• Title/Summary/Keyword: laser scanning confocal microscopy

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Effect of Potassium Iodide on Erythrosine-Mediated Photodynamic Therapy on Streptococcus Mutans Biofilms (Streptococcus mutans 바이오필름에 대한 에리스로신 매개 광역동 치료 시 potassium iodide의 효과)

  • Yongsoon, Kim;Howon, Park;Juhyun, Lee;Haeni, Kim;Siyoung, Lee
    • Journal of the korean academy of Pediatric Dentistry
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    • v.49 no.3
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    • pp.321-328
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    • 2022
  • The aim of this in-vitro study is to evaluate the effect of potassium iodide (KI) on erythrosine-mediated photodynamic therapy (PDT) against Streptococcus mutans biofilms. S. mutans ATCC 25175 was cultured to form a biofilm on a hydroxyapatite disk. After diluting erythrosine to 20 μM and KI to 10, 50, and 100 mM, respectively, PDT was performed. The number of surviving bacteria was calculated as colony forming units (CFU)/mL and the statistical significance of the difference between groups was confirmed by Bonferroni post-hoc analysis. Cell viability was visually evaluated using confocal laser scanning microscopy (CLSM). As a result of the experiment, a significant decrease (p < 0.05) in CFU was observed in the experimental groups in which PDT was performed after applying KI regardless of the concentration of KI. In addition, a significant reduction (p < 0.05) in CFU was observed in the experimental group to which 100 mM KI was applied compared to 10 mM KI. The same results were confirmed when observing CLSM. KI significantly improved the efficacy of erythrosine-mediated PDT on S. mutans biofilms at all concentrations. This may compensate for the low sensitivity of PDT to biofilm-state bacteria strains, but it is necessary to establish an optimal clinical protocol through further research.

Complete Genome and Calcium Carbonate Precipitation of Alkaliphilic Bacillus sp. AK13 for Self-Healing Concrete

  • Jung, Yoonhee;Kim, Wonjae;Kim, Wook;Park, Woojun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.3
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    • pp.404-416
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    • 2020
  • Bacteria that are resistant to high temperatures and alkaline environments are essential for the biological repair of damaged concrete. Alkaliphilic and halotolerant Bacillus sp. AK13 was isolated from the rhizosphere of Miscanthus sacchariflorus. Unlike other tested Bacillus species, the AK13 strain grows at pH 13 and withstands 11% (w/v) NaCl. Growth of the AK13 strain at elevated pH without urea promoted calcium carbonate (CaCO3) formation. Irregular vaterite-like CaCO3 minerals that were tightly attached to cells were observed using field-emission scanning electron microscopy. Energy-dispersive X-ray spectrometry, confocal laser scanning microscopy, and X-ray diffraction analyses confirmed the presence of CaCO3 around the cell. Isotope ration mass spectrometry analysis confirmed that the majority of CO32- ions in the CaCO3 were produced by cellular respiration rather than being derived from atmospheric carbon dioxide. The minerals produced from calcium acetate-added growth medium formed smaller crystals than those formed in calcium lactate-added medium. Strain AK13 appears to heal cracks on mortar specimens when applied as a pelletized spore powder. Alkaliphilic Bacillus sp. AK13 is a promising candidate for self-healing agents in concrete.

Initial adhesion of bone marrow stromal cells to various bone graft substitutes

  • Jo, Young-Jae;Kim, Kyoung-Hwa;Koo, Ki-Tae;Kim, Tae-Il;Seol, Yang-Jo;Lee, Yong-Moo;Ku, Young;Chung, Chong-Pyoung;Rhyu, In-Chul
    • Journal of Periodontal and Implant Science
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    • v.41 no.2
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    • pp.67-72
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    • 2011
  • Purpose: The aim of this study is to determine whether certain biomaterials have the potential to support cell attachment. After seeding bone marrow stromal cells onto the biomaterials, we investigated their responses to each material in vitro. Methods: Rat bone marrow derived stromal cells were used. The biomaterials were deproteinized bovine bone mineral (DBBM), DBBM coated with fibronectin (FN), synthetic hydroxyapatite (HA), HA coated with FN, HA coated with $\beta$-tricalcium phosphate (TCP), and pure $\beta$-TCP. With confocal laser scanning microscopy, actin filaments and vinculin were observed after 6, 12, and 24 hours of cell seeding. The morphological features of cells on each biomaterial were observed using scanning electron microscopy at day 1 and 7. Results: The cells on HA/FN and HA spread widely and showed better defined actin cytoskeletons than those on the other biomaterials. At the initial phase, FN seemed to have a favorable effect on cell adhesion. In DBBM, very few cells adhered to the surface. Conclusions: Within the limitations of this study, we can conclude that in contrast with DBBM not supporting cell attachment, HA provided a more favorable environment with respect to cell attachment.

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.

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.

