hee-Su;Kim, Hyeong-Seob; Baik, Jin;Woo, Yi-Hyung
    • The Journal of Korean Academy of Prosthodontics
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    • v.46 no.3
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    • pp.227-237
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    • 2008
  • STATEMENT OF PROBLEM: Zirconium oxide can be a substitute to titanium as implant materials to solve the esthetic problems of dark color in the gingival portion of implant restorations. PURPOSE: This study was performed to define attachment and growth behavior of osteoblast- like cells cultured on grooved surfaces of zirconium oxide and evaluate the genetic effect of zirconium oxide surfaces using the reverse transcriptase-polymerase chain reaction (RT-PCR). MATERIAL AND METHODS: MC3T3-E1 cells were cultured on (1) commercially pure titanium discs with smooth surface (T group), (2) yttrium-stabilized tetragonal zirconia polycrystal (Y-TZP) with machined surface (ZS group), and (3) Y-TZP with $100{\mu}m$ grooves (ZG group). Cell proliferation activity was evaluated through MTT assay and cell morphology was examined by SEM. The mRNA expression of Runx2, alkaline phosphatase, osteocalcin, TGF-${\beta}1$, IGF-1, G3PDH in E1 cells were evaluated by RT-PCR. RESULTS: From the MTT assay, after 48 hours of adhesion of MC3T3-E1 cells, the mean optical density value of T group and ZG group significantly increased compared to the ZS group. SEM images of osteoblast-like cells showed that significantly more cells were observed to attach to the grooves and appeared to follow the direction of the grooves. After 24 hours of cell adhesion, more spreading and flattening of cells with active filopodia formation occurred. Results of RT-PCR suggest that T group, ZS group, and ZG group showed comparable osteoblast-specific gene expression after 24 hours of cell incubation. CONCLUSION: Surface topography and material of implants can play an important role in expression of osteoblast phenotype markers. Zirconia ceramic showed comparable biological responses of osteoblast-like cells with titanium during a short-time cell culture period. Also, grooves influence cell spreading and guide the cells to be aligned within surface grooves.

Effects of Uncaria rhynchophylla Extracts on Differentiation and Bone Mineralized Formation in Human Osteoblast-like SaOS-2 cells

  • Huh, Jeong-Eun;Baek, Yong-Hyeon;Choi, Do-Young;Lee, Jae-Dong;Park, Dong-Suk
    • The Journal of Korean Medicine
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    • v.28 no.4
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    • pp.158-167
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    • 2007
  • Background & Objective : Uncaria rhynchophylla is traditional medicine herb used for enhancing body resistance against various diseases. The aim of this study was to identify if Uncaria rhynchophylla extracts induce osteogenic activity in human osteoblast-like SaOS-2 cells. Methods : The osteogenic activity of Uncaria rhynchophylla was evaluated on cell proliferation assay by WST-8, and osteoblast-specific genes, such as VEGF, type I collagen (Col I), osteocalcin (OCN), and osteopontin (OPN) by RT-PCR analysis and ELISA assay in osteoblasts-like SaOS-2 cells. Bone mineralization was stained with Alizalin red method. Results : Uncaria rhynchophylla had significantly increased cell proliferation at a dose dependent manner in human osteoblast-like SaOS-2 cells. Uncaria rhynchophylla markedly increased alkaline phosphatase (ALP), vascular endothelial growth factor (VEGF) mRNA expression at 7 days and dose dependently increased ALP activity and VEGF secretion in human osteoblast-like SaOS-2 cells. Also, Uncaria rhynchophylla time-dependently increased type I collagen (Col I), osteopontin (OPN), and osteocalcin (OCN) mRNA in SaOS-2 cells. Extracellular accumulation of proteins such as Col I and OCN was maximal increased by Uncaria rhynchophylla at 10 ${\mu}g/ml$. Also, Uncaria rhynchophylla significantly induced mineralization in the culture of SaOS-2 cells. Conclusion : This study showed that Uncaria rhynchophylla had enhanced proliferation, ALP activity, VEGF, bone matrix proteins such as OCN, OPN, and Col I, and mineralization in SaOS-2 cells. These results propose that Uncaria rhynchophylla can play an important role in osteoblastic bone formation, osteogenesis, and may possibly lead to the development of bone-forming drugs.

