• 대한소아치과학회지
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• 제47권2호
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• pp.120-127
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• 2020

이 연구의 목적은 치면착색제인 10 - 20 mM erythrosine을 광감각제로 사용한 광역동 치료의 항균 효과를 평가하는 것이다. Streptococcus mutans, Lactobacillus casei, Candida albicans를 포함하는 다종 우식원성 세균막을 hydroxyapatite disc에 형성하였다. 광감각제로 20 μM, 10 mM, 20 mM erythrosine을 3분간 적용 후 24초 동안 광조사를 시행하였다. 각 실험군의 Colony-forming unit (CFU)을 측정하고 세균막을 Confocal laser scanning microscopy (CLSM)을 이용하여 관찰하였다. 10 - 20 mM erythrosine을 광감각제로 사용한 광역동 치료 실험군에서 CFU의 현저한 감소가 관찰되었고, 그 결과는 CLSM 관찰에서도 확인되었다. 이번 연구를 통해 치과 임상에서 사용되는 치면착색제를 광감각제로 이용한 광역동 치료의 높은 항균효과를 확인할 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제9권2호
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• pp.168-172
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• 1999

The antifungal mechanism of the antifungal peptide against Aspergillus fumigatus, $K^{18,19}$-CA(l-8)-ME(l-12), derived from cecropin A(l-8)-melittin(l-12) was investigated by confocal laser scanning microscopy, cell wall regeneration, ATPase activity inhibition, and released potassium ion. By confocal laser scanning microscopy, $K^{18,19}$-CA(l-8)-ME(l-12) was detected on the surface of A. fumigatus, while cecropin A used as a negative control peptide was not detected. The protoplast of A. fumigatus treated with$K^{18,19}$-CA(1-8)-ME(1-12) failed to regenerate the fungal cell walls. Compared with cecropin A, the amount of potassium ion released by $K^{18,19}$-CA(l-8)-ME(l-12) was increased. Furthermore, $K^{18,19}$-CA(l-8)-ME(l-12) inhibited the ATPase activity on the plasma membrane. These results suggested that $K^{18,19}$-CA(l-8)-ME(1-12) acts on the plasma membrane of A. fumigatus and its antifungal action is due to the ion channel or pore formation on the plasma membrane.

• 펄프종이기술
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• 제38권3호
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• pp.47-60
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• 2006

This study was carried out to investigate the sheet formation properties of Morus Hanjis, made of bast and whole stalk pulps by different pulping methods, such as alkali, alkali-peroxide and sulfomethylated pulping. Two species of Morus, M. alba and M. lhou, were used. Effect of morphological properties of pulp stocks on the sheet formation and its gray levels based on optical property were evaluated using an Image analyzer. In addition, the effect of fiber distribution index(FDI) which was calculated from tile data of Confocal laser scanning microscopy(CLSM) on the sheet formation and optical properties of Morus Hanji were also discussed. On the sheet formation, Hanji from whole stalk pulp was superior than that of bast pulp. The more the sheet formation improved, the more paper opacity decreased. In the aspect of Hanji's surface characteristics analyzed by an Image analyzer, the average gray level and its standard deviation of Hanji from the whole stalk pulp were rather lower than those of bast pulp because of better sheet formation of the former. However, high brightness Hanji showed high value of gray level. The sheet formation and paper opacity were increased with the decrease of standard deviation of gray level. From these results, gray level measurement could be used to predict the paper opacity as well as sheet formation. Sheet formation of whole stalk Hanji and its FDI measured by CLSM were higher than those of bast fibers. In conclusion, the sheet formation and opacity of Hanji could be evaluated by standard deviation value of Hanji's gray level using an Image analyzer and by fiber distribution index using CLSM.

• Journal of Microbiology and Biotechnology
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• 제23권12호
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• pp.1673-1682
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• 2013

Citrobacter sp. is a cause of significant opportunistic nosocomial infection and is frequently found in human and animal feces, soil, and sewage water, and even in industrial waste or putrefaction. Biofilm formation is an important virulence trait of Citrobacter sp. pathogens but the process and characteristics of this formation are unclear. Therefore, we employed in vitro assays to study the nutritional and environmental parameters that might influence biofilm formation of C. werkmanii BF-6 using 96-well microtiter plates. In addition, we detected the relative transcript levels of biofilm formation genes by RT-PCR. Our results indicated that the capacity of C. werkmanii BF-6 to form biofilms was affected by culture temperature, media, time, pH, and the osmotic agents glucose, sucrose, NaCl, and KCl. Confocal laser scanning microscopy results illustrated that the structure of biofilms and extracellular polysaccharide was influenced by 100 mM NaCl or 100 mM KCl. In addition, nine biofilm formation genes (bsmA, bssR, bssS, csgD, csgE, csgF, mrkA, mrkB, and mrkE) were found to contribute to planktonic and biofilm growth. Our data suggest that biofilm formation by C. werkmanii BF-6 is affected by nutritional and environmental factors, which could pave the way to the prevention and elimination of biofilm formation using proper strategies.

