• Journal of Periodontal and Implant Science
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• v.33 no.1
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• pp.49-60
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• 2003

Several growth factors and polypeptides are not commonly yet used for regenerators of bone tissue or alveolar bone because of the insufficiency of studies on their side effects, genetic engineering for mass production and stability for clinical application. Recently, many herbal medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, antiinflammatory and regenerative potential of periodontal tissues. Morindae Radix, Cibotium Barometz (L.), Albizziae Cortex, Cistandhis Herba have been traditionally used as medicines for treatment of bone disease in Eastern medicine. The objective of the present study is to examine the ability of alkaline phosphatase (ALP) activity of human fetal osteoblast (hFOB1) when several natural medicines were supplemented. hFOB1 were cultured with Dulbecuo's Modified Eagle's Medium Nutrient Mixture F-12 HAM ( DMEM/F-12 1:1 Mixture, Sigma, USA) and negative control, dexamethasone (positive control), and each natural medicines for 3 days. And then ALP activity was measured by spectrophotometer for enzyme activity and Alizarin red S staining for morphometry. Among the natural medicines of this study, Morindae Radix, Cibotium Barometz (L.) and Cistanchis Herba induced higher activity of ALP synthesis than negative controls in all experimental group. Albizziae Cortex showed mild increases than negative control group. According to measurement of positively stained area, all of the natural medicines of this study increased compared to negative control. Especially, Cibotium Barometz (L.) and Cistanchis Herba showed statistical significance compared to negative control (p<0.05). These results indicate that Morindae Radix, Cibotium Barometz (L.), Albizziae Cortex, Cistandhis Herba have an inducing ability of ALP synthesis on osteoblast.

• Journal of Periodontal and Implant Science
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• v.33 no.3
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• pp.415-437
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• 2003

Ginseng Radix(GR) had been used widely from oriental medicine and the effects of it have been investigated by many researchers. The purpose of present study was to investigate the effects of GR on the cell cycle progression and its molecular mechanism in human fetal osteoblast. The results were as follows. Increased cell proliferation was observed in cells exposed to 100 ng/ml, 10 ng/ml of GR-1 at 12 hours and 24 hours, 1 ${\mu}g$/ml of GR-1 at 48 hours, and 100 ${\mu}g$/ml, 10 ${\mu}g$/ml of GK-2 at 12 hours, all treatment groups of GR-2 at 24 hours(p<0.05). S phase and G1 phase was increased in the group of treated with 100 ng/ml of GR-1, with 10 ${\mu}g$/ml and 1 ${\mu}g$/ml of GR-2, with 100 ${\mu}g$/ ml and 10 ${\mu}g$/ml of GR-3 in the cell cycle analysis. The cell cycle regulation protein levels of Cyclin D1, Cyclin E, CDK 2. CDK 4 and CDK 6 were increased in the group of treated with 1 ${\mu}g$/ml and 100 ng/ml of GR-1, with 10 ${\mu}g$/ml and 1 ${\mu}g$/ml of GR-2, with 100 ${\mu}g$/ ml and 10 ${\mu}g$/ml of GR-3. On the other hand, p21 was decreased in the treatment group with 1 ${\mu}g$/ml and 100 ng/ml of GR-1, with 10 ${\mu}g$/ml and 1 ${\mu}g$/ml of GR-2, 10 ${\mu}g$/ml of GR-3, and p53 and p16 was decreased in the treatment group with 100 ng/ml of GR-1, 100 ${\mu}g$/ml and GR-3 and pRb was decreased in the all treatment groups except 1 ${\mu}g$/ml of GR-1. These results suggested that GR increases the cell proliferation and the cell cycle progression in human fetal osteoblast, which is linked to increased cell cycle regulation protein levels of Cyclin D1 , Cyclin E, CDK 2, CDK 4, CDK 6 and decreased cell cycle regulation protein levels of p21, pRb.

