• The Journal of Korean Obstetrics and Gynecology
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• v.29 no.2
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• pp.15-28
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• 2016

Objectives : The purpose of this study is to evaluate the effect of water extract of Samki-eum Gamibang (SKG) on osteoblast proliferation in murine calvarial cells. Methods : The osteoblast separated from calvariae of murine was cultivated and evaluated the function of cell. After the addition of SKG on the culture medium, we investigated the effect of SKG on the cell viability, cell proliferation, alkaline phosphatase (ALP) activity, bone matrix protein synthesis and collagen synthesis of the cultivated osteoblast.Results : SKG increased the survival rate and proliferation of rat calvarial osteoblast. SKG increased ALP activity, bone matrix protein synthesis and collagen synthesis of rat calvarial osteoblast. Conclusions : This study suggests that SKG has effect on glucocorticoid-induced osteoporosis (GIO) resulting from increase of osteoblast function.

• Polymer(Korea)
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• v.29 no.5
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• pp.501-507
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• 2005

Small intestinal submucosa (SIS) had been widely used as a biomaterial without immune rejection responses. SIS sponges prepared by crosslinking with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). SIS powders dissolved in $3\%(v/v)$ acetic acid aqueous solution for 48hrs and freeze-dried. EDC solution ($H_2O$ : ethanol = 5 : 95) as a crosslink agent was used in concentration of 100mM. In vitro, rat-BMSCs seeded in SIS sponges and induced the osteogenesis for 28 days. We have characterized the osteogenic potential of rat-BMSCs in SIS sponges by alkaline phosphatase activity(ALP), n assay, SEM and RT-PCR for osteogenic phenotype. In SEM, all morphology of SIS sponges was regular and showed interconnected pore structure. By RT-PCR analysis, we observed type I collagen expression. These results demonstrate osteogenic differentiation of rat-BMSCs. In conclusion, we confirmed that the morphology of surface, cross-section, and side of SIS sponges were highly porous with good interconnections between each pores, which can support the surface of cell growth, proliferation, and differentiation. This result indicates that SIS sponge is useful for osteogenesis of BMSCs.

• Korean Journal of Veterinary Service
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• v.23 no.3
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• pp.247-254
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• 2000

