• Archives of Craniofacial Surgery
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• v.19 no.3
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• pp.181-189
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• 2018

Background: Autogenous bone grafts have several limitations including donor-site problems and insufficient bone volume. To address these limitations, research on bone regeneration is being conducted actively. In this study, we investigate the effects of a three-dimensionally (3D) printed polycaprolactone (PCL)/tricalcium phosphate (TCP) scaffold on the osteogenic differentiation potential of adipose tissue-derived stem cells (ADSCs) and bone marrow-derived stem cells (BMSCs). Methods: We investigated the extent of osteogenic differentiation on the first and tenth day and fourth week after cell culture. Cytotoxicity of the 3D printed $PCL/{\beta}-TCP$ scaffold was evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay, prior to osteogenic differentiation analysis. ADSCs and BMSCs were divided into three groups: C, only cultured cells; M, cells cultured in the 3D printed $PCL/{\beta}-TCP$ scaffold; D, cells cultured in the 3D printed $PCL/{\beta}-TCP$ scaffold with a bone differentiation medium. Alkaline phosphatase (ALP) activity assay, von Kossa staining, reverse transcription-polymerase chain reaction (RT-PCR), and Western blotting were performed for comparative analysis. Results: ALP assay and von Kossa staining revealed that group M had higher levels of osteogenic differentiation compared to group C. RT-PCR showed that gene expression was higher in group M than in group C, indicating that, compared to group C, osteogenic differentiation was more extensive in group M. Expression levels of proteins involved in ossification were higher in group M, as per the Western blotting results. Conclusion: Osteogenic differentiation was increased in mesenchymal stromal cells (MSCs) cultured in the 3D printed PCL/TCP scaffold compared to the control group. Osteogenic differentiation activity of MSCs cultured in the 3D printed PCL/TCP scaffold was lower than that of cells cultured on the scaffold in bone differentiation medium. Collectively, these results indicate that the 3D printed PCL/TCP scaffold promoted osteogenic differentiation of MSCs and may be widely used for bone tissue engineering.

• Journal of Physiology & Pathology in Korean Medicine
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• v.26 no.4
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• pp.511-518
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• 2012

The mechanisms of Toll-like receptor (TLR) signaling have been the focus of extensive studies because TLRs are the target of therapeutic intervention on multiple diseases. In this study, we investigated the inhibitory potential of Vigna angularis (azuki bean) on the TLR signaling. The effect of Vigna angularis extract (JSD) on TLR activation was investigated by assessing NF-${\kappa}B$ and AP-1 inducible secreted embryonic alkaline phosphatase (SEAP) activity. JSD significantly inhibited SEAP activity induced by poly I:C (TLR3 ligand) and poly I (TLR7 ligand) in a dose-dependent manner at concentration below 100 ${\mu}g/ml$ with no sign of cytotoxicity. Pretreatment of JSD markedly suppressed mRNA expressions of pro-inflammatory cytokines and adhesive molecules such as TNF-${\alpha}$, IL-6, RANTES, MCP-1 and ICAM-1 induced by TLR ligands. It also diminished the phosphorylation of $I{\kappa}B$ kinase and $I{\kappa}B$, and followed by $I{\kappa}B$-mediated nuclear translocation of p50, p65, and phosphorylation of p38, JNK, and IRF signaling pathway. In conclusion, our results suggest that Vigna angularis has inhibitory activity on TLR-3 and -7 signaling and it can be further developed as a remedy in curing TLR-related multiple diseases.

