• Title/Summary/Keyword: laser scanning confocal microscopy

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The effects of a sealant resin on enamel demineralization in orthodontic bracket bonding (브라켓 부착 시 실런트 레진이 법랑질 탈회에 미치는 영향)

  • Youn, Young-An;Cho, Jin-Hyoung;Lee, Ki-Heon
    • The korean journal of orthodontics
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    • v.35 no.1 s.108
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    • pp.43-50
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    • 2005
  • The purpose of this study was to evaluate the effects of a sealant resin on enamel demineralization In orthodontic bracket bonding. The forty eight extracted sound bovine teeth were subdivided into four groups and treated with Phase II (Reliance, itasca. III) on the surface Group 1 was not treated. Group 2 was acid etched with 37% phosphoric acid for 30 seconds. Group 3 was applied with sealant after acid etching. Group 4 was applied with resin paste after acid etching and sealant application. Each group was demineralized in artificial caries solution. Demineralized enamel depth was measured by confocal laser scanning microscopy. The results were as follows: the mean demineralized enamel depth was $47.4{\mu}m$. (Group 1), $61.8{\mu}m$ (Group 2), $13.9{\mu}m$ (Group 3). $8.2{\mu}m$ (Group 4) the demineralized enamel depth was increased in Group 2 than in Group 1 (p<0.05); the demineralized enamel depth was reduced in Group 3 than in Group 1 and Group 2 with statistically significant differences (p<0.05): and demineralization in Group 4 was very little. The results of the present study indicate that sealant application is useful for reducing enamel demineralization in orthodontic bracket bonding.

In Vitro Development of Porcine Oocytes Following Intracytoplasmic Injection of Round Spermatid (원형정자 미세주입에 의한 돼지 난자의 체외 배 발달)

  • Shin J. S.;Kim, C.;S. J. Uhm;Kim, N. H.;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.395-403
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    • 1998
  • In this study we determined fertilization processes and developmental ability of porcine oocytes following injection of round spermatid in the presence and absence of artificial activation. Electrical stimulation at 2 h before spermatid injection significantly increased the incidence of normal fertilization as compared to those following injection without stimulation or with stimulation immediately after injection. The incidences of two pronuclear formation and apposition were not different in oocytes between following intracytoplasmic spermatid and spermatid nucleus injection. Indirect immunocyto chemistry and laser scanning confocal microscopy study revealed that microtubuels were organized from the oocyte cortex following round spermatid injection, and this seemed to move both male and female pronuclei into the oocyte center. At 6 days following injection blastocoele formation was seen in the eggs following round spermatid (25%) and round spermatid nucleus injection (27%). However, none of oocytes developed to the blastocyst stage at 6 days following sham injection. The average cell numbers of blastocysts at 8 days following injection of spermatid and spermatid nucleus were 87 to 99. These results suggested that either round spermatid or it's nudeus can be used to produce viable embryos by injection into unfertilized oocytes in the pig.

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Preparation and Evaluation of a 4-Branched Polyethylene Glycol Derivative Modified with Exendin-4 and Stearylamine for Extended Hypoglycemic Action

  • Kim, In-Soo;Ma, Kyung-Wan;Bae, Sung-Ho;Yoon, Jeong-Hyun;Oh, Kyung-Taek;Lee, Eun-Seong;Lee, Don-Haeng;Lee, Kang-Choon;Youn, Yu-Seok
    • Journal of Pharmaceutical Investigation
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    • v.40 no.3
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    • pp.175-180
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    • 2010
  • Albumin-modification has been viewed as one of the most effective ways of extending the short in vivo lifetimes of peptide drugs by delaying glomerular filtration. In this study, we describe a new type 2 anti-diabetic exendin-4 (Ex4) peptide derivative with significant binding ability to human serum albumin (HSA). This exendin-4 derivative consists of a 4-branched polyethylene glycol $(PEG)_{5k}$ (Mw: 20 kDa) modified with three stearylamines ($C_{18}-NH_2$) and one exendin-4 on its branches. PEG and stearylamine were selected to provide functionality to increase molecular size and bind to albumin, respectively. This derivative ($3C_{18}-4PEG_{5k}$-Ex4) was shown to have larger molecular size (Ca. 152 kDa) than actual (25.0 kDa) when subjected to size-exclusion chromatography, and the fluorescein-tagged $3C_{18}-4PEG_{5k}$-Ex4 displayed significant binding to the HSA-immobilized Sepharose CL-4B resin using confocal laser scanning microscopy. Furthermore, $3C_{18}-4PEG_{5k}$-Ex4 was found to have acceptable anti-hyperglycemic efficacy via three consecutive oral glucose tolerance testings (OGTT) in fasted type 2 diabetic db/db mice. The $HD_{total}$ value ($57.6{\pm}12.3%$) of $3C_{18}-4PEG_{5k}$-Ex4 at a 50 nmol/kg dose was 2-fold greater than that ($31.0{\pm}8.7%$) of native exendin-4 in non-fasted db/db mice. Especially, the blood glucose levels in the mice group treated with $3C_{18}-4PEG_{5k}$-Ex4 did not rebound to ~150 mg/dL until 24 h after the injection, which obviously shows the extended hypoglycemia. We believe that this derivative has great pharmaceutical potential as a novel long-acting type 2 anti-diabetic injection treatment.

