Effects of Ipriflavone on bone remodeling in the rat calvarial cell / 대한치과교정학회지

Ipriflavone (isoprofoxyisoflavone), a synthetic derivative from soy isoflavone diazein, has been shown to inhibit bone resorption and perhaps stimulate bone formation. This study was performed to examine the effects of ipriflavone on the proliferation and bone remodeling in rat calvarial cells in vitro. The rat calvarial cells were isolated from fetus aged 20 to 21 days and cultured in BGJb media. The graded concentration of ipriflavone (10(-9)-10(-5) M) was administered into cultured cells. When the cell proliferation was estimated through the measurement of MTT assay, there was no increase in cellular proliferation of the rat calvarial cell at any ipriflavone concentration. The cellular activity was evaluated through the formation of mineralized nodules stained by alizarin red. The formation of mineralized nodules significantly increased at concentrations of 10(-8) M, 10(-7) M and 10(-6) M ipriflavone. Reverse transcription-polymerase chain reaction analyses (RT-PCR) were done at 7 and 14 days after culture to detect the expression of Bone Sialoprotein (BSP), Type I Collagen (COL I) and Osteocalcin (OCN). As a result, the expressions of BSP and COL I increased on the 7th day of culture and the expression of OCN increased on the 14th day of culture. These results indicate that ipriflavone facilitates the bone remodeling process by promoting rat calvarial cell differentiation and stimulating mineralization through increased expression of extracellular matrix genes, such as BSP, COL I and OCN.

Inhibition of MMP-13 mRNA expression by ginseng saponin in fetal rat calvarial cells / 대한치주과학회지

There is a potential role of collagenase-3 in alveolar bone loss and periodontal disease progression, we need to develope or find chemotherapeutic drugs or herbal agents which may regulate the expression of MMP-13. Ginseng saponin, one of the major components of Korea ginseng(panax ginseng) root, has many various biologic effects, such as cytotoxic effect, tumoricidal effects, cytokine regulations, and protein biosynthesis effect. The purpose of this study was to determine the effects of Korea red ginseng saponin on MMP-13 gene expression in osteoblasts. The experimental groups were cultured with ginseng saponin in concentration of 1.0, 10, 25, 50, 100, 250 and 500microgram/ml for MTT assay. Primary rat calvarial cells were pre-treated for 1 hour with ginseng saponin(100 microgram/ml) and then stimulated with IL-1beta(1.0ng/ml) and PTH (10 nM). MMP-13 gene expression was evaluated by RT-PCR. The results were as follows: Ginseng saponin was cytotoxic to osteoblast at concentration exceeding 250microgram/ml for longer than 24 hours in tissue culture(p<0.01). In RT-PCR analysis, steady state MMP-13 mRNA levels were increased approximately 350% by IL-1beta, and 400% by PTH when normalized to untreated control. IL-1beta-indued MMP-13 mRNA expression was reduced 50% by pre- treatment with ginseng saponin. But ginseng saponin didn't inhibit MMP-13 expression from PTH stimulated cells. This results suggest that ginseng saponin inhibit IL-1beta-indued MMP-13 mRNA expres- sion.

Effect of Manganese on primary rat calvarial cell / 대한치주과학회지

Chronic exposure to high levels of manganese leads a pronounce and debilitating disorder known as manganism. Research on the toxic manifestation of manganese have focused primarily on its neurological effects because exposure to high levels of the metal produces a distinct and irreversible extrapyramidal dysfunction resembling the dystonic movements associated with Parkinson's physiological and biochemical systems in the body. The purpose of this study was to evaluate the effect of manganeses on primary rat calvarial cell growth and toxicity. The experimental groups were in concentration of 0, 10, 30, 60, 100, 300 micrometer. Cell activity was assessed at day 1 and day 3 using a fluorescent molecular probe. Cell proliferation was evaluated at day 1 and day 3 by MTT assay. The amount of total protein synthesis was measured at day 3 and day 7. The results were as follows: The proliferation of primary rat calvarial cells were inhibited by MnCl2 in the concentration exceeding 100micrometer. The primary rat calvarial cells treated with MnCl2 showed similar protein synthesis to the control group except in 100 micrometer. These result suggest that manganese suppress the viability and protein synthesis of primary rat calvarial cells in concentration exceeding 100 micrometer.

