• Journal of Dairy Science and Biotechnology
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• v.23 no.2
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• pp.135-141
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• 2005

• 한국유가공학회:학술대회논문집
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• 2005.06a
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• pp.85-106
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• 2005

• Journal of Acupuncture Research
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• v.29 no.6
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• pp.11-21
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• 2012

Objectives : BHH10 is traditional medicine herb used for enhancing body resistance against various diseases. The aim of this study was to identify BHH10 extract induces osteogenic activity in human osteoblast-like MC3T3-E1 cells. Methods : MC3T3-E1, pre-osteoblast cell line, were treated with BHH10 of various concentrations($0.1{\mu}g/mL$, $1{\mu}g/mL$, $10{\mu}g/mL$). And then, the effect of BHH10 on osteoblast differentiation was examined by alkaline phosphatase(ALP) activity, von Kossa staining and RT-PCR for osteoblast differentiation markers such as osteocalcin(OCN), osteopontin(OPN). Results : BHH10 had dose-dependent effect on the viability of osteoblastic cells, and dose-dependently increased alkaline phosphatase(ALP) activity. BHH10 markedly increased mRNA expression for OCN, OPN in MC3T3-E1 cells. Also, BHH10 significantly induced mineralization in the culture of MC3T3-E1 cells. Conclusions : In conclusion, these results propose that BHH10 can play an important role in osteoblastic bone formation, osteogenesis, and may possibly lead to the development of bone-forming drugs.

• BMB Reports
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• v.50 no.2
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• pp.97-102
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• 2017

Patients with inflammatory bone disease or cancer exhibit an increased risk of fractures and delayed bone healing. The S100A4 protein is a member of the calcium-binding S100 protein family, which is abundantly expressed in inflammatory diseases and cancers. We investigated the effects of extracellular S100A4 on osteoblasts, which are cells responsible for bone formation. Treating primary calvarial osteoblasts with recombinant S100A4 resulted in matrix mineralization reductions. The expression of osteoblast marker genes including osteocalcin and osterix was also suppressed. Interestingly, S100A4 stimulated the nuclear factor-kappaB (NF-${\kappa}B$) signaling pathway in osteoblasts. More importantly, the ex vivo organ culture of mouse calvariae with recombinant S100A4 decreased the expression levels of osteocalcin, supporting the results of our in vitro experiments. This suggests that extracellular S100A4 is important for the regulation of bone formation by activating the NF-${\kappa}B$ signaling pathway in osteoblasts.

• Korean Journal of Medicinal Crop Science
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• v.19 no.6
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• pp.491-500
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• 2011

Ursolic acid, triterpenoid compound has been shown to stimulate osteoblast differentiation and enhance bone formation. In the present study, we examined the effects of similar triterpenoid compounds, oleanolic acid (OA) and its derivatives, such as oleanolic acid acetate (OAA) and oleanolic acetate methyl ester (OAM) on the bone formation in MC3T3-E1 osteoblast cells. We determined cellular proliferation, alkaline phosphatase (ALP) activity, mineralization, and expression of osteoblast specific genes and mitogen activated protein kinase phosphorylation. Treatment of $0.1-10{\mu}m$ OA, OAA, and OAM increased cellular proliferation, but not significantly increased as compared with dimethyl sulfoxide (DMSO). OA, OAA, and OAM at 5uM concentration enhanced ALP expression, mineralization, and osteocalcin (OCN) mRNA level. In conclusion, OA and its derivatives stimulated the osteoblast differentiation by increasing ALP, mineralization, and OCN mRNA expression. However, there were no significantly difference on osteoblast differentiation among treatment of OA, OAA, and OAM.

• Journal of Periodontal and Implant Science
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• v.28 no.3
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• pp.475-489
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• 1998

The main goal for the treatment of periodontal diseases is the regeneration of lost cementum, bone and connective tissue. Clinical and histological research suggests that it is possible to restore periodontal structures. Seeds of Carthamus tinctorius L. has been used for the treatment of bone fracture and osteoporosis in traditional Korean medicine. The purpose of this study is to examine the effect of extract of seeds of Carthamus tinctorius L. on mineralization in periodontal ligament cells and osteoblastic cells. Periodontal ligament cells were primarily obtained from a extracted premolars with non-periodontal diseases, Osteoblastic cells were obtained from calvariae of a fetal rat, Cells were cultured with DMEM at $37^{\circ}C$ with 5% $CO_2$ in 100% humidity incubator. Alkaline phosphatase(ALP) level and the number of calcification nodules were examined and western blot analysis using osteonectin was performed, Measurements of ALP levels and calcification nodules showed that extract of seeds of Carthamus tinctorius L. had significantly higher activity than control in all of both cells. In western blot analysis, protein expression of osteonectin indicated that extract of seeds of Carthamus tinctorius L. showed an increased pattern than control in all of both cells. From the above results, it seems that extract of seeds of Carthamus tinctorius L. has excellent effect on mineralization in periodontal ligament cells and osteoblastic cells.

