• Title/Summary/Keyword: scanning microscopy

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Tissue Engineered Cartilage Formation on Various PLGA Scaffolds (PLGA 종류와 담체의 형성 방법에 따른 인간의 조직공학적 연골형성)

  • 김유미;임종옥;정호윤;박태인;백운이
    • Journal of Biomedical Engineering Research
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    • v.23 no.2
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    • pp.147-153
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    • 2002
  • The purpose of this study was to evacuate the effect of different types of Poly(lactic-co-glycolic acid) (PLGA) scaffolds on the formation of human auricular and septal cartilages. All of the scaffolds were formed in a tubular shape for potential application for artificial trachea or esophagus with either 110,000 g/mol PLGA. 220,000 g/mol PLGA. or a combination of both. In order to maintain the tubular shape in vivo, two methods were used. One method was inserting polyethylene tube at the center of scaffolds made of 110,000 g/mol PLGA. The other method involved combination of the two different molecular weight PLGA's. The inner surface of tubular shaped scaffold made with 110,000 g/mol PLGA was coated with 220,000 9/mol PLGA to give more mechanical rigidity. Elastic cartilage was taken from the ear of a patient aged under 20 nears old and hyaline cartilage was taken from the nasal septum. The chondrocytes were then isolated. After second passage, the chondrocytes were seeded on the PLGA scaffolds followed by in vitro culture for one week. The cells-PLGA scaffold complex were implanted subcutaneously on the back of nude mice for 8 weeks. The tissue engineered cartilages were separated from nude mice and examined histologically after staining with the Hematoxylin Eosin. The morphology of the scaffolds were examined by scanning electron microscopy. The pores were well formed and uniformly distributed in the various PLGA scaffolds. After 8 weeks in vivo culture, cartilage was well formed with 110,000 g/mol PLGA. however lumen had collapsed. In contrast. a minimal amount of neocartilage was formed with 220,000 g/mol PLGA, while the architecture of scaffold and lumen were well preserved. Elastic cartilage formed more neocartilage than hyaline. Hyaline and elastic neocartilage were well formed on 110,000 g/mol PLGA with the polyethylene tube, exhibiting mature chondrocytes and preservation of the tubular shape. It was found that 110,000 g/mol PLGA was more appropriate for cartilage formation but higher molecular weight polymer was necessary to maintain the three dimensional shape of the scaffold.

Animal Experiments on an Antithrombogenic Small-Caliber Vascular Prostheses and Vascualr Patch : Observation in Canine Models (항혈전성 소구경 인조 혈관 및 봉합편에 대한 동물 실험)

  • 김수철;김원곤;유세영
    • Journal of Chest Surgery
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    • v.36 no.2
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    • pp.63-72
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    • 2003
  • Although a variety of synthetic vascular grafts are available in modern vascular surgery, no ideal prosthesis ha,4 yet been developed. Small-caliber vascular grafts with low flow, as used in the lower extremity, continue to become thrombosed at unacceptable rates. We have developed and evaluated the new antithrombogenic blood contacting surfaces in canine model. Material and Method: Two now antithrombogenic blood contacting surfaces(Polyvinylalcohol -Polyurethane(PVA-PU) blend and natural Graphite-polyurethane(G-PU) blend) have been developed and evaluated in canine model, using vascular grafts and patches. The luminal surfaces of the test vascular grafts(5 mm ID) were fabricated by dipping a glass rod in PVA-PU blend solution(50 % PVA) using phase separation method. Mongrel dogs of either sex weighing 18-22 kg were anesthetized by endotracheal intubation using halothane and their lungs were ventilated with a volume-cycled ventilator, Maintenance anesthesia with 0.5-1.0% halothane and supplemental oxygen was used. Two pairs were used for comparison in the bilateral femoral arteries for both vascular grafts(PVA-PU vs. PU) and vascular patches(G-PU vs. PU). Bilateral groin incisions were made and the arteries were exposed and clamped. After an excision of 1 cm of the artery between clamps, a grail of 2.5 cm in length was implanted end-to-end using 6-0 polypropylene suture. The vascular patch was implanted as a form of on-lay patch. Animals were sacrificed at 1, 2, 4, 6, 8 and 16 weeks for vascular grafts and 1, 2. 4 and 6 weeks for vascular patches. Result The vascular grafts of PVA-PU blends showed patent lumina in the 2 and 16 weeks animals, while those of PU showed a patent lumen in 2 weeks animal. PVA-PU graft of 16 weeks showed a fairly clean luminal surface. A light microscopic finding of this graft demonstrated good tissue infiltration through porosity, The animals with vascular patches showed patent arteries in both groups except 2 weeks animal. Scanning electron microscopy of the luminal surfaces of G-PU patches in 4 and 6 weeks animals showed endothelial cell covering with microvilli. PU patches showed qualitatively less endothelial cell covering. Conclusion: In conclusion, PVA-PU and G-PU blends can be a promising blood contacting surfaces for application in a synthetic vascualr graft. However, further animal study is needed to determine the real long-term effects of these methods of surface modifications.

