• Title/Summary/Keyword: Scanning Confocal Microscope

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Effect of Radiation Dosage Changes on the Cell Viability and the Apoptosis Induction on Normal and Tumorigenic Cells (방사선의 선량변화가 수종의 정상세포와 종양세포주의 세포활성도와 apoptosis 유발에 미치는 영향)

  • Park In-Woo;Lee Sam-Sun;Heo Min-Suk;Choi Soon-Chul
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.29 no.2
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    • pp.435-449
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    • 1999
  • Purpose : The study was aimed to detect the differences in the cell viability and the apoptosis induction after irradiation on normal and tumorigenic cells. Materials and Methods : The study. that was generated for two human normal cells(RHEK, HGF-l) and two human tumor cells(KB. HT-1080). was tested using MTT assay at 1 day and 3 day after irradiation and TUNEL assay under confocal laser scanning microscope at 1 day after irradiation. Single irradiation of 0.5. 1, 2. 4. and 8Gy were applied to the cells. The two fractions of 1. 2. 4. and 8Gy were separated with a 4-hour time interval. The irradiation was done with 5.38Gy/min dose rate using Cs-137 irradiator at room temperature. Results and Conclusions : 1. In 3-day group. the cell viability of HGF-1 cell was significantly decreased at 2. 4 and 8Gy irradiation, the cell viability of KB cell was significantly decreased at 8Gy irradiation and the cell viability of HT-I080 cell was significantly decreased at 4 and 8Gy irradiation. 2. There was significant difference between RHEK and KB cell line in the cell viability of 3-day group at 8Gy irradiation. There was significant difference between RHEK and HGF-1 cell line in the cell viability of 3-day group at 4 and 8Gy irradiation. 3. There was a significantly decreased cell viability in 3-day group than those in 1-day group at 2. 4 and 8Gy on HGF-1 cell. at 4 and 8Gy on HT-I080 cell. at 8Gy on KB cell. 4. We could detect DNA fragmented cells only on KB cell. Number of apoptotic cells of KB cell was significantly increased at 4 and 8Gy irradiation. However, there was no correlation between cell viability and apoptosis. 5. On all 4 cell lines, there were no differences between single and split irradiation method in cell viability and apoptosis.

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A Modified Pretreatment with Deproteinization for Resin Infiltration in Early Childhood Caries (유아기우식증 치료를 위한 레진침투법에서 제단백제재의 사용)

  • Nam, Siyeon;Shin, Jonghyun;Jeong, Taesung;Kim, Shin;Kim, Jiyeon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.45 no.3
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    • pp.290-298
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    • 2018
  • This study aimed to evaluate surface morphology and resin tag penetration of resin infiltration into primary anterior teeth after enamel deproteinization with sodium hypochlorite (NaOCl) prior to phosphoric acid ($H_3PO_4$) etching. Ninety primary anterior teeth with non-cavitated caries lesion were devided five groups according to enamel pretreatment as follows, group I-15% hydrochloric acid (HCl) 2min. ; group II-5.25% NaOCl 1min., 35% $H_3PO_4$ 1min. ; group III-5.25% NaOCl 2min., 35% $H_3PO_4$ 1min. ; group IV-5.25% NaOCl 1min., 35% $H_3PO_4$ 2min. ; group V-5.25% NaOCl 2min., 35% $H_3PO_4$ 2min. Fifteen teeth were examined etched surface structure using field emission-scanning electron microscope. Seventy five teeth were infiltrated with resin, maximum penetration depth and percentage penetration were analysed using dual fluorescence confocal microscopy. As the application time of NaOCl increased, ratio of enamel type I, II were increased. Percentage penetration (PP) was higher in group V than group II, III (p < 0.05). PP of group IV, V did not show any differences. Non-cavitated caries of primary anterior teeth can be treated with resin infiltration. Enamel deproteinization with NaOCl prior to 35% $H_3PO_4$ etching could be an alternative of 15% HCl etching in resin infiltration.

