• Title/Summary/Keyword: Scanning Confocal Microscope

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Analysis of osteogenic potential on 3mol% yttria-stabilized tetragonal zirconia polycrystals and two different niobium oxide containing zirconia ceramics

  • Hein, Aung Thu;Cho, Young-Dan;Jo, Ye-Hyeon;Kim, Dae-Joon;Han, Jung-Suk
    • The Journal of Advanced Prosthodontics
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    • v.10 no.2
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    • pp.147-154
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    • 2018
  • PURPOSE. This study was performed to evaluate the osteogenic potential of 3mol% yttria-stabilized tetragonal zirconia polycrystals (3Y-TZP) and niobium oxide containing Y-TZPs with specific ratios, new (Y,Nb)-TZPs, namely YN4533 and YN4533/Al20 discs. MATERIALS AND METHODS. 3Y-TZP, YN4533 and YN4533/Al20 discs (15 mm diameter and 1 mm thickness) were prepared and their average surface roughness ($R_a$) and surface topography were analyzed using 3-D confocal laser microscope (CLSM) and scanning electron microscope (SEM). Mouse pre-osteoblast MC3T3-E1 cells were seeded onto all zirconia discs and evaluated with regard to cell attachment and morphology by (CLSM), cell proliferation by PicoGreen assay, and cell differentiation by Reverse-Transcription PCR and Quantitative Real-Time PCR, and alkaline phosphatase (Alp) staining. RESULTS. The cellular morphology of MC3T3-E1 pre-osteoblasts was more stretched on a smooth surface than on a rough surface, regardless of the material. Cellular proliferation was higher on smooth surfaces, but there were no significant differences between 3Y-TZP, YN4533, and YN4533/Al20. Osteoblast differentiation patterns on YN4533 and YN4533/Al20 were similar to or slightly higher than seen in 3Y-TZP. Although there were no significant differences in bone marker gene expression (alkaline phosphatase and osteocalcin), Alp staining indicated better osteoblast differentiation on YN4533 and YN4533/Al20 compared to 3Y-TZP. CONCLUSION. Based on these results, niobium oxide containing Y-TZPs have comparable osteogenic potential to 3Y-TZP and are expected to be suitable alternative ceramics dental implant materials to titanium for aesthetically important areas.

Surface and Tribological Characteristics of Air-cooled and Oil-cooled AISI 4140 Steel (냉각공정에 따른 AISI 4140 강의 표면 및 트라이볼로지 특성)

  • Cho, Hak-Rae;Lee, Sang Don;Son, Jung Ho;Chung, Koo-Hyun
    • Tribology and Lubricants
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    • v.32 no.5
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    • pp.160-165
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    • 2016
  • AISI 4140 steel is widely used in various mechanical components owing to its superior mechanical properties. Surface hardening techniques are often used to further improve the properties, particularly for applications with moving components. The aim of this research is to understand the effect of heat treatment process on surface properties and tribological characteristics of AISI 4140 steel. In this work, we prepare two different AISI 4140 steel specimens- one cooled by air and one by oil- and determine surface properties such as surface topography and roughness using a confocal microscope. We also observe the cross-sections of the specimens using a scanning electron microscope to understand the difference in the material structure. In addition, we assess the hardness with respect to the distance from the surface using a micro-Vickers hardness tester. After characterizing the surfaces of the specimens, we investigate the wear characteristics of the specimens under hydrodynamic lubrication. The results show the presence of grooves on the surface of the oil-cooled specimens. It is likely that such grooves are formed during the cooling process using the oil. However, we observe no other significant differences in the surface properties of the specimens. The wear test results show the occurrence of severe wear on the oil-cooled specimens, which may be due to the groove formed on the surface. The results of this work may be helpful to improve surface properties using surface hardening techniques from a tribological perspective.