• Journal of Life Science
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• v.10 no.5
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• pp.529-532
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• 2000

Prostaglandin $E_2$(PG$E_2$) is an abundant eicosanoid in bone that has been implicated in a number of pathological states associated with bone loss, and is also known to stimulate matric metalloproteinase-1 systhesis and secretion in rat and human osteoblast cells, although the intracellular reaction remain unclear. Interleukin-1$\beta$ (IL-1$\beta$) is a cytokine that plays a critical role in bone remodelling and appears to act as a downstream effector of most bone-resorbing agents. However, it is still interesting to examine whether PG$E_2$ regulates IL-1$\beta$ expression by mouse osteoblasts or not. Here we demonstrate that PG$E_2$is a potent inducer of IL-1$\beta$ production by fetal osteoblasts.

• Journal of Periodontal and Implant Science
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• v.36 no.2
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• pp.435-448
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• 2006

The purpose of this study was to investigate the effects of ICB(Irradiated frozen allogenic bone, Rocky Mountain Tissue Bank, USA) and MTF(Decalcified freeze-dried bone allograft, Musculoskeletal Transplant Foundation, USA) on the cell proliferation and differentiation of human fetal osteoblasts. Human fetal osteoblasts (hFOB1) were cultured with $10\;ng/m{\ell}$of ICB and MTF. The negatvie control group was cultured with DMSO and positive control group was cultured with BMF ($2\;ng/m{\ell}$). MIT was performed to examine the viability of the cell, and alkaline phosphatase activity was analyzed to examine the mineralization. Calcium accumulation was also evaluated. ICB and MTF did not increase the rate of the cellular proliferation of hFOB1s while they enhanced ALP and calcium accumulation. The expression of osteocalcin (OC) and bone silaloprotein (BSP) increased in hFOB1 treated with ICB and MTF ($10\;ng/m{\ell}$). These results suggest that ICB and MTF stimulate osteoblastic activity of the hFOBl.

• Archives of Plastic Surgery
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• v.33 no.5
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• pp.612-615
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• 2006

Purpose: An ideal bony construct can be divided into two broad categories: (1) the design and fabrication of biodegradable, biomimetic scaffolds that provide correct signals to induce osteogenesis: (2) the identification of an ideal source of osteoprogenitor cells to seed onto the scaffold. We selected poly-glycolic acid as a synthetic scaffold among various scaffolds because of these properties. Meanwhile, culture medium is supplemented with fetal bovine serum(FBS): such serum contains essential elements such as proteins, hormones, growth factors and trace minerals. The composition of FBS can be ideal for various cell growth in vitro. We supposed that we could enhance bone growth at a fractured site if FBS was mixed with synthetic scaffold-PGA. Methods: We cultured human osteoblasts in five different prepared culture dishes made with FBS and PGA mixture. The mixtures contained different ratio of FBS, that is, 0, 1.5, 3, 7, and 10%. We cultured human osteoblasts for seven days and examined the growth and attachment of the cells at the 1st, 3rd, 5th, 7th days, respectively. Results: In the mixture of 0% FBS and PGA, the growth of the cells lasted for one day. In 1.5 and 3% FBS and PGA, the growth of the cells was examined at the 3rd day, then minimally declined at the 5th and 7th days. In 7% FBS and PGA, the growth of the cells lasted for 5 days, then declined at the 7th day. In 10% FBS and PGA, the growth of the cells lasted for 5 days, then declined at the 7th day. Staining status of the osteoblasts with alkaline phosphatase showed pale pink color in 0% FBS and PGA groups, but bright pink color in 1.5, 3, 7, 10% FBS and PGA groups, especially in 3%, 7%. Conclusion: In consequence, the growth of human osteoblast was higher in the mixture of FBS and PGA groups than in pure PGA ones. It is assumed that the mixture of FBS and PGA affects the proliferation of human osteoblasts.