Identification of Active Agents for Reductive Dechlorination in Cement/Fe(II) Systems (시멘트와 Fe(II)을 이용한 환원성 탈염소화반응의 유효반응성분 규명)

  • Kim, Hong-Seok;Lee, Yu-Jung;Kim, Ha-Yan;Hwang, In-Seong
    • Journal of Soil and Groundwater Environment
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    • v.11 no.6
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    • pp.35-42
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    • 2006
  • Experimental study was conducted to identify the active agent for reductive dechlorination of TCE in cement/Fe(II) systems. Several potential materials-hematite (${\alpha}-Fe_2O_3$), lepidocrocite (${\gamma}$-FeOOH), akaganeite (${\beta}$-FeOOH), ettringite ($Ca_6Al_2(SO_4)_3(OH)_{12}$)-that are cement components or parts of cement hydrates were tested if they could act as reducing agents by conducting TCE degradation experiments. From the initial degradation experiments, hematite was selected as a potential active agent. The pseudo-first-order degradation rate constant ($k\;=\;0.637\;day^{-1}$) for the system containing 200 mM Fe(II), hematite and CaO was close to that ($k\;=\;0.645\;day^{-1}$) obtained from the system containing cement and 200 mM Fe(II). CaO, which was originally added to simulate pH of the cement/Fe(II) system, was found to play an important role in degradation reactions. The reactivity of the hematite/CaO/Fe(II) system initially increased with increase of CaO dosage. However, the tendency declined in the higher CaO dosage region, implying a saturation type of behavior. The SEM analysis revealed that the hexagonal plane-shaped crystals were formed during the reaction with increasing degradation efficiency, which was brought about by increasing the CaO dosage. It was suspected that the crystals could be portlandite or green rust ($SO_4$) or Friedel's salt. The XRD analysis of the same sample identified the peaks of hematite, magnetite/maghemite, green rust ($SO_4$). Either instrumental analysis predicted the presence of the green rust ($SO_4$). Therefore, the green rust ($SO_4$) would potentially be a reactive agent for reductive dechlorination in cement/Fe(II) systems.

[ $Ca^{2+}\;and\;K^+$ ] Concentrations Change during Early Embryonic Development in Mouse (생쥐 초기 배 발달 동안 변화되는 칼슘과 포타슘 이온)

  • Kang D.W.;Hur C.G.;Choi C.R.;Park J.Y.;Hong S.G.;Han J.H.
    • Journal of Embryo Transfer
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    • v.21 no.1
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    • pp.35-43
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    • 2006
  • Ions play important roles in various cellular processes including fertilization and differentiation. However, it is little known whether how ions are regulated during early embryonic development in mammalian animals. In this study, we examined changes in $Ca^{2+}\;and\;K^+$ concentrations in embryos and oviduct during mouse early embryonic development using patch clamp technique and confocal laser scanning microscopy. The intracellular calcium concentration in each stage embryos did not markedly change. At 56h afier hCG injection when 8-cell embryos could be Isolated from oviduct, $K^+$ concentration in oviduct increased by 26% compared with that at 14h after injection of hCG During early embryonic development, membrane potential was depolarized (from -38 mV to -16 mV), and $Ca^{2+}$ currents decreased, indicating that some $K^+$ channel might control membrane potential in oocytes. To record the changes in membrane potential induced by influx of $Ca^{2+}$ in mouse oocytes, we applied 5 mM $Ca^{2+}$ to the bath solution. The membrane potential transiently hyperpolarized and then recovered. In order to classify $K^+$ channels that cause hyperpolarization, we first applied TEA and apamin, general $K^+$ channel blockers, to the bath solution. Interestingly, the hyperpolarization of membrane potential still appeared in oocytes pretreated with TEA and apamin. This result suggest that the $K^+$ channel that induces hyperpolarization could belong to another $K^+$ channel such as two-pore domain $K^+(K_{2P})$channel that a.e insensitive to TEA and apamin. From these results, we suggest that the changes in $Ca^{2+}\;and\;K^+$ concentrations play a critical role in cell proliferation, differentiation and reproduction as well as early embryonic development, and $K_{2P}$ channels could be involved in regulation of membrane potential in ovulated oocytes.

