• Journal of Periodontal and Implant Science
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• v.29 no.2
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• pp.277-289
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• 1999

The ultimate aim of periodontal treatment is periodontal regeneration, which necessiates the regeneration of bone tissues. To evaluate the effects of Dex growth and differentiation of MC3T3-E1 cells, cells were seeded in alpha-modified eagle medium containing 10% fetal bovine serum, 10mM beta-glycerophosphate , $50{\mu}g/ml$ of ascorbic acid, with or without $10^{-7}M$ Dex and examined cell proliferation activities, alkaline phosphatase activities, and bone nodule formation until 25days. The results were as follows : 1. In Dex group, cell proliferation activities were lower until 15 days compared to control group. Bone nodules formation were showed at 10 days. 2. In the time-response effect, ALP activities were increased until the 10 days in control groups thereafter decreased and ALP activities of Dex group were lower aspect than control group until the 10 days In this study, bone nodule formation of osteoblast-like cells were accelerated by Dex and cell proliferation activities, ALP activity of Dex group showed lower than control group. Dex was considered that it did suppress initial growth, but accerelate mineralization of osteoblast-like cells.

• Korean Journal of Food Science and Technology
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• v.39 no.6
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• pp.694-700
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• 2007

Acanthopanax senticosus is a common Asian herb also known as "Siberian Ginseng". It is often used as a traditional herbal medicine for reducing damage in the liver, kidney, bone and muscle. In the present study we investigated the ferric reducing/antioxidant power and total phenolic contents of the ethanol-/water-extracts obtained from the stems and leaves of Acanthopanax senticosus. Osteoblast cellular proliferation was evaluated using the MTT and alkaline phosphatase activity assays in the human osteoblast-like MG-63 cell line. Acanthopanax senticosus extracts exerted remarkable ferric reducing/antioxidant power and contained high amount of phenolics. Among the extracts the stem-/ethanol-extract showed the highest antioxidant activity and total phenol content. Interestingly a highly positive correlation was found between antioxidant activity and total phenol content (p < 0.01). Proliferation of MG-63 osteoblast cells was highest in the stem-/ethanol-extract and alkaline phosphatase activity significantly increased in the water-extract of the stems (p < 0.05). These findings suggest that Acanthopanax senticosus extracts have antioxidant activity for preventing oxidative stress-related diseases and may have beneficial effects on bone health through the proliferation of osteoblast cells.

• The Journal of Korean Academy of Prosthodontics
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• v.42 no.2
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• pp.175-191
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• 2004

Statement of problem: Platelet-rich plasma(PRP) is well known to be very effective method to stimulate and accelerate the healing of bone and soft tissue. However, there are few reports which deal with the mechanisms of the PRP on the activation of the osteoblasts. Purpose: This study was aimed to investigate the effect of growth factors in PRP on the activity of osteoblasts. Material and method: To evaluate the effect on human, human osteoblast cell line was cultured. PRP was extracted from the blood of a healthy volunteer. Using the recombinant growth factors of PDGF, $TGFT-\beta$, IGF-1, bFGF which are mainly found at bone matrix and their neutralizing antibody, the effect of PRP on the attachment and proliferation of osteoblasts was evaluated. To evaluate the autocrine and paracrine effects, conditioned media(CM) of PRP was made and compared with PRP. By the western blot analysis, the expression of growth factors in PRP, CM was examined. Cell morphology was compared by the light microscope. Results : 1) The effects of CM on osteoblast were similar to the effects of PRP. 2) PRP, CM, recombinant $TGF-\beta$, bFGF, IGF-1 showed significantly higher cellular attachment than control(p<0.05) in the cell attachment assay. In the cell proliferation assay, PRP, CM, recombinant $TGF-\beta$, IGF-1, bFGF, PDGF increased significantly cell proliferation(p<0.01). Among the recombinant growth factors, IGF-1 showed the highest cellular attachment and proliferation. 3) In the western blot assay, bFGF, IGF-1, PDGF weve equally expressed in PRP and CM. 4) The attachment of osteoblast cell decreased significantly after the addition of neutralizing antibody against $TGF-\beta$, IGF-1(p<0.05). In the cell proliferation assay, the addition of neutralizing antibody against $TGF-\beta$, bFGF, PDGF, IGF-1 decreased significantly the cellular proliferation(p<0.05). The amount of decreasing in the cell attachment and proliferation is the highest in at-lGF-1. 5) The cells in control group were flattened and elongated with a few cellular processes in the a light microscope. But, the cells appeared as spherical, plump cells with well developed cellular processes in experimental groups. The cells in PRP and CM had more prominent developed features than recombinant growth factor groups. Conclusions : These findings imply that PRP maximize the cellular activity in early healing period using the synergistic effect, autocrine, paracrine effects of growth factors and increase the rate and degree of bone formation.

