• YAKHAK HOEJI
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• v.58 no.5
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• pp.307-313
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• 2014

Rumex crispus (curled dock), which is a perennial wild plant, has long been used as a laxative, astringent, and medicine to treat blood and skin diseases. We recently reported that the roots of R. crispus are an effective nutraceutical for bone. This study prepared ethanol extracts of the leaves and roots of R. crispus, and analyzed the major constituents using liquid chromatography and mass spectrometry. In addition, their effects on the proliferation and differentiation of human osteoblast-like MG-63 cells, such as cell viability, alkaline phosphatase (ALP) activity, collagen content, and mineralization, were compared. The chromatograms of the chemical constituents of the two extracts exhibited quite different profiles: quercetin and quercitrin were identified as major peaks in the leaf extract, whereas cinnamtannin B1 and procyanidin isomers were the major peaks for the root extract. Neither extract was cytotoxic at concentrations of < $25{\mu}g/ml$. ALP activity and collagen synthesis-which are markers of the early stage of osteogenesis-in MG-63 cells were significantly increased upon the addition of the root extract compared with the addition of the leaf extract. In contrast, the leaf extract had a more stimulatory effect on mineralization-which is marker of the late stage of osteogenesis-in MG-63 cells than did the root extract. In conclusion, extracts of both leaves and roots of R. crispus stimulated the bone-forming activity of osteoblasts; in particular, the root extract was more effective in the early stage of osteoblast differentiation, while the leaf extract was more effective in the late stage. This difference in anabolic activity may be due to differences in the constituents of the leaves and roots. The leaves and roots of R. crispus appear to complement each other as stimulators of bone formation.

• Korean Journal of Materials Research
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• v.16 no.10
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• pp.606-613
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• 2006

This study was purposed to evaluate the influence of anodically oxidized film on titanium (Ti) onto MG-63 osteoblast-like cell attachment and activity. Only scratch lines created by polishing were seen in ASR and ANO-1 groups. About $1.5{\mu}m$-thick homogeneous oxide film which has pores of about $0.5{\mu}m$ diameter were formed in ANO-12. The crystalline structure of the oxide films formed by anodization in phosphoric acid electrolyte was $TiP_2O_7$. The total protein amounts of ANO-1 and ANO-12 groups showed higher values of maximum protein amount than that of AS-R group. At 3 days of incubation, total protein amount showed higher value in ANO-2 when comparing to that of AS-R (p<0.05). Based on the results of ALPase activity test, the degree of MG-63 cell differentiation for initial mineralization matrix formation was similar. For all the test groups after 1 day of incubation, MG-63 cells grew healthily in mono-layer with dendritic extensions. After incubation for 3 days, the specimen surfaces were covered more densely by cells, and numerous micro filaments were extruding to the extracellular matrix.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.1
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• pp.142-147
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• 2008

This study was purpose to investigate that Gucheokbogol-tang(GBT), mainly used in osteoporosis, has possible regulatory effect on bone regeneration by operating in proliferation and calcification of osteoblast. So that, their cytotoxicity and proliferation was investigated with MTT by ELISA, and morphological change was observed by optical microscope and electron microscope. And activation of alkaline phosphates(ALP) which is secreted in early stage of bone formation was measured, and accumulation of $Ca^{2+}$ in the process of calcification was investigated by alizaline red S assay(ARS). The results were as follows: The MG-63 cell, originated from human cell was activated in $10^{-5}$ treated group, and the level of ALP was also increased in the treated group, highest on the third day. And from the outcome of ARS assay of calcification process in the Gucheokbogol-tang(GBT) treated group for 21days. These results suggested that Gucheokbogol-tang(GBT) is effective in proliferation and calcification of osteoblast.

• Journal of Society of Preventive Korean Medicine
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• v.8 no.2
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• pp.13-30
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• 2004

This study was performed to evaluate the effect of Kangwhal-Sokdan tang(KS) on osteoblast function and gene expression. The osteoblast separated from the murine calvariae and MG-63 cell were cultivated to evaluate the cell function and gene expression. The results were summarized as followes. 1) KS increased cell proliferation of murine calvarial cell. 2) KS increased protein synthesis, collagen synthesis and ALP activity of murine calvarial cell. 3) KS increased the survival rate of murine calvarial cell. 4) KS increased the expression of calcitonin receptor and PTH receptor. 5) KS increased the expression of PKA and PKC. 6) KS decreased the expression of $PLA_2$, COX, $PGE_2$ synthase, but increased prostacyclin synthase. 7) KS increased the expression of collagen(type IV) gene. It is concluded that KS might improve the osteoporosis resulted from augumentation of osteoblast proliferation and gene expression.