The Effect of Surface Defects on the Cyclic Fatigue Fracture of HEROShaper Ni-Ti rotary files in a Dynamic Model: A Fractographic Analysis (Fractographic 분석을 통한 HEROShaper 니켈티타늄 전동 파일의 피로파절에 미치는 표면결함의 역할)

  • Lee, Jung-Kyu;Kim, Eui-Sung;Kang, Myoung-Whai;Kum, Kee-Yeon
    • Restorative Dentistry and Endodontics
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    • v.32 no.2
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    • pp.130-137
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    • 2007
  • This in vitro study examined the effect of surface defects on cutting blades on the extent of the cyclic fatigue fracture of HEROShaper Ni-Ti rotary files using fractographic analysis of the fractured surfaces. A total of 45 HEROShaper (MicroMega) Ni-Ti rotary flies with a #30/.04 taper were divided into three groups of 15 each. Group 1 contained new HEROShapers without any surface defects. Group 2 contained HEROShapers with manufacturing defects such as metal rollover and machining marks. Croup 3 contained HEROShapers that had been clinically used for the canal preparation of 4-6 molars A fatigue-testing device was designed to allow cyclic tension and compressive stress on the tip of the instrument whilst maintaining similar conditions to those experienced in a clinic. The level of fatigue fracture time was measured using a computer connected the system. Statistical analysis was performed using a Tukey's test. Scanning electron microscopy (SEM) was used for fractographic analysis of the fractured surfaces. The fatigue fracture time between groups 1 and 2, and between groups 1 and 3 was significantly different (p<0.05) but there was no significant difference between groups 2 and 3 (p>0.05). A low magnification SEM views show brittle fracture as the main initial failure mode At higher magnification, the brittle fracture region showed clusters of fatigue striations and a large number of secondary cracks. These fractures typically led to a central region of catastrophic ductile failure. Qualitatively, the ductile fracture region was characterized by the formation of microvoids and dimpling. The fractured surfaces of the HEROShapers in groups 2 and 3 were always associated with pre-existing surface defects. Typically, the fractured surface in the brittle fracture region showed evidence of cleavage (transgranular) facets across the grains, as well as intergranular facets along the grain boundaries. These results show that surface defects on cutting blades of Ni-Ti rotary files might be the preferred sites for the origin of fatigue fracture under experimental conditions. Furthermore this work demonstrates the utility of fractography in evaluating the failure of Ni-Ti rotary flies.

Optimization of Culture Conditions and Encapsulation of Lactobacillus fermentum YL-3 for Probiotics (가금류 생균제 개발을 위한 Lactobacillus fermentum YL-3의 배양조건 최적화 및 캡슐화)

  • Kim, Kyong;Jang, Keum-Il;Kim, Chung-Ho;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.255-262
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    • 2002
  • This experiment was performed to improve the stability of Lactobacillus fermentum YL-3 as a poultry probiotics. The culture conditions that improve acid tolerance of L. fermentum YL-3 were investigated by changing several factors such as medium composition, temperature, anaerobic incubation and culture time. Also, L. fermentum YL-3 was encapsulated with alginate, calcium chloride and chitosan. The stable culture conditions of L. fermentum YL-3 were obtained in anaerobic incubation using MRS media without tween 80 for 20 hour at $42^{\circ}C$. The capsule after treatment with 1% chitosan was formed close membrane by a bridge bond. Immobilization of L. fermentum YL-3 in capsule was observed by confocal laser scanning microscopy, and cell viability was $2.0{\times}10^9\;CFU/g$ above the average. L. fermentum YL-3 capsule after acid treated at pH 2.0 for 3 hour survived about 40%, but those encapsulated with 1% chitosan survived about 65%. Survival rate of capsule stored at room temperature decreased about $2{\sim}3$ log cycle during 3 weeks, but viability of capsule stored at $4^{\circ}C$ during 3 weeks maintained almost $10^8\;CFU/g$ levels.