Effect of chitosan in primary rat calvarial cell / 대한치주과학회지

The effect of chitosan, a carbohydrate biopolymer extracted from chitin, on periodontal regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on primary rat calvarial cells in vitro, with special focus on their proliferative properties by cell activity and the amount of total protein synthesis. The experimental groups were cultured with chitosan in concentration of 0.01, 0.1, 1.0, 2.0 and 5.0 mg/ml for MTT assay. In the experimental groups, cells were cultured with chitosan in concentration of 0.01, 0.1, 1.0 and 2.0 mg/ml. Each group was characterized by examining alkaline phosphatase activity at 3 and 7 days and the ability to produce mineralized nodules of rat calvarial cells at 14 and 21 days. The results were as follows: 1. The cell activity was not reduced in the concentration of 0.01~1.0 mg/ml whereas the cell activity was reduced in the concentration of 5.0 mg/ml than the control at day 1 and 3 (p<0.05). 2. Primary rat calvarial cells treated with chitosan in the concentration 0.01 mg/ml and 0.1 mg/ml showed more protein synthesis than the control at day 3 (p<0.01). But primary rat calvarial cells treated with chitosan showed more protein synthesis than in control but they didn't have statistically difference among groups at day 7. 3. At 3 and 7 days, alkaline phosphatase activity was significantly increased in the concentration of 0.01 mg/ml. 0.1 mg/ml and 1.0 mg/ml (p<0.05). 4. The percentage of mineralized bone nodule was more in the concentration of chitosan 0.1 mg/ml and 1.0 mg/ml than the control. These results suggested that chitosan has a positive effect on the bone formation of primary rat calvarial cells in the concentration of 0.1 mg/ml and 1.0 mg/ml.

Effect of chitosan on bone matrix expression and mineralization in primary rat calvarial cell / 대한치주과학회지

Periodontal therapy has dealt primarily with attempts at arresting progression of disease, however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. The effect of chitosan, a carbohydrate biopolymer extracted from chitin, on periodontal ligament regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on the expression of extracellular matrix proteins in primary rat calvarial cells in vitro. In the control group, cells was cultured with BGJb media. In the experimental groups, cells were cultured with chitosan in concentration of 0.01, 0.1, 1.0 and 2.0 mg/ml. Then each group was characterized by examining alkaline phosphatase activity at 3 and 7 days, and the ability to produce mineralized nodules of rat calvarial cells at 14 and 21 days. Synthesis of type I collagen (COL-I), osteocalcin (OCN), bone sialoprotein (BSP) was evaluated by RT-PCR at 14 days. The results were as follows: 1. Alkaline phosphatase activity was significantly higher in the concentration of chitosan 0.01mg/ml, 0.1mg/ml and 1.0mg/ml compared to control (p<0.05). 2. The percentage of mineralized bone nodule was more in the concentration of chitosan 0.1mg/ml and 1.0 mg/ml than the control. 3. At 14 day culture, the expression of OCN was increased by chitosan in concentration of 1.0 mg/ml and 2.0 mg/ml. These results suggested that chitosan in concentration of 0.1 and 1.0 mg/ml stimulate the extracellular matrix of primary rat calvarial cells and may facilitate the formation of bone.

Effects of the MnCl2 on bone formation in fetal rat calvarial cell / 대한치주과학회지

Chronic exposure to high levels of manganese (Mn) leads a pronounced and debilitating disorder known as manganism. Research on the toxic manifestation of manganese have focused primarily on its neurological effects because exposure to high levels of the metal produces a distinct and irreversible extrapyramidal dysfunction resembling the dystonic movements associated with Parkinson's physiological and biochemical systems in the body. The purpose of this study is to determine the effects of Mn on mineralization in primary rat calvarial cells. The experimental groups were in concentration of 0, 10, 30 and 60 micrometer. The results were as follows: 1. ALP activity was decreased in concentration of 30 and 60 micrometer (p<0.01). 2. Bone nodule formation was depressed in concentration of 30 and 60 micrometer at day 14 and 21 (p<0.01). 3. RT-PCR results showed an altered expression of bone matrix proteins. These result suggested that manganese might decrease or alter the expression of the osteoblast phenotype.

