• Journal of dental hygiene science
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• v.21 no.4
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• pp.243-250
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• 2021

Background: Endodontic sealers or their toxic components may become inflamed and lead to delayed wound healing when in direct contact with periapical tissues over an extended period. Moreover, an overfilled sealer can directly interact with adjacent tissues and may cause immediate necrosis or further resorption. Therefore, the treatment outcome conceivably depends on the endodontic sealer's biocompatibility and osteogenic potential. This study aimed to evaluate the cell viability and osteogenic effects of four different sealers in osteoblastic cells. Methods: AH Plus (resin-based sealer), Pulp Canal Sealer EWT (zinc oxide-eugenol sealer), BioRoot RCS (calcium silicate-based sealer), and Well-Root ST (MTA-based calcium silicate sealer) were mixed strictly according to the manufacturer's instructions, and dilutions of sealer extracts (1/2, 1/5 and 1/10) were determined. Cell viability was measured using the water-soluble tetrazolium-8 (WST-8) assay. Differentiation was assessed by alkaline phosphatase (ALP) activity and mineralized nodule formation by Alizarin Red S staining. Results: The cell viability of the extracts derived from the sealers excluding Well-Root ST was concentration dependent, with sealer extracts having the least viability at a 1/2 dilution. At sealer extract dilution of 1/10, the test groups showed the same survival rate as that control group, with the exception of BioRoot RCS. Among all experimental groups, BioRoot RCS showed the highest cell viability after 48 hours. The ALP activity was significantly higher in a concentration-dependent manner. Furthemore, all four materials promoted ALP activity and mineralized nodule formation compared to the control at 1/10 dilutions. Conclusion: This is the first study to highlight the differences in biological activity of these four materials. These results suggest that the composition of root canal sealers appears to alter the form of biocompatibility and osteoblastic differentiation.

• Preventive Nutrition and Food Science
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• v.12 no.1
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• pp.1-6
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• 2007

Colostrum basic proteins (CBP) (MW 1$\sim$30 kDa) were isolated from bovine colostrum using a series of ultrafiltration processes and their effects on osteoblast cell proliferation and bone metabolism were investigated in cell line and animal models. Treatments with CBP (1, 10, 100 $\mu$g/mL) dose-dependently increased cell proliferation of osteoblastic MC3T3 cells. Alkaline phosphatase activity, a marker of osteoblastic phenotype, in the cells was also increased after treatments with CBP in a dose-dependent manner. Significant increases in bone density were observed in femur of ovariectomized rats which were fed a diet with 1% and 10% CBP, compared to rats fed a normal diet. These results suggest that CBP may increase bone mass and density and be useful for the prevention of bone-related diseases.

• Imaging Science in Dentistry
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• v.34 no.2
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• pp.99-106
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• 2004

Purpose: To investigate the effects of irradiation on the phenotypic expression of the MC3T3-El osteoblastic cell line, especially on the osteonectin and bone sialoprotein. Materials and Methods: Cells were irradiated with a single dose of 0.5, 1, 4 and 8 Gy at a dose rate of 5.38 Gy/min using Cs-I37 irradiator. After specimens were harvested, total RNA was extracted on the 3rd, 7th, 14th, 21st day after irradiation. The total RNA was reverse-transcribed and the resulting cDNAs were subjected to amplification by PCR with a pair of primers. Results: The irradiated cells showed a dose-dependent increase in osteonectin mRNA expression when compared with the unirradiated control group. The irradiated cells showed no difference in bone sialoprotein mRNA expression when compared with the unirradiated control group. In accordance with the duration of culture period after irradiation, the level of osteonectin mRNA expression showed no difference, but it increased a little at the 21st day in the 4 and 8 Gy exposure groups. In the case of bone sialoprotein, however, the level of mRNA expression increased significantly at the 3rd and 7th day after irradiation, but it showed no difference at the 14th and 21st day when compared with the control group. Conclusion: These results showed that each single dose of 0.5, 1, 4 and 8 Gy influenced the mRNA expression of osteonectin and bone sialoprotein at the calcification stage of osteoblastic cells, suggesting that single dose of irradiation affected the osteoblastic bone formation at the cell level.

