• Title/Summary/Keyword: Human Osteoblast

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A STUDY OF $TGF-{\beta}$ EXPRESSION DURING PALATOGENESIS IN RATS WITH CLEFT PALATE INDUCED BY BAPN (($TGF-{\beta}$ 발현이 BAPN으로 유도된 구개열 백서의 구개 형성에 미치는 영향에 대한 실험적 연구)

  • Tae, Ki-Chul;Lee, Dong-Kun;Kim, Jeng-Ghee
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.23 no.3
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    • pp.205-211
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    • 2001
  • Cleft palate is one of the most serious congenital anomalies in human that causes a sucking problem in newborn babies and morphologic deformity that usually leads to death in newborn mouse offspring due to an insufficient ability to suck milk. Therefore cleft palate had been researched with epidemiologic and molecular methods, and many etiologic factors were examined closely. Among of the research methods, biologic molecule researches have been more important method for cleft palate formation study. The $TGF-{\beta}$ had an important role in the cell migration, epithelial-mesenchymal transdifferentiation, extracellular matrix synthesis and deposition. But there was a little research which was study about correlation cleft palate induced by beta-aminonitroproprionitrile(BAPN) with $TGF-{\beta}$ expression. A purpose of this presented study was examed how $TGF-{\beta}$ expression in cleft palate mice. At gestation days 13, BAPN-monofumarate salts($(C_3H_6N_2)_2$ ${\cdot}$ $C_4H_4O_4$, Sigma Co.) was single oral administered to 4 pregnant rats according to 1g/kg body weight. And pregnant rats were sacrificed on day 20 post coitus(p.c.), The $TGF-{\beta}$ expression patterns of cleft formed fetus mice was followed that; 1.Osteoblast, mesenchymal cell and epithelial cell of cleft mice were low expression compare to control mice. 2.There was no $TGF-{\beta}$ difference expression pattern of osteocyte of cleft mice compare to control mice. 3. In western blot analysis, thickness of band of $TGF-{\beta}$ in cleft mice was thin and dilute compare to control mice.

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Anthraquinone Glycoside Aloin Induces Osteogenic Initiation of MC3T3-E1 Cells: Involvement of MAPK Mediated Wnt and Bmp Signaling

  • Pengjam, Yutthana;Madhyastha, Harishkumar;Madhyastha, Radha;Yamaguchi, Yuya;Nakajima, Yuichi;Maruyama, Masugi
    • Biomolecules & Therapeutics
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    • v.24 no.2
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    • pp.123-131
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    • 2016
  • Osteoporosis is a bone pathology leading to increased fracture risk and challenging the quality of life. The aim of this study was to evaluate the effect of an anthraquinone glycoside, aloin, on osteogenic induction of MC3T3-E1 cells. Aloin increased alkaline phosphatase (ALP) activity, an early differentiation marker of osteoblasts. Aloin also increased the ALP activity in adult human adipose-derived stem cells (hADSC), indicating that the action of aloin was not cell-type specific. Alizarin red S staining revealed a significant amount of calcium deposition in cells treated with aloin. Aloin enhanced the expression of osteoblast differentiation genes, Bmp-2, Runx2 and collagen 1a, in a dose-dependent manner. Western blot analysis revealed that noggin and inhibitors of p38 MAPK and SAPK/JNK signals attenuated aloin-promoted expressions of Bmp-2 and Runx2 proteins. siRNA mediated blocking of Wnt-5a signaling pathway also annulled the influence of aloin, indicating Wnt-5a dependent activity. Inhibition of the different signal pathways abrogated the influence of aloin on ALP activity, confirming that aloin induced MC3T3-E1 cells into osteoblasts through MAPK mediated Wnt and Bmp signaling pathway.

Preparation and Characterization of Natural Material Extracted from Germinated Brown Rice

