• Clinical and Experimental Reproductive Medicine
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• v.36 no.1
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• pp.23-34
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• 2009

Objectives: Mesenchymal stem cells (MSC) comprise a promising tool for cellular therapy. It is known that long-term in vitro culture of human bone marrow and adipose tissue derived-MSCs lead to a reduction of life span and a change of stem-like characters. The aim of our study was to examine whether stem cell properties of human umbilical cord-derived stem cells (HUC) could be affected by in vitro expansion. Methods: HUC were isolated from human umbilical cord and cultured for 10 passages in vitro. Morphology and population doubling time (PDT) were investigated, and changes of stem cell properties were examined using RT-PCR and immunocytochemistry during serial subcultures. Results: Morphology and PDT of HUC began to change slightly from the 7th passage (p7). Expression level of nestin and vimentin mRNAs increased along with the culture period from p4 until p10. In contrast, expression level of SCF mRNA decreased during the same culture period. Expression level of Oct-4 and HNF-4${\alpha}$ mRNAs was not significantly changed throughout the culture period until p10. Expression level of BMP-4, FGF-5, NCAM and HLA-ABC mRNAs appeared to increase as the culture continued, however, the difference was not significant. Immunocytochemical studies showed that HUC at p3, p6 and p9 positively were stained with antibodies against SSEA-3 and SSEA-4 proteins. Interestingly, staining intensity of HUC for ICAM-1 and HLA-ABC gradually increased throughout the culture period. Intensity against thy-1 and fibronectin antibodies increased at p9 while that against TRA-1-60 and VCAM-1 antibodies began to decrease at p6 until p9. Conclusions: These results suggest that HUC change some of their stem cell characteristics during in vitro culture. Development of culture system might be needed for the maintenance of characteristics.

• Journal of Applied Biological Chemistry
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• v.57 no.1
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• pp.95-101
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• 2014

Nonpoint pollution sources from agricultural activities are a major cause of water quality impairment. A nutrient management program utilizes farm practices that maintain efficient crop production systems and control agricultural nonpoint pollution sources. The objectives of present study were to identify appropriate best management practices (BMPs) according to changes of cropping system of main upland crop for reducing AGNPs loadings and to simulate the effects of the application of the several BMPs scenarios in Saemangeum watershed. The selected BMP scenarios were: 1) to convert naked barley and hulled barley to hairy vetch or chinese milk vetch, 2) to convert red pepper to soybean crop, and 3) to combine two scenarios, converting naked barley and hulled barley to hairy vetch or chinese milk vetch + converting red pepper to soybean crop. As a result of BMPs application, the crop requirement of nitrogen and phosphorus for upland crop reduced nitrogen by 41% and phosphorus by 47% in scenario 1, whereas scenario 2 reduced nitrogen by 30% and phosphorus by 23%. Overall, scenario 3 reduced nitrogen by 72% and phosphorus by 70% in agricultural non-point pollution sources associated with chemical fertilizer and livestock manure in Saemangeum watershed.

• Journal of Periodontal and Implant Science
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• v.34 no.2
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• pp.269-279
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• 2004

백악질 세포의 분리 및 배양방법을 확립하고, 이를 이용하여 백악질 세포의 형질특성을 알아보고자 하였다. 교정목적으로 발거된 소구치를 이용하여, 치은섬유아세포, 치주인대 세포 및 백악질 유래세포를 분리, 배양하였다. 백악질 유래 세포 배양시에는 백악질을 절제한 후 Collagenase P를 이용하여 백악질 유래 세포 외의 다른 세포의 개제를 배제하였고, 기질을 분해하여 세포의 분리 및 배양이 용이하도록 하였다. 분리 및 배양시기의 세포의 형태를 광화현미정을 이용하여 관찰하였다. 조골세포의 특성을 가지는 SaOs-2 세포를 대조군으로 이용하여 분리 및 배양된 세포군들을 동일한 조건으로 배양하였다. 3일 및 7일째에 세포증식도를 측정하였고 7일째에 ALPase 효소 활성도를 측정하였다. 각 세포의 형질 특성을 알아보기 위해 RT-PCR을 실시하여 조골세포 분화 표식자와 연관된 osteopontin(OPN), Alkaline phosphatase(ALP), type I collagen(COL-I), Bone sialoprotein(BSP), BMP-2 및 osteocalcin(OC)의 발현을 비교 관찰하였다. 백악질 유래 세포의 분리 및 배양을 시도한 5명의 치아 중에서 3명의 치아에서 세포군을 배양해 낼 수 있었다. 배양한 백악질 유래 세포는 섬유아세포와 유사한 형태와 증식을 보였다. ALPase 효소 활성도 검사 결과 백악질 유래 세포는 SaOs-2 세포보다 낮은 활성도를 나타내었으며, 배양된 세포의 RT-PCR 결과 백악질 유래 세포군에서는 ALPase의 발현이 나타나지 않았고, 다른 조골세포 표식자의 발현도 낮게 나타났다. 이는 백악질 유래 세포가 조골세포 및 다른 대조군의 세포와는 다른 형질 특성을 가지고 있다는 것을 시사한다. 이상의 관찰결과로 사람의 백악질 유래 세포롤 백악질의 절제 및 효소처리 방법으로 효과적인 분리 및 배양이 가능하며, 이는 향후 백악질 세포의 형질 특성 및 백악질 형성의 분자적 기전을 파악하는 중요한 연구자료로 활용 될 수 있을 것으로 사료된다.

