• Title/Summary/Keyword: confocal microscopy

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In vivo comparison between the effects of chemically modified hydrophilic and anodically oxidized titanium surfaces on initial bone healing

  • Lee, Hyo-Jung;Yang, Il-Hyung;Kim, Seong-Kyun;Yeo, In-Sung;Kwon, Taek-Ka
    • Journal of Periodontal and Implant Science
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    • v.45 no.3
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    • pp.94-100
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    • 2015
  • Purpose: The aim of this study was to investigate the combined effects of physical and chemical surface factors on in vivo bone responses by comparing chemically modified hydrophilic sandblasted, large-grit, acid-etched (modSLA) and anodically oxidized hydrophobic implant surfaces. Methods: Five modSLA implants and five anodized implants were inserted into the tibiae of five New Zealand white rabbits (one implant for each tibia). The characteristics of each surface were determined using field emission scanning electron microscopy, energy dispersive spectroscopy, and confocal laser scanning microscopy before the installation. The experimental animals were sacrificed after 1 week of healing and histologic slides were prepared from the implant-tibial bone blocks removed from the animals. Histomorphometric analyses were performed on the light microscopic images, and bone-to-implant contact (BIC) and bone area (BA) ratios were measured. Nonparametric comparison tests were applied to find any significant differences (P<0.05) between the modSLA and anodized surfaces. Results: The roughness of the anodized surface was $1.22{\pm}0.17{\mu}m$ in Sa, which was within the optimal range of $1.0-2.0{\mu}m$ for a bone response. The modSLA surface was significantly rougher at $2.53{\pm}0.07{\mu}m$ in Sa. However, the modSLA implant had significantly higher BIC than the anodized implant (P=0.02). Furthermore, BA ratios did not significantly differ between the two implants, although the anodized implant had a higher mean value of BA (P>0.05). Conclusions: Within the limitations of this study, the hydrophilicity of the modSLA surface may have a stronger effect on in vivo bone healing than optimal surface roughness and surface chemistry of the anodized surface.

Analysis of the Molecular Event of ICAM-1 Interaction with LFA-1 During Leukocyte Adhesion Using a Reconstituted Mammalian Cell Expression Model

  • Han, Weon-Cheol;Kim, Kwon-Seop;Park, Jae-Seung;Hwang, Sung-Yeoun;Moon, Hyung-Bae;Chung, Hun-Taeg;Jun, Chang-Duk
    • Animal cells and systems
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    • v.5 no.3
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    • pp.253-262
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    • 2001
  • Ligand-receptor clustering event is the most important step in leukocyte adhesion and spreading on endothelial cells. Intercellular adhesion molecule-1 (ICAM-1) has been shown to enhance leukocyte adhesion, but the molecular event during the process of adhesion is unclear. To visualize the dynamics of ICAM-1 movement during adhesion, we have engineered stable Chinese hamster ovary cell lines expressing ICAM-1 fused to a green fluorescent protein (IC1_GFP/CHO) and examined them under the fluorescence microscopy. The transfection of IC1_GFP alone in these cells was sufficient to support the adhesion of K562 cells that express $\alpha$L$\beta$2 (LFA-1) integrin stimulated by CBR LFA-1/2 mAb. This phenomenon was mediated by ICAM-1-LFA-1 interactions, as an mAb that specifically inhibits ICAM-1-LFA-1 interaction (RRl/l) completely abolished the adhesion of LFA-1* cells to IC1_ GFP/CHO cells. We found that the characteristic of adhesion was followed almost immediately (~10 min) by the rapid accumulation of ICAM-1 on CHO cells at a tight interface between the two cells. Interestingly, ICI_GFP/CHO cells projected plasma membrane and encircled approximately half surface of LFA-1+ cells, as defined by confocal microscopy. This unusual phenomenon was also confirmed on HUVEC after adhesion of LFA-1* cells. Neither cytochalasin D nor 2,3-butanedione 2-monoxime an inhibitor of myosin light chain kinase blocked LFA-1-mediated ICAM-1 clustering, indicating that actin cytoskeleton and myosin-dependent contractility are not necessary for ICAM-1 clustering. Taken together, we suggest that leukocyte adhesion to endothelial cells induces specialized form of ICAM-1 clustering that is distinct from immunological synapse mediated by T cell interaction with antigen presenting cells.