• Journal of Periodontal and Implant Science
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• v.31 no.2
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• pp.287-298
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• 2001

Recently, many natural medicines, which have advantage of less side effects and possibility of long-term use have been studied for their capacity of anti-bacterial, anti-inflammatory and regenerative potential of periodontal tissues. Olibanum has the effects to hemostasis, analgesic and anti-inflammatory, and it also has been traditionally used as a drug for the treatment of bone disease in oriental medicine. The purpose of the present study was to investigate the effects of Olibanum extracts on the activity and differentiation of MC3T3-E1 cells, alkaline phosphatase(ALP) synthesis, formation of bone nodules and expression of type I collagen of MC3T3-E1 cells. To examine the cellular activity, MC3T3-E1 cells were cultured with ${\alpha}-MEM(control)$ and each concentration of Olibanum for 2 days and 4 days. To compare the ALP synthesis, MC3T3-E1 cells were cultured with ${\alpha}-MEM(negative\; control)$, dexamethasone(positive control), and each concentration of Olibanum for 2 days and 4 days. To compare the bone nodule formation, MC3T3-E1 ells were cultured for 21 days, and to compare the type I collagen expression, MC3T3-E1 cells were cultured for 4 days. The cellular activity of MC3T3-E1 cells treated with $1{\mu}g/ml$ of Olibanum extracts was significantly increased at 4-day(p<0.05) to control. The activity of ALP in MC3T3-E1 cells treated with $1{\mu}g/ml$ Olibanum extracts was significantly increased at 4-day(p<0.05). All the experimental groups showed much more bone nodule formation than control groups. The group treated with $1{\mu}g/ml$ of Olibanum extracts was the highest bone nodule formation, and showed much more type I collagen expression than negative control. These results indicate that Olibanum extracts may be considered effective in the activity and differentiation of MC3T3-E1 cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.3
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• pp.516-520
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• 2002

This study was designed to investigate the hepatoprotective effects of Semisulcospira libertina and garlic on the acute hepatotoxicity induced by carbon tetrachloride (CCl$_4$) of rats. Male Sprague-Dawley rats weighing 200∼220g were pretreated with dehydrated powder of Semisulcospira libertina (2.1 g/kg, po; SL) and dehydrated powder mixture of Semisulcospira libertina and garlic (3g/kg, 7:3 ratio, po; SG) once daily for 3 consecutive days, and then given a single dose of CCl$_4$(1g/kg in 5ml/kg corn oil, po) and liver function was determined 24 hrs later. Liver damage was assessed by quantitating activities of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), sorbitol dehydrogenase (SDH) and alkaline phosphatase (ALP) as well as by histopathological examination. Pretreatments with SL and SG significantly decleased CCl$_4$-elevated ALT (48% and 61% respectively), AST (32% and 47%) and SDH (51% and 76%), but had no effect on ALP. SL and SG had revealed hepatoprotective effects against CCl$_4$-induced histopathological changes such as severe necrosis, inflammatory cell infiltration and congestion in the central gene of hepatic lobule. These findings demonstrate that SL and SG may haute the hepatoprotective effect on CCl$_4$-induced liver damage.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.6
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• pp.887-892
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• 2003

The effects of liquid culture of Agaricus blazei Murill on the body weight gain, lipid metabolism, protein levels, and enzyme activities were studied in growing male rats. Sprague-Dawley rats were divided into three groups ; control group (A. blazei free water), 20 or 30% A. blazei groups (20 or 30% A. blazei in water) according to the levels of A. blazei supplementation. The rats were fed ad libitum each of the experimental beverage for 5 weeks. The body weight gain, food intake, food efficiency ratio, and organ weight of the 20 or 30% A. blazei groups were similar to those of the control group. There were no differences in the concentrations of triglyceride, total cholesterol, HDL-cholesterol, total protein and albumin, and the activities of GOT, GPT, LDH and alkaline phosphatase of the serum among the experimental groups. The calcium and phosphorus concentrations of serum in the 20 or 30% A. blazei group were significantly decreased compared with those of the control group. The concentrations of LDL-cholesterol and the atherogenic index of the 20 or 30% A. blazei groups were significantly lower than those of the control group, but the serum HDL-cholesterol/total cholesterol ratio was higher. These results showed that the 20 or 30% A. blazei feeding decreased the LDL-cholesterol and the atherogenic index, and increased HDL-cholesterol/total cholesterol ratio in serum of rats.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.243-248
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• 2003