Experimental Study of Tribological Properties According to Oil Grade (오일 등급에 따른 트라이볼로지 특성의 관한 실험적 고찰)

  • Lee, Jong-Ho;Seo, Kuk-Jin;Hwang, Youn-Hoo;Han, Jae-Ho;Kim, Dae-Eun
    • Tribology and Lubricants
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    • v.37 no.6
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    • pp.246-252
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    • 2021
  • Among the engine components of an internal combustion engine, the valve train is a series of systems that supply intake gas to the combustion chamber and operate intake and exhaust valves that discharge exhaust gas. If excessive wear occurs in the valve train system, the suction and exhaust valves do not open and close on time, which leads to abnormal combustion and exhaust gas. In this study, we conduct experiments and analyses on friction and wear characteristics of the valve train system. Moreover, we experimentally study the correlation between the pinball and pinball cap on engine oil lubrication, friction experiment, wear amount analysis, and surface analysis. Specifically, we experiment using Ball on reciprocating tribo-tester and apply commercial engine oil sold on the market engine oil. We construct the experimental conditions for each new oil and oil. Accordingly, the completed specimen was subjected to a confocal microscope to check the wear volume, observe the surface of the specimen, and confirm the elemental components using a scanning microscope (SEM) and an energy dispersion X-ray spectrometer (EDS). Through this experiment, we analyze the friction and wear characteristics of valve train components according to engine oil grade, and the obtained data serve as an effective engine oil management method.

Development of Hair Keratin Protein to Accelerate Oral Mucosal Regeneration

  • So-Yeon Kim
    • Journal of dental hygiene science
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    • v.23 no.4
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    • pp.369-377
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    • 2023
  • Background: In this study, we investigated the potential use of keratin for oral tissue regeneration. Keratin is well-known for its effectiveness in skin regeneration by promoting keratinization and enhancing the elasticity and activity of fibroblasts. Because of its structural stability, high storability, biocompatibility, and safety in humans, existing research has predominantly focused on its role in skin wound healing. Herein, we propose using keratin proteins as biocompatible materials for dental applications. Methods: To assess the suitability of alpha-keratin protein as a substrate for cell culture, keratin was extracted from human hair via PEGylation. Viabilities of primary human gingival fibroblasts (HGFs) and human oral keratinocytes (HOKs) were assessed. Fluorescence immunostaining and migration assays were conducted using a fluorescence microscope and confocal laser scanning microscope. Wound healing and migration assays were performed using automated software to analyze the experimental readout and gap closure rate. Results: We confirmed the extraction of alpha-keratin and formation of the PEG-g-keratin complex. Treatment of HGFs with keratin protein at a concentration of 5 mg/ml promoted proliferation and maintained cell viability in the test group compared to the control group. HOKs treated with 5 mg/ml keratin exhibited a slight decrease in cell proliferation and activity after 48 hours compared to the untreated group, followed by an increase after 72 hours. Wound healing and migration assays revealed rapid closure of the area covered by HOKs over time following keratin treatment. Additionally, HOKs exhibited changes in cell morphology and increased the expression of the mesenchymal marker vimentin. Conclusion: Our study demonstrated the potential of hair keratin for soft tissue regeneration, with potential future applications in clinical settings for wound healing.

The Surface Characteristic Changes of Hydroxyapatite Coated Ti Disc When Immersed in NaCl Solution (NaCl 수용액에 담근 Hydroxyapatite 코팅된 타이타늄 시편의 표면 변화)

  • Baek, Yeon-Wha;Kim, Myung-Joo;Kwon, Ho-Beom;Lim, Young-Jun
    • Journal of Dental Rehabilitation and Applied Science
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    • v.28 no.4
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    • pp.339-347
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    • 2012
  • In previous studies, methods for enhancing cellular response on the Hydroxyapatite coated implant surface were described. In this study, the changes of surface characteristics such as surface roughness, contact angle, surface energy and surface morphology were observed when Hydroxyapatite coated Ti discs were immersed in NaCl solution for various time. Hydroxyapatite coated Ti discs were immersed in 0.9% NaCl solution for 7, 14 and 21 days at $37^{\circ}C$. The control group comprises dry identical discs not immersed in a solution. (n=3) All discs were dried in air completely and the surface roughness was measured using confocal laser scanning microscopy(CLSM). Static contact angle was recorded by video contact angle analyzer after dropping distilled water on the surface. The surface energy was calculated from contact angles of the three liquids. Surface was observed using a field emission-scanning electron microscope(FE-SEM). As a result, the surface roughness of immersed Hydroxyapatite coated Ti discs increased significantly and the contact angle decreased comparing with control group discs. The surface energy of immersed discs increased except for discs immersed for 14 days.

