• Title/Summary/Keyword: regeneration medium

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Effects of three-dimensionally printed polycaprolactone/β-tricalcium phosphate scaffold on osteogenic differentiation of adipose tissue- and bone marrow-derived stem cells

  • Park, Hannara;Kim, Jin Soo;Oh, Eun Jung;Kim, Tae Jung;Kim, Hyun Mi;Shim, Jin Hyung;Yoon, Won Soo;Huh, Jung Bo;Moon, Sung Hwan;Kang, Seong Soo;Chung, Ho Yun
    • 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.

Studies on the Application of Tissue Culture for Plant Breeding I. Effect of genotype and other combined factors on the callus induction and plant regeneration in anther culture of rice (농작물 조직배양의 육종학적 이용에 관한 연구 제1보 벼 약배양효율에 대한 유전자형과 다른 요인간의 복합효과)

  • Kim, K.H.;Rush, M.C.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.28 no.4
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    • pp.425-430
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    • 1983
  • Rice (Oryza sativa L.) anthers collected from different genotypes were cultured to investigate the priority to increase callus production rate among several factors affecting on callus formation. Rice varieties, F$_1$ hybrids and F$_2$ plants differed greatly in their abilities to produce callus from anthers, and it was confirmed that the culturability of rice anther was a heritable characteristic. Cold shock treatment before plating anthers promoted callus formation rate, but combined effect with genotype having high culturability was more significant. The response to sucrose concentration in culture medium in callus induction rate was different between rice genotypes, but combined effect with genotype was not significant. Single supplement of NAA to R$_2$ medium increased callus production rate remarkably only in rice genotype having high culturability. Conclusively selection of genotypes is most important to increase callus initiation frequency from rice anthers.

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Effects Of Cultured Bone Cell On The Regeneration Of Alveolar Bone (배양골세포 이식이 치조골재생에 미치는 영향)

  • Jeong, Soon-Joon;Herr, Yeek;Park, Joon-Bong;Lee, Man-Sup;Kwon, Young-Hyuk
    • Journal of Periodontal and Implant Science
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    • v.26 no.1
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    • pp.1-26
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    • 1996
  • This study was performed to estimate the effects of cultured bone cell inoculated on porous type hydroxyaptite for the regeneration of the artificial alveolar bone defect. In this experiment 3 beagle dogs were used, and each of them were divided into right and left mandible. Every surgical intervention were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). To reduce the gingival bleeding during surgery, operative site was injected with Lidocaine hydrochloride(l:80,000 Epinephrine) as local anesthesia. After surgery experimental animal were feeded with soft dietl Mighty dog, Frisies Co., U.S.A.) for 1 weeks to avoid irritaion to soft tissue by food. 2 months before surgery both side of mandibular 1st premolar were extracted and bone chips from mandibular body were obtained from all animals. Bone cells were cultured from bone chips obtained from mandible with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Porous type hydroxyapatite were immerse into the high concentrated cell suspension solution, and put 4 hours for attachin the cells on the surface of hydroxyapatite. Graft material were inserted on the artificial bone defect after 3 days of culture. Before insertion of cellinoculated graft material, scanning electronic microscopic observation were performed to confirm the attachment and spreading of cell on the hydroxyapatite surface. 3 artificial bone defects were made with bone trephine drill on the both side of mandible of the experimental animal. First defect was designed without insertion of graft material as negative control, second was filled with porous replamineform hydroxyapatite inoculated with cultured bone marrow cells as expermiental site, and third was filled with graft materials only as positive control. The size of every artificial bone defect was 3mm in diameter and 3mm in depth. After the every surgical intervention of animals, oral hygiene program were performed with 1.0% chlorhexidine digluconate. All of the animals were sacrificed at 2, 4, 6 weeks after surgery. For obtaining histological section, tissus were fixed in 10% Buffered formalin and decalcified with Planko - Rycho Solution for 72hr. Tissue embeding was performed in paraffin and cut parallel to the surface of mandibular body. Section in 8um thickness of tissue was done and stained with Hematoxylin - Eosin. All the specimens were observed under the light microscopy. The following results were obtained : 1. In the case of control site which has no graft material, less inflammatory cell infiltration and rapid new bone forming tendency were revealed compared with experimental groups. But bone surface were observed depression pattern on defect area because of soft tissue invasion into the artificial bone defect during the experimental period. 2. In the porous hydroxyapatite only group, inflammatory cell infiltration was prominet and dense connective tissue were encapsulated around grafted materials. osteoblastic activity in the early stage after surgery was low to compared with grafted with bone cells. 3. In the case of porous hydroxyapatite inoculated with bone cell, less inflammatory cell infiltration and rapid new bone formation activity was revealed than hydroxyapatite only group. Active new bone formation were observed in the early stage of control group. 4. The origin of new bone forming was revealed not from the center of defected area but from the surface of preexisting bony wall on every specimen. 5. In this experiment, osteoclastic cell was not found around grafted materials, and fibrovascular invasion into regions with no noticeable foreign body reaction. Conclusively, the cultured bone cell inoculated onto the porous hydroxyapatite may have an important role of regeneration of artificial bone defects of alveolar bone.