  • Lim, Ki-Taek;Choi, Jeong Moon;Lim, Won-Chul;Kim, Jangho;Cho, Hong-Yon;Chung, Jong Hoon
    • Journal of Biosystems Engineering
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    • v.39 no.3
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    • pp.235-243
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    • 2014
  • Purpose: The aim of this study was to prepare and evaluate a natural material extracted from germinated brown rice (GBR). Herein, we evaluated whether the natural material could positively activate the biological effects seen during bone formation, including enhancement of metabolic activity, osteogenesis, and the expression of vascular endothelial growth factor (VEGF), one of the growth factors in human osteoblast-like cells. Methods: The natural material was created by a hot water extraction process after being soaked for 2~3 days in tap water and dried at $50^{\circ}C$. The material was characterized using field emission scanning electron microscopy (FE-SEM), energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and Fourier transformed infrared (FTIR) spectroscopy. The biological behaviors of the material were also investigated; we performed tests to assess cell cytotoxicity, metabolic activity, osteogenic markers related to bone formation, and VEGF. Results: The EDX, XRD, and FTIR results for the natural material indicated the presence of organic compounds. The natural material caused positive increases in cell metabolic activity and mineralized bone formation without cytotoxicity. The protein levels in the extract for the $6.25{\mu}g/mL$, $12.25{\mu}g/mL$, $25{\mu}g/mL$, $50{\mu}g/mL$, and $100{\mu}g/mL$ groups were significantly different from that for the control. Conclusions: The GBR-based natural material was easy to prepare and had characteristics of a potential biomaterial. The biocompatibility of this natural material was evaluated using in vitro techniques; our findings indicate that this novel material is promising for agricultural and biological applications.

Effect of Jeungikgwiryon-tang (Tsengikueijung-tang) on Bone Density and Bone Biochemical Markers in Osteoporotic Rats (난소적출 흰쥐의 골밀도 및 골대사에 증익귀용탕이 미치는 영향)

  • 송영상;임형호
    • The Journal of Korean Medicine
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    • v.24 no.3
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    • pp.11-22
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    • 2003
  • Objective : As the average span of human life extends, more and more people are at risk of developing osteoporosis, one of the typical diseases of the aged. This thesis presents the effects of Jeungikgwiryon-tang (Tsengikueijung-tang) on bone density, bone biochemical markers, and fetal calvarial cells (FCC) of Sprague Dawleys (S.D.) rats that have induced osteoporosis. The purpose is to see how Jeungikgwiryon-tang (Tsengikueijung-tang) reduces osteoporosis symptoms. Methods : In the first experiment Sprague Dawleys rats were administered Jeungikgwiryon-tang (Tsengikueijung-tang) for 70 days, once a day. Two different doses were used, creating high-dosed and low-dosed groups. The results were compared with a control group. In the second experiment, Jeungikgwiryon-tang (Tsengikueijung-tang) was applied to fetal calvarial cells (FCC) obtained from fetuses inside pregnant Sprague Dawleys rats. The FCCs from high-dosed and low-dosed groups were compared with those from a control group. Results : 1. Bone densities in Groups A and B increased significantly from a control group. 2. Bone ash densities in Group A showed substantial increase. 3. Calcium and phosphorus in bones in Group A increased significantly. 4. Activity of fetal calvarial cells' division in Groups A and B increased significantly from a control group, and ALP of fetal calvarial cells' formation in Group A increased significantly. 5. Protein and collagen levels of fetal calvarial cells in Group A increased significantly. Conclusion : It was found that Jeungikgwiryon-tang (Tsengikueijung-tang) has a tendency to make significant increases in bone densities by enhancing bone formation and by retarding bone absorption. It was concluded that Jeungikgwiryon-tang (Tsengikueijung-tang) activates osteoblast cells effectively.

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Implant surface treatments affect gene expression of Runx2, osteogenic key marker

  • Na, Young;Heo, Seong-Joo;Kim, Seong-Kyun;Koak, Jai-Young
    • The Journal of Advanced Prosthodontics
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    • v.1 no.2
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    • pp.91-96
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    • 2009
  • STATEMENT OF PROBLEM. The aim of this study was to study the effects of various surface treatments to a titanium surface on the expression of Runx2 in vitro. MATERIAL AND METHODS. Human Osteosarcoma TE-85 cells were cultured on machined, sandblasted, or anodic oxidized cpTi discs. At various times of incubation, the cells were collected and then processed for the analysis of mRNA expression of Runx2 using reverse transcription-PCR. RESULTS. The expression pattern of Runx2 mRNA was differed according to the types of surface treatment. When the cells were cultured on the untreated control culture plates, the gene expression of Runx2 was not increased during the experiments. In the case of that the cells were cultured on the machined cpTI discs, the expression level was intermediate at the first day, but increased constitutively to day 5. In cells on sandblasted cpTi discs, the expression level was highest in the first day sample and the level was maintained to 5 days. In cells on anodized cpTi discs, the expression level increased rapidly to 3 days, but decreased slightly in the 5-th day sample. CONCLUSION. Different surface treatments may contribute to the regulation of osteoblast function by influencing the level of gene expression of key osteogenic factors.