• The korean journal of orthodontics
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• v.28 no.6 s.71
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• pp.915-926
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• 1998

The cytoskeleton has been shown to form a network, connecting the extracelluar matrix via integrin with the nucleus and the cytoplasmic constituents of the cell. It is therefore assumed that the cytoskeleton may mediate signals generated by perturbations originating in the matrix. The purpose of this study is to examine the effect of cytoskeletal change on osteoblastic cell activities. The author cultured osteoblastic cells obtained from neonatal mouse calvaria. The cells were teated with cytochalasin B(CB) or colchicine (COL) at four concentrations for 3 hours and after another 24 hours the conditioned media was collected and assayed for prostaglandin $E_2\;(PGE_2)$, interleukin-6(IL-6), tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and matrix metalloproteinase-1(MMP-1). In addition, the cytoskeletal protein actin were observed by immuno-fluorescence. The results were as follows: 1. The production of $PGE_2$ showed the tendency to be increased in CB-treated group. $PGE_2$ was increased in COL-treated group dose-dependantly, 2. IL-6 production, in CB-treated group, was increased, except at 1.0 ${\mu}g/ml$. IL-6 was induced in COL-treated group. 3. TNF-$\alpha$ production was increased in CB-treated group, except at 1.0 ${\mu}g/ml$, and in COL-treated group, that was increased. 4. The MMP-1 production was decreased in CB-treated soup and was not changed in COL-treated group, which could be selectively visualized by immunoblotting with monospecific antibody. 5. The cytoskeletal actin stress fibers were disappeared and the cells showed to be rounded in CB-treated group. These results indicated that there are a relationship between the cytoskeletal rearrangements and osteoblastic cell activities, especially in release of paracrine/autocrine factors, such as $PGE_2$, IL-6, and TNF-$\alpha$.

• Imaging Science in Dentistry
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• v.35 no.1
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• pp.1-8
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• 2005

Purpose : To investigate the effects of irradiation on the calcium content and calcific nodule formation in the MC3T3-E1 osteoblastic cell line. Materials and Methods : Cells were irradiated with a single dose of 2,4 and 8 Gy at a dose rate of 5.38 Gy/min using a Cs-137 irradiator. After irradiation, the calcium content and calcific nodule formation were examined on the 1 st, 2nd, 3rd and 4th week. Results : A decreasing dose-dependent tendency of the cell proliferation rate was found in all irradiated groups of this experiment when compared with the unirradiated control group. In accordance with the duration of culture, there was no significant difference in the cell proliferation rate after irradiation of 2 Gy when compared with the unirradiated group, however a decreasing tendency was found in 4 Gy- and 8 Gy-irradiated groups. While an increase in total calcium content after irradiation of 2 Gy was found at week 1, week 2, and week 4, there was a decrease in calcium content at week 1 through 4 in the 8 Gy- irradiated group. Calcific nodule formation was increased in irradiated experimental groups when compared with the unirradiated control group in the 2 Gy-irradiated group, but decreased in the 4 Gy- and 8 Gy-irradiated groups at the same stage. Conclusion : The results showed a mild increasing tendency of the calcific nodule formation after irradiation of 2 Gy. However, a decreased calcific nodule formation in 4 Gy- and 8 Gy-irradiated groups was found. Taken together, the irradiation of 2 Gy mildly activated bone formation, however 4 Gy or 8 Gy suppressed bone formation by decreasing cell numbers in the MC3T3-El osteoblastic cell line.

• Bulletin of the Korean Chemical Society
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• v.31 no.4
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• pp.929-933
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• 2010

A new compound, 4-methoxyl 5-hydroxymethyl benzoic 3-O-$\beta$-D-glucopyranoside (1), has been isolated from the leaves and stems of Acer mandshuricum, along with nine known compounds (2-10). Their structures were determined by a variety of spectroscopic analyses. The effect of compounds 1-10 on the function of osteoblastic MC3T3-E1 cells was examined by determining alkaline phosphatase (ALP) activity, collagen synthesis, and mineralization. Compound 1 significantly increased the function of osteoblastic MC3T3-E1 cells; $5.0\;{\mu}M$ of 1 increased ALP activity, collagen synthesis, and mineralization of MC3T3-E1 cells to 114.7, 119.5, and 108.2% (P < 0.05) of the basal value, respectively. In addition, compounds 2-10 also potently increased the function of osteoblastic MC3T3-E1 cells.