• Journal of Physiology & Pathology in Korean Medicine
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• v.31 no.6
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• pp.374-379
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• 2017

Hair loss affects interpersonal relationships and causes psychological stress. In this study, we investigated the antioxidant activity of Cynanchi Wilfordii Radix (CWR) and its effects on dermal papilla (DP) cells. Antioxidant efficacy was examined by ABTS assay. To confirm the effect on cell activity, MTS assay was performed and cell count was directly measured by hemocytometer. The mRNA expression of genes involved in hair formation and hair loss formation was confirmed by quantitative RT-PCR. CWR has a strong antioxidant activity. Cell viability of DP cells was increased to 118.5% by treatment of 0.5 mg/ml CWR for 24 hours, but the effect on the cell number was insignificant. These results suggest that CWR increases mitochondrial activity without promoting cell proliferation. Treatment of DP cells with 0.5 mg/ml CWR resulted in 48.5% reduction of mRNA expression of type 2 $5{\alpha}$-reductase, a major cause of male hair loss. In addition, mRNA expression of bone morphogenetic pretein (BMP), fibroblast growth factor (FGF)7, and FGF10, which are closely related to hair growth, was also decreased. Reactive oxygen species (ROS) acts as a cause of hair loss. The excellent antioxidant efficacy of CWR is thought to be able to effectively remove ROS. The dihydrotestosterone produced by type 2 $5{\alpha}$-reductase in DP cells is a potent inducer of male pattern hair loss. The inhibitory effect of type 2 $5{\alpha}$-reductase mRNA on DP cells induced by CWR may induce a positive therapeutic effect of male pattern hair loss.

• 한국신재생에너지학회:학술대회논문집
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• 2010.11a
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• pp.101-101
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• 2010

Anaerobic digestion(AD) is the most promising method for treating and recycling of different organic wastes, such as organic fraction of municipal solid waste, household wastes, animal manure, agro-industrial wastes, industrial organic wastes and sewage sludge. During AD, i.e. organic materials are decomposed by anaerobic forming bacteria and fina1ly converted to excellent fertilizer and biogas which is a mixture of carbon dioxide and methane. AD has been one of the leading technologies that can make a large contribution to produce renewable energy and to reduce $CO_2$ and other green-house gas(GHG) emission, it is becoming a key method for both waste treatment and recovery of a renewable fuel and other valuable co-products. Currently some 80% of the world's overall energy supply of about 400 EJ per year in derived from fossil fuels. Nevertheless roughly 10~15% of this demand is covered by biomass resources, making biomass by far the most important renewable energy source used to date. The representative biofuels produced from the biomass are bioethanol, biodiesel and biogas, and currently biogas plays a smaller than other biofuels but steadily growing role. Traditionally anaerobic digestion applied for different biowaste e.g. sewage sludge, manure, other organic wastes treatment and stabilization, biogas has become a well established energy resource. However, the biowaste are fairly limited in respect to the production and utilization as renewable source, but the plant biomass, the so called "energy crops" are used for more biogas production in EU countries and the investigation on the biomethane potential of different crops and plant materials have been carried out. In Korea, with steadily increasing oil prices and improved environmental regulations, since 2005 anaerobic digestion was again stimulated, especially on the biogasification of different biowastes and agro-industrial biomass including "energy crops". This study have been carried out to investigate anaerobic biodegradability by the biochemical methane potential(BMP) test of animal manures, different forage crops i.e. "energy crops", plant and industrial organic wastes in the condition of thermophilic temperature, The biodegradability of animal manure were 63.2% and 58.2% with $315m^3CH_4/tonVS$ of cattle slurry and $370m^3CH_4/tonVS$ of pig slurry in ultimate methane yields. Those of winter forage crops were the range 75% to 87% with ultimate methane yield of $378m^3CH_4/tonVS$ to $450m^3CH_4/tonVS$ and those of summer forage crops were the range 81% to 85% with ultimate methane yield of $392m^3CH_4/tonVS$ to $415m^3CH_4/tonVS$. The forge crops as "energy crops" could be used as good renewable energy source to increase methane production and to improve biodegradability in co-digestion with animal manure or only energy crop digestion.

