• Molecules and Cells
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• v.39 no.6
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• pp.439-446
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• 2016

Clearing and labeling techniques for large-scale biological tissues enable simultaneous extraction of molecular and structural information with minimal disassembly of the sample, facilitating the integration of molecular, cellular and systems biology across different scales. Recent years have witnessed an explosive increase in the number of such methods and their applications, reflecting heightened interest in organ-wide clearing and labeling across many fields of biology and medicine. In this review, we provide an overview and comparison of existing clearing and labeling techniques and discuss challenges and opportunities in the investigations of large-scale biological systems.

• Journal of Periodontal and Implant Science
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• v.28 no.4
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• pp.601-611
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• 1998

Ion irradiation is a very promising tool to modify the chemical structure and physical properities of polymers. This study was aimed to evaluate the cellular adhesion to ion beam-irradiated surface of biodegradable poly-l-lactide(PLLA) membrane. The PLLA membrane samples were irradiated by using 35 KeV $Ar^+$ to fluence of $5{\times}10^{13}$, $5{\times}10^{14}$ and $5{\times}10^{15}\;ion/cm^2$. Water contact angles to control and each dose of ion beam-irradiated PLLA membranes were measured. Cultured fetal rat calvarial osteoblasts were seeded onto control and each dose of ion beam-irradiated PLLA membranes and cultured. After 24 hours, each PLLA membranes onto which osteoblasts attached were examined by scanning electron microscopy(SEM). Osteoblasts were removed from each PLLA membrane and then, the vitality and the number of cells were calibrated. Alkaline phosphatase of detached cells from each PLLA membranes were measured. Ion beam-irradiated PLLA membranes showed no significantly morphological change from control PLLA membranes. In the measurement of water contact angle to each membrane, the dose range of ion beam employed in this study reduced significantly contact angles. Among them, $5{\times}10^{14}\;ion/cm^2$ showed the least contact angle. The vitalities of osteoblastes detached from each membranes were confirmed by flow cytometer and well attached cells with their own morphology onto each membranes were observed by SEM. A very strong improvement of the cell adhesion and proliferation was observed for ion beam-irradiated surfaces of PLLA membranes. $5{\times}10^{15}\;ion/cm^2$ exhibited the most strong effect also in cellular adherence. ALPase activities also tended to increase in ion beam-irradiated membranes but statistical differences were not found. These results suggested that ion beam irradiation is an effective tool to improve the adhesion and spreading behaviour of the cells onto the biodegradable PLLA membranes for the promotion of membrane-tissue integration.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.5
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• pp.720-732
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• 2001

Skeletal muscle is of major economic importance since it is finally converted to meat for consumers. The increase in meat production with low costs of production may be achieved by optimizing muscle growth, whereas a high meat quality requires, among other factors, the optimization of intramuscular glycogen and fat stores. Thus, research in energy metabolism aims at controling muscle metabolism, but also liver and adipose tissue metabolism in order to optimize energy partitioning in favour of muscles. Liver is characterized by high anabolic and catabolic rates. Metabolic enzymes are regulated by nutrients through short-term regulation of their activities and long-term regulation of expression of their genes. Consequences of liver metabolic regulation on energy supply to muscles may affect protein deposition (and hence growth) as well as intramuscular energy stores. Adipose tissues are important body reserves of triglycerides, which result from the balance between lipogenesis and lipolysis. Both processes depend on the feeding level and on the nature of nutrients, which indirectly affect energy delivery to muscles. In muscles, the regulation of rate-limiting nutrient transporters, of metabolic enzyme activities and of ATP production, as well as the interactions between nutrients affect free energy availability for muscle growth and modify muscle metabolic characteristics which determine meat quality. The growth of tissues and organs, the number and the characteristics of muscle fibers depend, for a great part, on early events during the fetal life. They include variations in quantitative and qualitative nutrient supply to the fetus, and hence in maternal nutrition. During the postnatal life, muscle growth and characteristics are affected by the age and the genetic type of the animals, the feeding level and the diet composition. The latter determines the nature of available nutrients and the rate of nutrient delivery to tissues, thereby regulating metabolism. Physical activity at pasture also favours the orientation of muscle metabolism, towards the oxidative type. Consequently, breeding systems may be of a great importance during the postnatal life. Research is now directed towards the determination of individual tissue and organ energy requirements, a better knowledge of nutrient partitioning between and within organs and tissues. The discovery of new molecules (e. g. leptin), of new molecular mechanisms and of more powerful techniques (DNA chips) will help to achieve these objectives. The integration of the different levels of knowledge will finally allow scientists to formulate new types of diets adapted to sustain a production of high quality meat with lower costs of production.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.5
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• pp.405-418
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• 2007