• Journal of Periodontal and Implant Science
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• v.36 no.2
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• pp.449-459
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• 2006

DFDBA(Decalcified freeze-dried bone allograft) is one of the allograft materials for periodontal bone regeneration. DFDBA provides an osteoconductive surface and osteoinductive factors. Therefore, DFDBA have been used successfully to regenerate the attachment apparatus during periodontal treatment. But recent studies was reported that wide variations in commercial bone bank preparations of DFDBA do exist, including the ability to induce new bone formation. DFDBA was experimental materials that was recovered, processed, tested, shipped and invoiced through Musculoskeletal Transplant Foundation. MTF(Musculoskeletal Transplant Foundation) is the world largest, non-profit, AATB(American Association of Tissue Banks) accredited tissue bank. The objective of this study was to determine the effects of serial dilutions of a DFDBA on human fetal osteoblastic cell proliferation and their potential to form and mineralize bone nodules. Human fetal osteoblastic cell line(hFOB 1.19) was cultured with DMEM and SSE($1{\mu}g/m{\ell}$,$10{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $1mg/m{\ell}$) at $34^{\circ}C$ with 5% CO2 in 100% humidity. Cell proliferation was significantly increased at $1mg/m{\ell}$, $100{\mu}g$, $10{\mu}g/m{\ell}$, $1{\mu}g/m{\ell}$, $100ng/m{\ell}$, $10ng/m{\ell}$, $1ng/m{\ell}$ of DFDBA after 5 days incubation (p<0.05). Alkaline Phosphatase(ALP) level was significantly increased in $100ng/m{\ell}$, $10ng/m{\ell}$, $1ng/m{\ell}$ of DFDABA(p<0.05). A quantified calcium accumulation was significantly increased at $1ng/m{\ell}$, $10ng/m{\ell}$ of MTF(p<0.05). These results indicated that DFDBA has an inductive effect on bone formation in vitro.

• The Journal of Korean Medicine
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• v.24 no.3
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• pp.35-44
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• 2003

Objective : This study was performed to evaluate the effects of Palmijihwang-hwan (Baweidehuang-wan) and Obaeja (Galla Rhois) on the regeneration of periodontal tissue. Methods : In this study, we used MC3T3-El cells, such as osteoblast-like cell lines and human periodontal ligament fibroblasts, for experimental material. We separated each type of cells into a control group and an experimental group. In the control group, the cells were cultivated for 48 hours with distilled water and media which contained 10% fetal bovine serum (FBS) and penicillin (l00unit/ml)-streptomycin ($l00{\mu\textrm{g}}/ml$) at $37^{\circ}$ in 5% $CO_2$ gas. In the experimental group, the cells were cultivated for 48 hours with Palmijihwang-hwan extract and Obaeja extract (concentrations $1{\mu\textrm{g}}/ml,{\;}25{\mu\textrm{g}}/ml,{\;}50{\mu\textrm{g}}/ml$) under the same conditions as the control group. Investigating the regeneration of periodontal tissue was performed by evaluating proliferation, the activity of alkaline phosphatase and the synthetic ability of proteins using those cultivated cells by means of microculture tetrazolium (MTT) assay, alkaline phosphatase substrate kit and protein assay kit. Results : 1. In vitro, Palmijihwang-hwan extract increased the proliferation of MC3T3-El cells. 2. In vitro, Obaeja extract increased the activity of alkaline phosphatase and the synthetic ability of protein in MC3T3-El cells and human periodontal ligament fibroblasts depending on Obaeja extract's concentration. Conclusion : Obaeja extract can be developed as a subsidiary medicine for the regeneration of periodontal tissue. Further studies to evaluate the different concentrations the Obaeja extract and clinical trials in vivo are suggested.