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Physicochemical Characterization and Carcinoma Cell Interaction of Self-Organized Nanogels Prepared from Polysaccharide/Biotin Conjugates for Development of Anticancer Drug Carrier

  • Park Keun-Hong;Kang Dong-Min;Na Kun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1369-1376
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    • 2006
  • Self-organized nanogels were prepared from pullulan/biotin conjugates (PU/Bio) for the development of an effective anticancer drug delivery system. The degree of biotin substitution was 11, 19, and 24 biotin groups per 100 anhydroglucose units of pullulan. The physicochemical properties of the nanogels (PU/Bio1, 2 and 3) in aqueous media were characterized by dynamic light scattering, transmission electron microscopy, and fluorescence spectroscopy. The mean diameter of all the samples was less than 300 nm with a unimodal size distribution. The critical aggregation concentrations (CACs) of the nanoparticles in distilled water were $2.8{\times}10^{-2},\;1.6{\times}10^{-2}$, and $0.7{\times}10^{-2}mg/ml$ for the PU/Bio1, 2, and 3, respectively. The aggregation behavior of the nanogels indicated that biotin can perform as a hydrophobic moiety. To observe the specific interaction with a hepatic carcinoma cell line (HepG2), the conjugates were labeled with rhodamine B isothiocyanate (RITC) and their intensities measured using a fluorescence microplate reader. The HepG2 cells treated with the fluorescence-labeled PU/Bio nanoparticles were strongly luminated compared with the control (pullulan). Confocal laser microscopy also confirmed internalization of the PU/Bio nanogels into the cancer cells. Such results demonstrated that the biotin in the conjugate acted as both a hydrophobic moiety for self-assembly and a tumor-targeting moiety for specific interaction with tumor cells. Consequently, PU/Bio nanogels would appear to be a useful drug carrier for the treatment of liver cancer.

Anti-cariogenic Properties of α-Pinene, a Monoterpene in Plant Essential Oil

  • Park, Bog-Im;You, Yong-Ouk;Mo, Ji-Su;An, So-Youn;Choi, Na-Young;Kim, Kang-Ju
    • International Journal of Oral Biology
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    • v.42 no.1
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    • pp.25-31
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    • 2017
  • Dental caries is the most common chronic disease in the dental field. Streptococcus mutans (S. mutans) is the most important bacteria in the formation of dental plaque and dental caries. In a previous study, we confirmed that the essential oil of Chrysanthemum boreale has antibacterial activity against S. mutans. Alpha-pinene is one of the major chemical components of Chrysanthemum boreale essential oil. In the present study, we investigated the inhibitory effects of ${\alpha}-pinene$ on cariogenic properties such as growth, acid production, biofilm formation, and bactericidal activity on S. mutans. Alpha-pinene at a concentration range of 0.25-0.5 mg/mL significantly inhibited the growth of S. mutans and acid production of S. mutans. Biofilm formation was significantly inhibited at > 0.0625 mg/mL ${\alpha}-pinene$, similar to the data from scanning electronic microscopy. Under confocal laser scanning microscopy, the bacterial viability was decreased by ${\alpha}-pinene$ in a dose-dependent manner. These results suggested that ${\alpha}-pinene$ may be a useful agent for inhibiting the cariogenic properties of S. mutans.

Chelating and antibacterial properties of chitosan nanoparticles on dentin

  • del Carpio-Perochena, Aldo;Bramante, Clovis Monteiro;Duarte, Marco Antonio Hungaro;de Moura, Marcia Regina;Aouada, Fauze Ahmad;Kishen, Anil
    • Restorative Dentistry and Endodontics
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    • v.40 no.3
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    • pp.195-201
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    • 2015
  • Objectives: The use of chitosan nanoparticles (CNPs) in endodontics is of interest due to their antibiofilm properties. This study was to investigate the ability of bioactive CNPs to remove the smear layer and inhibit bacterial recolonization on dentin. Materials and Methods: One hundred bovine dentin sections were divided into five groups (n = 20 per group) according to the treatment. The irrigating solutions used were 2.5% sodium hypochlorite (NaOCl) for 20 min, 17% ethylenediaminetetraacetic acid (EDTA) for 3 min and 1.29 mg/mL CNPs for 3 min. The samples were irrigated with either distilled water (control), NaOCl, NaOCl-EDTA, NaOCl-EDTA-CNPs or NaOCl-CNPs. After the treatment, half of the samples (n = 50) were used to assess the chelating effect of the solutions using portable scanning electronic microscopy, while the other half (n = 50) were infected intra-orally to examine the post-treatment bacterial biofilm forming capacity. The biovolume and cellular viability of the biofilms were analysed under confocal laser scanning microscopy. The Kappa test was performed for examiner calibration, and the non-parametric Kruskal-Wallis and Dunn tests (p < 0.05) were used for comparisons among the groups. Results: The smear layer was significantly reduced in all of the groups except the control and NaOCl groups (p < 0.05). The CNPs-treated samples were able to resist biofilm formation significantly better than other treatment groups (p < 0.05). Conclusions: CNPs could be used as a final irrigant during root canal treatment with the dual benefit of removing the smear layer and inhibiting bacterial recolonization on root dentin.