• Journal of the Korean Applied Science and Technology
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• v.38 no.1
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• pp.168-177
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• 2021

The effects of the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture on the activities of osteoblast differentiation and the restraint of osteoclast formation were investigated. the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture in the human osteoblast "MG-63" cell, was examined in relation to alkaline phosphatase (ALP) activity and alizarin red stains. In order to observe the effects of osteoclasts formation, we analyzed RAW 264.7 cell tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains. In cytotoxicity testing, it was confirmed that apple extract is safe at a concentration of 50 ㎍/㎖ or less. The ALP activity and Bone calcification formation ability were the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture had a lower activity than that of control group. However the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture significantly reduced activity of TRAP in the RAW 264.7 osteoclastic cell generation and effectively Inhibited the TRAP(+) multinuclear cells. Thus, our results demonstrate that the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture enhances the inhibitory activity of bone-resorption in RAW 264.7 cells. In conclusion, the Perilla frutescens var. crispa and Atractylodes macrocephala Koidzumi mixture seem to be effective in the prevention and treatment of bone related disorders.

• The Journal of Korean Academy of Prosthodontics
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• v.52 no.3
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• pp.211-221
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• 2014

Purpose: The purpose of this study was to evaluate the surface characteristics and response of osteoblast-like cell at SLA surface treated with NaOH. Materials and methods: Three kinds of specimens were fabricated for the experiment groups. Control group was a machined surface, SLA group was a conventionally SLA treated surface, and SLA/NaOH gorup was SLA surface treated with NaOH. To evaluate the surface characteristics, the surface elemental composition (XPS), surface roughness and surface contact angle were evaluated in each group. And the cytotoxicity, cell adhesion, cell proliferation and ATP activity of osteoblast-like cells (MG-63 cells) were compared in each group for evaluatation of the cell responses. Statistical comparisons between groups were carried out via one-way ANOVA using the SPSS software (SPSS Inc., Chicago, USA), and then performed multiple comparisons. The differences were considered statistically significant at P<.05. Results: SLA surface treated with NaOH (SLA / NaOH group) was changed to hydrophilic surface. All groups did not show the cytotoxicity to the MG-63. In cell adhesion studies, SLA / NaOH group showed the higher degree of adhesion than anothers (P<.05), Up to 7 days of incubation, the proliferation was showed the increasing tendency in all groups but SLA / NaOH group showed the highest cell proliferation between the three groups (P<.05). At 7 days of incubation, there was no difference in ALP activities between the three groups, but at 14 days, SLA / NaOH group showed significant increase in ALP activities (P<.05). Conclusion: In this study, SLA surface treated with NaOH promoted cell adhesion, proliferation and differentiation. It means that SLA/NaOH group is possible to promote osseointegration of implants.

• Microbiology and Biotechnology Letters
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• v.43 no.4
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• pp.322-329
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• 2015

The effects of water extracts of Gallus gallus var. domesticus (Yeonsan ogolgye, GD) on the activities of osteoblast differentiation and the restraint of osteoclast formation were investigated. The water extract of GD in the human osteoblast "MG-63" cell, was examined in relation to alkaline phosphatase (ALP) activity and alizarin red stains. In order to observe the effects of osteoclasts formation, we analyzed RAW 264.7 cell tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains. The ALP activity of the water extract of hen and cock flesh (3 years) were 133.8% and 129.6%, respectively. The ALP activity of flesh extracts was also higher than that of the skin extracts. Concerning the effects of age, the 3 years old flesh extracts had a higher activity than that of the one year old extracts. However the activity of the 3 years old skin extracts was lower than that of the one year old extracts. For gender conditions, the ALP activity of the hen extract was higher than that of the cock. The degree bone mineralization in the three years old hen flesh exhibited the highest rate, at 124.3%, amongst all the groups. The TRAP activity of the flesh extracts of the three years old cock revealed the lowest rate, at 31.8%, compared to the control. Our results demonstrate that the water extract of GD increases bone mineralization and osteoblast differentiation activity in MG-63 cells and enhances the inhibitory activity of bone-resorption in RAW 264.7 cells. In conclusion, the water extracts of GD seem to be effective in the prevention and treatment of bone related disorders.