Evaluation of shear-bond strength between different self-adhesive resin cements with phosphate monomer and zirconia ceramic before and after thermocycling (인산염계 기능성 단량체가 첨가된 수종의 자가 접착 레진시멘트와 지르코니아 세라믹 사이 열순환 전후 전단결합강도 비교)

  • Lee, Ji-Hun;Kim, Min-Kyung;Lee, Jung-Jin;Ahn, Seung-Geun;Park, Ju-Mi;Seo, Jae-Min
    • The Journal of Korean Academy of Prosthodontics
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    • v.53 no.4
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    • pp.318-324
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    • 2015
  • Purpose: This study compared shear bond strengths of five self-adhesive cements with phosphate monomer to zirconium oxide ceramic with and without airborn particle abrasion. Materials and methods: One hundred zirconia samples were air-abraded ($50{\mu}mAl_2O_3$). One hundred composite resin cylinders were fabricated. Composite cylinders were bonded to the zirconia samples with either Permacem 2.0 (P), $Clearfil^{TM}$ SA Luting (C), $Multilink^{(R)}$ Speed (M), $RelyX^{TM}$ U200 Automix (R), G-Cem $LinkAce^{TM}$ (G). All bonded specimens were stored in distilled water ($37^{\circ}C$) for 24 h and half of them were additionally aged by thermocycling ($5^{\circ}C$, $55^{\circ}C$, 5,000 times). The bonded specimens were loaded in shear force until fracture (1 mm/min) by using Universal Testing Machine (Model 4201, Instron Co, Canton, MA, USA). The failure sites were inspected under field-emission scanning electron microscopy. The data was analyzed with ANOVA, Tukey HSD post-hoc test and paired samples t-test ($\alpha$=.05). Results: Before and after thermocycling, $Multilink^{(R)}$ Speed (M) revealed higher shear-bond strength than the other cements. G-Cem $LinkAce^{TM}$ (G) showed significantly lower bond strengths after thermocycling than before treatment (P<.05), but the other groups were not significantly different (P>.05). Conclusion: Most self-adhesive cements with phosphate monomer showed high shear bond strength with zirconia ceramic and weren't influenced by thermocycling, so they seem to valuable to zirconia ceramic bonding.

Effects of Flushing, Preservation and Reperfusion in the Canine Transplanted Lung Tissue (관류, 보존 및 재관류 과정이 이식된 개의 폐조직에 미치는 영향)

  • Lim, Young-Keun;Park, Chang-Kwon;Kwon, Kun-Young
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.4
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    • pp.512-522
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    • 1999
  • Background: Due to the paucity of suitable donor organs for lung allotransplantation, a number of techniques have been developed to improve the lung preservation. Ultrastructural studies of the morphologic changes of the flushing, preservation and reperfusion injury in donor lungs have rarely been reported. Methods: Adult dogs (n=46) were matched as donors and recipients for the single lung transplantation. The donor lungs were preserved after flushing with preservation solution and transplanted after 20-hours of preservation at $10^{\circ}C$. Ultrastructural features of the lung were examined after flushing, preservation and 2 hours after lung transplantation (reperfusion) respectively. Results: Electron microscopy after flushing showed focal alveolar collapse and mild swelling of type I epithelial cells. After preservation both type I epithelial cells and endothelial cells were swollen and destroyed focally. The endothelial cells showed protrusion of tactile-like structures into the lumina, blebs or vacuoles of the cytoplasm After reperfusion the lung tissue showed fibrin material in the alveoli, prominent type I epithelial cell swelling with fragmented cytoplasmic debris and marked endothelial cell swelling with vacuoles or tactile-like projections. The alveolar macrophages showed active phagocytosis. Scanning electron microscopic examination of the pulmonary parenchyma showed focally alveolar collapse and focal consolidation after the preservation and more prominent changes after the reperfusion procedure. The lungs preserved with low potassium dextran glucose solution, with additional prostaglandin $E_1(PGE_1)$ and verapamil(VP) showed relatively well preserved ultrastructures compared with those which were preserved with modified Euro-Collins or University of Wisconsin, and with additional $PGE_1$ and/or VP. Conclusion: The ultrastructural changes associated with flushing were mild in severity, the donor lungs were injured during the preservation, and further damage was occurred during the reperfusion. The reperfusion injury resulted in prominent pulmonary parenchymal alterations with a pattern of acute lung injury.