Development of Probiotic Microcapsules for the Preservation of Cell Viability (생균활성 보존을 위한 유산균 미세캡슐 개발)

  • Lee, Kang-Whi;Jang, Keum-Il;Lee, Yoon-Bok;Sohn, Heon-Soo;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.39 no.1
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    • pp.66-70
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    • 2007
  • Lactobacillus fermentum YL-3 was encapsulated to increase acid tolerance and its total viability. After micro-encapsulation of L. fermentum YL-3 cells with sodium alginate and soybean oil, the morphology of the microcapsule was observed using confocal laser scanning microscopy (CLSM) after staining with pyronin Y and fluorescein isothiocyanate. The sizes of the microcapsules were 120-126 ${\mu}m$, 444-486 ${\mu}m$ and 401-463 ${\mu}m$ when manufactured at pH 2, 3 and 4, respectively. The microcapsule could hold live cells of L. fermentum YL-3 up to $1.2{\times}10^{7}$, $8.1{\times}10^{7}$ and $1.1{\times}10^{8}$ CFU/mL at pH 2, 3 and 4, respectively. The acid tolerance and preservative ability of L. fermentum YL-3 in microcapsule and macrocapsule at $4^{\circ}C$ and $25^{\circ}C$ were tested. L. fermentum YL-3 cells were evenly located in the alginate capsule matrix structure and the firmness of microcapsule was highest at pH 2. Micro-encapsulation showed the most effective acid tolerance at pH 2.0 and preservation of viability at $4^{\circ}C$. However, at $25^{\circ}C$, the macrocapsules showed more effective cell protection than the microcapsules. The application range for microcapsules could be wider than for macrocapsules in the food industry.

3D Quantitative Analysis of Cell Nuclei Based on Digital Image Cytometry (디지털 영상 세포 측정법에 기반한 세포핵의 3차원 정량적 분석)

  • Kim, Tae-Yun;Choi, Hyun-Ju;Choi, Heung-Kook
    • Journal of Korea Multimedia Society
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    • v.10 no.7
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    • pp.846-855
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    • 2007
  • Significant feature extraction in cancer cell image analysis is an important process for grading cell carcinoma. In this study, we propose a method for 3D quantitative analysis of cell nuclei based upon digital image cytometry. First, we acquired volumetric renal cell carcinoma data for each grade using confocal laser scanning microscopy and segmented cell nuclei employing color features based upon a supervised teaming scheme. For 3D visualization, we used a contour-based method for surface rendering and a 3D texture mapping method for volume rendering. We then defined and extracted the 3D morphological features of cell nuclei. To evaluate what quantitative features of 3D analysis could contribute to diagnostic information, we analyzed the statistical significance of the extracted 3D features in each grade using an analysis of variance (ANOVA). Finally, we compared the 2D with the 3D features of cell nuclei and analyzed the correlations between them. We found statistically significant correlations between nuclear grade and 3D morphological features. The proposed method has potential for use as fundamental research in developing a new nuclear grading system for accurate diagnosis and prediction of prognosis.