Surface roughness and surface free energy components of various orthodontic adhesives (다양한 교정용 접착제의 표면거칠기와 표면에너지 요소 분석)

  • Ahn, Hyo-Beom;Ahn, Sug-Joon;Nahm, Dong-Seok
    • The korean journal of orthodontics
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    • v.36 no.5
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    • pp.360-368
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    • 2006
  • Objective: Surface characteristics of dental materials play an important role in bacterial adhesion. The purpose of this study was to investigate surface characteristics of 5 different light-cured orthodontic adhesives (1 fluoride-releasing composite, 3 non-fluoride-releasing composites, and f resin-modified glass ionomer). Methods: Surface roughness was measured using a confocal laser scanning microscope. Contact angle and surface free energy components were analyzed using the sessile drop method. Results: Surface roughness was significantly different between adhesives despite a relatively small variation (less than $0.05\;{\mu}m$). Lightbond and Monolok2 were rougher than Enlight and Transbond XT. There were also significant differences in contact angles and surface free energy components between adhesives. In particular, considerable differences in contact angles and surface free energy components were found between resin modified glass ionomer and the composites. Resin modified glass ionomer showed significantly smaller contact angles in 3 different probe liquids and had higher total surface free energy and stronger polarity, with notably stronger basic property than the composites. Conclusion: Resin modified glass ionomer may provide a more favourable environment for bacterial adhesion than composite adhesives.

Study of Applicability for Removing Contaminants on Surface of Color Pigment Using the Laser Cleaning Technique -Focus on Analysis Method of Confocal Laser Scanning Microscope- (채색 안료 층의 표면오염물에 대한 레이저클리닝기법의 적용성 평가 연구 -공초점레이저주사현미경(CLSM)을 통한 분석평가 중심으로-)

  • Kim, Jin-Hyoung
    • Journal of Conservation Science
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    • v.30 no.2
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    • pp.123-136
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    • 2014
  • Considering that decision in conservation treatment for damaged objects should consider not only various options of methodology of intervention but also possible consequences of different types of intervention, it is a difficult task to decide and propose clear and safest solution of preserving an object. In addition, it should be constantly challenged by conservators even if it is proved technique or methodology in a past treatment. Therefore, there is no absolute solution which can be applied to all practice but each decision can be different case by case. It is not possible to estimate the way how the present condition of material and environmental aspects would affect to the condition of an object in future. However if conservators keep trying to set out various ways of analysing pro and against effect of past treatments, it would be able to provide useful logics of proving efficiency and appropriateness of a certain treatment. Understanding that the advantage of laser technique is to adopt a way of cleaning an object without making a direct contact, which is different from other techniques, this paper aims at securing stability of laser techniques, although it remains a limitation in the compatibility to all other materials. This study has examined reacting process on the painted pigments against laser beam by using CLSM in order for it to display both the problems from such reacting process and the efficiency of it as a cleaning methodology. It has intended to estimate the result of laser techniques and propose the range of applicability.

The Effect of Silver Diamine Fluoride on Salivary Biofilm (Silver diamine fluoride가 타액 생물막에 미치는 영향)

  • Seo, Meekyung;Song, Ji-Soo;Shin, Teo Jeon;Hyun, Hong-Keun;Kim, Jung-Wook;Jang, Ki-Taeg;Lee, Sang-Hoon;Kim, Young-Jae
    • Journal of the korean academy of Pediatric Dentistry
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    • v.47 no.4
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    • pp.406-415
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    • 2020
  • Silver diamine fluoride (SDF) is an effective and efficient agent for arresting dental caries. It can be useful in treating children with behavioral or medical limitations. The purpose of this study was to evaluate the antimicrobial effect of SDF by using salivary biofilm. Pellicle-like saliva coated structure was prepared by using unstimulated saliva. For developing cariogenic biofilm, Streptococcus mutans was added to the mixture of pooled saliva and inoculated into a saliva coated glass or chamber. SDF was applied to cariogenic biofilm to evaluate the antimicrobial effect of SDF. As time passed, total bacteria and S. mutans were reduced after application of SDF (p < 0.000). Confocal laser scanning microscope also showed the increment of the ratio of dead cell. As a result of experiment using enamel and dentin of primary teeth, it was confirmed that the growth of cariogenic biofilm was inhibited when the SDF was treated (p = 0.029 each). This study showed excellent anti-microbial effect of SDF. And anti-caries effect in clinical practice can be expected.

Crocetin Induces Cytotoxicity in Colon Cancer Cells Via p53-independent Mechanisms