• Journal of Periodontal and Implant Science
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• v.24 no.3
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• pp.618-632
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• 1994

The ultimate goal of periodontal therapy is promoting the regeneration of lost periodontal tissue. The purpose of this study is to evaluate the effect of treatment using decalcified freeze dried bone allograft as a bone graft material. 47 intrabony defects from 27 patients with clinical diagnosis of chronic periodontitis were selected among those 24 defects were treated via flap operation only and designated as the control group, the other 23 defects were treated with decalcified freeze dired bone allografting via flap operation and designated as the experimental group. Clinical parameters including probing depth, loss of attachment, probing bone level and gingival recession have been recorded at 6th months, and the significance of the changes has been analyzed. The results are as follows : 1. Probing depths were reduced significantly in both control group($2.75{\pm}0.99mm$) and experimental group($3.69{\pm}0.97mm$) postoperatively(p<0.01). Experimental group showed significantly higher decrease compared to the control group(p]0.01). 2. Loss of attachment showed statistically significant decrease in both control group($1.77{\pm}1.08mm$) and experimental group postoperatively($2.70{\pm}1.55mm$). Experimental group showed significantly higher decrease compared to the control group(p]0.05). 3. Probing bone levels were reduced with statistically significance in both control group($1.08{\pm}0.97mm$) and experimental group($4.00{\pm}1.41mm$) postoperatively(p<0.01). Experimental group showed significantly higher decrease compared to the control group(p<0.01). 4. Gingival recession showed statistically significant increase in the control group($1.21{\pm}0.72mm$) and experimental group($1.00{\pm}1.09mm$) postoperatively(p<0.01). There was no statistical significance between the control group and the experimental group. On the basis of these results, treatment using allogenic decalcified freeze dried bone is effective in reducing probing depth, loss of attachment and probing bone level. Therefore allogenic decalcified freeze dried bone is an effective bone graft material in periodontal regeneration.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.5
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• pp.635-647
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• 2014

Unfertilized oocytes age inevitably after ovulation, which limits their fertilizable life span and embryonic development. Rapamycin affects mammalian target of rapamycin (mTOR) expression and cytoskeleton reorganization during oocyte meiotic maturation. The goal of this study was to examine the effects of rapamycin treatment on aged porcine oocytes and their in vitro development. Rapamycin treatment of aged oocytes for 24 h (68 h in vitro maturation [IVM]; $44h+10{\mu}M$ rapamycin/24 h, $47.52{\pm}5.68$) or control oocytes (44 h IVM; $42.14{\pm}4.40$) significantly increased the development rate and total cell number compared with untreated aged oocytes (68 h IVM, $22.04{\pm}5.68$) (p<0.05). Rapamycin treatment of aged IVM oocytes for 24 h also rescued aberrant spindle organization and chromosomal misalignment, blocked the decrease in the level of phosphorylated-p44/42 mitogen-activated protein kinase (MAPK), and increased the mRNA expression of cytoplasmic maturation factor genes (MOS, BMP15, GDF9, and CCNB1) compared with untreated, 24 h-aged IVM oocytes (p<0.05). Furthermore, rapamycin treatment of aged oocytes decreased reactive oxygen species (ROS) activity and DNA fragmentation (p<0.05), and downregulated the mRNA expression of mTOR compared with control or untreated aged oocytes. By contrast, rapamycin treatment of aged oocytes increased mitochondrial localization (p<0.05) and upregulated the mRNA expression of autophagy (BECN1, ATG7, MAP1LC3B, ATG12, GABARAP, and GABARAPL1), anti-apoptosis (BCL2L1 and BIRC5; p<0.05), and development (NANOG and SOX2; p<0.05) genes, but it did not affect the mRNA expression of pro-apoptosis genes (FAS and CASP3) compared with the control. This study demonstrates that rapamycin treatment can rescue the poor developmental capacity of aged porcine oocytes.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.2
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• pp.97-106
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• 2010