• Restorative Dentistry and Endodontics
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• 제43권4호
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• pp.46.1-46.10
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• 2018

Objectives: This study was to evaluate the antibacterial effect of different instrumentation and irrigation techniques using confocal laser scanning microscopy (CLSM) after root canal inoculation with Enterococcus faecalis (E. faecalis). Materials and Methods: Mesiobuccal and mesiolingual canals of extracted mandibular molars were apically enlarged up to a size 25 hand K-file, then autoclaved and inoculated with E. faecalis. The samples were randomly divided into 4 main groups according to the system of instrumentation and irrigation: an XP-endo Shaper (XPS) combined with conventional irrigation (XPS/C) or an XP-endo Finisher (XPF) (XPS/XPF), and iRaCe combined with conventional irrigation (iRaCe/C) or combined with an XPF (iRaCe/XPF). A middle-third sample was taken from each group, and then the bacterial reduction was evaluated using CLSM at a depth of $50{\mu}m$ inside the dentinal tubules. The ratio of red fluorescence (dead cells) to green-and-red fluorescence (live and dead cells) represented the percentage of bacterial reduction. The data were then statistically analyzed using the Kruskal-Wallis test for comparisons across the groups and the Dunn test was used for pairwise comparisons. Results: The instrumentation and irrigation techniques had a significant effect on bacterial reduction (p < 0.05). The iRaCe/XPF group showed the strongest effect, followed by the XPS/XPF and XPS/C group, while the iRaCe/C group had the weakest effect. Conclusions: Combining iRaCe with XPF improved its bacterial reduction effect, while combining XPS with XPF did not yield a significant improvement in its ability to reduce bacteria at a depth of $50{\mu}m$ in the dentinal tubules.

• 대한소아치과학회지
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• 제34권2호
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• pp.183-191
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• 2007

본 연구는 Digital Imaging Fiber-Optic Trans-Illumination(DIFOTI)이나 Laser Fluorescence(LF)를 이용한 방법이 평활면 법랑질 초기우식증의 재광화 정도를 정확하게 monitoring 할 수 있는지의 여부를 평가하기 위하여 진행되었다. 우치 법랑질로부터 얻어진 인공 우식 절편을 특별히 제작된 의치에 식립한 다음, 10명의 실험 참가자들이 구강 내에 장착하고 3주 동안 불소 양치용액을 사용하도록 한 다음 Confocal Laser Scanning Microscopy(CLSM)로 측정된 병소 깊이를 gold standard로 사용하여 DIFOTI와 LF로 측정 된 우식 절편의 재광화 정도를 비교 평가한 후 다음과 같은 결론을 얻었다. 1. DIFOTI에 측정된 광도 백분율이 재광화 기간이 지남에 따라 증가하였으며, CLSM에서 측정된 병소 깊이와 유의한 역상관관계를 보였다(r=-0.683, p<0.01). 2. LF 측정치는 재광화 기간이 경과함에 따라 증가하였으며 CLSM에서 측정된 병소 깊이와 유의한 상관관계를 보였다(p<0.05). 3. CLSM 촬영 결과 500 ppm 불소 양치군이 0 ppm 불소 양치군보다 빠른 속도로 병소 깊이가 감소하는 양상을 보여 주었다.

• International Journal of Oral Biology
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• 제40권2호
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• pp.103-109
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• 2015

Growing evidence suggests that mitochondrial reactive oxygen species (ROS) are involved in various pain states. This study was performed to investigate whether ROS-induced changes in neuronal excitability in trigeminal subnucleus caudalis are related to ROS generation in mitochondria. Confocal scanning laser microscopy was used to measure ROS-induced fluorescence intensity in live rat trigeminal caudalis slices. The ROS level increased during the perfusion of malate, a mitochondrial substrate, after loading of 2',7'-dichlorofluorescin diacetate ($H_2DCF-DA$), an indicator of the intracellular ROS; the ROS level recovered to the control condition after washout. When pre-treated with phenyl N-tert-butylnitrone (PBN) and 4-hydroxy-2,2,6,6-tetramethylpiperidene-1-oxyl (TEMPOL), malate-induced increase of ROS level was suppressed. To identify the direct relation between elevated ROS levels and mitochondria, we applied the malate after double-loading of $H_2DCF-DA$ and chloromethyl-X-rosamine (CMXRos; MitoTracker Red), which is a mitochondria-specific fluorescent probe. As a result, increase of both intracellular ROS and mitochondrial ROS were observed simultaneously. This study demonstrated that elevated ROS in trigeminal subnucleus caudalis neuron can be induced through mitochondrial-ROS pathway, primarily by the leakage of ROS from the mitochondrial electron transport chain.

