• 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 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.

• The Journal of Korean Obstetrics and Gynecology
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• v.25 no.2
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• pp.23-42
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• 2012

Objectives: This study was performed to evaluate the effect of Samkieumgamibang (SKG) on osteoporosis. Methods: The osteoclastogenesis and gene expression were determined in RANKL-stimulated RAW 264.7 cell. And, osteoblastogenesis was also determined in rat calvarial cell. Results: SKG decreased the number of TRAP positive cell in osteoclast. It also decreased the expression of Cathepsin K, MMP-9, TRAP, c-fos, NAFTc1 and JNK1 in osteoclast. SKG increased the expression of iNOS in RANKL-stimulated in osteoclast. Otherwise, SKG inhibited TRAP activity in osteoclast. SKG increased cell proliferation, ALP activity, bone martix protein, collagen and nodule in osteoblast. Conclusions: It is concluded that SKG might decrease the bone resorption resulted from decrease of osteoclast differentiation and it's related gene expression. And, SKG might increase the bone formation resulted from increase of osteoblast function.

• 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.

• Journal of Periodontal and Implant Science
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• v.35 no.4
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• pp.851-861
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• 2005

The nature of the implant surface can directly influence cellular response, ultimately affecting the rate and quality of new bone tissue formation. The aim of this in vitro study was to investigate if human osteoblast-like cells, Saos-2, would respond differently when plated on disks of magnesium titanate and machined titanium. Magnesium titanate disks were prepared using Micro Arc Oxidation(MAO) methods. Control samples were machined commercially pure titanium disks. The cell adhesion, proliferation and differentiation were evaluated by measuring cell number, and alkaline phosphatase(ALPase) activity at 1 day and 6 day after plating on the titanium disks. Measurement of cell number and ALPase activity in Saos-2 cells at 1 day did not demonstrate any difference between machined titanium and magnesium titanate. When compared to machined titanium disks, the number of cells was reduced on the magnesium titanate disks at 6 day, while ALPase activity was more pronounced on the magnesium titanate. Enhanced differentiation of cells grown on magnesium titanate samples was indicated by decreased cell proliferation and increased ALPase activity.

• YAKHAK HOEJI
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• v.57 no.6
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• pp.406-411
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• 2013

Rumex crispus (Curled Dock, Polygonaceae) is a perennial wild plant used in traditional medicine as a laxative, astringent, and to treat blood and skin disease. The ethanol extract of R. crispus was obtained and its carbohydrate contents were analyzed using high-performance anion-exchange chromatography with pulsed amperometric detection. The anabolic effects of R. crispus in human osteoblastic MG-63 cells were investigated using the WST-8 assay, alkalinephosphatase (ALP) assay, and mineralization assay. The ethanol extract increased the proliferation of MG-63 cells and stimulated ALP activity in a dose-dependent manner over a 72-hrs period. Additionally, the ethanol extract dose-dependently stimulated the formation of bone nodules in MG-63 cells treated for 12 days. The ethyl acetate fraction from the ethanol extract did not affect osteoblast viability but induced an increase in ALP activity. In conclusion, the ethanol extract of R. crispus increases the proliferation and bone-forming activity of osteoblasts, and hence it could be used in the development of bone-forming stimulatory nutraceuticals and osteoporosis-related medicines.

• Archives of Plastic Surgery
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• v.36 no.3
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• pp.247-253
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• 2009

Purpose: Hydroxyapatite(HA) has been widely used due to its chemical similarity to bone and good biocompatibility. HA is composed of macropores and micropores. Too much irregularities of the micropores are ineffective against the adhesion and proliferation of osteoblast. Many efforts have been tried to overcome these drawbacks. HA crystal coating on the irregular surface of HA scaffold, crystallized HA, is one of the method to improve cell adhesion. Meanwhile, the collagen has been incorporated with HA to create composite scaffold that chemically resembles the natural extracellular matrix components of bone. The authors proposed to examine the effect of collagen - coated crystallized HA on the adhesion and proliferation of osteoblast. Method: HA powder containing $10{\mu}m$ pore size was manufactured as 1 cm pellet size. For the making crystallized HA, 0.1 M EDTA solution was used to dissolve HA powder and heated $100^{\circ}C$ for 48 hours. Next, the crystallized HA pellets were coated with collagen (0.1, 0.5, and 1%). The osteoblasts were seeded into HA pellets and incubated for the various times (1, 5, and 9 days). After the indicating days, methylthiazol tetrazolium (MTT) assay was performed for cell proliferation and alkaline phosphatase (ALP) activty was measured for bone formation. Result: In SEM study, the surface of crystallized HA pellet was more regular than HA pellet. MTT assay showed that the proliferation of osteoblasts increased in a collagen dose - dependent and time - dependent manner and had a maximum effect at 1% collagen concentration. ALP activity also increased in a collagen dose - dependent manner and had a highest effect at 1% collagen concentration. Conclusion: These data showed that crystallization and collagen coating of HA was effective for osteoblast proliferation and ALP activity. Therefore, our results suggest that crystallized - HA scaffold with collagen coating is may be a good strategy for tissue engineering application for bone formation.

