• 제목/요약/키워드: confocal microscopy

검색결과 436건 처리시간 0.033초

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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    • 제45권3호
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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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    • 제5권3호
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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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    • 제16권9호
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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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    • 제42권1호
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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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    • 제40권3호
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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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    • 제25권3호
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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
    • 한국표면공학회지
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    • 제49권4호
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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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    • 제14권6호
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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.

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

  • 이희상;김대진;손동섭;정봉수;최형택;심규민;이금정;조혜진;김석중;이종찬;정윤희;김성수;이원복
    • Applied Microscopy
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    • 제34권4호
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    • pp.231-239
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    • 2004
  • 뇌에서 혈액뇌장벽을 형성하는 내피세포는 치밀이음부를 통해 뇌의 항상성을 유지하고 있다. 치밀이음부의 단백질 중의 하나인 occludin은 뇌혈관장벽(BBB)의 기능을 유지하는 중요한 단백질로 인식되고 있다. 본 실험에서는 소의 뇌에서 배양된 BBB 내피세포에서 활성산소종의 하나인 $H_2O_2$에 의해 일어나는 occludin 단백질의 변화를 관찰하였다. $H_2O_2$에 의해 TEER가 감소하는 것은 occludin의 재분포에 의한 것이었다. 세포독성은 4시간내에서는 1mM $H_2O_2$ 이하에서는 나타나지 않았다. Confocal laser microscope으로 관찰한 결과, $H_2O_2$에 의해 occludin은 치밀이음부에서 중간중간이 사라져 감소해 있었고, 이러한 양상은 $H_2O_2$의 용량과 노출시간에 비례하였다. 그러나 Western blot 결과, occludin의 총량은 증가하였다. 투과전자현미경 관찰을 통해 $H_2O_2$가 세포사이의 결합의 구조에 뚜렷한 변화를 미치지 않는 것을 알 수 있었다. 이를 통해 $H_2O_2$에 의한 BBB 기능소실은 occludin이 치밀이음부에서 부분적으로 사라지는 것에 의하지만, 세포는 기능손상을 복구하기위한 방편으로 이 단백질의 생산을 더욱 증가시키는 것으로 생각된다.

Mitochondrial oxidative phosphorylation complexes exist in the sarcolemma of skeletal muscle

  • Lee, Hyun;Kim, Seung-Hyeob;Lee, Jae-Seon;Yang, Yun-Hee;Nam, Jwa-Min;Kim, Bong-Woo;Ko, Young-Gyu
    • BMB Reports
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    • 제49권2호
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    • pp.116-121
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    • 2016
  • Although proteomic analyses have revealed the presence of mitochondrial oxidative phosphorylation (OXPHOS) proteins in the plasma membrane, there have been no in-depth evaluations of the presence or function of OXPHOS I-V in the plasma membrane. Here, we demonstrate the in situ localization of OXPHOS I-V complexes to the sarcolemma of skeletal muscle by immunofluorescence and immunohistochemistry. A portion of the OXPHOS I-V complex proteins was not co-stained with MitoTracker but co-localized with caveolin-3 in the sarcolemma of mouse gastrocnemius. Mitochondrial matrix-facing OXPHOS complex subunits were ectopically expressed in the sarcolemma of the non-permeabilized muscle fibers and C2C12 myotubes. The sarcolemmal localization of cytochrome c was also observed from mouse gastrocnemius muscles and C2C12 myotubes, as determined by confocal and total internal resonance fluorescence (TIRF) microscopy. Based on these data, we conclude that a portion of OXPHOS complexes is localized in the sarcolemma of skeletal muscle and may have non-canonical functions.