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Characteristics of Sintered Bodies Made from the System of Paper Sludge Ash - Fly Ash - Clay (종이재-석탄회-점토계 소지를 이용한 소결체의 특성 연구)

  • Hong, Jin-Ok;Kang, Seung-Gu;Lee, Ki-Gang;Kim, Yoo-Taek;Kim, Young-Jin;Kim, Jung-Hwan;Park, Myoung-Sik
    • Journal of the Korean Ceramic Society
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    • v.38 no.10
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    • pp.908-913
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    • 2001
  • Paper sludge Ash (PA) and Fly Ash (FA) wastes are usually land-filled for reclamation or substituted for cements as a resource. It could also offer some advantages when they are substituted for clay to preserve the environment. To recycle those wastes, the sintered specimen made of PA-FA-Clay system were examined to find the microstructure and physical properties. The ratio of clay to wastes was fixed as 30:70 by wt%, while PA to FA within waste portion were varied in the range of $1:6{\sim}7:0$. Those specimens were fired in $1150{\sim}1350^{\circ}C$. It was found that the relative density of sintered specimen was increased with amount of PA added at low sintering temperature (i.e, $1150{\sim}1200^{\circ}C$). This is due to increased amount of liquid during sintering. It is shown, however that at high sintering temperature ($1250{\sim}1350^{\circ}C$), the relative density of specimens was decreased with amount of PA added. This is because of overfiring phenomenon which may be able to induce an inhomogeneous microstructure and increased porosity. The mechanical properties of sintered specimen were depended upon the homogeneity of microstructure in accordance with SEM (Scanning Electron Microscopy) and pore size distribution analysis. For example, the compressive strength of 10PA-60FA-30Clay specimen sintered at $1225^{\circ}C$ was twice higher than that of 70PA-30Clay specimen even thought the relative density of those specimen was similar. This decreased strength of 70PA-30Clay specimen appears to be an inhomogeneity of microstructure due to overfiring.

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Evaluation of In-Vitro Efficacy of Active Ingredients in Dentifrice Used for Different Treatment Times (치약용 약효제의 적용시간에 따른 실험실적 효능 연구)

  • Ahn, Jae-Hyun;Kim, Ji-Hye;Kim, Ji-Young
    • Journal of dental hygiene science
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    • v.16 no.2
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    • pp.176-182
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    • 2016
  • The purpose of this study was to evaluate the in-vitro efficacy of the active ingredients of dentifrice following treatment time. The whitening effect was evaluated by a change in lightness value relative to the contact time of hydrogen peroxide, by using artificially stained hydroxyapatite discs. The anti-calculus effect was assessed based on the amount of calcium eluted from the human dental calculus by sodium pyrophosphate. Remineralization was evaluated by the Vickers hardness test following the application of sodium fluoride to bovine enamel. In order to view dentinal tubules occlusion, the formation of insoluble calcium salts by bovine dentin specimens was observed using scanning electron microscopy. Change in lightness value (${\Delta}L$) was $5.50{\pm}1.51$ after 1 min of treatment, $5.73{\pm}0.43$ after 3 min, $8.64{\pm}0.24$ after 10 min, $18.93{\pm}0.76$ after 30 min, and $27.35{\pm}0.54$ after 60 min. The amount of calcium eluted from the human dental calculus was $4.23{\pm}0.14ppm$ after 1 min of treatment, $4.51{\pm}0.04ppm$ after 3 min, $12.12{\pm}0.16ppm$ after 10 min, $17.85{\pm}0.81ppm$ after 30 min, and $25.15{\pm}0.32ppm$ after 60 min. The Vickers hardness change value (${\Delta}VHN$) was $1.96{\pm}1.44$ after 1 min, $1.52{\pm}1.06$ after 3 min, $9.06{\pm}0.15$ after 10 min, $10.83{\pm}5.13$ after 30 min, and $12.55{\pm}2.09$ after 60 min. Partial dentinal tubules occlusion was observed at 10 min and complete occlusion was evident at 60 min. In summary, the use of patch type dentifrices for 10, 30, or 60 min were 1.57 to 8.26 times more effective than using the paste type dentifrices for 1 to 3 min. Based on these findings, it is reasonable to expect that the use of patch type dentifrices for 10 min would lead to remineralization, anti-calculus and dentinal tubules occlusion effects, and that use for 30 min would result in a whitening effect.