• 제목/요약/키워드: Confocal laser scanning microscopy(CLSM)

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Effect of three different irrigation solutions applied by passive ultrasonic irrigation

  • Llena, Carmen;Forner, Leopoldo;Cambralla, Raquel;Lozano, Adrian
    • Restorative Dentistry and Endodontics
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    • 제40권2호
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    • pp.143-148
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    • 2015
  • Objectives: This study evaluated the maximum depth and percentage of irrigant penetration into dentinal tubules by passive ultrasonic irrigation (PUI). Materials and Methods: Thirty extracted human teeth were instrumented and divided into three groups. According to final irrigation regimen, 5.25% sodium hypochlorite (Group A, NaOCl), 2% chlorhexidine (Group B, CHX) and saline solution (Group C, control group) were applied with Irrisafe 20 tips (Acteon) and PUI. Irrigant was mixed with 0.1% rhodamine B. Sections at 2 mm, 5 mm, and 8 mm from the apex were examined with confocal laser scanning microscopy (CLSM). The percentage and maximum depth of irrigant penetration were measured. Kruskal-Wallis test and Mann-Whitney test were performed for overall comparison between groups at each level and for pairwise comparison, respectively. Within a group, Wilcoxon test was performed among different levels. p values less than 0.05 were considered significant. Results: In all groups, highest penetration depth and percentage of penetration were observed at the 8 mm level. At 2 mm level, Groups A and B had significantly greater depths and percentages in penetration than Group C (p < 0.05), but there were no significant differences between Groups A and B. At 5 mm level, penetration depths and percentage of penetration was not significantly different among the groups. Conclusions: NaOCl and CHX applied by PUI showed similar depth and percentage of penetration at all evaluated levels.

글리세릴 베이스의 신규베지클 이용 캡슐화를 통한 항산화성 아스타잔틴의 성질 강화 (The Cosmeceutical Property of Antioxidant Astaxanthin is Enhanced by Encapsulation Using Glyceryl Based New Vesicle)

  • 김동명;홍원기;공수성;이정현
    • 대한화장품학회지
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    • 제40권3호
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    • pp.247-257
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    • 2014
  • 새로운 베지클인 glyceryl citrate/ lactate/ linoleate/ oleate를 이용한 수중유형 형태의 아스타잔틴 나노에멀젼에 대해 항산화 효과, 세포 생존력, 단백질과 관련한 효소의 영향, 피부 침투도 그리고 피부에 대한 보습 및 탄력 등의 약용화장품적인 측면에서의 전반적 연구를 실시하였다. 항산화력 및 세포 생존력에 대해선 각각 DPPH법과 MMT assay를 이용하여 측정하였다. 아스타잔틴 나노에멀젼에 대한 또 다른 성질은 2D-Page를 이용한 단백질 분석 및 컨포칼, in-vivo 테스트를 통해 측정하였다. 본 연구를 통해, 아스타잔틴을 포함하는 나노에멀젼은 MMP발현에 관련한 단백질 억제 및 세포외 기질의 분해를 막고 라디칼의 소거에 매우 우수한 결과를 보였다. 종전의 레시친을 이용한 나노에멀젼 보다는 새로운 베지클을 이용한 아스타잔틴 나노에멀젼의 피부 침투가 매우 효과적임을 CLSM을 통해 측정하였다. 또한 28일 동안의 한국 성인 여성 11명을 통한 보습 및 탄력 인비보 테스트에서 우수한 효과를 확인할 수 있었다.

