• Journal of Biosystems Engineering
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• 제39권3호
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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.

• 대한한의학회지
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• 제24권3호
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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.

• The Journal of Advanced Prosthodontics
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• 제1권2호
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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.

• Journal of Periodontal and Implant Science
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• 제42권6호
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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.

• 한국세라믹학회지
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• 제43권8호
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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.

• International Journal of Industrial Entomology and Biomaterials
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• 제27권2호
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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.

• Journal of Periodontal and Implant Science
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• 제30권1호
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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.

• Journal of Periodontal and Implant Science
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• 제35권3호
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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.

• Tissue Engineering and Regenerative Medicine
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• 제15권6호
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• pp.793-801
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• 2018

BACKGROUND: The aim of this study was to evaluate the combined effect of low-level laser treatment (LLLT) and recombinant human bone morphological protein-2 (rhBMP-2) applied to hypoxic-cultured MC3T3-E1 osteoblastic cells and to determine possible signaling pathways underlying differentiation and mineralization of osteoblasts under hypoxia. METHODS: MC3T3-E1 cells were cultured under 1% oxygen tension for 72 h. Cell cultures were divided into four groups: normoxia control, low-level laser (LLL) alone, rhBMP-2 combined with LLLT, and rhBMP-2 under hypoxia. Laser irradiation was applied at 0, 24, and 48 h. Cells were treated with rhBMP-2 at 50 ng/mL. Alkaline phosphatase activity was measured at 3, 7, and 14 days to evaluate osteoblastic differentiation. Cell mineralization was determined with Alizarin red S staining at 7 and 14 days. Western blot assays were performed to evaluate whether p38/protein kinase D (PKD) signaling was involved. RESULTS: The results indicate that LLLT and rhBMP-2 synergistically increased alkaline phosphatase (ALP) activity and mineralization. Western blot analyses showed that expression of type I collagen, runt-related transcription factor 2 (RUNX2), and Osterix (Osx), increased and expression of hypoxia-inducible factor 1-alpha ($HIF-1{\alpha}$), decreased more in the LLLT and rhBMP-2 combined group than in the rhBMP-2 or LLL alone groups. Moreover, LLLT and rhBMP-2 stimulated p38 phosphorylation and rhBMP-2 and LLLT increased Prkd1 phosphorylation. CONCLUSION: Combined treatment with rhBMP-2 and LLL induced differentiation and mineralization of hypoxic-cultured MC3T3-E1 osteoblasts by activating p38/PKD signaling in vitro.

• Journal of Dental Anesthesia and Pain Medicine
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• 제16권4호
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• pp.263-271
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• 2016

Background: Bone injury is common in many clinical situations, such as surgery or trauma. During surgery, excessive reactive oxygen species (ROS) production decreases the quality and quantity of osteoblasts. Remifentanil decreases ROS production, reducing oxidative stress and the inflammatory response. We investigated remifentanil's protective effects against $H_2O_2$-induced oxidative stress in osteoblasts. Methods: To investigate the effect of remifentanil on human fetal osteoblast (hFOB) cells, the cells were incubated with 1 ng/ml of remifentanil for 2 h before exposure to $H_2O_2$. For induction of oxidative stress, hFOB cells were then treated with $200{\mu}M$ $H_2O_2$ for 2 h. To evaluate the effect on autophagy, a separate group of cells were incubated with 1 mM 3-methyladenine (3-MA) before treatment with remifentanil and $H_2O_2$. Cell viability and apoptotic cell death were determined via MTT assay and Hoechst staining, respectively. Mineralized matrix formation was visualized using alizarin red S staining. Western blot analysis was used to determine the expression levels of bone-related genes. Results: Cell viability and mineralized matrix formation increased on remifentanil pretreatment before exposure to $H_2O_2$-induced oxidative stress. As determined via western blot analysis, remifentanil pretreatment increased the expression of bone-related genes (Col I, BMP-2, osterix, and $TGF-{\beta}$). However, pretreatment with 3-MA before exposure to remifentanil and $H_2O_2$ inhibited remifentanil's protective effects on hFOB cells during oxidative stress. Conclusions: We showed that remifentanil prevents oxidative damage in hFOB cells via a mechanism that may be highly related to autophagy. Further clinical studies are required to investigate its potential as a therapeutic agent.