  • Li, Cai-Yan;Huang, Wen-Feng;Wang, Qun-Li;Wang, Fan;Cai, E.;Hu, Bing;Du, Jia-Cheng;Wang, Jing;Chen, Rong;Cai, Xiao-Jing;Feng, Jing;Li, Hui-Hui
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.3757-3761
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    • 2012
  • Objective: Crocin has been proposed as a promising candidate for cancer chemoprevention. The purpose of this investigation was to investigate the chemopreventive action and the possible mechanisms of crocin against human colon cancer cells in vitro. Methods: Cell proliferation was examined using MTT assay and the cell cycle distribution fractions were analyzed using fow cytometric analysis after propidium iodide staining. Apoptosis was detected using theTUNEL Apoptosis Detection Kit with laser scanning confocal microscope. DNA damage was assessed using the alkaline single-cell gel electrophoresis assay, while expression levels of p53, cdk2, cyclinA and P21 were examined by Western blot analysis. Results: Treatment of SW480 cells with crocetin (0.2, 0.4, 0.8 mmol/L) for 48 h signifcantly inhibited their proliferation in a concentration-dependent manner. Crocetin (0.8 mmol/L) signifcantly induced cell cycle arrest through p53-independent mechanisms accompanied by P21 induction. Crocetin (0.8 mmol/L) caused cytotoxicity in the SW480 cells by enhancing apoptosis and decreasing DNA repair capacity in a time-dependent manner. Conclusions: This report provides evidence that crocetin is a potential anticancer agent, which may be used as a chemotherapeutic drug.

Effects of Whitmania pigra on the expressions of cytokine genes in human THP-1 cells (수질(水蛭)이 THP-1세포의 사이토카인 유전자 발현에 미치는 영향)

  • Kim, Hyun-Ji;Kim, Gyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.26 no.1
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    • pp.19-34
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    • 2013
  • Objective : This study was performed to evaluate the effect of immune reaction inductive substances such as PMA, LPS, DPE, DNCB and WP, the whitman pigra extracting substance at simultaneously on the translocation of $NF{\kappa}B$ towards to the nucleus and the mRNA expression patterns of various cytokine genes in THP-1 cells, monocytes of human. Methods : To analyze the cytokine genes expressions, the RT-PCR method was used, and measuring TNF-${\alpha}$ that had been secreted during cell culture by the ELISA method. The morphological changes were observed during THP-1 cell by a scanning electron microscope and the quantitative distribution of $NF{\kappa}B$ in the cell that was analyzed through immunocytochemistry and a confocal microscopy. Results : WP showed different influences onto the mRNA expression patterns of cytokine genes with PMA, LPS. DPE and DNCB according to the types of immune inductive substances in the THP-1 cells. Upon treating PMA and DPE on the THP-1 cells at the same time or either additionally treating WP thereon, the movement of $NF{\kappa}B$ increase towards the nucleus from cell cytoplasm was able to be observed. The expressions of IL-$1{\alpha}$ and IFN-${\gamma}$ induced by PMA and PMA+DNCB were suppressed by WP while the expression of TGF-${\beta}$ was promoted. Regarding the secretion pattern of TNF-${\alpha}$ according to the treatment of PMA, its secretion amount was incredibly increased by concurrent treatment of WP, however, in case of co-treatment of WP with PMA and DNCB, it was found that the secretion amount of TNF-${\alpha}$ decreased. Conclusions : In this study, the WP extracting substance was confirmed that it had an influence on expression patterns of cytokine genes according to the actions of a variety kinds of immune reaction inductive substances treated on the THP-1 cells. Especially, WP co-treatment with PMA and DNCB was suppressed the expression of inflammatory cytokines, such as IL-$1{\alpha}$, IFN-${\gamma}$ and TNF-${\alpha}$.

Effect of Ruthenium Red and Ryanodine on Calcium Ion Metabolism in Oocyte and Early Embryo of Mouse (생쥐의 난자와 초기배아의 칼슘이온 대사에 미치는 Ruthenium Red와 Ryanodine의 영향)

  • Lee Joon Yeong;Hong Soon Cap;Kim Tae Sik;Min Byeong Yeol;Kim Haekwon;Yoon Yong-Dal
    • Development and Reproduction
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    • v.7 no.2
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    • pp.95-103
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    • 2003
  • Intracellular calcium is an important physiological factor in most cells, and ruthenium red and ryanodine play an important role as calcium modulators. Ruthenium red inhibits calcium-induced calcium release(CICR) from the intracellular calcium store. Ryanodine activates calcium release through ryanodine channel. The present experiment was performed to investigate the effects of two modulators on calcium ion metabolism and to determine their dose-dependency in oocyte and early embryo of mouse. Intracellular calcium ion concentration was measured in realtime by using confocal laser scanning microscope(CLSM) after loading of Fluo-3/AM in mouse oocytes and early embryos. Ruthenium red decreased intracellular calcium ion concentration in oocytes and early embryos at its high concentration(30, 300 $\mu$M). Ryanodine increased intracellular calcium ion concentration in oocytes and early embryos in low concentration(0.01 $\mu$M) but decreased that at higher concentrations(1, 10 $\mu$M). These results indicate that two modulators affected calcium ion metabolism in oocyte and early embryo of mouse, and their dose-dependency was different from somatic cell including myocytes.