Aim of the study: An alternative source of adult stem cells that could be obtained in large quantities, under local anesthesia, with minimal discomfort would be advantageous. Adipose tissue could be processed to obtain a fibroblast-like population of cells or adipose tissue-derived stromal cells (ATSCs). This study was performed to confirm the availability of ATSCs in bone tissue engineering. Materials amp; Methods: In this study, adipose tissue-derived mesenchymal stem cell was extracted from the liposuctioned abdominal fat of 24-old human and cultivated, and the stem cell surface markers of CD 105 and SCF-R were confirmed by immunofluorescent staining. The proliferation of bone marrow mesenchymal stem cell and ATSCs were compared, and evaluated the osteogenic differentiation of ATSCs in a specific osteogenic induction medium. Osteogenic differentiation was assessed by von Kossa and alkaline phosphatase staining. Expression of osteocyte specific BMP-2, ALP, Cbfa-1, Osteopontin and osteocalcin were confirmed by RT-PCR. With differentiation of ATSCs, calcium concentration was assayed, and osteocalcin was evaluated by ELISA (Enzyme-linked immunosorbant assay). The bone formation by 5-week implantation of HA/TCP block loaded with bone marrow mesenchymal stem cells and ATSCs in the subcutaneous pocket of nude mouse was evaluated by histologic analysis. Results: ATSCs incubated in the osteogenic medium were stained positively for von Kossa and alkaline phosphatase staining. Expression of osteocyte specific genes was also detected. ATSCs could be easily identified through fluorescence microscopy, and bone formation in vivo was confirmed by using ATSC-loaded HA/TCP scaffold. Conclusions: The present results show that ATSCs have an ability to differentiate into osteoblasts and formed bone in vitro and in vivo. So ATSCs may be an ideal source for further experiments on stem cell biology and bone tissue engineering.

• Journal of Life Science
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• v.28 no.7
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• pp.874-883
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• 2018

Osteoporosis is a disease that increases the risk of fracture by decreasing the mass and strength of bone. It is caused by imbalance of osteoclast bone formation and osteoclast bone resorption. Bone formation by osteoblast is activated via bone morphogenetic proteins and runt-related transcription factor 2. $Wnt/{\beta}-catenin$ signaling and bone resorption by osteoclast are initiated by the binding of receptor activator of nuclear $factor-{\kappa}B$ ligand and receptor activator of nuclear $factor-{\kappa}B$. Menopausal women are at risk for many diseases due to hormonal imbalances, and osteoporosis is the most common metabolic disorder in 30% of postmenopausal women. When estrogen is deficient, bone resorption of osteoclasts is promoted, and the risk of osteoporosis especially increases in postmenopausal women. Hormone replacement therapy has been widely used to relieve or treat the symptoms of menopausal syndrome. However, long-term administration of hormone therapy has been associated with a high risk of side effects, such as breast cancer, ovarian cancer, and uterine cancer. Recently, phytochemicals have been actively studied as a phytoestrogen, which has an estrogen-like activity to cope with symptoms of menopausal syndrome. Therefore, in this review, we investigated the differentiation mechanism of osteoblast and osteoclast and the role of estrogen and phytoestrogen in bone metabolism in relation to previous studies.

• BMB Reports
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• v.49 no.10
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• pp.548-553
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• 2016

Mesenchymal stem cells (MSCs) are widely used in cartilage tissue engineering to repair articular cartilage defects. However, hypertrophy of chondrocytes derived from MSCs might hinder the stabilization of hyaline cartilage. Thus, it is very important to find a suitable way to maintain the chondrogenic phenotype of chondrocytes. It has been reported that cordycepin has anti-inflammatory and anti-tumor functions. However, the role of cordycepin in chondrocyte hypertrophy remains unclear. Therefore, the objective of this study was to determine the effect of cordycepin on chondrogenesis and chondrocyte hypertrophy in MSCs and ATDC5 cells. Cordycepin upregulated chondrogenic markers including Sox9 and collagen type II while down-regulated hypertrophic markers including Runx2 and collagen type X. Further exploration showed that cordycepin promoted chondrogenesis through inhibiting Nrf2 while activating BMP signaling. Besides, cordycepin suppressed chondrocyte hypertrophy through PI3K/Bapx1 pathway and Notch signaling. Our results indicated cordycepin had the potential to maintain chondrocyte phenotype and reconstruct engineered cartilage.