Effects of puromycin aminonucleoside on the cytoskeletal changes of glomerular epithelial cells (Puromycin aminonucleoside의 사구체 상피세포에 대한 영향)

  • Lee, Jun Ho;Ha, Tae Sun
    • Clinical and Experimental Pediatrics
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    • v.51 no.1
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    • pp.54-61
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    • 2008
  • Purpose : This study was designed to clarify the mechanism of proteinuria in nephrotic syndrome patients by using puromycin aminonucleoside (PAN) nephrosis model. Methods : Following administration of various concentrations of PAN and antioxidants we observed the changes of podocyte cytoskeletons in cultured rat glomerular epithelial cells (GEpC) by method of scanning electron microscope, reactive oxyten species (ROS) analysis, permeability assay, confocal microscope, and Western blot assay. Results : PAN not only induced the ultrastructural changes of GEpC, such as shortening and fusion of microvilli, but also separated the intercellular gaps and linear ZO-1. PAN induced oxidative stresses in time and dose dependent manners and increases of intercellular permeability in anti-oxidants inhibitable manners. High concentration of PAN induced not only actin polymerization and disorganization, but also the conglomerulation and internal dislocation of ${\alpha}-actinin$ protein. The intensities of fluorescences of ZO-1 protein were diminished and internalized by PAN in a dose-dependent manner, which were inhibited by anti anti-oxidants. Conclusion : PAN induced the changes of podocytes cytoskeleton and junctional barriers by way of increasing ROS in GEpC that resulted in increasing their permeability in a antioxidatn-inhibitable manner. Glomerular hyperpermeability induced by PAN mediateing through oxidative stresses is thought to take part in the mechanism of proteinuria in nephrotic syndrome.

Aberrant Microtubule Assembly and Chromatin Configuration of Homan Oocytes Which Failed to Complete Fertilization Following In Vitro Fertilization and Intracytoplasmic Sperm Injection (일반적 수정과 세포질내 정자주입법에 의해 수정에 실패한 인간난자의 미세소관과 염색체의 형태이상)

  • Chung, H. M.;Kim, N. H.;Kim, J. W.;J. M. Lim;Park, C.;J. J. Ko;K. Y. Cha;Kim, J. M.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.24 no.2
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    • pp.143-154
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    • 2000
  • Most eggs initiated the fertilization processes but arrested at specific stages. The stages included failure of the oocyte to exit from the meiotic metaphase-II with or without sperm penetration, failure of appropriate sperm aster formation, inability to form proper male and female pronuclei, failure of suitable pronuclear apposition, and failure to form proper number of either male or female pronuclei. Various images of defective microtubule organization and chromatin configuration during IVF and ICSI procedures were observed. We discussed the data with previous research results during normal fertilization in humans and other mammals. In conclusion, various aberrant patterns in microtubule assembly and chromatin configuration, which were assessed in the present study, could be used as criteria to improve assisted reproductive technology in clinics. However, further cellular and molecular characterization is needed to clarify these aberrant patterns of cytoskeletal assembly.

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Colonization and Population Changes of a Biocontrol Agent, Paenibacillus polymyxa E681, in Seeds and Roots

  • Park, Okhee;Kim, Jinwoo;Ryu, Choong-Min;Park, Chang-Seuk
    • The Plant Pathology Journal
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    • v.20 no.2
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    • pp.97-102
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    • 2004
  • Paenibacillus polymyxa E681, with its plant growth promotion and root colonization ability, has been proven to be a promising biocontrol agent of cucumber and barley. This study investigated the attributes related to the movement of bacteria from the seed to the radicle and to the whole root system. It also illustrated the existing form and population changes of the bacteria on seed and root using the scanning electron microscope and confocal laser scanning microscopy. The bacteria invaded and colonized the inside of the seed coat while the seeds were soaked in bacterial suspension. Almost the same number of bacteria on seed surface invaded the inside of the seed coat right after seed soaking. The population densities of E681 increased greatly inside as well as on the surface of the seed before the radicle emerged. The bacteria attached on the emerging radicle directly affected the initial population of newly emerg-ing root. The colonized cells on the root were arranged linearly toward the elongation of the root axis. In addition to colonizing the root surface, strain E681 was found inside the roots, where cells colonized the inter-cellular space between certain epidermal and cortical cells. When the cucumber seeds were soaked in bacterial suspension and sown in pot, the bacterial populations attached on both the surface and inside of the root were sustained up to harvesting time. This means that E681 successfully colonized the root of cucumber and sustained its population density up to harvesting time through seed treatment.