Mesenchymal stem cells(MSCs) have been though to be multipotent cells that can replicate that have the potential to differentiate into lineages of mesenchymal tissue including the bone, cartilage, fat, tendon, muscle, and marrow stroma. Especially, scaffolds to support cell-based tissue engineering are critical determinants of clinical efforts to regenerate and repair the body. Selection of a matrix carrier imvolves consideration of the matrix's role as a scaffold for physical support and host tissue integration as well as its ability to support of synergize the osteoinductive program of the implanted mesenchymal stem cell. The aim of this study is to evaluate the effect of autobone and Bio-$Oss^{(R)}$ to adherent mesenchymal stem cells as scaffolds on sinus augmentation with fibrin glue mixture in a rabbit model. 16 New Zealand White rabbits were divided randomly into 4 groups based on their time of sacrifice(1, 2, 4 and 8 weeks). First, mesenchymal stem cells were isolated from iliac crest marrow of rabbits and expanded in vitro. Cell culture was performed in accordance with the technique described by Tsutsumi et al. In the present study, the animals were sacrificed at 1, 2, 4 and 8 weeks after transplantation, and the bone formation ability of each sides was evaluated clinically, radiologically, histologically and histomorphologically. According to the histological observations, autobone scaffolds group showed integrated graft bone with host bone from sinus wall. At 2 and 4 weeks, it showed active newly formed bone and neovascularization. At 8 weeks, lamellae bone was observed in sinus graft material area. Radiologically, autobone with stem cell showed more radiopaque than Bio-$Oss^{(R)}$ scaffolds group. there were significant differences in bone volume between 4 and 8 weeks(p<0.05).

• Reproductive and Developmental Biology
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• v.33 no.2
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• pp.113-117
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• 2009

Animals produced by somatic cell nuclear transfer (SCNT) using genetically modified cells are almost always transgenic, implying that this method is more efficient than the traditional pronuclear microinjection method. Most somatic cells for SCNT in animals are fetus-derived primary cells and successful gene integration in somatic cells will depend on transfection condition. The objective of this study is to evaluate the efficiency of electroporation (Microporator) and liposome reagents (F-6, F-HD, W-EX, W-Q, W-M) for tissue-type plasminogen activator (tPA) gene transfection and to estimate the overall efficiency of transfection of Korean native pig fetal fibroblast cells (KNPFF). Electroporation showed significantly higher transfection efficiency than liposome reagents with regard to the transfection of in vitro cultures in the early stages of development (41.7% with Microporator vs. 18.3% with F-6, 20.0% with F-HD 18.5% with W-EX, 5.0% with W-M and 6.3% W-Q,). Colonies identified as tPA-positives were treated once more with G418 for 10 to 14 days and growing colonies were selected again. When the cells of newly selected colonies were subjected to single-cell PCR, reselection of colonies following second round of G418 selection increased the rate of transgene integration per each colony. These results suggest that transfection with electroporation is the most efficient and the second rounds of G418 selection may be an effective method for transfection of porcine fetal fibroblast cells.