This study was performed to elucidate the effects of antler, red ginseng, safflower seed, ipriflavone and estrogen on ovariectomized rats. The rats were fed with Ca and P deficient diet for five weeks to induce osteoporosis. After this period, these animals were fed with normal feed and treated every other day with antler(600 mg/kg, p.o), red ginseng(200 mg/kg, p.o), safflower(200 mg/kg, p.o), ipriflavone(80 mg/kg, p.o) and estrogene(400$\mu\textrm{g}$/kg, i.m) for 5 weeks. During the treatment, the rats were examined for serum concentrations of estradiol, calcitonin, Ca, p and alkaline phosphatase(ALP) activities. The results are summarized as follows 1. The levels of serum 17 $\beta$-estradiol after five weeks of treatment were showed 39.6$\pm$3.0 pg/ml by antler, 33.2$\pm$2.5pg/ml by red ginseng, 34.9$\pm$2.4pg/ml by safflower, 28.1$\pm$3.1 pg/ml by ipriflavone and 40.6$\pm$3.0pg/ml by estrogen-treated group. They were lower than 50.8$\pm$3.1pg/ml of normal control group which had not received ovariectomy. They, however, were significantly higher than 26.8$\pm$1.8pg/ml of ovariectomized non-treatment group(p<0.05). 2. The levels of serum calcitonin after five weeks of treatment were showed 0.60$\pm$0.02 ng/ml by antler, 0.55$\pm$0.04ng/ml by red ginseng, 0.59$\pm$0.02ng/ml by safflower, 0.56$\pm$0.04ng/ml by ipriflavone and 0.62$\pm$0.02ng/ml by estrogen-treated group. They were lower than 0.67$\pm$0.03pg/ml of normal control group. However, they were significantly higher than 0.45$\pm$0.05ng/ml of ovariectomized non-treatment group(p<0.05). 3. The levels of serum Ca of the rats after five weeks of treatment with antler, red ginseng, safflower, ipriflavone and estrogen were 23.51$\pm$2.19$\mu\textrm{g}$/$m\ell$, 25.22$\pm$3.44$\mu\textrm{g}$/$m\ell$, 23.20$\pm$0.02$\mu\textrm{g}$/$m\ell$, 24.76$\pm$3.57$\mu\textrm{g}$/$m\ell$, 23.07$\pm$3.66$\mu\textrm{g}$/$m\ell$, respectively. They were a bit higher than 21.43$\pm$2.22$\mu\textrm{g}$/$m\ell$ of normal control group. And non-treatment group showed 26.12$\pm$0.29$\mu\textrm{g}$/$m\ell$ which was significantly higher than that of control group(P<0.05). 4. The serum P concentraions after five weeks of treatment were showed 12.11$\pm$2.14$\mu\textrm{g}$/$m\ell$ by antler, 13.18$\pm$1.64u91m4 by red ginseng, 12.67 $\pm$2.31$\mu\textrm{g}$/$m\ell$ by safflower, 12.38$\pm$2.07$\mu\textrm{g}$/$m\ell$ by ipriflavone, 11.86$\pm$1.93$\mu\textrm{g}$/$m\ell$ by estrogen-treated group. They were a bit higher than 11.29$\pm$1.23$\mu\textrm{g}$/$m\ell$ of normal control group. And non-treatment group showed 13.42$\pm$1.87$\mu\textrm{g}$/$m\ell$ which was higher than that of control group but not significant. 5. The levels of serum ALP after five weeks of treatment were showed 164.8$\pm$3.8IU/ml by antler, 277.7$\pm$4.8IU/ml by red ginseng, 288.5$\pm$4.5IU/ml by safflower, 214.7$\pm$5.7IU/ml by ipriflavone and 159.4$\pm$5.4IU/ml by estrogen-treated group. They were significantly higher than 144.1$\pm$3.5IU/ml of normal control group(p<0.05). However, they were significantly lower than 336.9 $\pm$12.7IU/ml of ovariectomized non-treatment group(p<0.05). Antler and safflower elevated serum estradiol and calcitonin, and decreased serum ALP significantly. Therefore they were thought to have therapeutic effect on osteoposis by making inhibitory effect on osteoclasts rather than activating osteoblasts.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.29 no.4
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• pp.279-288
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• 2007

In the present study, we focused on stem cells in the dental papilla of the tooth germ. The tooth germ, sometimes called the tooth bud, is the primordial structure from which a tooth is formed. The tooth germ consists of the enamel organ, the dental papilla, and the dental follicle. The dental papilla lies below a cellular aggregation of the enamel organ. Mesenchymal cells within the dental papilla are responsible for formation of dentin and pulp of a tooth. Tooth germ disappears as a tooth is formed, but that of a third molar stays in the jawbone of a human until the age of 10 to 16, because third molars grow slowly. Impacted third molar tooth germs from young adults are sometimes extracted for orthodontic treatment. In the present study, we evaluated the osteogenic activity and mineralization of cultured human dental papilla-derived cells. Dental papillas were harvested from mandible during surgical extraction of lower impacted third molar from 3 patients aged 13-15 years. After passage 3, the dental papilla-derived cells were trypsinized and subsequently suspended in the osteogenic induction DMEM medium supplemented with 10% fetal bovine serum, 50 g/ml L-ascorbic acid 2-phosphate, 10 nM dexamethasone and 10 mM -glycerophosphate at a density of $1\;{\times}10^6\;cells/dish$ in a 100-mm culture dish. The dental papilla-derived cells were then cultured for 6 weeks and the medium was changes every 3 days during the incubation period. Dental papilla-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 7 of culture period, then decreased in intensity during the culture period. ALP mRNA level was largely elevated at 1 weeks and gradually decreased with culture time. Osteocalcin mRNA expression appeared at day 14 in culture, after that its expression continuously increased in a time-dependent manner up to day 28. The expression remained constant thereafter. Runx2 expression appeared at day 7 with no detection thereafter. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. Osteocalcin secretion was detectable in the culture medium from 1 week. The secretion of osteocalcin from dental papilla-derived cells into the medium greatly increased after 3 weeks although it showed a shallow increase by then. In conclusion, our study showed that cultured human dental papilla-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.577-598
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• 1996