• Nutrition Research and Practice
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• v.10 no.2
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• pp.148-153
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• 2016

BACKGROUND/OBJECTIVES: Bone formation and bone resorption continuously occur in bone tissue to prevent the accumulation of old bone, this being called bone remodeling. Osteoblasts especially play a crucial role in bone formation through the differentiation and proliferation. Therefore, in this study, we investigated the effects of Scytosiphon lomentaria extract (SLE) on osteoblastic proliferation and differentiation in MC3T3-E1 cells. MATERIALS/METHODS: A cell proliferation assay, alkaline phosphatase (ALP) activity assay, alizarin red staining and protein expression analysis of osteoblastic genes were carried out to assess the osteoblastic proliferation and differentiation. RESULTS: The results indicated that treatment of SLE promoted the proliferation of MC3T3-E1 cells and improved ALP activity. And, SLE treatment significantly promoted mineralized nodule formation compared with control. In addition, cells treated with SLE significantly upregulated protein expression of ALP, type 1 collagen, bone morphogenetic protein 2, runt-related transcription factor 2, osterix, and osteoprotegerin. CONCLUSIONS: The results demonstrate that SLE promote differentiation inducement and proliferation of osteoblasts and, therefore may help to elucidate the transcriptional mechanism of bone formation and possibly lead to the development of bone-forming drugs.

• 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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.4
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• pp.802-809
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• 2008

Gamijangsing-tang (GMJST) has been used for treatment of bone formation in traditional korean medicine. The purpose of this study is to examine effects of GMJST on bone metabolism. The effects on the osteoblasts were determined by measuring (1) cell proliferation, (2) alkaline phosphatase (ALP) activity, (3) osteoprotegerin (OPG) secretion. (4) The morphologic changes of cells were observed by light microscopy and electron microscopy. Mineralization of calcium was determined by quantitative alizarin red-S assay and mineralization of phosphate was observed by von kossa staining. The morphologic changes of mineralization on the cells were observed by transmission electron microscopy (TEM). The effects on the osteoclast were investigated by tartrate-resistant acid phosphatase (TRAP) staining. Following results were obtained: Celluar activity of osteoblastic cells (MG-63) was significantly increased in 10-5 of dilution of GMJST. ALP and OPG activity of osteoblastic cells were increased in GMJST than normal MG-63 cell. Mineralization of osteoblastic cells were increased in GMJST than normal MG-63 cell. The activity of osteoclast cells (RAW 264.7) was significantly decreased in GMJST than normal MG-63 cell. From the results, GMJST stimulated the proliferation and mineralization of bone-forming osteoblast and inhibited by bone- lysis osteoclast.

• Nutrition Research and Practice
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• v.4 no.5
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• pp.356-361
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• 2010

Zinc is an essential trace element required for bone formation, however not much has been clarified yet for its role in osteoblast. We hypothesized that zinc would increase osteogenetic function in osteoblasts. To test this, we investigated whether zinc treatment enhances bone formation by stimulating osteoblast proliferation, bone marker protein alkaline phosphatase activity and collagen synthesis in osteoblastic MC3T3-E1 cells. MC3T3-E1 cells were cultured and treated with various concentrations of zinc (0, 1, 3, 15, 25 uM) along with a normal osteogenic medium (OSM) as control for 1, 5, 10 days. As measured by MTT assay for mitochondrial metabolic activity, cell proliferation was stimulated even at low zinc treatment (1-3 ${\mu}M$) compared to OSM, and it was stimulated in a zinc concentration-dependent manner during 5 and 10 days, with the most pronounced effect at 15 and 25 uM Zn. Cellular (synthesized) alkaline phosphatase (ALP) activity was increased in a zinc concentration-dependent manner, so did medium (secreted) ALP activity. Cellular collagen concentration was increased by zinc as time went by, therefore with the maximum zinc stimulatory effect in 10 days, and medium collagen concentration showed the same pattern even on 1 and 5 day. This zinc stimulatory effect of collagen synthesis was observed in cell matrix collagen staining. The study results imply that zinc can increase osteogenic effect by stimulating cell proliferation, ALP activity and collagen synthesis in osteoblastic cells.