The remineralization effect of topical fluoride agents using confocal laser scanning microscope on artificial enamel caries aspects of convergence observation (공초점 레이저 주사 현미경을 이용한 우치 인공우식법랑질에서 불소도포 전·후의 재광화 효과에 대한 융합적 관찰)

  • Lee, Min-Sun;Kim, Hyo-Jin
    • Journal of the Korea Convergence Society
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    • v.8 no.6
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    • pp.139-145
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    • 2017
  • The aim of this study was to measure the remineralization effect of APF gel and fluride varnish on artificial enamel caries using CLSM in vitro. The samples were divided into 3 groups: control, 1.23% APF gel, 5% NaF varnish. The specimen surfaces were observed by CLSM and measured average fluorescence of the lesion(AFL). The results were analyzed using one-way ANOVA and Pearson's correlation analysis at a significance level off 0.05(PSWA 18.0, SPSS Inc., USA). There were significant differences between AFL at baseline and 1 day after fluoride application(p<0.05) but there are no significant differences between ${\Delta}$ AFL of all groups (p=0.222). Result of Pearson's correlation analysis, there are no significant correlation between VHN and AFL, but there were significant correlation between AFL at baseline and 1 day after fluoride application(r=0.811, p<0.001). Although AFL decreased after fluoride application, but there was no difference between the groups. In the future, it is necessary to test the oral environment model or in situ experiment supplemented the limitations of this study.

A study on the differentiation of MC3T3-E1 incubated on the layer-built silica/polycaprolactone non-woven fabric produced by electrospinning (전기방사법으로 제조된 실리카/폴리카프로락톤 적층형 부직포에 배양한 골아 세포의 중식, 분화에 관한 연구)

  • AN, Min-Kuk;Kim, Kyoung-Hwa;Kim, Tae-II;Lee, Yong-Moo;Rhee, Sang-Hoon;Ku, Young;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Seol, Yang-Jo
    • Journal of Periodontal and Implant Science
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    • v.37 no.1
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    • pp.115-124
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    • 2007
  • Silica is known as a promising osteoconductive material, and polycaprolactone is a bioactive and degradable material. The purpose of this study was to monitor the differentiation of MC3T3-E1 cells cultured on the layer-built silica/poly caprolactone non-woven fabric produced by electrospinning. Non-woven fabric (silica, polycaprolactone, PSP, SPS) was made by electrospinning and they were inserted in the 48 well cell culture plate. MC3T3-E1 cells were prepared by subculture. Cells were seeded to each well $1{\times}10^5$ concentration per well. Dulbecco's modified eagle medium with 10% FBS and 1% antibiotic-antimycotic solution was used. Confocal laser scanning microscope was taken 4 hours after incubation (95% air. 5% $CO_2$, $37^{\circ}C$). Cell proliferation was monitored by spectrophotometer on 1, 7, 14 days, and the morphology of the growing cells was observed by field emission scanning electron microscope. To monitor the differentiation of osteoblasts on the materials, MC3T3-E1 cells were incubated in 48 well culture plate after seeding with the density of $1{\times}10^5$ concentration. Then ELISA kit & EIA kit were used on to assess osteocalcin and osteopontin expression respectively. The other conditions were the same as above. MC3T3-E1 cells were proliferated well on all of the materials. There were no statistical differences among them. The osteopontin expression of silica, PSP, SPS was significantly higher than other groups on day 3 (p/0,05), but after that time, there were no statistically signigicant differences. The osteocalcin expression was significantly higher in silica and PSP than other groups on day 14. These findings show that PSP was as good as silica on the effect of osteoblast differentiation. The PSP non-woven fabric may have the possibility as bone graft materials.