In vitro assay for osteoinductive activity of different demineralized freeze-dried bone allograft

  • Vaziri, Shahram;Vahabi, Surena;Torshabi, Maryam;Hematzadeh, Somayeh
    • Journal of Periodontal and Implant Science
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    • v.42 no.6
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    • pp.224-230
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    • 2012
  • Purpose: Various bone graft materials have been used for periodontal tissue regeneration. Demineralized freeze-dried bone allograft (DFDBA) is a widely used bone substitute. The current widespread use of DFDBA is based on its potential osteoinductive ability. Due to the lack of verifiable data, the purpose of this study was to assess the osteoinductive activity of different DFDBAs in vitro. Methods: Sarcoma osteogenic (SaOS-2) cells (human osteoblast-like cells) were exposed to 8 mg/mL and 16 mg/mL concentrations of three commercial types of DFDBA: Osseo+, AlloOss, and Cenobone. The effect of these materials on cell proliferation was determined using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The osteoinductive ability was evaluated using alizarin red staining, and the results were confirmed by evaluating osteogenic gene expression using reverse transcription polymerase chain reaction (RT-PCR). Results: In the SaOS-2 cells, an 8 mg/mL concentration of Osseo+ and Cenobone significantly increased cell proliferation in 48 hours after exposure (P<0.001); however, in these two bone materials, the proliferation of cells was significantly decreased after 48 hours of exposure with a 16 mg/mL concentration (P<0.001). The alizarin red staining results demonstrated that the 16 mg/mL concentration of all three tested DFDBA induced complete morphologic differentiation and mineralized nodule production of the SaOS-2 cells. The RT-PCR results revealed osteopontin gene expression at a 16 mg/mL concentration of all three test groups, but not at an 8 mg/mL concentration. Conclusions: These commercial types of DFDBA are capable of decreasing proliferation and increasing osteogenic differentiation of the SaOS-2 cell line and have osteoinductive activity in vitro.

Fabrication of Functionally Gradient Porous Al2O3-(t-ZrO2)/HAp Composites and their In-Vitro Study (조성 경사구조를 갖는 다공질 Al2O3-(t-ZrO2)/HAp 복합체의 제조 및 In-Vitro 실험)

  • Kim, Ki-Ho;Kim, Young-Hee;Song, Ho-Yeon;Lee, Byong-Taek
    • Journal of the Korean Ceramic Society
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    • v.43 no.8 s.291
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    • pp.504-508
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    • 2006
  • Functionally gradient porous $Al_2O_3-(t-ZrO_2)/HAp$ composites consist of 3 layers were fabricated using the multi-pass extrusion process at the various temperatures. The continuous pores were homogeneously formed in the $2^{nd}$ passed samples and their size was about $180{\mu}m$ in diameter. In the porous composites sintered at $1200-1400^{\circ}C$, the relative density and bending strength increased with the sintering temperature. The maximum values of relative density and bending strength in the $2^{nd}$ passed $Al_2O_3-(t-ZrO_2)/HAp$ composites were 62.2% and 107.8 MPa, respectively. In order to investigate the growth behavior of osteogenic cells on the functionally gradient porous $Al_2O_3-(t-ZrO_2)/HAp$ composites, an in vitro test was performed, using human osteoblast-like MG-63 cells. The cells were well attached and grown on the rough surface of the inside of the functionally gradient porous body.

Growth and Osteoblastic Differentiation of Mesenchymal Stem Cells on Silk Scaffolds

  • Cho, Hee-Yeon;Baik, Young-Ae;Jeon, Suyeon;Kwak, Yoon-Hae;Kweon, Hae Yong;Jo, You Young;Lee, Kwang Gill;Park, Young Hwan;Kang, Dongchul
    • International Journal of Industrial Entomology and Biomaterials
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    • v.27 no.2
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    • pp.303-311
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    • 2013
  • In this study, we compared the efficiency of osteoblast differentiation media (ODM) containing three distinct reagent combinations in osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) in monolayer culture. In addition, we analyzed growth and differentiation of hBMSCs on silk scaffolds and examined the bone-forming activity of a nanofibrous silk scaffold in a tibia diaphysis defect model of a rat hind limb with intramedullary nailing. Although all three ODM increased alkaline phosphatase activity to a comparable extent, the ODM containing bone morphogenetic protein-2 (BMP-2) was found to be significantly less effective in promoting mineral deposition than the others. Growth of hBMSCs on sponge-form silk scaffolds was faster than on nanofibrous ones, while osteoblastic differentiation was apparent in the cells grown on either type of scaffold. By contrast, bone formation was observed only at the edge of the nanofibrous scaffold implanted in the tibia diaphysis defect, suggesting that use of the silk scaffold alone is not sufficient for the reconstitution of the long bone defect. Since silk scaffolds can support cell growth and differentiation in vitro, loading MSCs on scaffolds might be necessary to improve the bone-forming activity of the scaffold in the long bone defect model.

