• Title/Summary/Keyword: Laser Scanning 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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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.

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.

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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The Effects of Size Reduction and pH on Dispersion Characteristics of ATO (Antimony-Doped Tin Oxide) (입자 저감 및 pH가 ATO (안티몬도핑 산화주석)의 분산 특성에 미치는 영향)

  • Kim, Jin-Hoon;Jeong, Eui-Gyung;Lee, Sang-Hun;Han, Won-Hee;Lee, Young-Seak
    • Applied Chemistry for Engineering
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    • v.21 no.3
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    • pp.311-316
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    • 2010
  • This study aimed to prepare antimony-doped tin oxide (ATO) dispersion with high stability. The methods to achieve this goal were sought by investigating the changes of ATO particle size, size distribution, dispersion property as wet ball milling treatment time increased. And the changes of wet ball milled ATO dispersion property were also investigated, as pH increased. The changes of ATO particle size and size distribution, according to wet ball milling treatment time were evaluated with laser diffraction particle size analyzer and scanning electron microscope (SEM). The changes of ATO dispersion property, as wet ball milling treatment time and pH increased, were evaluated with zeta potential analysis and Turbiscan. By 60 min wet ball milling treatment time, ATO particle size decreased and size distribution became narrower, as the treatment time increased. After 60 min milling, the ATO particle size decreased to less than 30% of the initial size and the size distribution was narrowed to $0.1{\sim}5{\mu}m$ from $1{\sim}35{\mu}m$. However, more than 60 min milling, ATO particles aggregated and the particle size increased. ATO dispersion stability also increased as the treatment time and pH increased because the reduced particle size increased particle surface energy and repulsion between particles and the increased pH enhanced particle surface ionization. Hence, after proper length of wet ball milling treatment, highly stable ATO dispersion can be prepared, as increasing pH of the dispersion.

A Study on the Stabilization of the Papain Enzyme in the Moderately Concentrated Anionic Surfactant System (음이온 계면활성제에서 파파인 효소의 안정도에 관한 연구)

  • Kim, Ji-Yeong;Kim, Jin-Woo;Kim, Yong-Jin;Lee, Jae-Wook;Lee, Hae-Kwang;Kang, Hak-Hee
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.2
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    • pp.93-97
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    • 2007
  • Even in the moderately concentrated anionic surfactant system, some special encapsulation method can shield the papain enzyme from proteolytic attacks. The stabilization of enzyme has been a major issue for successful therapies. In this study, we first stabilized an enzyme, papain in the microcapsules by using polyols, polyethyleneglycol (PEG), poly-propyleneglycol (PPG), and PEG-PPG-PEG block copolymer. In the analysis of EDS and CLSM, it was demonstrated that polyols are effectively located in the interface of papain and polymer. Polyols located in the interface had an ability to buffer the external triggers by hydrophobic partitioning, preventing consequently the catalytic activity of papain in the micro-capsules. Second. we introduced multi-layer capsulation methods containing ion complex. Such a moderately concentrated anionic surfactant system as wash-off cleansers, surfactants and waters can cause instability of entrapped enzymes. Surfactants and water in our final products swell the surface of enzyme capsules and penetrate into the core so easily that we can not achieve the effect of enzyme, papain. In this case, the ion complex multi-layer capsule composed of sodium lauroyl sarcosinate and polyquaternium-6 could effectively prevent water from penetration into the core enzyme, followed by in vivo test, and evaluate the stratum corneum (SC) turn-over speed.