• 대한소아치과학회지
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• 제48권1호
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• pp.21-30
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• 2021

이 연구의 목적은 indocyanine green (ICG)과 근적외선 diode 레이저의 사용이 다종 우식원성 바이오필름에 미치는 항균 효과를 조사하는 것이다. Streptococcus mutans, Lactobacillus casei, Candida albicans를 포함하는 다종 바이오필름이 ICG와 808 nm 근적외선 diode 레이저를 사용하여 서로 다른 조사 시간에 따라 처리되었다. Colony-forming unit (CFU)을 측정하였고, 바이오필름의 정성적 평가를 위해 공초점 레이저 주사 현미경(Confocal laser scanning microscopy, CLSM) 관찰을 시행하였다. 또한 광열 치료의 효과를 평가하기 위해 온도 측정이 시행되었다. ICG와 근적외선 diode 레이저를 사용한 군에서 CFU의 감소량이 통계적으로 유의하였으나, L. casei와 C. albicans에서는 시간에 따른 항균 효과의 유의한 차이는 관찰되지 않았다. CLSM 관찰에서도 유사한 세균 감소를 확인할 수 있었다. ICG와 근적외선 diode 레이저를 사용한 군은 ICG 없이 광조사를 시행한 군보다 더 높은 온도 상승을 보였으며, 측정된 온도는 열 치료의 온도 범위와 유사하였다. 결론적으로, ICG와 근적외선 diode 레이저는 다종 우식원성 바이오필름에 항균 효과를 보였다. 우식 예방을 위한 보조 수단으로 사용될 가능성을 가지나, 임상적인 적용을 위해서는 적용 프로토콜에 관한 연구가 필요하다.

• 지질공학
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• 제25권2호
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• pp.165-178
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• 2015

Physical and chemical weathering degrades rock, affecting its structural properties and thus the stability of stone buildings or other structures. Confocal laser scan microscopy (CLSM) is used here to observe temporal changes in the surface roughness of rock samples under simulated accelerated weathering. Samples were pressurized to 50, 55, or 70 MPa using a pressure frame, and subjected to freeze/thaw cycling controlled by a thermostat. The temperature was cycled from -20℃ to 40℃ and back. After each 20 cycles, CLSM was used to assess the change in surface roughness, and roughness factors were calculated to quantify the progression of the surface condition over time. Variations in cross-section line-roughness parameters and surface-roughness parameters were analyzed for specific parts of the sample surfaces at 5× and 50× magnification. The result reveals that the highest and lowest values of the roughness factors are changed according to elapsed time. Freezing/thawing at high pressure caused larger changes in the roughness factor than at low pressure.

• Restorative Dentistry and Endodontics
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• 제33권1호
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• pp.1-8
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• 2008

본 연구는 공초점 현미경을 이용하여 새로운 단량체와 filler의 크기, 연마방법이 실험용 복합레진의 표면조도에 미치는 영향을 연구하였다. 단점이 많은 희석재인 TEGDMA의 사용을 줄이기 위해, Bis-GMA의 유도체로서 중합수축이 적고 점도가 낮은 새로운 단량체인 methoxlyated Bis-GMA (Bis-M-CMA)를 첨가하고 다른 크기의 filer를 갖는 2종의 실험용 복합레진과 TEGDMA를 함유한 1종의 실험용 복합레진을 제작하였다. EX1; 실험용 복합레진 1 (Bis-M-GMA/TEGDMA = 95/5 wt%, 40 nm 나노필러 함유), EX2; 실험용 복합레진 2 (Bis-M-GMA/TEGDMA = 95/5wt%, 20 nm 나노필러 함유), EX3; 실험용 복합레진 3 (Bis-GMA/TEGDMA = 70/30 wt%, 40 nm 나노필러 함유). 테프론 몰드를 이용하여 지름 6 mm 두께 2 mm의 시편을 각 실험용 복합레진과 Filtek Z250으로 9개씩 만들고 3군으로 분류하였다. Mylar strip 군은 연마를 하지 않았고, Sof-lex군은 #1000 SiC paper로 연마한 뒤 501-lex disc로 연마하였다. Diapolisher 군은 #1000 SiC paper로 연마한 뒤 DiaPolisher polishing point로 연마하였다. 공초점 현미경을 이용하여 각 시편당 7군데에서 Rq (Root mean square roughness), Rv (Valley roughness), Rp (Peak roughness), Rc (2D Roughness), Sc (3D Roughness) 값을 측정하였고, Two-way ANOVA와 Tukey multiple comparisons test로 유의수준 0.05로 통계처리 하였다. 복합레진의 종류 (p < 0.001), 연마 방법 (p < 0.001)은 각각 모두 표면조도 값에 영향을 미치며, 복합레진의 종류와 연마 방법 간에는 교호 작용이 관찰되었다 (p < 0.001). 복합레진의 종류에 따른 표면조도는 EX2가 가장 거친 표면을 보였고, EX3이 가장 평활한 면을 형성하였으며 (p < 0.05). 연마 방법에서는 연마하지 않은 Mylar strip 군이 가장 평활한 면을 형성하였다 (p < 0.05). 본 연구 결과를 종합하여보면 연마하지 않고 Mylar strip하에서 복합레진이 중합된 경우 가장 낮은 표면조도와 평활한 표면을 보였으며, 새로운 레진 단량체인 Bis-M-GMA를 함유한 복합레진이 수복물의 표면조도 측면에서는 필러 크기에 관계없이 기존의 Bis-GMA/TEGDMA를 기질단량체로 사용하는 복합레진에 비교하여 우수하지 못한 것을 확인하였다.