The Effects of Dichloromethane fraction of Phlomodis Radix(DFPR) on differentiation of Mouse Calvarial Cell / 대한치주과학회지

The purpose of this study was to evaluate the effects of DFPR on differentiation of mouse calvarial cell in vitro, to examine the possibility for periodontal regeneration. 10microgram/ml of DFPR was used as experimental concentration. osteogenic medium only was assigned as control, Experimental 1 was supplemented with 10nM dexamethasone, Experimental 2 with 10microgram/ml DFPR and Experimental 3 with 10nM dexamethasone + 10microgram/ml DFPR. cellular activity was evaluated by MTT method at 8, 12, 16 days, expression of mRNA of ALP, osteopontin, osteocalcin, collagen type-1 was detected by RT-PCR method at 4, 8, 12, 16 days of culture . extent of mineralization was observed by Von Kossa staining at 16 day of culture. The results are as follows 1)Any acceleration of differentiation was not observed at expression of differentiation marker, 2) Decrease in expression of extracelluar matrix and in bone nodule formation was observed The results suggested that DFPR have negative effect on the rate of differentiation on rat calvarial cell, decrease extracelluar matrix formation ,decrease bone nodule formation. Ongoing studies are necessary in order to determine effect of DFPR on periodontal regeneration.

The Effects of Calcium Phosphate Glass on Mouse Calvarial Cell / 대한치주과학회지

The goal of periodontal treatment is not only to arrest the progression of the disease but also to promote the functional, esthetic regeneration of the periodontium. Flap operation, bone graft, guided tissue regeneration, growth factors and bone morphogenetic protein have been used for this purpose. Among these techniques of regeneration, alloplastic graft, especially calcium phosphate is getting more attention recently. The purpose of this study was to evaluate the effects of calcium phosphate glass on mouse calvarial cell in vitro. The toxicity of calcium phosphate glass was measured using MTT assay, the synthesis of collagen was measured using collagen assay, and ALP activity was measured. The experimental groups were cultured with calcium phosphate glass(both AQ-, and HT-CPG) in concentration of 0.01, 0.02, 0.1, 0.2g/ml. The results are as follows 1. In concentrations not exceeding 0.02g/ml, both the groups(AQ-CPG, HT-CPG) didn't show any toxicity on mouse calvarial cell(p<0.05). 2. In both the experimental groups are the concentration of 0.02g/ml, collagen expressions were significantly up-regulated (p<0.05). 3. In both the experimental groups are the concentration of 0.02g/ml, ALP activity was not significantly upregulated, but ALP activity in both experimental groups were greater than control group(p<0.05). The results suggested that the use of calcium phosphate glass may promotes periodontal regeneration. Ongoing studies are necessary in order to determine their regeneration effects.

The Effect of Cyclosporin A on Osteoblast in vitro / 대한치주과학회지

Cyclosporin A(CsA) is an immunosuppressive agent widely used for preventing graft rejecting response in organ transplantation. The basic properties of CsA to osteoblast has not been well known yet. A better understanding of the mechanisms of CsA function on bone could provide valuable information regarding basic properties of bone remodeling, pharmacotherapeutic intervention in metabolic bone disease, and the consequences of immunosuppression in bone physiology. The purpose of this study was to investigate the effect of CsA on osteoblast by evaluating parameters of proliferation, collagen synthetic activity, alkaline phosphatase activity, and ALP mRNA expression in mouse calvarial cell. 1. CsA(3microgram/ml) treated mouse calvarial cell showed statistically significant increase in cell proliferation.(P<0.05) 2. CsA(1, 3microgram/ml) treated MC3T3 cell line showed statistically significant increase in cell proliferation. 3. The amount of collagen of CsA(3microgram/ml) treated mouse calvarial cell was decreased statistically significantly. 4. Alkaline phosphatase activity was increased statistically significantly in CsA treated group(1microgram/ml). 5. mRNA expression of ALP was increased in CsA treated group These results suggest that CsA could affect bone remodeling by modulating proliferation & differentiation of osteoblast.