Reduced Autophagy in 5-Fluorouracil Resistant Colon Cancer Cells

  • Yao, Cheng Wen;Kang, Kyoung Ah;Piao, Mei Jing;Ryu, Yea Seong;Fernando, Pattage Madushan Dilhara Jayatissa;Oh, Min Chang;Park, Jeong Eon;Shilnikova, Kristina;Na, Soo-Young;Jeong, Seung Uk;Boo, Sun-Jin;Hyun, Jin Won
    • Biomolecules & Therapeutics
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    • v.25 no.3
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    • pp.315-320
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    • 2017
  • We investigated the role of autophagy in SNUC5/5-FUR, 5-fluorouracil (5-FU) resistant SNUC5 colon cancer cells. SNUC5/5-FUR cells exhibited low level of autophagy, as determined by light microscopy, confocal microscopy, and flow cytometry following acridine orange staining, and the decreased level of GFP-LC3 puncta. In addition, expression of critical autophagic proteins such as Atg5, Beclin-1 and LC3-II and autophagic flux was diminished in SNUC5/5-FUR cells. Whereas production of reactive oxygen species (ROS) was significantly elevated in SNUC5/5-FUR cells, treatment with the ROS inhibitor N-acetyl cysteine further reduced the level of autophagy. Taken together, these results indicate that decreased autophagy is linked to 5-FU resistance in SNUC5 colon cancer cells.

Anodic Oxidation Behavior of AZ31 Magnesium Alloy in Aqueous Electrolyte Containing Various Na2CO3 Concentrations

  • Moon, Sungmo;Kim, Yeajin
    • Journal of the Korean institute of surface engineering
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    • v.49 no.4
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    • pp.331-338
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    • 2016
  • In this work, anodic oxidation behavior of AZ31 Mg alloy was studied as a function of $Na_2CO_3$ concentration in electrolyte by voltage-time curves and observation of surface appearances and morphologies after the anodic treatments, using optical microscopy and confocal scanning laser microscopy (CSLM). The voltage-time curves of AZ31 Mg alloy surface and surface appearances after the anodic treatments showed three different regions with $Na_2CO_3$ concentration : region I, below 0.2 M $Na_2CO_3$ where shiny surface with a number of small size pits; region II, between 0.4 M and 0.6 M $Na_2CO_3$ where dark surface with relatively low number of large size burned or dark spots; region III, more than 0.8 M $Na_2CO_3$ where bright surface with or without large size dark spots were obtained. The anodically treated AZ31 Mg alloy surface became significantly brightened with increasing $Na_2CO_3$ concentration from 0.5 M to 0.8 M which was attribute to the formation of denser and smoother surface films. Pits and porous protruding reaction products were found at relatively large size and small size spots, respectively, on the AZ31 Mg alloy surface in low concentration of $Na_2CO_3$ less than 0.2 M. The formation of pits is attributed to the result of repetition of the formation and detachment of porous anodic reaction products. Based on the experimental results obtained in this work, it is concluded that more uniform, denser and smoother surface of AZ31 Mg alloy could be obtained at more than 0.8 M $Na_2CO_3$ concentration if there is no other oxide forming agent.

Analysis of surface characteristics of (Y, Nb)-TZP after finishing and polishing

  • Seong-keun, Yoo;Ye-Hyeon, Jo;In-Sung Luke, Yeo;Hyung-In, Yoon;Jae-Hyun, Lee;Jin-Soo, Ahn;Jung-Suk, Han
    • The Journal of Advanced Prosthodontics
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    • v.14 no.6
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    • pp.335-345
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    • 2022
  • PURPOSE. This in vitro study aimed to evaluate the surface characteristics of a full veneer crown fabricated chairside (CS) from a (Y, Nb)-TZP zirconia block in response to conventional zirconia grinding and polishing. MATERIALS AND METHODS. Zirconia crowns (n = 40) were first prepared and divided into two groups of materials: Labside (LS) and CS, after which each specimen went through a five-step grinding and polishing procedure. Following each surface treatment, surface characteristics were analyzed using confocal laser microscopy (CLSM), average surface roughness (Ra) values were processed from the profile data through Gaussian filtering, and X-ray diffraction pattern analysis was performed to evaluate the monoclinic (M) phase content. Then, a representative specimen was selected for field-emission scanning electron microscopy (FE-SEM), followed by a final analysis of the roughness and X-ray diffraction of the specimens using the independent t-test and repeated measures analysis of variance (RM-ANOVA). RESULTS. In every group, polishing significantly reduced the Ra values (P < .001). There was no significant difference in Ra between the polished state CS and LS. Furthermore, CLSM and FE-SEM investigations revealed that even though grain exposure was visible in CS specimens throughout the as-delivered and ground states, the exposure was reduced after polishing. Moreover, while no phase transformation was visible in the LS, phase transformation was visible in CS after every surface treatment, with the M phase content of the CS group showing a significant reduction after polishing (P < .001). CONCLUSION. Within the limits of this study, clinically acceptable level of surface finishing of (Y, Nb)-TZP can be achieved after conventional zirconia polishing sequence.