Antibacterial Effects and Cytotoxicity of Crassirhizomae Rhizoma (관중의 항균작용 및 세포독성에 관한 연구)

  • Kim, Seung-Nam;Ku, Young;Rhyu, In-Cheol;Hahm, Byung-Do;Bae, Ki-Hwan;Han, Soo-Boo;Chung, Chong-Pyoung;Choi, Sang-Mook
    • Journal of Periodontal and Implant Science
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    • v.30 no.1
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    • pp.65-77
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    • 2000
  • The purpose of this study was to evaluate the antimicrobial activity of Crassirhzimae rhizoma and its possible use as an oral antiseptics for prevention of periodontitis. Its antibacterial activity against periodontopathic microorganisms including Actinobacillus actiomycetem - comitans, Capnocytophaga ochracea, Streptococcus mutans, Porphyromonas gingivalis, Prevotella intermedia, Actinomyces viscosus, Fusobacterium nucleatumwas evaluated via modified stab culture method. The cytotoxicity against gingival fibroblasts and rat osteoblasts was investigated via [$^3H$]thymidine incorporation and cellular activity was investigated via MTT assay. Chlorhexidine was used as control group. Crassirhizomae rhizoma was prepared at concentrations of 0.2, 0.15, 0.1, 0.05%. Chlorhexidine was also prepared at the same concentration. Crassirhizomae rhizoma showed lower antimicrobial antivity against these microorganism than chlorhexidine, but this difference was not significant. And, Crassirhzomae rhizoma showed more cellular activity and less cytotoxicity than chlorhexidine on human gingival fibrablast and rat osteoblast. This study suggests that Crassirhzomae rhizoma might be a candidate for a safe oral antiseptic for the prevention and treatment of periodontal disease.

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Effect of BMP-7 on osteoblastic differentiation of rat periodontal ligament cells (백서 치주인대세포의 분화에 대한 Bone morphogenetic protein-7의 영향)

  • Lee, Ho-Jae;Kim, Young-Jun;Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.35 no.3
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    • pp.747-760
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    • 2005
  • Periodontal therapy has dealt primarily with attempts at arresting progression of disease. however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. Recombinant human bone morphogenetic protein-7(rhBMP-7) can differentiate the osteoprogenitor cells and induce bone formation. The purpose of this study was to evaluate the effect of BMP-7 on rat periodontal ligament cells differentiation, in vitro. In the control group, cells was cultured with DMEM media. In the experimental groups, cells were cultured with rhBMP-7 in concentration of 10, 25, 50 and 100 ng/ml. Each group was characterized by examining alkaline phosphatase activity at 3 and 5 days of culture and the ability to produce mineralized nodules of rat calvarial cells at 14 days of culture. Synthesis of type I collagen(COL-I), osteocalcin(OCN), and bone sialoprotein(BSP) was evaluated by RT-PCR at 7 days of culture. Activation of Smad proteins and p38 MAP kinase was determined by western blot analysis of the cell lysates. Alkaline phosphatase activity was significantly increased in the concentration of BMP-7 50 ng/ml and 100 ng/ml compared to the control(p<0.05). The mineralized bone nodule formation was greater with addition of 50 ng/ml and 100 ng/ml BMP-7 than the control(p<0.01). In 7 days' culture, the expressions of COL-I, BSP, and OCN was increased by BMP-7 in concentration of 10 $ng/ml{\sim}100$ ng/ml. In western blot analysis, BMP-7 treated culture cells expressed Smad 1,5,8 in dose-dependent manner, whereas BMP-7 did not activate phosphorylated form of p38 MAP kinase. These result suggested that BMP-7 stimulate rat periodontal ligament cells to differentiate toward osteoblast phenotype and increase bone matrix production by activation of BMP-Smad pathway.