Sperrnatogenesis, the process by which the male germ-line stem cells(GSCs; type A spermatogonia) divide and differentiate to produce the mature spermatozoa, occurs in the seminiferous tubules of the testis. The GSCs proliferate actively to produce two types of cells: other GSCs and differentiating spermatogonia. GSCs have unipotentcy, devoted solely to the generation of sperm. The function of GSCs has broad implications for development, disease, and evolution. Spermatogenesis is fundamental for propagation of species and the defects of this system can result in infertility or disease. The ability to identify, isolate, culture, and alter GSCs will allow powerful new approaches in animal transgenesis and human gene therapy relating to infertility. Until recently, research on stem cells in the testis has been limited because of technical difficulties in isolating and identifying these cell populations. Here, we were trying to find out optimal conditions for in vitro culture of GSCs for identifying and isolating GSCs. We collected mouse GSCs from 3-days old mouse by two-step enzyme digestion method. GSCs were plated and grown on mouse embryonic fibroblasts in Dulbecco's modified Eagle's medium (DMEM) containing 15％ fatal bovine serum, 10 mM 2-mercaptoethanol, 1％ non-essential amino acids, 1 ng/$m\ell$ bFGF, 10 $\mu$M forskolin, 1500 U/$m\ell$ human recombinant leukemia inhibitory factor (LIF). Over a period 3∼5 days, GSCs gave rise to large multicellular colonies resembling those of mouse pluripotent stem cells. After 5th passages, cells within the colonies continued to be alkaline phosphatase and Oct-4 positive and tested positive against a panel of two immunological markers(Integrin $\alpha$ 6 and Integrin $\beta$ 1) that have been recognized generally to characterize GSCs. SSEA-1, SSEA-3, and SSEA-4 also showed positive signals. Based on our data, these GSCs-derived cultures meet the criteria for GSCs itself and even other pluripotent stem cells. We reported here the establishment of in vitro cultures from mouse male GSCs.

• Reproductive and Developmental Biology
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• v.37 no.4
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• pp.219-224
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• 2013

Muscular satellite cell (SC), which is stem cell of postnatal pig, is an important for study of differentiation into adipogenesis, myogenesis, and osteoblastogenesis. In this study, we isolated and examined from pig muscle tissue to determine capacity in proliferate, differentiate, and expression of various genes. Porcine satellite cells (PSC) were isolated from semimembranosus (SM) muscles of 90~100 days old pigs according to standard conditions. The cell proliferation increased in multi-potent cell by Masson's, oil red O, and Alizarin red staining respectively. We performed the expression levels of differentiation related genes using real-time PCR. We found that the differentiation into adipocyte increased expression levels of both fatty acid binding protein 4 (FABP4) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) genes (p<0.01). Myocyte increased the expression levels of the myosin heavy chain (MHC), myogenic factor 5 (Myf5), myogenic regulatory factor (MyoD), and Myogenic factor 4 (myogenin) (p<0.01). Osteoblast increased the expression levels of alkaline phosphatase (ALP) (p<0.01). Finally, porcine satellite cells were induced to differentiate towards adipogenic, myogenic, and osteoblastogenic lineages. Our results suggest that muscle satellite cell in porcine may influence cell fate. Understanding the progression of PSC may lead to improved strategies for augmenting meat quality.

• Korean Journal of Pharmacognosy
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• v.46 no.2
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• pp.167-172
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• 2015

Aleurone layer of Black rice (Oryza sativa L.) is enriched with anthocyanin that could increase bone density. This study was conducted to investigate the osteoporosis-preventing effects of the aleurone layer extract (BRE) on bone loss of ovariectomized (OVX) rats. OVX (or sham-operated) rats were assigned to three groups (n=8 per group): sham operated group (Sham); OVX control group (OVX); OVX-BRE group, OVX rats treated with BRE at 90 mg/kg B.W. The deionized water alone or deionized water with BRE was orally administrated to Sham, OVX or OVX-BRE groups, respectively for 12 weeks. High fat diet with 45 kcal% fat and water were fed to all rats ad libitum. Body weight was significantly decreased in the OVXBRE group compared to the OVX group (p<0.05). The bone mineral density and bone length of tibia were significantly higher in the OVX-BRE group compared to the OVX group and breaking force was significantly higher for the both tibia and femur bones. Serum estradiol concentration and calcium concentration of femur were higher in the OVX-BRE group than those of OVX group. However, serum alkaline phosphatase activity and parathyroid hormone concentration were decreased in the OVXBRE group compared to the OVX group. The results suggest that aleurone layer of Black rice is a potentially useful ingredient to protect against estrogen deficiency or menopause related osteoporosis.