The Antitumor Effects of Selenium Compound $Na_5SeV_5O_{18}{\cdot}3H_2O$ in K562 Cell

  • Yang, Jun-Ying;Wang, Zi-Ren
    • Archives of Pharmacal Research
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    • v.29 no.10
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    • pp.859-865
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    • 2006
  • With an approach to study the anti-tumor effects and mechanism of selenium compound, we investigated the anti-tumor activity and mechanism of $Na_5SeV_5O_{18}{\cdot}3H_2O$ (NaSeVO) in K562 cells. The results showed that $0.625{\sim}20\;mg/L$ NaSeVO could significantly inhibit the proliferation of K562 cells in vitro in a time- and concentration-dependent manner as determined by microculture tetrazolium (MTT) assay, the IC50 values were 14.41 (4.45-46.60) and 3.45 (2.29-5.22) mg/L after 48 hand 72 h treatment with NaSeVO respectively. In vivo experiments demonstrated that i.p. administration of 5, 10 mg/kg NaSeVO exhibited an significant inhibitory effect on the growth of transplantation tumor sarcoma 180 (S180) and hepatoma 22 (H22) in mice, with inhibition rate 26.8% and 58.4% on S180 and 31.3% and 47.4% on H22, respectively. Cell cycle studies indicated that the proportion of G0/G1 phase was increased at 2.5 mg/L while decreased at 10 mg/L after treatment for 24, 48 h. Whereas S phase was decreased at 2.5-5 mg/L and markedly increased at 10 mg/L after treatment for 48 h. After treatment for 24 h, 10 mg/L NaSeVO also markedly increased S and G2/M phases. Take together, the result clearly showed that NaSeVO markedly increased S and G2/M phases at 10 mg/L. The study of immunocytochemistry showed that the expression bcl-2 is significantly inhibited by 10 mg/L NaSeVO, and bax increased. Morphology observation also revealed typical apoptotic features. NaSeVO also significantly caused the accumulation of $Ca^{2+}$ and $Mg^{2+}$, reactive oxygen species (ROS) and the reduction of pH value and mitochondrial membrane potential in K562 cells as compared with control by confocal laser scanning microscope. These results suggest that NaSeVO has anti-tumor effects and its mechanism is attributed partially to apoptosis induced by the elevation of intracellular $Ca^{2+}$, $Mg^{2+}$ and ROS concentration, and a reduction of pH value and mitochondria membrane potential (MMP).

THE EFFECT OF VARIOUS SURFACE TREATMENT METHODS ON THE OSSEOINTEGRATION (임플랜트의 표면처리 방법이 골유착에 미치는 영향에 관한 연구)

  • Choi Jeong-Won;Kim Kwang-Nam;Heo Seong-Joo;Chang Ik-Tae;Han Chong-Hyun;Baek Hong-Gu;Choi Yong-Chang;Wennerberg Ann
    • The Journal of Korean Academy of Prosthodontics
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    • v.39 no.1
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    • pp.71-83
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    • 2001
  • The purpose of this study was to compare the effects of various surface treatments by measuring removal torque on bone healing around titanium implants. 40 Screw-shaped cp titanium implants with length of 4mm, outer diameter of 3.75mm, and pitch-height of 0.5mm were used Group 1 was left as machined(control), Group 2 was blasted with $50{\mu}m\;Al_2O_3$, group 3 was blasted and etched in etching solution($NH_4OH : H_2O_2:H_2O= 1 : 1 : 5$) at $90^{\circ}C$ for 1 minute group 4 was blasted and oxidated under pure oxygen at $800^{\circ}C$. The implant surface roughness was analyzed with SEM and CLSM(Confocal Laser Scanning Microscope) and implants were placed in proximal tibial metaphysis of 10 New Zealand White rabbits. After 3 months of healing period, removal torque of each implant was measured to compare bone healing around implant. The results obtained were as follows 1. In SEM view, blasting increased the roughness of the surface, but etching of that rough surface decreased the roughness due to the removal of the tip of the peak. Oxidation also decreased the roughness due to formation of needle-like oxide grains on the implant surface. 2. The Sa value from CLSM was least in the machined group($0.47{\mu}m$), greatest in blasted group($1.25{\mu}m$), and the value decreased after etching($0.91{\mu}m$) and oxidation($0.94{\mu}m$). 3. The removal torque of etched group(24.5Ncm) was greater than that of machined group(16.7Ncm) (P<0.05), and was greatest in the oxidated group(40.3Ncm) and the blasted group(34.7Ncm).

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