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Transformation of Populus nigra × P. maximowiczii Using Agrobacterium tumefaciens vectors (Agrobacterium tumefaciens vector를 이용(利用)한 양황철의 형질전환(形質轉換))

  • Son, Suk Gyu;Hyu, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.87 no.2
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    • pp.164-172
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    • 1998
  • This study was conducted to find the optimum transformation condition using Agrobacterium harboring promoterless GUS gene. The optimal medium for shoot induction from leaves of Populus nigra${\times}$P. maximowiczii was MS medium supplemented with $0.1mg/{\ell}$ NAA, $0.5mg/{\ell}$ BAP(94% regeneration frequency and 11.5 average number of shoot) According to the test using pBI121, the concentration of antibiotics for selection marker gene was $100mg/{\ell}$ kanamycin or $60mg/{\ell}$ geneticin in the SIM(shoot inducing medium) 3. Two weeks later, callus was induced in the SIM 3 and this callus grew up to 0.5-1cm shoots after 6 weeks in the new SIM 3. And the treatment with methylation inhibitor(5-azacytidine) led to a dramatic increase in foreign gene expression rate from 5.7% to 26.7%. The vector systems showed. different transformation efficiencies based on the fluorometric and histochemical GUS assay. In this study the vector systems used for transformation seemed to affect transformation frequency, in which pEHA101 yielded more transformants(35.9%) than LBA4404/pBI121 did(5.7%). This result indicated that pEHA101 was effective to insert the promoterless foreign gene into a poplar genome.

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Exploring an Integrated Garden City Theory Based on East Asian Garden Culture - Centering on Community and Integration - (동아시아 정원문화에 기반한 통합적 정원도시론의 모색 - 공동체성과 통합성을 중심으로 -)

  • Ahn, Myung-June
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.41 no.3
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    • pp.13-26
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    • 2023
  • Landscapes and gardens have emerged as an important medium of practice in contemporary cities. Among them, this paper examines the city through the frame of gardens. This is because gardens are being reconceptualized as a medium of activity for urban residents and have become an important subject of action in urban regeneration and the creation of urban villages. From this perspective, this paper examines and proposes an "integrated garden city theory" as a landscape theory suitable for the contemporary era by focusing on the urban structure and the behavior of urban residents through the medium of gardens, as well as the process and results. This is both a process and a result of looking back at the evolution of landscape for over a century and rethinking the identity of landscape. We first examined garden city theory, noting that Ebenezer Howard and Frederick Law Olmsted's positions on the relationship between gardens and cities were not so different, and that "working and responsive landscapes" were fundamental to cities and the beginning of landscape theory. We also examine how their ideals have not been fully realized in cities over the past century, but the prototype of gardens based on traditional garden culture is now being formed in East Asian cities, and the evolution of landscape theory in response. The conclusion is that a new version of the garden concept should be reestablished as a living infrastructure in our cities, and a new garden city theory is needed to make it work. To this end, each chapter examines three arguments, as follows First, the values of gardens and East Asian garden cultures in contemporary cities are shaped by the themes of community and integrity. Second, Korean communality, represented by apartments, is expressed through gardening and requires the reconciliation of city and life and the role of landscape architecture as a specialized field to support it. Third, we examine and consider an integrated garden city theory as a theory of practice in which city-based, everyday life, and garden mediums, i.e., city, life, and garden, are organic, based on an oriental view of nature. As a result, it is confirmed that contemporary gardens and cities are looking for important elements and values that still need to be rediscovered in East Asian landscape and garden cultures. Although the proposal of an integrated garden city theory cannot guarantee the continuation of landscaping, it can be an opportunity for all fields related to cities, not just landscaping, to collaborate and consider garden cities. Through this, it is hoped that "the concept of garden and city suitable for metropolitan or dense cities, ways to spread and support garden culture based on community, evolution of landscape theory/design theory suitable for lifestyle and terrain conditions, search for sustainable/resilient garden city theory that can respond to climate change, and establishing a new role for landscape in the 21st century" will be seriously considered.

