• Title/Summary/Keyword: Confocal Laser Scanning Microscope

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Antimicrobial effect of infrared diode laser utilizing indocyanine green against Staphylococcus aureus biofilm on titanium surface (티타늄 표면에 형성한 Staphylococcus aureus 바이오필름에 대한 인도시아닌 그린을 활용한 광역학치료의 항미생물 효과)

  • Seung Gi Kim;Si-Young Lee;Jong-Bin Lee;Heung-Sik Um;Jae-Kwan Lee
    • Journal of Dental Rehabilitation and Applied Science
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    • v.40 no.2
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    • pp.55-63
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    • 2024
  • Purpose: This study aimed to assess the antimicrobial efficacy of an 810-nm infrared diode laser with indocyanine green (ICG) against Staphylococcus aureus on sandblasted, large grit, and acid-etched (SLA) titanium surfaces, comparing its effectiveness with alternative chemical decontamination modalities. Materials and Methods: Biofilms of S. aureus ATCC 25923 were cultured on SLA titanium disks for 48 hours. The biofilms were divided into five treatment groups: control, chlorhexidine gluconate (CHX), tetracycline (TC), ICG, and 810-nm infrared diode laser with ICG (ICG-PDT). After treatment, colony-forming units were quantified to assess surviving bacteria, and viability was confirmed through confocal laser-scanning microscope (CLSM) imaging. Results: All treated groups exhibited a statistically significant reduction in S. aureus (P < 0.05), with notable efficacy in the CHX, TC, and ICG-PDT groups (P < 0.01). While no statistical difference was observed between TC and CHX, the ICG-PDT group demonstrated superior bacterial reduction. CLSM images revealed a higher proportion of dead bacteria stained in red within the ICG-PDT groups. Conclusion: Within the limitations, ICG-PDT effectively reduced S. aureus biofilms on SLA titanium surfaces. Further investigations into alternative decontamination methods and the clinical impact of ICG-PDT on peri-implant diseases are warranted.

Effects of acid-treatment conditions on the surface properties of the RBM treated titanium implants (산-처리 조건이 RBM처리한 티타늄 임플란트의 표면 특성에 주는 영향)

  • Lee, Han-Ah;Seok, Soohwang;Lee, Sang-Hyeok;Lim, Bum-Soon
    • Korean Journal of Dental Materials
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    • v.45 no.4
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    • pp.257-274
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    • 2018
  • The purpose of this study was to evaluate the effect of acid-treatment conditions on the surface properties of the RBM (Resorbable Blast Media) treated titanium. Disk typed cp-titanium specimens were prepared and RBM treatments was performed with calcium phosphate ceramic powder. Acid solution was mixed using HCl, $H_2SO_4$ and deionized water with 4 different volume fraction. The RBM treated titanium was acid treated with different acid solutions at 3 different temperatures and for 3 different periods. After acid-treatments, samples were cleaned with 1 % Solujet solution for 30 min and deionized water for 30 min using ultrasonic cleanser, then dried in the electrical oven ($37^{\circ}C$). Weight of samples before and after acid-treatment were measured using electric balance. Surface roughness was estimated using a confocal laser scanning microscopy, crystal phase in the surface of sample was analyzed using X-ray diffractometer. Surface morphology and components were evaluated using Scanning Electron Microscope (SEM) with Energy Dispersive X-ray spectroscopy (EDX) and X-ray Photoemission Spectroscopy (XPS). Values of the weight changes and surface roughness were statistically analyzed using Tukey-multiple comparison test (p=0.05). Weight change after acid treatments were significantly increased with increasing the concentration of $H_2SO_4$ and temperature of acid-solution. Acid-treatment conditions (concentration of $H_2SO_4$, temperature and time) did not produce consistent effects on the surface roughness, it showed the scattered results. From XRD analysis, formation of titanium hydrides in the titanium surface were observed in all specimens treated with acid-solutions. From XPS analysis, thin titanium oxide layer in the acid-treated specimens could be evaluated. Acid solution with $90^{\circ}C$ showed the strong effect on the titanium surface, it should be treated with caution to avoid the over-etching process.

The Study of anti-inflammatory Mechanism with Cobra Venom on Astrocytes of Rats (뇌(腦) 성상세포(星狀細胞)를 대상으로 한 Cobrotoxin의 염증(炎症) 치료(治療) 기전(機轉) 연구(硏究))