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

  • 김용순;박호원;이주현;김해니;이시영
    • 대한소아치과학회지
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    • 제49권3호
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    • pp.321-328
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    • 2022
  • 이 연구는 Streptococcus mutans 바이오필름에 대한 에리스로신 매개 광역동 치료에서 potassium iodide (KI)의 효과를 평가하는 In vitro 실험이다. S. mutans ATCC 25175를 배양하여 hydroxyapatite disk에 바이오필름을 형성하였다. 광감각제인 에리스로신을 20 μM, KI를 각각 10, 50, 100 mM로 희석하여 적용한 뒤 광역동 치료를 시행하였다. 생존한 세균의 수는 colony forming units (CFU)/mL로 산정하였으며 Bonferroni 사후 분석을 통해 그룹 간 차이의 유의성을 확인하였다. Confocal laser scanning microscopy (CLSM)을 이용하여 세포 생존율을 시각적으로 평가하였다. 실험 결과 KI를 적용한 후 광역동 치료를 시행한 실험군에서 KI의 농도의 관계 없이 현저한 CFU의 감소가 관찰되었다(p < 0.05). 또한 10 mM KI에 비해 100 mM KI를 적용한 실험군에서 유의한 CFU의 감소가 관찰되었다(p < 0.05). CLSM 관찰 시에도 동일한 결과를 확인하였다. KI는 모든 농도에서 S. mutans 바이오필름에 대한 에리스로신 매개 광역동 치료의 효과를 유의하게 향상시켰다. 이는 바이오필름 상태의 균주에 대한 광역동 치료의 낮은 감수성을 보완할 수 있지만, 추가적인 연구를 통한 최적의 임상 프로토콜 확립이 필요하다.

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

  • 맹명호;김승오;김종수
    • 대한소아치과학회지
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    • 제33권2호
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    • pp.207-220
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    • 2006
  • 새로 개발된 조기 진단 장비들로는 laser fluorescence device(LFD), 초음파 진단 시스템, confocal laser scanning microscopy(CLSM), quantitative light-induced fluorescence(QLF) 시스템 그리고 digital imaging fiber-optic trans-illumination(DIFOTI) 시스템 등이 있다. 본 연구는 임상에서 사용되고 있는 DIFOTI시스템과 LFD를 이용하여 유치 교환 시기에 있는 환아 21명, 25개의 유치를 대상으로 각 치아당 $1{\sim}3$점을 선정하여 구강내에서 초기 우식법랑질에 대한 DIFOTI 이미지 촬영 결과와 LFD 계측값을 3회 측정하고 그 평균을 대표값으로 한 결과를 CLSM과 비교하여 진단 능력을 평가하였다. 실험실 연구에서는 인공우식 용액을 이용하여 수거된 40개의 유치를 협설면에 $2{\times}3mm$ 크기의 창을 형성하고 4, 8, 12, 16일간 탈회시키면서 그 변화를 DIFOTI 시스템과 LFD를 이용하여 측정하고 이를 CLSM과 비교 분석하여 다음과 같은 결론을 얻었다. 1. 구강내에서 촬영된 DIFOTI의 민감도는 88.2%이었고, 특이도는 76.9%이었다. 2. 구강내에서 측정된 LDD의 민감도는 76.5%이었고, 특이도는 69.2%이었다. 3. 실험실 연구에서 유치 법랑질의 탈회 기간에 따른 DIFOTI 광투과율의 회귀 분석한 결과, 탈회 시간에 따라 광투과율은 감소하였다(r=-0.96, p<0.05). 4. 실험실에서 탈회 기간에 따른 LFD의 측정값의 회귀 분석 결과 통계적 유의성을 보이지 않았다(p>0.05). 5. DIFOTI 이미지의 광투과율과 CLMS의 병소 깊이에 대한 상관 계수는 -0.688이었으나(p<0.05), LFD의 측정값은 유의성을 보이지 않았다.