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Response of osteoblast-like cells cultured on zirconia to bone morphogenetic protein-2

  • Han, Seung-Hee;Kim, Kyoung-Hwa;Han, Jung-Seok;Koo, Ki-Tae;Kim, Tae-Il;Seol, Yang-Jo;Lee, Yong-Moo;Ku, Young;Rhyu, In-Chul
    • Journal of Periodontal and Implant Science
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    • v.41 no.5
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    • pp.227-233
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    • 2011
  • Purpose: The aim of this study was to compare osteoblast behavior on zirconia and titanium under conditions cultured with bone morphogenetic protein-2. Methods: MC3T3-E1 cells were cultured on sandblasted zirconia and sandblasted/etched titanium discs. At 24 hours after seeding MC3T3-E1, the demineralized bone matrix (DBM) gel alone and the DBM gel with bone morphogenetic protein-2 (BMP-2) were added to the culture medium. The surface topography was examined by confocal laser scanning microscopy. Cellular proliferation was measured at 1, 4, and 7 days after gel loading. Alkaline phosphatase activity was measured at 7 days after gel loading. The mRNA expression of ALPase, bone sialoprotein, type I collagen, runt-related transcription factor 2 (Runx-2), osteocalcin, and osterix were evaluated by real-time polymerase chain reaction at 4 days and 7 days. Results: At 1, 4, and 7 days after loading the DBM gel alone and the DBM gel with BMP-2, cellular proliferation on the zirconia and titanium discs was similar and that of the groups cultured with the DBM gel alone and the DBM gel with BMP-2 was not significantly different, except for titanium with BMP-2 gel. ALPase activity was higher in the cells cultured with BMP-2 than in the other groups, but there was no difference between the zirconia and titanium. In ALPase, bone sialoprotein, osteocalcin, Runx-2 and osterix gene expression, that of cells on zirconia or titanium with BMP-2 gel was much more highly increased than titanium without gel at day 7. The gene expression level of cells cultured on zirconia with BMP-2 was higher than that on titanium with BMP-2 at day 7. Conclusions: The data in this study demonstrate that the osteoblastic cell attachment and proliferation of zirconia were comparable to those of titanium. With the stimulation of BMP-2, zirconia has a more pronounced effect on the proliferation and differentiation of the osteoblastic cells compared with titanium.

Application of Teeth Whitening LED for Prevention of Dental Caries : Antimicrobial Photodynamic Therapy Approach (치아우식 예방을 위한 치아미백기의 활용 : 광역동 치료로서의 접근)

  • Park, Choa;Park, Howon;Lee, Juhyun;Seo, Hyunwoo;Lee, Siyoung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.47 no.1
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    • pp.70-77
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    • 2020
  • The present study is aimed to assess the effect of antimicrobial photodynamic therapy (aPDT) on Streptococcus mutans biofilm through teeth whitening light emitting diode (LED). Planktonic and dynamic biofilm state cultures of S. mutans were used. Erythrosine 20 μM/L was used as the photosensitizer. Irradiation was performed by exposing cultures to clinic and homecare whitening LEDs for 15 minutes. The viability was measured through Colony Forming Unit counts and confocal laser scanning microscopy. aPDT using whitening LEDs and erythrosine significantly decreased the CFU count of S. mutans compared to that in the control group. Dynamic biofilm group showed more resistant features to aPDT compared with planktonic state. Clinic and homecare whitening LED device showed similar antimicrobial effect. The whitening LED, which could irradiate the entire oral arch, showed a significant photodynamic effect on cariogenic S. mutans biofilm. aPDT mediated by erythrosine and LEDs used for teeth whitening exhibited promising antimicrobial activity.

Comparison of Cariogenic Characteristics between Fluoride-sensitive and Fluoride-resistant Streptococcus mutans (불소 민감성 Streptococcus mutans와 불소 저항성Streptococcus mutans의 우식원성 특성 비교)