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Effects of Radix Achyranthis Bidentatae Extract on Proliferation and Differentiation in Human Osteoblast-like Cells (우슬 추출물이 골아세포 증식과 분화에 미치는 효과)

  • Seo Eun A;Moon Hyung Cheal
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.6
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    • pp.1821-1824
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    • 2004
  • In order to investigate the effects of Radix Achyranthis Bidentatae (RAB) on the growth and differentiation of human osteoblast-like cells, we supplemented the culture medium of MG-63 cells with various concentrations of RAB water extracts. RAB extracts significantly stimulated cell growth, as confirmed by the colorimetric MTT (3-dimethylthiazol-2,5-diphenyltetrazolium bromide) assay. RAB extracts also increased the alkaline phosphatase (ALP) activity, which is a osteoblast differentiation marker. These results suggest that RAB can stimulate osteoblastic activity and may represent new pharmacological tools for the treatment of osteoporosis.

EFFECT OF TITANIUM SURFACE ROUGHNESS ON CELL ADHESION OF HUMAN OSTEOBLAST-LIKE CELLS (MG63)

  • Yim Soon-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.42 no.3
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    • pp.261-266
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    • 2004
  • Statement of problem. The effects of surface roughness have not or insufficiently been analyzed on earlier events such as cell adhesion though cell behavior most germane to implant performance is cell adhesion. Purpose. The purpose of this study was to evaluate cell adhesion of osteoblast-like cells (MG63) onto three types of titanium disks with varying roughness using the Elisa assay. Materials and methods. Representative disks from each group (SLA, HA, machined) were subjected to surface analysis and surface roughness was measured by the optical interferometer (Accura 2000, Intekplus Co., Seoul, Korea). Following this, MG63 cells were cultured on the titanium disks and released. Cell adhesion measurements using the Elisa assay were performed specifically at three points: after 24, 48, and 72 hours of culture. Results. Among the 3 types of surface analyzed, the SLA surface was the roughest with a Ra value of $1.114{\mu}m$ followed by HA coated surface and machined surface, consecutively. The optical density values for the SLA surface group was significantly higher than that of the machined and HA coated surface groups following 24 and 48 hours of culture. The cell culture on HA coated surface showed significantly higher values compared to the machined surface following 24, 48 and 72 hours of culture. Conclusion. The results suggest that surface treatment of titanium surfaces enhanced cell adhesion of human osteoblast-like cells (MG63).

BIOCHEMICAL CHARACTERIZATION OF EMBRYONIC CHICK CALVARIAL CELLS

  • Yu, Jae-Hyung;Kim, Jung-Kun;Cha, Kyung-Suk
    • The korean journal of orthodontics
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    • v.25 no.6 s.53
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    • pp.697-704
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    • 1995
  • Chicken calvarial bone is known to contain various cell types, but their exact composition is unknown. By characterizing the chicken calvarial bone biochemically, it can be used to study biochemical, histochemical actions of bone cells in general. Calvaria of 18-day-old white leg horn embryo was aseptically dissected and bone cell populations were isolated by sequential enzymatic digestion. Histochemical study for osteoclast-like bone cell. population was performed with tartrate resistant acid phosphatase(TRAP) stain and for osteoblast-like bone cell population, alkaline phosphatase(ALP) stain was performed. Biochemical study for osteoblast-like bone cell population was performed using alkaline phosphatase(ALP) assay. Following conclusions were obtained from this study. 1. TRAP positive multi and mononuclear cells were mostly observed in group I and II, indicating that osteoclast-like bone cell population is mostly found in these groups. 2. All the cultured groups showed almost equal ALP activities and were positive for ALP stain, indicating that osteoblast-like bone cell population is evenly dispersed in all culture groups. 3. Experimental group treated with $1,25(OH)_{2}D_3$ showed increase in ALP activity in contrast to the control group, confirming previous studies that $1,25(OH)_{2}D_3$ increases ALP activities in in vitro bone cultures. 4. Results from von Kossa's stain indicated that in vitro bone formation had occured after 3 weeks of culture with beta-glycero phosphate.

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Potential Effect of Monascus-fermented Soybean Extracts on Alkaline Phosphatase Activity of Human Osteoblast-like Cells

  • Pyo, Young-Hee;Kwon, Mi-Ja;Kim, In-Ho
    • Food Science and Biotechnology
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    • v.17 no.2
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    • pp.434-437
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    • 2008
  • The aim of this study was to investigate whether Monascus-fermented soybean extracts (MFSE) containing natural estrogen-like compounds such as isoflavones and mevinolins has potential effects on human osteoblast-like SaOS2 cells using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and alkaline phophatase (ALP) assaies. MFSE exerted biphasic dose-dependent effect; stimulating osteoblastic activity at low concentrations and inhibiting SaOS2 cells viability at high concentrations. At $10^{-8}-10^{-4}\;mg/mL$, MFSE is not only non-cytotoxic but also induced comparatively high ALP activity on SaOS2 cells. ALP activity (%) significantly increased (220.1%, p<0.05) when SaOS2 cells were treated with MFSE at a concentration of $10^{-5}\;mg/mL$, whereas slowly increased (185.6%, p<0.05) in unfermented soybean extracts (UFSE) at $10^{-3}\;mg/mL$. The potentially greater ALP activity of MFSE compared to the UFSE might partially be caused by its mevinolin, which was derived from the soybean during Monascus-fermentation. Our findings indicate that supplementation of MFSE may accelerate the speed of intracellular ALP synthesis by the bone cells when provided at optimal dosages.