• Journal of Biomedical Engineering Research
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• v.16 no.2
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• pp.223-230
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• 1995

This study investigates a combined mathematical model for the quantitative estimation of regional myocardial blood flow in experimental canine coronary artery occlusion and in patients with ischemic myocardial diseases using Rb-82 dynamic myocardial positron emission tomography. The coronary thrombosis was induced using the new catheter technique by narrowing the lumen of coronary vessel gradually, which finally led to partial obstruction of coronary artery. Thirty four Rb-82 dynamic myocardial PET scans were performed sequentially for each experiment using our 5, 10 and 20 second acquisition protocol, respectively, and six to seven regions of interest were drawn on each transaxial slices, one on left ventricular chamber for input function and the others on normal and decreased perfusion myocardial segments for the flow estimation in those regions. Two compartment model and graphical analysis method have been applied to the measured sets of regional PET data, and the rate constants of influx to myocardial tissue were calculated for regional myocardial flow estimates with the two parameter fits of raw data by the Levenberg-Marquardt method. The results showed that, (I) two compartment model suggested by Kety-Schmidt, with proper modification of the measured data and volume of distribution, could be used for the simple estimation of regional myocardial blood flow, (2) the calculated regional myocardial blood flow estimates were dependent on the selection of input function, which reflected partial volume effect and left ventricular wall motion in previously used graphical analysis, and (3) mathematically fitted input and tissue time activity curves were more suitable than the direct application of the measured data in terms of convergence.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.475-478
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• 2000

The cytosolic glutathione reductase(GR) gene of chinese cabbage was introduced into Lettuce (Lactuca sativa L.) with particle bombardment method. When cotyledon explants were treated with osmoticum-conditioning medium (0.6 M sorbitol/mannitol) 4 hours prior to and 16 hours after bombardment, it was identified by GUS assay that this condition was the most efficient one for introduction of foreign genes into cotyledon tissue of lettuce with particle bombardment. The stable integration of a GR gene was confirmed by the PCR analysis. 0.3, 0.6, 1.5 kbp PCR fragments of transgenes were obtained by three types of primers designed on the basis of GR sequence. To know whether the expression of the GR gene of pBKs-GR 1 can be stably maintained in the next generation, T$_2$selfing seeds obtained from the transformed mother plants were sowed on MS medium supplemented with 200 mg/L kanamycin sulfate. 70% of seedlings showed resistance to kanamycin.

• Proceedings of the Korean Institute of Surface Engineering Conference
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• 2017.05a
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• pp.79-79
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• 2017

Metallic biomaterials have been mainly used for the fabrication of medical devices for the replacement of hard tissue such as artificial hip joints, bone plates, and dental implants. Because they are very reliable on the viewpoint of mechanical performance. This trend is expected to continue. Especially, Ti and Ti alloys are bioinert. So, they do not chemically bond to the bone, whereas they physically bond with bone tissue. For their poor surface biocompatibility, the surface of Ti alloys has to be modified to improve the surface osteoinductivity. Recently, ceramic-like coatings on titanium, produced by plasma electrolytic oxidation (PEO), have been developed with calciumand phosphorus-enriched surfaces. A lso included the influences of coatings, which can accelerate healing and cell integration, as well as improve tribological properties. However, the adhesions of these coatings to the Ti surface need to be improved for clinical use. Particularly Silicon (Si) has been found to be essential for normal bone, cartilage growth and development. This hydroxyapatite, modified with the inclusion of small concentrations of silicon has been demonstrating to improve the osteoblast proliferation and the bone extracellular matrix production. Strontium-containing hydroxyapatite (Sr-HA) was designed as a filling material to improve the biocompatibility of bone cement. In vitro, the presence of strontium in the coating enhances osteoblast activity and differentiation, whereas it inhibits osteoclast production and proliferation. The objective of this work was to study Morphology of bone-like apatite formation on Sr and Si-doped hydroxyapatite surface of Ti-6Al-4V alloy after plasma electrolytic oxidation. Anodized alloys was prepared at 270V~300V voltages with various concentrations of Si and Sr ions. Bone-like apatite formation was carried out in SBF solution. The morphology of PEO, phase and composition of oxide surface of Ti-6Al-4V alloys were examined by FE-SEM, EDS, and XRD.