• The Journal of Korean Medicine
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• v.24 no.3
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• pp.11-22
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• 2003

Objective : As the average span of human life extends, more and more people are at risk of developing osteoporosis, one of the typical diseases of the aged. This thesis presents the effects of Jeungikgwiryon-tang (Tsengikueijung-tang) on bone density, bone biochemical markers, and fetal calvarial cells (FCC) of Sprague Dawleys (S.D.) rats that have induced osteoporosis. The purpose is to see how Jeungikgwiryon-tang (Tsengikueijung-tang) reduces osteoporosis symptoms. Methods : In the first experiment Sprague Dawleys rats were administered Jeungikgwiryon-tang (Tsengikueijung-tang) for 70 days, once a day. Two different doses were used, creating high-dosed and low-dosed groups. The results were compared with a control group. In the second experiment, Jeungikgwiryon-tang (Tsengikueijung-tang) was applied to fetal calvarial cells (FCC) obtained from fetuses inside pregnant Sprague Dawleys rats. The FCCs from high-dosed and low-dosed groups were compared with those from a control group. Results : 1. Bone densities in Groups A and B increased significantly from a control group. 2. Bone ash densities in Group A showed substantial increase. 3. Calcium and phosphorus in bones in Group A increased significantly. 4. Activity of fetal calvarial cells' division in Groups A and B increased significantly from a control group, and ALP of fetal calvarial cells' formation in Group A increased significantly. 5. Protein and collagen levels of fetal calvarial cells in Group A increased significantly. Conclusion : It was found that Jeungikgwiryon-tang (Tsengikueijung-tang) has a tendency to make significant increases in bone densities by enhancing bone formation and by retarding bone absorption. It was concluded that Jeungikgwiryon-tang (Tsengikueijung-tang) activates osteoblast cells effectively.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.40 no.6
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• pp.291-296
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• 2014

Objectives: Bisphosphonate-related osteonecrosis of the jaw (BRONJ) is a side effect of bisphophonate therapy that has been reported in recent years. Osteoclastic inactivity by bisphosphonate is the known cause of BRONJ. Bone morphogenetic protein-2 (BMP-2) plays an important role in the development of bone. Recombinant human BMP-2 (rhBMP-2) is potentially useful as an activation factor for bone repair. We hypothesized that rhBMP-2 would enhance the osteoclast-osteoblast interaction related to bone remodeling. Materials and Methods: Human fetal osteoblast cells (hFOB 1.19) were treated with $100{\mu}M$ alendronate, and 100 ng/mL rhBMP-2 was added. Cells were incubated for a further 48 hours, and cell viability was measured using an MTT assay. Expression of the three cytokines from osteoblasts, receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL), osteoprotegerin (OPG), and macrophage colony-stimulating factor (M-CSF), were analyzed by real-time polymerase chain reaction and enzyme-linked immunosorbent assay. Results: Cell viability was decreased to $82.75%{\pm}1.00%$ by alendronate and then increased to $110.43%{\pm}1.35%$ after treatment with rhBMP-2 (P<0.05, respectively). OPG, RANKL, and M-CSF expression were all decreased by alendronate treatment. RANKL and M-CSF expression were increased, but OPG was not significantly affected by rhBMP-2. Conclusion: rhBMP2 does not affect OPG gene expression in hFOB, but it may increase RANKL and M-CSF gene expression.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.5
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• pp.1042-1047
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• 2002

Recently, many natural medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential for periodontal tissues. Cortex Eucommiae, Eupoly phaga, Semen Cuscutae, Halloysitum Rubrum have been traditionally used as medicines for treatment of bone disease in Korea. The objective of the present study is to examine the ability of alkaline phosphatase (ALP) activity in human fetal osteoblast cell line (hFOB1) with several natural medicines. hFOB1 added DMEM/F-12 were cultured with dexamethasone as a positive control, and with each natural medicine. ALP activity was measured by spectrophotometer for enzyme activity and naphthol AS-Bl staining was performed for morphometry. All of the natural medicines induced a higher ALP activity compared to negative control, especially, Cortex Eucommiae increased an ALP activity in all experimental groups (p<0.05). In naphthol AS-Bl staining, all of the natural medicines of this study increased the stained area compared to negative control. Especially, Cortex Eucommiae and Eupoly phaga showed statistical significance compared to negative control (p<0.05). These results indicate that Cortex Eucommiae, Eupoly phaga, Semen Cuscutae, Halloysitum Rubrum have an inducing ability of ALP synthesis on osteoblasts.