• The Journal of Korean Academy of Prosthodontics
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• v.46 no.3
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• pp.227-237
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• 2008

STATEMENT OF PROBLEM: Zirconium oxide can be a substitute to titanium as implant materials to solve the esthetic problems of dark color in the gingival portion of implant restorations. PURPOSE: This study was performed to define attachment and growth behavior of osteoblast- like cells cultured on grooved surfaces of zirconium oxide and evaluate the genetic effect of zirconium oxide surfaces using the reverse transcriptase-polymerase chain reaction (RT-PCR). MATERIAL AND METHODS: MC3T3-E1 cells were cultured on (1) commercially pure titanium discs with smooth surface (T group), (2) yttrium-stabilized tetragonal zirconia polycrystal (Y-TZP) with machined surface (ZS group), and (3) Y-TZP with $100{\mu}m$ grooves (ZG group). Cell proliferation activity was evaluated through MTT assay and cell morphology was examined by SEM. The mRNA expression of Runx2, alkaline phosphatase, osteocalcin, TGF-${\beta}1$, IGF-1, G3PDH in E1 cells were evaluated by RT-PCR. RESULTS: From the MTT assay, after 48 hours of adhesion of MC3T3-E1 cells, the mean optical density value of T group and ZG group significantly increased compared to the ZS group. SEM images of osteoblast-like cells showed that significantly more cells were observed to attach to the grooves and appeared to follow the direction of the grooves. After 24 hours of cell adhesion, more spreading and flattening of cells with active filopodia formation occurred. Results of RT-PCR suggest that T group, ZS group, and ZG group showed comparable osteoblast-specific gene expression after 24 hours of cell incubation. CONCLUSION: Surface topography and material of implants can play an important role in expression of osteoblast phenotype markers. Zirconia ceramic showed comparable biological responses of osteoblast-like cells with titanium during a short-time cell culture period. Also, grooves influence cell spreading and guide the cells to be aligned within surface grooves.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.4
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• pp.243-249
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• 2010

Tissue engineered bone (TEB) can replace an autogenous bone graft requiring an secondary operation site as well as avoid complications like inflammation or infection from xenogenic or synthetic bone graft. Adult mesenchymal stem cells (MSC) for TEB are considered to have various ranges of differentiation capacity or multipotency by the donor site and age. This study examined the effect of age on proliferation capacity, differentiation capacity and bone morphogenetic protein-2 (BMP-2) responsiveness of human bone marrow stromal cells (hBMSC) according to the age. In addition, to evaluate the effect on enhancement for osteoblast differentiation, the hBMSC were treated with Trichostatin A (TSA) and 5-Azacitidine (5-AZC) which was HDAC inhibitors and methyltransferase inhibitors respectively affecting chromatin remodeling temporarily and reversibly. The young and old group of hBMSC obtained from the iliac crest from total 9 healthy patients, showed similar proliferation capacity. Cell surface markers such as CD34, CD45, CD90 and CD105 showed uniform expression regardless of age. However, the young group showed more prominent transdifferentiation capacity with adipogenic differentiation. The osteoblast differentiation capacity or BMP responsiveness was low and similar between young and old group. TSA and 5-AZC showed potential for enhancing the BMP effect on osteoblast differentiation by increasing the expression level of osteogenic master gene, such as DLX5, ALP. More study will be needed to determine the positive effect of the reversible function of HDAC inhibitors or methyltransferase inhibitors on enhancing the low osteoblast differentiation capacity of hBMSC.

• Preventive Nutrition and Food Science
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• v.19 no.4
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• pp.353-357
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• 2014

Phytic acid (myo-inositol hexakisphosphate) or phytate is considered an anti-nutrient due to the formation of precipitated complexes that strongly reduces the absorption of essential dietary minerals. In this study, brown rice with reduced phytate was made by inoculation with Lactobacillus sakei Wikim001 having high phytase activity. The effects of brown rice extract treated with L. sakei Wikim001 (BR-WK) on osteoblast differentiation and osteoclast formation were investigated. The proliferation of SaOS-2 cells was measured by the MTT assay. Treatment with BR-WK increased cell proliferation by 136% at a concentration of $100{\mu}g/mL$. The Alkaline phosphate activity in SaOS-2 cells was 129% higher when BR-WK was processed at a concentration of $100{\mu}g/mL$. The proliferation of bone marrow macrophages decreased by nearly 60% in response to treatment with BR-WK. In addition, BR-WK reduced the number of tartrate-resistant acid phosphatase-positive ($TRAP^+$) multinucleated cells from bone marrow macrophages. These results indicate that BR-WK stimulates bone formation through its positive action on osteoblast differentiation and function and furthermore, decreases osteoclast differentiation.