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Evaluation of Alginate Microspheres Prepared by Emulsion and Spray Method for Oral Vaccine Delivery System (유화법과 분무법에 의해 제조된 경구백신용 알긴산 마이크로스피어의 평가)

  • Jiang, Ge;Jee, Ung-Kil;Maeng, Pil-Jae;Hwang, Sung-Joo
    • Journal of Pharmaceutical Investigation
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    • v.31 no.4
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    • pp.241-256
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    • 2001
  • Alginate microspheres, containing fluorescein isothiocyanate-bovine serum albumin (FITC-BSA) or green fluorescent protein (GFP) were prepared and used as a model drug to develop the oral vaccine delivery system. The alginate microspheres were coated with poly-L-lysine or chitosan. Two methods, w/o-emulsion and spray, were used to prepare alginate microspheres. To optimize preparation conditions, effects of several factors on the particle size and particle morphology of microsphere, and loading efficiency of model antigen were investigated. In both preparation methods, the particle size and the loading efficiency were enhanced when the concentration of sodium alginate increased. In the w/o-emulsion preparation method, as the concentration of Span 80 was increased from 0.5% to 2%, the particle size was decreased, but the loading efficiency was increased. The higher the emulsification speed was, the smaller the particle size and loading efficiency were. The concentration of calcium chloride did not show any effect on the particle size and loading efficiency. In the spray preparation method, the particle size was increased as the nozzle pressure $(from\;1\;kgf/m^2\;to\;3\;kgf/m^2)$ and spray rate was raised. Increasing calcium chloride concentration (<7%) decreased the particle size, in contrast to no effect of calcium chloride concentration on the w/o-emulsion preparation method. Alginate microspheres prepared by two methods were different in the particle size and loading efficiency, the particle size of microspheres prepared by the spray method was about $2-6\;{\mu}m$, larger than that prepared by the w/o emulsion method $(about\;2{\mu}m)$, and the loading efficiency was also higher with spray method. Furthermore, drying process for the microspheres prepared by the spray was simpler and easier, compared with the w/o emulsion preparation. Therefore, the spray method was chosen to prepare alginate microspheres for further experiments. Release pattern of FITC-BSA in alginate microspheres was evaluated in simulated intestinal fluid and PBS (phosphate buffered saline). Dissolution rate of FITC-BSA from alginate/chitosan microsphere was lower than that from alginate microsphere and alginate/poly-L-lysine microsphere. By confocal laser scanning microscope, it was revealed that alginate/FITC-poly-L-lysine microspheres were present in close apposition epithelium of the Peyer's patches of rabbits following inoculation into lumen of intestine, which proved that microspheres could be taken up by Peyer's patch. In conclusion, it is suggested that alginate microsphere prepared by spray method, showing a particle size of & $10\;{\mu}m$ and a high loading efficiency, can be used as a model drug for the development of oral vaccine delivery system.

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Overview of Technology for Fixation of Carbon Dioxide Using Microalgae (미세조류를 이용한 이산화탄소 고정화 기술 현황)

  • Jeon, Seon-Mi;Kim, In Hae;Ha, Jong-Myung;Lee, Jae-Hwa
    • Applied Chemistry for Engineering
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    • v.19 no.2
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    • pp.145-150
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    • 2008
  • In this work we have studied the antifouling properties of the hydrophobic sol-gel modified sensing membrane and its optical properties for sensor application. E. coli JM109, B. cereus 318 and P. pastoris X-33 were cultivated in confocal cultivation dishes with glass surface, respectively. The glass surface was coated with the hydrophobic sol-gels prepared by the dimethoxy-dimethyl-silane (DiMe-DMOS) and tetramethyl-orthosilicate (TMOS). After cultivation, microorganisms adhered on the surface coated with sol-gels and glass surface were dyed by gram-staining method and the numbers of microorganisms were analyzed based on the image data of the scanning electronic microscope (SEM). A great number of microorganisms, about $2{\sim}3{\times}10^4/mm^2$, was adhered on the glass surfaces which no hydrophobic sol-gels were coated. But, the antifouling effect of the hydrophobic sol-gels was large, that microorganisms of less than $200{\sim}300/mm^2$ were adhered on the coated glass surface. The performance of the sensing membranes for detection of pH and dissolved oxygen was enhanced by recoating the light insulation layer prepared with the mixture of the hydrophobic sol-gel and graphite particles.