Clinical comparison of marginal fit of ceramic inlays between digital and conventional impressions

  • Franklin Guillermo Vargas-Corral;Americo Ernesto Vargas-Corral;Miguel Angel Rodríguez Valverde;Manuel Bravo;Juan Ignacio Rosales Leal
    • The Journal of Advanced Prosthodontics
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    • v.16 no.1
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    • pp.57-65
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    • 2024
  • PURPOSE. The aim of this stuldy was to compare the clinical marginal fit of CAD-CAM inlays obtained from intraoral digital impression or addition silicone impression techniques. MATERIALS AND METHODS. The study included 31 inlays for prosthodontics purposes of 31 patients: 15 based on intraoral digital impressions (DI group); and 16 based on a conventional impression technique (CI group). Inlays included occlusal and a non-occlusal surface. Inlays were milled in ceramic. The inlay-teeth interface was replicated by placing each inlay in its corresponding uncemented clinical preparation and taking interface impressions with silicone material from occlusal and free surfaces. Interface analysis was made using white light confocal microscopy (WLCM) (scanning area: 694 × 510 ㎛2) from the impression samples. The gap size and the inlay overextension were measured from the microscopy topographies. For analytical purposes (i.e., 95-%-confidence intervals calculations and P-value calculations), the procedure REGRESS in SUDAAN was used to account for clustering (i.e., multiple measurements). For p-value calculation, the log transformation of the dependent variables was used to normalize the distributions. RESULTS. Marginal fit values for occlusal and free surfaces were affected by the type of impression. There were no differences between surfaces (occlusal vs. free). Gap obtained for DI group was 164 ± 84 ㎛ and that for CI group was 209 ± 104 ㎛, and there were statistical differences between them (p = .041). Mean overextension values were 60 ± 59 ㎛ for DI group and 67 ± 73 ㎛ for CI group, and there were no differences between then (p = .553). CONCLUSION. Digital impression achieved inlays with higher clinical marginal fit and performed better than the conventional silicone materials.

The Changes of Occludin in Tight Junction of Blood-Brain Barrier by ROS (치밀이음부 구조단백질인 Occludin에 대한 활성산소종의 영향)

  • Lee, Hee-Sang;Kim, Dae-Jin;Sohn, Dong-Suep;Jeong, Bong-Su;Choi, Hyung-Taek;Sim, Kyu-Min;Lee, Keum-Jeong;Cho, Hye-Jin;Kim, Suk-Joong;Lee, Jong-Chan;Jeong, Yoon-Hee;Kim, Sung-Su;Lee, Won-Bok
    • Applied Microscopy
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    • v.34 no.4
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    • pp.231-239
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    • 2004
  • Cerebral microvessel endothelial cells that form blood-brain barrier (BBB) have tight junction for maintaining brain homeostasis. Occludin, one of tight junction protein, is crucial for BBB function. $H_2O_2$ induced occludin changes and effects in bovine brain BBB endothelial cells were examined in this study. The decrease of transendothelial electrical resistance (TEER) by $H_2O_2$ was due to disruption of occludin localization. Cytotoxicity test revealed that $H_2O_2$ did not cause cell death below 1 mM $H_2O_2$ within 4 hr. $H_2O_2$ caused intermittent disruption and loss of occludin at tight junctions and occludin disappeared with dose dependent manner from tight junction in confocal laser microscopy. But Western blot revealed that the total amounts of occludin increased by $H_2O_2$ administration. Transmission electron microscopy revealed that the ultrastructure of tight junction was not changed by $H_2O_2$. These data suggest that functional disruption of BBB by $H_2O_2$ was due to the localized loss of occludin in tight junction, but the expression of occludin increased in order to compensate the disrupted function in BBB.