• The Korean Journal of Physiology and Pharmacology
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• v.18 no.4
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• pp.347-352
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• 2014

Most known osteoporosis medicines are effective for bone resorption, and so there is an increasing demand for medicines that stimulate bone formation. Watercress (N. officinale R. Br.) is widely used as a salad green and herbal remedy. This study analyzed a watercress extract using ultra-performance liquid chromatography/mass spectrometry, and identified a rutin as one of its major constituents. Osteogenic-related assays were used to compare the effects of watercress containing rutin (WCR) and rutin alone on the proliferation and differentiation of human osteoblast-like MG-63 cells. The reported data are expressed as percentages relative to the control value (medium alone; assigned as 100%). WCR increased cell proliferation to $125.0{\pm}4.0%$ ($mean{\pm}SD$), as assessed using a cell viability assay, and increased the activity of alkaline phosphatase, an early differentiation marker, to $222.3{\pm}33.8%$. In addition, WCR increased the expression of collagen type I, another early differentiation marker, to $149.2{\pm}2.8%$, and increased the degree of mineralization, a marker of the late process of differentiation, to $122.9{\pm}3.9%$. Rutin alone also increased the activity of ALP (to $154.4{\pm}12.2%$), the expression of collagen type I (to $126.6{\pm}6.2%$), and the degree of mineralization (to $112.3{\pm}5.0%$). Daidzein, which is reported to stimulate bone formation, was used as a positive control; the effects of WCR on proliferation and differentiation were significantly greater than those of daidzein. These results indicate that WCR and rutin can both induce bone formation via the differentiation of MG-63 cells. This is the first study demonstrating the effectiveness of either WCR or rutin as an osteoblast stimulant.

• International Journal of Oral Biology
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• v.33 no.4
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• pp.179-186
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• 2008

Several lines of evidence suggest that osteocytes play a critical role in bone remodeling. Both healthy and apoptotic osteocytes can send signals to other bone surface cells such as osteoblasts, osteoclasts, osteoclast precursors, and bone lining cells through canalicular networks. Osteocytes responding to mechanical strain may also send signals to other cells. To determine the role for osteocytes an mechanical strain in bone remodeling, we examined the effects of fluid flow shear stress on osteoclast precursor cell and osteoblast proliferation and recruitment induced by osteocytes. In addition, the effects of fluid flow shear stress on osteocyte M-CSF, RANKL, and OPG mRNA expression were also examined. MLO-Y4 cells were used as an in vitro model for osteocytes, RAW 264.7 cells and MOCP-5 cells as osteoclast precursors, and 2T3 cells as osteoblasts. MLO-Y4 cells conditioned medium (Y4-CM) was collected after 24h culture. For fluid flow experiments, MLO-Y4 cells were exposed to 2h of pulsatile fluid flow (PFF) at 2, 4, 8, $16{\pm}0.6\;dynes/cm^2$ using the Flexcell $Streamer^{TM}$ system. For proliferation assays, MOCP-5, RAW 264.7, and 2T3 cells were cultured with control media or 10-100% Y4 CM. Cells were cultured for 3d, and then cells were counted. RAW 264.7 and 2T3 cell migration was assayed using transwells with control media or 10-100% Y4-CM. M-CSF, RANKL and OPG in MLO-Y4 mRNA expression was determined by semiquantitative RT-PCR. Y4-CM increased osteoclast precursor proliferation and migration, but decreased 2T3 cell proliferation and migration. CM from MLO-Y4 cells exposed to PFF caused decreased RAW 267.4 cell proliferation and migration and 2T3 migration compared to control Y4-CM. However, Y4-CM from cells exposed to PFF had no effect on 2T3 osteoblastic cell proliferation. PFF decreased RNAKL mRNA and increased OPG mRNA in MLO-Y4 cells compared to control(without PFF). PFF had no effect on M-CSF mRNA expression in MLO-Y4 cells. These results suggest that osteocytes can regulate bone remodeling by communication with osteoclast precursors and osteoblasts and that osteocytes can communicate mechanical signals to other cells.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.387.1-387.1
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• 2002

Aging and estrogen cleficiency after menopause induce bone loss and result in osteoporosis. This study was investigated effects of n-hexane fraction (Hx) extracted from Astragali Radix on osteoporosis with osteoblast-like cell line (MG-63 and Saos-2) and an ovariectomized (OVX) rat model. Proliferation of osteoblast-like cells. MG-63 and Saos-2. was tested with MTT and alkaline phosphatase (ALP) assays. (omitted)