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Morphology and Adhesion of Campylobacter jejuni to Chicken Skin Under Varying Conditions

  • Jang, Keum-Il;Kim, Min-Gon;Ha, Sang-Do;Kim, Keun-Sung;Lee, Kyu-Ho;Chung, Duck-Hwa;Kim, Cheorl-Ho;Kim, Kwang-Yup
    • Journal of Microbiology and Biotechnology
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    • 제17권2호
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    • pp.202-206
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    • 2007
  • The adhesion of Campylobacter jejuni to chicken skin, along with the associated morphological changes under aerobic conditions at 4, 25, and $37^{\circ}C$ and microaerobic ($O_2\;5%,\;CO_2\;10%,\;N_2\;85%$) conditions, were investigated using confocal laser scanning microscopy (CLSM), flow cytometry, and plate counting. The morphological change of C. jejuni from a spiral shape to a coccoid form or VBNC form (viable but nonculturable form) progressed rapidly under aerobic conditions at 25, 37, and $4^{\circ}C$. As regards adhesion, the C. jejuni cells were mostly located in the crevices and feather follicles of the chicken skin, where the cells in the feather follicles floated freely in the entrapped water, even after the skin was rinsed quite thoroughly. CLSM also revealed the penetration of some spiral-shaped C. jejuni cells into the chicken skin. Even after changing their shape at various temperatures, coccoid-form C. jejuni cells were still found in the crevices and feather follicles of the chicken skin.

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

  • 이민선;김효진
    • 한국융합학회논문지
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    • 제8권6호
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    • pp.139-145
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    • 2017
  • 본 연구는 우치의 인공우식법랑질에 대한 불소도포 후 재광화 효과를 공초점레이저주사현미경을 통해 비교하고자 진행되었다. 시편에 인공우식을 형성한 후, 대조군, 1.23% APF 겔 도포군, 그리고 5% NaF 바니쉬 도포군으로 구성하여 실험을 진행하였으며, 불소도포 전후의 양상은 CLSM을 이용하여 평균형광강도로 확인하였다. 불소도포 전 후 평균형광강도는 모든 군에서 유의하게 감소하는 양상을 보였으나, 평균형광강도변화량은 실험군간의 통계적으로 유의한 차이는 발견되지 않았다(p=0.222). 탈회된 표면의 표면미세경도와 평균형광강도 간에는 음의 상관관계를 보였고(p=0.941), 탈회표면의 평균형광강도가 클수록 불소도포 1일 경과 후 평균형광강도 증가하는 양의 상관관계를 보였다(r=0.811, p<0.001). 차 후 구강 내 환경을 반영한 모델이나 실제 in-situ 또는 임상실험을 통해 불소도포제제의 재광화 양상에 대한 연구가 필요할 것이다.

Antibacterial and Antibiofilm Effect of Cell-Free Supernatant of Lactobacillus brevis KCCM 202399 Isolated from Korean Fermented Food against Streptococcus mutans KCTC 5458

  • Kim, Jong Ha;Jang, Hye Ji;Lee, Na-Kyoung;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • 제32권1호
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    • pp.56-63
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    • 2022
  • This study aims to determine the antibiofilm effect of cell-free supernatant (CFS) of Lactobacillus brevis strains against Streptococcus mutans strains. To study the antibiofilm mechanism against S. mutans strains, antibacterial effects, cell surface properties (auto-aggregation and cell surface hydrophobicity), exopolysaccharide (EPS) production, and morphological changes were examined. The antibiofilm effect of L. brevis KCCM 202399 CFS as morphological changes were evaluated by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM), compared with the control treatment. Among the L. brevis strains, L. brevis KCCM 202399 showed the highest antibiofilm effect on S. mutans KCTC 5458. The antibacterial effect of L. brevis KCCM 202399 against S. mutans KCTC 5458 was investigated using the deferred method (16.00 mm). The minimum inhibitory concentration of L. brevis KCCM 202399 against S. mutans KCTC 5458 was 25.00%. Compared with the control treatment, L. brevis KCCM 202399 CFS inhibited the bacterial adhesion of S. mutans KCTC 5458 by decreasing auto-aggregation, cell surface hydrophobicity, and EPS production (45.91%, 40.51%, and 67.44%, respectively). L. brevis KCCM 202399 CFS inhibited and eradicated the S. mutans KCTC 5458 biofilm. Therefore, these results suggest that L. brevis KCCM 202399 CFS may be used to develop oral health in the probiotic industry.