• BMB Reports
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• v.52 no.7
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• pp.469-474
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• 2019

Kruppel-like factor 2 (KLF2) has been implicated in the regulation of cell proliferation, differentiation, and survival in a variety of cells. Recently, it has been reported that KLF2 regulates the p65-mediated transactivation of $NF-{\kappa}B$. Although the $NF-{\kappa}B$ pathway plays an important role in the differentiation of osteoclasts and osteoblasts, the role of KLF2 in these bone cells has not yet been fully elucidated. In this study, we demonstrated that KLF2 regulates osteoclast and osteoblast differentiation. The overexpression of KLF2 in osteoclast precursor cells inhibited osteoclast differentiation by downregulating c-Fos, NFATc1, and TRAP expression, while KLF2 overexpression in osteoblasts enhanced osteoblast differentiation and function by upregulating Runx2, ALP, and BSP expression. Conversely, the downregulation of KLF2 with KLF2-specific siRNA increased osteoclast differentiation and inhibited osteoblast differentiation. Moreover, the overexpression of interferon regulatory protein 2-binding protein 2 (IRF2BP2), a regulator of KLF2, suppressed osteoclast differentiation and enhanced osteoblast differentiation and function. These effects were reversed by downregulating KLF2. Collectively, our data provide new insights and evidence to suggest that the IRF2BP2/KLF2 axis mediates osteoclast and osteoblast differentiation, thereby affecting bone homeostasis.

• Journal of Periodontal and Implant Science
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• v.33 no.3
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• pp.499-518
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• 2003

The surface characteristics of titanium have been shown to have an important role in contact ossseointegration around the implant. Anodizing at high voltage produces microporous structure and increases thickness of surface titanium dioxide layer. The aim of present study was to analyse the response of rat calvarial osteoblast cell to commercially pure titanium and Ti-6A1-4V anodized in 0.06 mol/l ${\beta}$-glycerophosphate and 0.03 mol/l sodium acetate. In this study, rat calvarial osteoblasts were used to assay for cell viability and cell proliferation on the implant surface at 1,2,4,7 days. 1. Surface roughness was 1.256${\mu}m$ at 200V, and 1.745${\mu}m$ at 300V. 2. The thickness of titanium oxide layer was increased 1 ${\mu}m$ with the increase of 50V. 3. The proliferation rate of osteoblastic cells was increased with the increase of the surface roughness and the thickness of titanium oxide layer. 4. There was no difference in cell viability and cell proliferation between commercially pure titanium and Ti-6A1-4V anodized at the same condition. In conclusion, the titanium surface modified by anodizing was biocompatible, produced enhanced osteoblastic response. The reasons of enhanced osteoblast response might be due to reduced metal ion release by thickened and stabilized titanium dioxide layer and microporous rough structures.

• Journal of Periodontal and Implant Science
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• v.30 no.3
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• pp.491-504
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• 2000

In clinical therapy, the current goal of dental implants is to enhance quantity and quality of osseointegration. Surface roughness and oxide structure are considered to influence the behavior of adherent cells. The purpose of this study is to evaluate the effect of different surface treatment on cellular response. The attachment and proliferation of osteoblast-like cell on sandblasted, sandblasted and etched, thermal oxidated surfaces have been compared. Sandblasting was done with $Al_2O_3$ particles(grain size of $50{\mu}m$), etching was processed with $NH_4OH$ : $H_2O_2$ : $H_2O(1:1:5)$ at $90^{\circ}C$ for 1 minute. Thermal oxidation was followed sandblasting and etching at $400^{\circ}C$, $600^{\circ}C$, $800^{\circ}C$ for 2 hours. Measurement of surface roughness after the different treatment did not show any differences of Ra value between terated surfaces. Cell attachment and proliferation were increased during experiment period, but no difference was observed. SEM evaluation revealed a similar pattern of osteoblast-like cells, well attached with dendritic extension and producing numerous matrix vesicles on cell surface. The results of this study showed that oxide layer alteration by thermal oxidation did not affect the attachment and proliferation of osteoblast-like cells. This suggests the possibility that the cellular responses are further influenced by surface roughness than titaniun oxide structure.

• International Journal of Oral Biology
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• v.30 no.2
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• pp.39-45
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• 2005

The present study was performed to investigate whether Achyranthes Radix extracts play roles in the bone metabolism. Three kinds of Achyranthes Radix extracts (methylene chloride (MC), ethylacetate (Ea), and water (W)) were used for bioassay. We examined cellular activities of osteoblasts by measurement of cell proliferation rate, alkaline phosphatase (ALP) activity, and calcified nodule formation. Osteoclast generation was assayed by measuring the number of tartrate-resistant acid phosphatase (TRAP) (+) multinucleated cells after culture of osteoclast precursor cells. There was a maximum 20% increase in proliferation rate of osteoblastic cells after treatment with MC. First and second subfraction of MC layer increased proliferation of osteoblast. Ea layer and second subfraction of MC layer increased ALP activity. Also MC layer and second subfraction of MC layer from Achyranthes Radix extracts increased the calcified nodule. MC layer and second subfraction of MC layer from Achyranthes Radix extracts significantly decreased in the number of TRAP (+) multinucleated cells. Taken together, Achyranthes Radix stimulates the proliferation and bioactivities of bone-forming osteoblasts, and inhibits activities of bone-resorbing osteoclasts.