• Radiation Oncology Journal
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• v.22 no.4
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• pp.280-287
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• 2004

Purpose: This study was carried out to determine the protective effects of vitamin C on the hepatotoxicily induced by radiation. Materials and Methods : The Spraque Dawley rats were randomly divided into 3 groups; the control group, the radiation exposed group, aud the radiation and vitamin C-treated group. SOD activity, ca-talase, malondialdehyde and liver enzymes were analyzed to assess the antioxidant effects of vitamin C. Results: The increased level of malondialdehyde and the decreased catalase activity that were induced by radiation were improved after vitamin C but there was no statistical significance among three groups. The superoxide dismutase activity of the liver was increased by vitamin C, but there were no statistically significant differences between the vitamin C-treated group and the non vitamin C-treated group. The level of liver enzymes in sera such as glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, lactate dehyrogenase and alkaline phosphatase were remarkably elevated by radiation. The levels of those enzymes were decreased in the vitamin C-treated group and statistical significance was noted for the GPT level ($p<0.01$). On the electromicrographic findings, the hepatic cell destruction was considerably decreased in the vitamin C-treated group. Conclusion: Vitamin C is thought to be an effective antioxidant against the hepatotoxicity induced by radiation.

• Journal of Veterinary Clinics
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• v.19 no.3
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• pp.342-349
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• 2002

Vitamin D is one of important factors involved in the regulation of bone metabolism. In osteoporosis, the therapeutic effect of vitamin D on the healing process has still been controversial. To conform the effect of $1{\alpha}$, 25 dihydroxy-vitamin $D_3$ on osteoporosis, the change of serum calcium, serum phosphorus, serum alkaline phosphatase, bone mineral density and bone mineral content of osteoporotic tibia were examined comparatively in normal control group(positive control), CFA control(negative control), CFA+$1{\alpha}$, 25 dihydroxy-vitamin $D_3$ 0.01 ug/kg group and CFA+$1{\alpha}$, 25 dihydroxy-vitamin $D_3$ 0.1 ug/kg group after osteoporosis was induced by single injection of complete Freund's adjuvant(CFA) in rats. In change of serum calcium. the significantly increased value was shown on 2nd and 5th week(P< 0.05) after treatment in $1{\alpha}$, 25 dihydroxy-vitamin D$_3$ 0.01 ug/kg group and on 3rd week(P<0.05) after treatment in $1{\alpha}$, 25 dihydroxy-vitamin $D_3$ 0.1 ug/kg group than CFA control. In change of serum phosphorus, the significantly increased value was shown on 2nd week(P<0.05) after treatment in $1{\alpha}$, 25 dihydroxy-vitamin $D_3$ 0.01 ug/kg group and on 3rd and 4th week(P<0.05) after treatment in $1{\alpha}$, ,25 dihydroxy-vitamin $D_3$ 0.1 ug/kg group than CFA control. The value of bone mineral density and bone mineral content of tibia was increased in both $1{\alpha}$,25 dihydroxy-vitamin $D_3$ 0.01 ug/kg group and $1{\alpha}$, 25 dihydroxy-vitamin $D_3$ 0.1 ug/kg group than CFA control, and the increase rate of that was higher in $1{\alpha}$, 25 dihydroxy-vitamin $D_3$ 0.1 ug/kg group than $1{\alpha}$, 25 dihydroxy-vitamin $D_3$ 0.01 ug/kg group. Considering above findings collectively, it was considered that $1{\alpha}$, 25 dihydroxy-vitamin $D_3$ was effective in preventing the complete Freund's adjuvant-induced osteoporotic decrease of bone mass.