Histological studies on in vitro Propagation of Pulsatilla koreana Nakai (할미꽃 기내증식(器內增殖)에 관(關)한 조직학적(組織學的) 연구(硏究))

  • Lee, Man-Sang;Oh, Ki-Hong
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.2
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    • pp.137-157
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    • 1993
  • This study was carried out to investigate the optimal condition for multiple propagation through leaf tissue culture and to apply anther culture techniques to Pulsatilla koreana Nakai breeding. Leaf and anther of Pulsatilla koreana Nakai were cultured on MS, MT, LS and $B_5$ media supplemented with several growth regulators and nitrogen sources under various conditions. For callus induction and differentiation from the Pulsatilla koreana leaf segments were more effective in the combination of zeatin and auxin than auxin alone. The color of the callus was green when treated with IBA alone. Shoot differentiation was more effective when treated with zeatin than auxin alone, especially the best hormoal combination for shoot differentiation was zeatin 1.0mg/l +NAA 0.1mg/l, while 2,4-D inhibited shoot differentiation. The appeared rate of S pollen was 35% in vivo, while that of S pollen by low temperature$(4^{\circ}C)$ pretreatment for 4 days was increased by 53% and the optimum culture time for callus induction from anther was uni-nucleate stage. $B_5$ basal medium supplemented with NAA 0.5mg/l and zeatin 1 mg/l was the most effective on callus formation and the best results of plant regeneration were obtained from combination of NAA 0.5mg/l and zeatin 0.5mg/l in anther culture. $NH_{4}NO_3$ as more effectives as the nitrogen source than $KNO_3$ and the combination with zeatin 2.0mg /L was the best effective. The best combination for plant regeneration in callus induced from anther was $NH_{4}NO_3$ 1650mg/l + $KNO_3$ 3800mg/l + zeatin 2.0mg/l. Ploidy level of anther-derived plants appeared 28% haploid, 47% diploid and the others were triploid, tetraploid and mixploid. In compare with E.S.T, M.D.H and P.X banding patterns were distinguished among callus, haploid and diploid plants in electrophoresis.

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Development of Agrobacterium-mediated Transformation Method for Domestically Bred Chrysanthemum Cultivar 'Moulinrouge' and Genetic Change of Leaf Morphology Using AtSICKLE Gene (아그로박테리움를 이용한 국내개발 국화품종 '무랑루즈'의 형질전환 기술 및 AtSICKLE 유전자를 이용한 엽형 변화 국화 형질전환체 개발)

  • Kim, Yun-Hye;Park, Hyun-Myung;Jung, Ji-Yong;Kwon, Tack-Min;Jeung, Soon-Jae;Yi, Young-Byung;Kim, Gyung-Tae;Nam, Jae-Sung
    • Horticultural Science & Technology
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    • v.28 no.3
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    • pp.449-455
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    • 2010
  • 'Moulinrouge' was selected as the best regenerating cultivar among 18 different spray-type chrysanthemum cultivars bred in the Gyeongnam Flowers Breeding Research Institute. When the leaf explants from standard- and spray-type chrysanthemum 'Jinba' and 'Moulinrouge' were incubated on MS basal medium supplemented with $0.5mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA, both 'Jinba' and 'Moulinrouge' induced adventitious shoots that can be regenerated into plantlets. Based on these regeneration conditions, we developed an efficient $Agrobacterium$-mediated chrysanthemum 'Moulinrouge' transformation method by using sequential selection of shoots from low ($10mg{\cdot}L^{-1}$) to high ($30mg{\cdot}L^{-1}$) concentrations of kanamycin after co-cultivation of leaf explants with $Agrobacterium$ for 10 days and induction of shoots. All kanamycin resistant plants investigated with genomic PCR analysis carried the report gene, $AtSICKLE$, in their genome. Although expression levels of the report gene in the transgenic plants investigated with RT-PCR were relatively low because of inefficiency of CaMV 35S promoter in chrysanthemum, transgenic lines expressing $AtSICKLE$ efficiently showed leaf epinasty phenotype. We expect that our results will provide a useful method that can perform a high-throughput investigation of genes isolated and studied well in model plants for molecular breeding of chrysanthemum.