  • Yoo, Jae-ryong;Song, Ho-sueb
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.155-167
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    • 2005
  • Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Cobrotoxin on binding affinity of cobrotoxin with P50, $IKK{\alpa}$ and $IKK{\beta}$, activities of NF-${\kappa}B$, Cell viability of astrocyte, expressions of protein molecules of NF-${\kappa}B$ such as P50, P-$1{kappa}B$, $1{\kappa}B$ and iflammation related genes such as Cox-2, iNOS, cPLA2 in the SNP or LPS induced Inflammatory pathway of Rats' astrocytes. Methods : In this study, The expression of cytosolic phospholipase A2, Nitric oxcide, Cyclooxygenase-2 and inducible nitrogen oxide synthase was determined by western blotting with corresponding antibodies, and the generation of NF-${\kappa}B$ was assayed by EMSA method in astrocytes of rats. The Cell viability of astrocytes was determined by MTT assay, and Binding affinity of Cobrotoxin with P50, $IKK{\alpha}$ and $IKK{\beta}$ was assayed by Surface plasmon resonance analysis, and NF-${\kappa}B$ dependent luciferase activity was determined by luciferase analysis, and Uptake of cobrotoxin in astrocytes was identified by Confocal laser scanning microscope Results : 1. Compared with control, LPS-induced NF-${\kappa}B$ DNA binding activity was decreased significantly by 0.1, $0.5{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 2. Compared with control, LPS-induced NF-kB dependent luciferase expression was decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 3. Compared with control, SNP induced P50, $I{\kappa}B$ expressions in astrocyte were decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin and P-$1{\kappa}B$ expression was decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 4. Compared with control, LPS induced P50, $1{\kappa}B$ expressions in astrocyte were decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 5. Compared with control, SNP induced Cox-2, iNOS, CPLA2 expressions in astrocyte were decreased significantly by $1{\mu}g/m{\ell}$ of Cobrotoxin. 6. Compared with control, LPS induced Cox-2, cPLA2 expressions in astrocyte were decreased significantly by 0.1, 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin and iNOS expression was decreased significantly by 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin. 7. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, SNP-induced NF-${\kappa}B$ DNA bindins activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM. 8. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, LPS-induced NF-${\kappa}B$ DNA binding activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM, Cobrotoxin $0.5{\mu}g/m{\ell}$with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM 9. Compared with $0.1{\mu}g/m{\ell}$ of cobrotoxin, SNP induced P50 expressions in astrocyte were increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM. 10. The uptake of the labeled cobrotoxin into the cells was shown under a confocal laser scanning microscope. cobrotoxin was uptaken into the membrane and nucleus of astrocytes. Conclusions : In summary, the present results demonstrate that cobrotoxin directly binds to sulfhydryl group of p50 and IKKS resulting In the reduction of translocation of p50 and IkB release, thereby inhibits activation of NF-${\kappa}B$, and suggest that pico to nanomolar range of cobrotoxin could inhibit the expression of genes in the NF-${\kappa}B$ signal pathway.

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Cellular Localization and Translocation of Duplication and Alternative Splicing Variants of Olive Flounder Phospholipase C-δ1 (넙치 3가지 타입 인지질가수분해효소(PLC-δ1)의 세포 내 위치 및 이동)

  • Kim, Na Young;Kim, Moo-Sang;Jung, Sung Hee;Kim, Myoung Sug;Cho, Mi Young;Chung, oon Ki;Ahn, Sang Jung
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1369-1375
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    • 2017
  • The purpose of this study was to investigate the cellular characterization of phospholipase C-${\delta}1$ in olive flounders (Paralichthys olivaceus). In general, phospholipase C signaling pathways are distributed in nuclei at plasma membranes and in cytoplasms, although the pathways' nuclear localization mechanisms are unclear. P. olivaceus duplicates type-A PoPLC-${\delta}1$ (PoPLC-${\delta}1A$), which has a high similarity to the human isoform PLC-${\delta}$; type-B PoPLC-${\delta}1$ (PoPLC-${\delta}1B$ [Sf]), which has a low similarity to the human isoform PLC-${\delta}$ and the alternative splice variant PoPLC-${\delta}1B$ (Lf), which has a nuclear localization signal (NLS) and a nuclear export signal (NES) for nuclear imports and exports, respectively. This study confirmed the effects of the cellular localization and translocation of GFP-tagged PoPLC-${\delta}1A$, PoPLC-${\delta}1B$ (Sf) and PoPLC-${\delta}1B$ (Lf). It administered treatments of $Ca^{2+}$ ionophore ionomycin and endoplasmic reticulum (ER)-$Ca^{2+}$ pump inhibitor thapsigargin to hirame natural-embryo (HINAE) cells. A laser-scanning confocal microscope was used. GFP-tagged PoPLC-${\delta}1A$ was distributed to the cellular organelles, rather than to the cytoplasms and cytomembranes, when PoPLC-${\delta}1B$ (Lf) and PoPLC-${\delta}1B$ (Sf) were localized at the plasma membranes. The treatments of ionomycin and thapsigargin showed the accumulation of PoPLC-${\delta}1A$ in the nuclei when PoPLC-${\delta}1B$ (Lf) nucleocytoplasmic shuttling and PoPLC-${\delta}1B$ (Sf) nucleocytoplasmic shuttling were not observed. The results were the first evidence that PoPLC-${\delta}1A$, which contains functional, intact NES sequences, has a main role in nucleocytoplasmic shuttling and translocation in fish.

Components and Biological Activity of Aqueous Extract Isolated from Winged Stem of Euonymus alatus (화살나무 물 추출물의 구성성분과 생리활성)

  • Oh, Bong-Yun;Hwang, Soo-Kyung;Cheong, Mi-Young;Sin, Hong-Sig;Park, Bock-Hee;Lee, Jeong-Ho;Kim, Soo-Hyun
    • Korean Journal of Food Science and Technology
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    • v.37 no.6
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    • pp.898-904
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    • 2005
  • Although Euonymus alatus (EA) has been used as traditional medicine for cancer treatment, exact substances involved in curing of the disease are not yet known. Free radical scavenging and reactive oxygen species (ROS) removal activities of aqueous extract components isolated from winged stem of EA in animal cell line were investigated. Aqueous extract of EA (AEEA) was fractionated by ultrafiltration. All fractions mainly consisted of polysaccharide (44.8%), protein (2.1%), small amounts of phenol compounds and organic acids. Antioxidant activity of AEEA increased depending on concentration fractions, as determined by 1,1-diphenyl-2-picrylhydrazyl method. ROS removal activity was visualized in Chinese hamster ovary cell line using laser scanning confocal microscope, and AEEA activity increased in order of F IV>F III>F I>F II. These results suggest AETA has bioactive carbohydrates with potentials as functional foods and antioxidants.