This study was conducted to identify the effects of caffeine or combinations of caffeine and iron or vitamin E on the lipid and protein components or blood chemistry levels of the serum as well as the total homogenate, mitochondrial and microsomal fraction of the rat(Sprague-Dawley, female) liver. Chronic test were conducted to determine those effects. The chronic test was conducted by dividing rats into 5 groups according to the type of drugs and dosages administrated as follows; the control(group A), and group B was given 25mg/kg caffeine orally once daily for 30 days, group C was given 50mg/kg caffeine orally once daily for 30 days, group D was given 25mg/kg caffeine and orally ferric chloride once daily for 30 days and group E was given 25mg/kg caffeine and 25mg/kg vitamin E once daily for 30 days. The concentrations of glucose, urea nitrogen, uric acid, creatinine, total protein, albumin, A/G ratio, triglyceride, total cholesterol, HDL-cholesterol, free fatty acid, phospholipid as well as the activities of alanine aminotransferase(ALT), aspartate aminotransferase(AST) and alkaline phosphatase(ALP) were measured in the serum of each experimental groups. The concentrations of the carbonyl group and malondiaidehyde(MDA) and the patterns of the SDS-PAGE(Sodium Dodecyl Sulfate - Polyacrylamide Gel Electrophoresis) and fatty acid compositions in free fatty acids and phospholipids were analyzed to determine the oxidative damages and metabolic changes on the lipid and protein components in the serum, and total homogenate, mitochondrial and microsomal fractions of the rat liver. The results obtained from this study were summarized as follows; 1. Body weights of groups B, C, D and E were significantly decreased(p < 0.01) in comparison with that of the control in the chronic test. 2. The concentrations of serum glucose in groups B(124.5mg/dl), C(130.1mg/dl), D(122.1mg/dl), E(119.3mg/dl) were significantly higher(p < 0.01) in comparison to that of the control(101.5mg/dl). But, there were no significant differences in the concentrations of urea nitrogen, uric acid, creatinine, total protein, albumin and A/G ratio in comparison to that of the control. 3. The concentrations of total cholesterol and HDL-cholesterol in serum of groups B(69.6, 53.4mg/dl), C(73.0, 56.3mg/dl), D(68.9, 51.1mg/dl) and E(68.2, 51.3mg/dl) were significantly higher(p < 0.01) in comparison to that of the control(52.6, 38.8mg/dl). On the other hand, the concentrations of triglyceride in serum of groups B(45.0mg/dl), C(40.4mg/dl), D(33.8mg/dl) and E(47.2mg/dl) were significantly lower(p < 0.01) in comparison to that of the control(66.2mg/dl). There were no significant differences in the activities of ALT, AST and ALP in comparison to that of the control. 4. The concentrations of free fatty acid and phospholipid in serum of groups B(45.7, 154.4mg/dl), C(50.0, 167.2mg/dl), D(52.5, 148.4mg/dl) and E(41.1, 159.2mg/dl) were higher(p < 0.01) in comparison to that of the control(35.2, 125.3mg/dl). And the concentrations of the carbonyl group and malondialdehyde in serum of group D(1.82, 0.52nM/mg protein) were significantly higher(p < 0.01) in comparison to the control(1.53nM/mg protein). 5. The concentrations of carbonyl group in total homogenate, mitochondrial and microsomal fraction of group D(1.45, 0.94, 1.67nM/mg protein) were significantly higher (p < 0.01) in comparison to the control(1.16, 0.66, 1.27nM/mg protein). And the concentrations of malondialdehyde in the total homogenate, mitochondrial and microsomal fraction of group D(6.70, 6.10, 1.36nM/mg protein) were significantly higher(p < 0.01) in comparison to the control(5.17, 3.64, 0.68nM/mg protein). 