A Study of Ground Tire as a Sorption Media for the Passive Treatment Wall: Sorption of MTBE (Methyl tertiary Butyl Ether) (파쇄 폐타이어를 이용한 반응벽체에 관한 연구: 폐타이어 내의 MTBE(Methyl tertiary Butyl Ether)흡착 중심)

  • 박상현;이재영;최상일
    • Journal of Soil and Groundwater Environment
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    • v.8 no.3
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    • pp.37-44
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    • 2003
  • Fuel oxygenates, such as Methyl tertiary Butyl Ether (MTBE) is additive in gasoline used to reduce air pollution. Gasoline components and fuel additives can leak: form underground storage tanks. MTBE is far more water soluble than gasoline hydrocarbons like BTEX then it travels at essentially the same velocity as groundwater. MTBE in drinking water causes taste and odor problems. Therefore, the purpose of the this study is to examine the ability of ground tire to sorb MTBE in water. The study consisted of running both batch and column tests to determine the sorption capacity, the required sorption equilibration time, and the flow through utilization efficiency of ground tire. The batch test result indicated that ground tire can attain equilibrium sorption capacities about 0.5 mg of MTBE. The result of column test indicate that ground tire has on the 36% utilization rate. Finally, it is clear that ground tire represented an attractive and relatively inexpensive sorption medium for a MTBE. Authors thought that to determine the economic costs of ground tire utilization, the cost to sorb a given mass of contaminant by ground tire will have to be compared to currently accepted sorption media. The cost comparison will also have to include regeneration and disposal cost.

The Effect of EGF, T3 and HB-EGF on Human Periodontal Fibroblasts (EGF, T3, HB-EGF 가 치주인대섬유모세포에 미치는 영향)

  • Hong, Eun-Kyoung;Cha, Jeong-Heon;Kim, Yun-Tae;Choi, Byung-Jai;Kim, Seong-Oh
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.3
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    • pp.438-446
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    • 2007
  • Viable cells of periodontal ligament would be an important factor for the successful replantation of an avulsed tooth. Therefore, it is critical to choose the storage medium for the preservation of traumatically avulsed teeth. Growth factors and hormones could be considered for the therapeutic application of the maintenance of viable periodontal ligament fibroblasts (PDLFs). Epidermal growth factor (EGF) has been suggested as an important player for the regeneration and wound healing process on other tissues. Therefore, EGF was evaluated for the therapeutic application on avulsed teeth. In addition, the synergic effect of EGF with tri-iodothyronine (T3) and heparin-binding epidermal growth factor-like growth factor (HB-EGF). The cell proliferation of PDLFs was determined by MTT assay and increased dose-dependently up to 10 ng/ml in the presence of EGF. Maximum cellular growth was shown at the concentration of 10 ng/ml EGF. Also, EGF promoted the wound healing of PDLFs examined by in vitro wound healing assay. Combined effects of EGF with T3 or HB-EGF on the proliferation of PDLFs were also studied. Interestingly, EGF showed the synergic effect on the proliferation of PDLFs with T3 and HB-EGF. To find out the mechanism of the synergic effect of EGF and T3, the effect of T3 on the expression of endogenous EGF receptor was determined by RT-PCR. The result was that T3 enhanced the expression of EGF receptor in PDLFs. It suggested that EGF might be a good choice for a therapeutic application, which can be used as combination with T3 and HB-EGF.

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Regeneration and Tending Practices for Natural Mixed Stands of Quercus mongolica-Abies holophylla II. Prediction of Futures Stand Structure using Rate of Increment (신갈나무-전나무 천연(天然) 혼효임분(混淆林分)의 갱신(更新) 및 무육방법(撫育方法) II. 생장률(生長率)을 이용(利用)한 미래(未來) 임분구조(林分構造)의 예측(豫測))

  • Shin, Man Yong;Lim, Joo Hoon;Chun, Young Woo;Ko, Yung Zu
    • Journal of Korean Society of Forest Science
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    • v.81 no.2
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    • pp.146-155
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    • 1992
  • The natural mixed stand in Jindong-Ri, according to a recent study, showed the possibility of selection cutting as a silvicultural system based on the stand structure such as DBH distribution and height distribution. However, volume structure per DBH class of this stand had not a mature stand of selection cutting. In this study, wherefore, the rate of increment for DBH and volume was used to predict the future stand structure including volume distribution per DBH class. The possibility of selection cutting was then discussed using the future stand structure. From the prediction of future stand structure for 30 years per every 10 years, it could be concluded that the stand will be induced to selection cutting forest because of enough number of trees in objective DBH class and above, total volume per hectare, and the volume of large DBH class which can be harvested every year. However, this stand still did not show the structure of typical selection cutting which has the rate of 1 : 2 : 7 in the number of trees per hectare and the volume rate of 5 : 3 : 2 for large, medium, and small DBH class. This problem could be improved by appropriate silvicultural treatments.

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