• Proceedings of the Korean Institute of Surface Engineering Conference
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• 2017.05a
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• pp.159-159
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• 2017

Ti-6Al-4V alloy have been used for dental implant because of its excellent biocompatibility, corrosion resistance, and mechanical properties. However, the integration of such implant in bone was not in good condition to achieve improved osseointergraiton. For solving this problem, calcium phosphate (CaP) has been applied as coating materials on Ti alloy implants for hard tissue applications because its chemical similarity to the inorganic component of human bone, capability of conducting bone formation and strong affinity to the surrounding bone tissue. Various metallic elements are known to play an important role in the bone formation and also affect bone mineral characteristics. Especially, Zn is essential for the growth of the human and Zn coating has a major impact on the improvement of corrosion resistance. Plasma electrolytic oxidation (PEO) is a promising technology to produce porous and firmly adherent inorganic Zn containing TiO2(Zn-TiO2)coatings on Ti surface, and the a mount of Zn introduced in to the coatings can be optimized by altering the electrolyte composition. In this study, effect of Zn content on the corrosion behavior of Ti-6Al-4V alloy after plasma electrolytic oxidation were studied by SEM, EDS, XRD, AC impedance, and potentiodynamic polarization test. The potentiodynamic polarization and AC impedance tests for corrosion behaviors were carried out in 0.9% NaCl solution at similar body temperature using a potentiostat with a scan rate of 1.67 mV/s and potential range from -1500 mV to +2000 mV. Also, AC impedance was performed at frequencies ranging from 10 MHz to 100 kHz for corrosion resistance.

• Proceedings of the Korean Institute of Surface Engineering Conference
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• 2016.11a
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• pp.120-120
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• 2016

Ti-6Al-4V alloy have been used for dental implant because of its excellent biocompatibility, corrosion resistance, and mechanical properties. However, the integration of such implant in bone was not in good condition to achieve improved osseointergraiton. For solving this problem, calcium phosphate (CaP) has been applied as coating materials on Ti alloy implants for hard tissue applications because its chemical similarity to the inorganic component of human bone, capability of conducting bone formation and strong affinity to the surrounding bone tissue. Various metallic elements, such as strontium (Sr), magnesium (Mg), zinc (Zn), sodium (Na), silicon (Si), silver (Ag), and yttrium (Y) are known to play an important role in the bone formation and also affect bone mineral characteristics, such as crystallinity, degradation behavior, and mechanical properties. Especially, Zn is essential for the growth of the human and Zn coating has a major impact on the improvement of corrosion resistance. Plasma electrolytic oxidation (PEO) is a promising technology to produce porous and firmly adherent inorganic Zn containing $TiO_2(Zn-TiO_2)$coatings on Ti surface, and the a mount of Zn introduced in to the coatings can be optimized by altering the electrolyte composition. In this study, corrosion behavior of Ti-6Al-4V alloy after plasma electrolytic oxidation in solutions containing Ca, P and Zn were studied by scanning electron microscopy (SEM), AC impedance, and potentiodynamic polarization test. A series of $Zn-TiO_2$ coatings are produced on Ti dental implant using PEO, with the substitution degree, respectively, at 0, 5, 10 and 20%. The potentiodynamic polarization and AC impedance tests for corrosion behaviors were carried out in 0.9% NaCl solution at similar body temperature using a potentiostat with a scan rate of 1.67mV/s and potential range from -1500mV to +2000mV. Also, AC impedance was performed at frequencies ranging from 10MHz to 100kHz for corrosion resistance.