• Journal of Periodontal and Implant Science
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• v.49 no.2
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• pp.114-126
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• 2019

Purpose: The aim of this study was to evaluate the enhancement of osteogenic potential of biphasic calcium phosphate (BCP) bone substitute coated with Escherichia coli-derived recombinant human bone morphogenetic protein-2 (ErhBMP-2) and epigallocatechin-3-gallate (EGCG). Methods: The cell viability, differentiation, and mineralization of osteoblasts was tested with ErhBMP-2-/EGCG solution. Coated BCP surfaces were also investigated. Standardized, 6-mm diameter defects were created bilaterally on the maxillary sinus of 10 male New Zealand white rabbits. After removal of the bony windows and elevation of sinus membranes, ErhBMP-2-/EGCG-coated BCP was applied on one defect in the test group. BCP was applied on the other defect to form the control group. The animals were sacrificed at 4 or 8 weeks after surgery. Histologic and histometric analyses of the augmented graft and surrounding tissue were performed. Results: The 4-week and 8-week test groups showed more new bone (%) than the corresponding control groups (P<0.05). The 8-week test group showed more new bone (%) than the 4-week test group (P<0.05). Conclusions: ErhBMP-2-/EGCG-coated BCP was effective as a bone graft material, showing enhanced osteogenic potential and minimal side effects in a rabbit sinus augmentation model.

• Tissue Engineering and Regenerative Medicine
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• v.15 no.6
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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.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.27 no.1
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• pp.16-23
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• 2005

It is commonly acknowledged that bone morphogenic protein (BMP-2) functions as a potential osteogenic inducer in bone formation. Recently, several papers reported that bone marrow-derived stem cell (BMSC) from human is not responsive to BMP-2 in comparison to high capacity of BMP-2 in the osteoinduction of stromal cell derived from bone marrow of rodent animals such as rat or mouse. In this study, we characterized BMSC derived from 11 years old donor for the responsiveness to rhBMP-2, dexamethasone (Dex) and 1,25-dihydroxyvitamin D (vitamin D), in order to analyze their function in the early osteogenesis. The effect of over mentioned agents was evaluated by means of assessing alkaline phosphatase (ALP) activity/staining, RT-PCR analysis and von Kossa staining. In addition, we analyzed the meaning of expressed several osteoblastic markers such as alkaline phosphatase, collagen typeI, osteopontin, bone sialoprotein and osteocalcin with relation to either differentiation or mineralization. Only in the presence of Dex, human BMSC could commit osteoblastic differentiation and matrix mineralization, and either BMP-2 or vitamin D treatment was not able to induce. But BMP-2 or Vitamin D showed potential synergy effect with Dex. ALP and bone sialoprotein were clearly expressed in response of Dex treatment compared to weak expression of osteopontin in early osteogenesis. Therefore, we expect that this study will contribute partly to elucidiating early osteogenesis mechanism in human, but variations among bone marrow donors must be considered through further study.

• Journal of Periodontal and Implant Science
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• v.33 no.2
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• pp.259-269
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• 2003

The ideal goal of periodontal therapy is the regeneration of periodontal tissue repair of function. Although is very difficult to attain the goal, recent advances in periodontal wound healing concepts encourage hope reaching it. Recently many efforts are concentrated on the regeneration potential of material used in traditional Korean medicine. Phlomidis Radix has been used for the treatment of blood stasis, bone fracture and osteoporosis in traditional Korean medicine. The purpose of this study is to examine effects of dichloromethane fraction Phlomidis Radix on Bone Formation in Human Fetal Osteoblasts. Human fetal osteoblastic cell line(hFOB1 1.19 ;American Type Culture Collection, Manassas, VA) were used and cells were cultured containing DMEM and dichloromethane fraction Phlomidis Radix(100 ng/ml , 1 ${\mu}$/ml, 10 ${\mu}$/ml) at 34$^{\circ}C$ with 5% $CO_2$ in 100% humidity. MTT was performed to examine the viability of the cell, and alkaline phosphatase activity was analyzed to examine the mineralization. Also bone calcification nodules were evaluated. The cellular activity of hFOB1 was increased in 100 ng/ml, 1 ${\mu}$/ml , 10 ${\mu}$/ml of dichloromethane fraction of Phlomidis Radix and especially significant increation was showed in 100 ng/ml of dichloromethane fraction of Phlomidis Radix at 6days (p <0.05). ALP level of hFOB1 was significantly increased in 100 ng/ml , 1 ${\mu}$/ml, 10 ${\mu}$/ml of dichloromethane fraction of Phlomidis Radix and especially more increation was showed in 10 ${\mu}$/ml of dichloromethane fraction of Phlomidis Radix (p <0,05). Calcification nodules of hFOB1 significantly increased in 10 ${\mu]$/ml of dichloromethane fraction of Phlomidis Radix at 21 days of incubation(p<0.05). The results indicate that dicholoromethane fraction of Phlomidis Radix has excellent effects on mineralization of hFOB1.