COMPARATIVE STUDY ON THE EARLY DETECTION OF ENAMEL LESIONS USING DIFOTI AND LASER FLUORESCENCE (Digital Imaging Fiber-Optic Trans-Illumination과 Laser Fluorescence를 이용한 법랑질 우식증의 조기 진단에 관한 비교 연구)

  • Maeng, Myoung-Ho;Kim, Seung-Oh;Kim, Jong-Soo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.2
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    • pp.207-220
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    • 2006
  • The newly developed equipments for the early detection of carious lesion are LFD (laser fluorescence device), Ultrasonic diagnostic system, CLSM(confocal laser scanning microscopy), QLF(quantitative light-induced fluorescence) and DIFOTI (digital imaging fiber-optic trans-illumination) system. In this study, DIFOTI system and LFD were used for the detection of early enamel caries. Twenty five primary teeth extracted from twenty one children at around the dentitional exchanging period were selected as samples. The results obtained from DIFOTI imaging and LFD measurement were compared with those of CLSM and comprehensive evaluations were made for the diagnostic capacity of each device. In vitro test, 40 sample teeth with their buccal & lingual surface formed by a window of $2{\times}3mm$ in diameter were immersed in artificial demineralizing solution for the period of 4, 8, 12 and 16 days. The results obtained from the experimental groups (DIFOTI, LFD) were compared to control group (CLSM) and we have reached to the following conclusions. 1. The sensitivity and specificity of DIFOTI system operated in oral environment was 88.2% and 76.9% respectively. 2. The sensitivity and specificity of LFD measured in oral environment was 76.5% and 69.2% respectively. 3, Regression analysis on the light transparent rate of DIFOTI showed its decrease according to the length of primary enamel decalcification performed in vitro(r=-0.96, p<0.05). 4. No statistically significant difference between LFT measurement and the length of in vitro decalcification was found in regression analysis (p>0.05). 5. The correlation coefficient of DIFOTI image transparent rate and the lesion depth of CLMS was -0.6988 (p<0.05), whereas no statistically significant difference was found for LFD measurement.

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Effect of Probiotic Clostridium butyricum NCTC 7423 Supernatant on Biofilm Formation and Gene Expression of Bacteroides fragilis

  • Shi, Da-Seul;Rhee, Ki-Jong;Eom, Yong-Bin
    • Journal of Microbiology and Biotechnology
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    • v.30 no.3
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    • pp.368-377
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    • 2020
  • Enterotoxigenic Bacteroides fragilis (ETBF) is the main pathogen causing severe inflammatory diseases and colorectal cancer. Its biofilm plays a key role in the development of colorectal cancer. The objective of this study was to determine the antagonistic effects of cell-free supernatants (CFS) derived from Clostridium butyricum against the growth and biofilm of ETBF. Our data showed that C. butyricum CFS inhibited the growth of B. fragilis in planktonic culture. In addition, C. butyricum CFS exhibited an antibiofilm effect by inhibiting biofilm development, disassembling preformed biofilms and reducing the metabolic activity of cells in biofilms. Using confocal laser scanning microscopy, we found that C. butyricum CFS significantly suppressed the proteins and extracellular nucleic acids among the basic biofilm components. Furthermore, C. butyricum CFS significantly downregulated the expression of virulence- and efflux pump-related genes including ompA and bmeB3 in B. fragilis. Our findings suggest that C. butyricum can be used as biotherapeutic agent by inhibiting the growth and biofilm of ETBF.

Community structure analysis of nitrifying biofilms by 16S rRNA targeted probe and fluorescence in situ hybridization (FISH)

  • Han, Dong-U;Kim, Dong-Jin
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.282-285
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    • 2001
  • The microbial community structure and in situ spatial distribution of ammonia oxidizing and nitrite oxidizing bacteria in nitrifying biofilm of an upflow biological aerated filter system were investigated. The reactor had been continuously operated under high free ammonia concentration and low DO concentration for nitrite accumulation more than 2 years before the experiment. Fluorescence in situ hybridization

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Visualization of periodontopathic bacteria within crevicular epithelial cells with fluorescence in situ hybridization (형광제자리부합법을 이용한 치은열구세포 내의 치주염 유발 세균의 관찰)

  • Ko, Young-Kyung
    • Journal of Periodontal and Implant Science
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    • v.38 no.4
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    • pp.691-698
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    • 2008
  • Purpose: Periodontal pathogens can invade the host tissue. Morphologic studies have revealed bacteria within the pocket epithelium, gingival connective tissues, alveolar bone, and oral epithelium. The objective of this study was to visualize and evaluate presence of Porphyromonas gingivalis and Tannerella forsythia in crevicular epithelial cells of periodontally healthy subjects and chronic periodontitis patients. Materials and Methods: A total of 666 crevicular epithelial cells in the samples obtained from 27 chronic periodontitis patients and 9 healthy volunteers were examined. Specific probes for P. gingivalis and T. forsythia and a universal probe for detection of all eubacteria targeting 168 rRNA for fluorescence in situ hybridization was used in conjunction with confocal laser scanning microscopy. Results: 98.99% of sulcular epithelial cells from healthy volunteers and 84.40% of pocket epithelial cells from periodontitis patients were found to harbor bacteria. P. gingivalis and T. forsythia were discovered more often in crevicular epithelial cells from periodontitis patients. Conclusion: P. gingivalis and T. forsythia can invade crevicular epithelial cells and intracellular bacteria may act as a source of bacteria for persistent infection.