  • Ong, Seung-Hwan;Kim, Jongsoo;Baek, Dong-Heon;Yoo, Seunghoon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.47 no.4
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    • pp.397-405
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    • 2020
  • The aim of this study is to compare cariogenic characteristics of fluoride-sensitive Streptococcus mutans [fluoride-sensitive (FS) S. mutans ] and fluoride-resistant Streptococcus mutans [fluoride-resistant (FR) S. mutans] in the presence of sucrose, and to evaluate its effect on cariogenic biofilm formation. S. mutans ATCC 25175 was continuously cultured in trypticase soy broth (TSB) containing NaF (70 ppm) for 40 days to generate FR S. mutans. FS and FR S. mutans were inoculated in TSB with or without 2% sucrose, and optical density and pH were measured every hour. An oral biofilm was formed using saliva bacteria and analyzed through confocal laser scanning microscopy and CFU count. Finally, the expression of glucosyltransferases genes of both S. mutans was investigated through RT-PCR. FR S. mutans exhibited slower growth and lower acidogenicity in the presence of sucrose compared to FS S. mutans . Both cariogenic and single species biofilm formation was lower in the presence of FR S. mutans, along with reduced number of bacteria. FR S. mutans showed significantly low levels of gtfB, gtfC, and gtfD expression compared to FS S. mutans . On the basis of results, FR S. mutans may be less virulent in the induction of dental caries.

Effect of cultivar and ascorbic acid on in vitro shoot regeneration and development of bombardment-mediated plastid transformation of tomato (Lycopersicon esculentum) (토마토 재분화 효율 향상 및 엽록체 형질전환 조건)

  • Roh, Kyung-Hee;Lee, Ki-Jong;Park, Jong-Sug;Kim, Jong-Bum;Lee, Seung-Bum;Suh, Seok-Cheol
    • Journal of Plant Biotechnology
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    • v.37 no.1
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    • pp.77-83
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    • 2010
  • Eighteen cultivars of tomato were tested for their regeneration response. Lycopersicon esculentum cv. 2001-58 showed a very high frequency of regeneration (93%). We evaluated the effect of two compounds with known antioxidant activity (ascorbic acid and cystein). The use of ascorbic acid ($200\;-\;300\;{\mu}M/L$) has a positive effect on shoot regeneration. To develope a system for plastid transformation in tomato via homologous recombination, we constructed the tomato plastid expression vector (pKRT22-AG) harboring 2.2 kb flanking sequences cloned from intact plastid genome and gfp gene. To investigate the factors affecting the delivery system of the pKRT22-AG into chloroplast using bombardment, We assessed the optimal DNA concentration, gold particle volume and target distance. Expression of the GFP protein was observed within chloroplast on protoplast of cotyledon explant by confocal laser scanning microscopy, which indicates that the protocol developed in this study be useful for the production of plastid transgenic plants in tomato.

Microgrooves on titanium surface affect peri-implant cell adhesion and soft tissue sealing; an in vitro and in vivo study

  • Lee, Hyo-Jung;Lee, Jaden;Lee, Jung-Tae;Hong, Ji-Soo;Lim, Bum-Soon;Park, Hee-Jung;Kim, Young-Kwang;Kim, Tae-Il
    • Journal of Periodontal and Implant Science
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    • v.45 no.3
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    • pp.120-126
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    • 2015
  • Purpose: With the significance of stable adhesion of alveolar bone and peri-implant soft tissue on the surface of titanium for successful dental implantation procedure, the purpose of this study was to apply microgrooves on the titanium surface and investigate their effects on peri-implant cells and tissues. Methods: Three types of commercially pure titanium discs were prepared; machined-surface discs (A), sandblasted, large-grit, acid-etched (SLA)-treated discs (B), SLA and microgroove-formed discs (C). After surface topography of the discs was examined by confocal laser scanning electron microscopy, water contact angle and surface energy were measured. Human gingival fibroblasts (hGFs) and murine osteoblastic cells (MC3T3-E1) were seeded onto the titanium discs for immunofluorescence assay of adhesion proteins. Commercially pure titanium implants with microgrooves on the coronal microthreads design were inserted into the edentulous mandible of beagle dogs. After 2 weeks and 6 weeks of implant insertion, the animal subjects were euthanized to confirm peri-implant tissue healing pattern in histologic specimens. Results: Group C presented the lowest water contact angle ($62.89{\pm}5.66{\theta}$), highest surface energy ($45{\pm}1.2mN/m$), and highest surface roughness ($Ra=22.351{\pm}2.766{\mu}m$). The expression of adhesion molecules of hGFs and MC3T30E1 cells was prominent in group C. Titanium implants with microgrooves on the coronal portion showed firm adhesion to peri-implant soft tissue. Conclusions: Microgrooves on the titanium surface promoted the adhesion of gingival fibroblasts and osteoblastic cells, as well as favorable peri-implant soft tissue sealing.