6. As the analytical results of the fatty acid compositions of free fatty acid in serum, the proportions of stearic acid and arachidonic acid of groups B(16.52, 12.62%), C(17.52, 15.18%), D(19.73, 13.47%) and E(17.62, 13.28%) were significantly higher(p < 0.01) in comparison to the control(14.75, 7.88%), but the proportions of oleic acid and linoleic acid of groups B(12.97, 32.59%), C(10.88, 31.23%), D(12.37, 30.66%) and E(11.95, 32.41%) were significantly lower(p < 0.01) in comparison to the control(16.44, 35.12%). Otherwise, as the results of the fatty acid compositions of phospholipid in serum, the proportions of stearic acid and arachidonic acid of groups B(39.37, 16.39%), C(40.63, 17.83%), D(42.73, 15.39%) and E(39.16, 15.70%) were significantly higher(p < 0.01) in comparison to the control(37.74, 14.24%), but the proportions of oleic acid and linoleic acid of groups B(4.03, 14.38%), C(3.54, 12.38%), D(4.52, 11.68%) and E(4.29, 13.64%) were significantly lower(p < 0.01) in comparison to the control(5.53, 16.14%). 7. As the analytical results of the fatty acid compositions of free fatty acid in total homogenate, mitochondrial and microsomal fraction of liver, the proportions of oleic acid of groups B(7.8**, 8.73**, 6.88%) and C(6.89**, 7.75**, 6.58%) were lower(**:p < 0.01) in comparison to the control(8.67, 10.08, 7.81%), but the proportions of arachidonic acid of group C(22.62, 19.79, 23.71%) were significantly higher(p < 0.01) in comparison to the control(20.93, 18.47, 22.24%). And the proportions of palmitic acid of group D(25.95**, 26.16, 26.34**%) were significantly higher(**:p < 0.01) in comparison to the control(24.43, 25.42, 23.34%). In addition, the proportions of linoleic acid of group D(23.43, 25.02, 23.95%) were also significantly higher(p < 0.01) in comparison to the control(22.17, 23.75, 21.26%). The proportions of stearic acid of group D(19.87, 19.76**%) in mitochondrial and microsomal fraction were lower(**:p < 0.01) in comparison to the control(21.01, 24.18%), and the proportions of stearic acid of group E(16.71*, 19.65**%) in mitochondrial and microsomal fraction were significantly lower(**:p < 0.01, *:p < 0.05) in comparison to the control(21.01, 24.18%), and the proportions of linoleic acid of group E(25.04, 29.20, 26.48%) in total homogenate, mitochondria and microsome were significantly higher(p < 0.01) in comparison to the control(22.17, 23.75, 21.26%). 8. As the results of the fatty acid compositions of phospholipid in total homogenate, mitochondrial and microsomal fraction of liver, the proportions of palmitic acid of group D(17.58**, 18.78*, 18.23%**) were significantly higher(**:p < 0.01, *:p < 0.05) in comparison to the control(16.28, 17.22, 16.38%), and the proportions of stearic acid of group D(36.41, 37.23, 39.53%) were also significantly higher(p < 0.01) in comparison to the control(34.18, 34.16, 36.04%). But the proportions of oleic acid(3.41*, 3.11**, 3.12**%) and linoleic acid (18.03**, 15.79**, 14.74**%) of group D were significantly lower(**:p < 0.01, *:p < 0.05) in comparison to the control(oleic : 3.63, 3.72, 3.79%, linoleic : 20.03, 18.71, 18.48%). 9. In order to determine the oxidative damages to the protein in serum, mitochondrial and microsomal fraction of the rat liver, the patterns of the SDS-PAGE were identified, but the results of SDS-PAGE were not significantly different between the control and experimental groups.