Comparative lectin binding patterns of Cochlodinium polykrikoides Margalef

  • Rhodes, Lesley L.;Cho, Yong-Chul;Cho, Eun-Seob
    • Journal of the korean society of oceanography
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    • 제35권3호
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    • pp.153-157
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    • 2000
  • Four different FITC-conjugated lectins were used to visually evaluate lectin binding activity by optical staining quality using confocal laser scanning microscopy (CLSM) of Cochzodinium polykrikoides in nature (wild type) and culture (cultured type). Cells from the field and cultures treated with ConA fluoresced only at the outer cell wall, and the abundance and distribution of the fluorescent signal were similar. Treatment with PWM and HPA did not elicit fluorescence at the cell surface, but the wild type exposed to HPA showed greater binding than did the cultured cells, possibly due to greater concentrations of glucosamine. The wild type cells treated with LBL lectin showed a strong green fluorescence on the cell surface, whereas cultured cells did not. Signal intensity and abundance were greater than for any other lectins tested in this study. These results suggest that wild type and cultured type are significantly different based on surface sugar production. In particular, the wild type cells apear richer in galactosamine-like moieties. Neither glucose nor mannose-like moieties were present in either wild types or cultured cells.

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Photoresponsive Behavior of Liquid-Crystalline Networks

  • Yu, Yanlei;Ikeda, Tomiki;Nakano, Makoto
    • 한국정보디스플레이학회:학술대회논문집
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    • 한국정보디스플레이학회 2002년도 International Meeting on Information Display
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    • pp.34-37
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    • 2002
  • Freestanding azobenzene-containing liquidcrystalline network (LCN) films. with macroscopic uniaxial molecular alignment were prepared by insitu photopolymerization. By polarizing microscopy, fiber-like structures aligned in one direction were observed. Furthermore, with a confocal laser scanning microscope (CLSM), it was confirmed that the fiber-like structures were formed even in the bulk of the LCNs. Upon UV light irradiation to cause trans-cis photoisomerization of the azobenzene molecules, the LCNfilms underwent a significant and anisotropic bending toward the irradiation direction of UV light. When the bent LCNfilms were exposed to Vis light, unbending of the LCN films immediately took place and the initial flat LCN films were restored. This bending and unbending behavior of the LCN films could be repeated just by changing the wavelength of the irradiation light. It was suggested that the bending was induced by an absorption gradient which produced a volume difference between the front surface area and the bulk of the network films.

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Anti-biofilm Activity of Xanthorrhizol Isolated from Curcuma xanthorrhiza Roxb. against Bacterial Biofilms Formed by Saliva and Artificial Multi-species Oral Strains

  • Yanti, Yanti;Rukayadi, Yaya;Lee, Kwan-Hyoung;Han, Sung-Hwa;Hwang, Jae-Kwan
    • Food Science and Biotechnology
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    • 제18권2호
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    • pp.556-560
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    • 2009
  • Xanthorrhizol, a sesquiterpene isolated from Curcuma xanthorrhiza Roxb., was used to investigate its effect on reducing the saliva and multi-species oral biofilms consisting of Streptococcus mutans, Streptococcus sanguis, and Actinomyces viscosus by anti-biofilm and confocal laser scanning microscopy (CLSM) assays. Xanthorrhizol exhibited significant antibiofilm activity in the dose- and time-dependent manners. Exposure to 2 and $5{\mu}g/mL$ xanthorrhizol for 30 min remained <50% of saliva and multi-species biofilms formed for 24 hr. In addition, exposure to $10{\mu}g/mL$ xanthorrhizol for 30 min reduced 65 and 77% of 24 hr saliva and multi-species oral biofilms, respectively. CLSM results visually demonstrated that xanthorrhizol reduced bacterial viability in the saliva and multi-species oral biofilms. These results suggest that C. xanthorrhiza Roxb. containing xanthorrhizol with strong anti-biofilm activity can be employed as a plant source for oral care functional foods.