• Korean Journal of Oriental Medicine
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• v.6 no.1
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• pp.123-132
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• 2000

Bone is continuously remodeled during adult life with the resorption of old bone by osteoclasts and its subsequent replacement by osteoblast. Bone homeostasis is maintained by a balance between activities of osteoblasts and osteoclasts, but an imbalance between resorption and formation results in bone diseases including osteoporosis. Osteoblasts line up on the bone surface, especially regions of new bone formation, lay down bone matrix (osteoid) in orderly lamellae and induce its mineralization. Thus, the increased activity of osteoblasts is helpful to treat and prevent osteoporosis. In this study, we examined whether 80% EtOH extract of yukmijiwhang-whan is capable of affecting osteoblast proliferation using human osteoblast-like cell line, MG-63 and Saos-2. In an in vivo experiment, extract of yukmijiwhang-whan was administered for 9 weeks to ovariectomized (OVX) rats. At necropsy, uterus weights were measured, and trabecular bone areas (TBAS) of tibia and the sixth lumbar vertebra were measured by bone histomorphology. The maximum cell proliferation of MG-63 caused by extract of yukmijiwhang-whan at $5\;{\times}\;10^{-6}\;mg/ml$ was approximately 115% compared with control. In Saos-2, cell proliferation was approximately 145% of control at $5\;{\times}\;10^{-4}\;mg/ml$ and maximum alkaline phosphatase (ALP) activity was approximately 143% of control at $5\;{\times}\;10^{-5}\;mg/ml$. In animal study, however, the tibia and lumbar TBAS of the yukmijiwhang-whan group did not increased than the OVX control group. In conclusion, the 80% EtOH extract of yukmijiwhang-whan increased proliferation of osteoblasts but did not prevent bone loss in OVX rats.

• Animal cells and systems
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• v.13 no.4
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• pp.363-370
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• 2009

Since vitamin $D_3$ is an important regulator of osteoblastic differentiation, a presently-established vitamin $D_3$-entrapped calcium phosphate film (VCPF) was evaluated for hard tissue engineering. The entrapped vitamin $D_3$ more rapidly induced bone nodule formation. To characterize the cellular events leading to regulations including faster differentiation, signal transduction pathways were investigated in osteoblastic MG63 cells at a molecular level. Major signaling pathways for MG63 cell proliferation including phosphatidylinositol-3-kinase, extracellular signal-regulated kinase, c-Jun N-terminal kinase and focal adhesion kinase pathways were markedly down-regulated when cells were cultured on calcium phosphate film (CPF) and VCPF. This agreed with our earlier observations of the immediate delay in proliferation of MG63 cells upon culture on CPF and VCPF. On the other hand, the p38 mitogen-activated protein kinase (p38 MAPK) and protein kinase A (PKA) pathways were significantly up-regulated on both CPF and VCPF. CPF alone could simulate differential behaviors of MG63 cells even in the absence of osteogenic stimulation and entrapment of vitamin $D_3$ within CPF further amplified the signal pathways, resulting in continued promotion of MG63 cell differentiation. Interplay of p38 MAPK and PKA signaling pathways likely is a significant event for the promotion of differentiation and mineralization of MG63 cells.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.33 no.4
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• pp.308-313
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• 2011

Purpose: Beta-tricalcium phosphate (${\beta}$-TCP) is a synthetic calcium phosphate ceramic that has widely been used as a bone material to repair bone defects. Despite many clinical studies, the molecular mechanism whereby this biomaterial alters the gene expression in osteoblasts to promote bone formation is poorly understood. Thus, we attempted to address this question by using microarray techniques to identify the genes that are differentially regulated in osteoblasts exposed to ${\beta}$-TCP. Methods: By using DNA microarrays, we identified several genes whose expression levels were significantly up- or down-regulated in osteoblast-likeMG-63cells cultured with ${\beta}$-TCP at a concentration of 100 mg/10 ml for 24 hours. Results: The differentially expressed genes covered a broad range of functional activities: signal transduction, transcription, cell cycle regulation, vesicular transport, apoptosis, immunity, cytoskeletal elements and cell proliferation and differentiation. Conclusion: The gene expression changes related to cell proliferation and differentiation, vesicle transport, immunity and defense could affect the osteogenic activities of osteoblasts for bone regeneration. However, further studies will be required to verify the relative importance of these genes in bone formation, their temporal and spatial expression patterns and their interactions with each other.