Insulin as a Potent Stimulator of Akt, ERK and Inhibin-βE Signaling in Osteoblast-Like UMR-106 Cells

  • Ramalingam, Mahesh;Kwon, Yong-Dae;Kim, Sung-Jin
    • Biomolecules & Therapeutics
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    • v.24 no.6
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    • pp.589-594
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    • 2016
  • Insulin is a peptide hormone of the endocrine pancreas and exerts a wide variety of physiological actions in insulin sensitive tissues, such as regulation of glucose homeostasis, cell growth, differentiation, learning and memory. However, the role of insulin in osteoblast cells remains to be fully characterized. In this study, we demonstrated that the insulin (100 nM) has the ability to stimulate the phosphorylation of protein kinase B (Akt/PKB) and extracellular signal-regulated kinase (ERK) and the levels of inhibin-${\beta}E$ in the osteoblast-like UMR-106 cells. This insulin-stimulated activities were abolished by the PI3K and MEK1 inhibitors LY294002 and PD98059, respectively. This is the first report proving that insulin is a potential candidate that enables the actions of inhibin-${\beta}E$ subunit of the TGF-${\beta}$ family. The current investigation provides a foundation for the realization of insulin as a potential stimulator in survival signaling pathways in osteoblast-like UMR-106 cells.

Effects of Alkaline Phosphatase Activity on the Extract of Carthami Semen and Eucommiae Cortex in Human Osteoblast-like MG-63 Cell Line (홍화자와 두충 혼합 추출물이 MG-63 조골세포의 Alkaline Phosphatase 활성에 미치는 영향)

  • Sim, Jae-Geun;Lee, Jae-Hyeok;Yeo, Myeong Gu;Park, Jeong-Suk
    • Journal of the East Asian Society of Dietary Life
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    • v.23 no.1
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    • pp.39-43
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    • 2013
  • Carthamus tinctorius L. and Eucommia umoides Oliver are often used in traditional herbal medicines for reducing damage to the liver, kidney, bone and muscle. In the present study, we investigated cell viability and alkaline phosphatase activity in the human osteoblast-like MG-63 cell line with methanol extracts of Carthami Semen (CS) and Eucommiae Cortex (EC) alone or in a mixture (CS+EC). Osteoblast cell viability was evaluated using the MTS assay and alkaline phosphatase activity assays. The cell viability and alkaline phosphatase activity significantly increased in MG-63 osteoblast cells treated with the CS+EC mixture. These findings suggest the CS+EC mixture may have beneficial effects on bone health through the proliferation of osteoblast cells.

The Effect of Osteoblast like Cell on Low Intensity Pulsed Ultrasound Used in Current Clinical Practice (임상에서 쓰이는 저 출력 초음파(Low Intensity Pulsed Ultrasound)가 조골세포에 미치는 영향)

  • Park, Sang-Rye;Kim, Ji-Young;Choi, Byul Bo-Ra
    • Journal of dental hygiene science
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    • v.14 no.2
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    • pp.101-106
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    • 2014
  • In this study, we investigated the proliferative and adhesional effect of human osteoblast like MG-63 cell treated with low intensity pulsed ultrasound (LIPUS). We tested the effectiveness of LIPUS in human osteoblast like MG-63 cells. Cell proliferation was measured using a water soluble tetrazolium salts-1 assay. The mRNA expression of alkaline phosphate, vascular endothelial growth factor (VEGF), integrin alpha 2, colla 1A1 were performed by reverse transcription-polymerase chain reaction. LIPUS was no cytotoxicity in human osteoblast like MG-63 cells. In addition, the data show that treatment with 1 MHz and 3 MHz LIPUS on increased proliferation 7 days after. There were significant increased in mRNA expression of alkaline phosphatase, osteocalcin, VEGF, integrin alpha 2 and colla 1A1 (p<0.05). Therefore, the LIPUS significantly increased differential expression of mRNA levels in osteoblast like MG-63 cell and new possibilities in dental clinical practice.