• Restorative Dentistry and Endodontics
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• 제25권2호
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• pp.225-234
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• 2000

The purpose of this study was to investigate the distribution and fluorescene intensity of vasoactive intestinal polypeptide(VIP) immunoreactive cells in rat trigeminal ganglion after inferior alveolar nerve axotomy. The animals were divided into normal and two experimental groups. The experimental animals were sacrificed at 14th and 28th day after inferior alveolar nerve axotomy. The trigeminal ganglion was removed and immersed in the 4% paraformaldehyde-0.2% picric acid in 0.1M phosphate buffer. Serial frozon sections about $16{\mu}m$ in thickness were cut with a cryostat. The immunofluorescence staining was performed. The rabbit anti-VIP(1 : 8,000) was used as primary antibody and fluorescene isothiocynate(FITC)-conjugated anti-rabbit IgG(1 : 80) as secondary antibody. The slides were observed under confocal laser scanning microscope. Three-dimensional images were constructed from 9 serial images(each $1{\mu}m$ in thickness) made by automatic optical sectioning. Unprocessed optical sections were obtained and stored on a optical disk. Color picture were printed by a video copy processor. The results were as follows; 1. The appearance of VIP immunoreactive cells in the mandibular part of trigeminal ganglion was 8.79${\pm}$1.99% in normal group and 39.16${\pm}$5.62% in 14 days, 16.25${\pm}$2.39% in 28 days after inferior alveolar nerve axotomy groups. 2. The relative fluorescence intensity of VIP immunoreactive cell bodies in the mandibular part of trigeminal ganglion was 134.40${\pm}$10.39 in normal group and 192.88${\pm}$14.06 in 14 days, 143.10${\pm}$5.02 in 28 days after nerve axotomy groups. Therefore, the relative fluorescence intensity of 14 days after nerve axotomy group was 43.3% higher than intensity of normal group. 3. In optical single section analysis of VIP immunoreactive cell bodies, white cell bodies(moderate fluorescence intensity) were the most abundant in normal and 28 days after nerve axotomy groups. Whereas, in 14 days after nerve axotomy group, red cell bodies(high fluorescence intensity) were the most abundant. 4. In optical serial section analysis of VIP immunoreactive cell bodies, red cell bodies(high fluorescence intensity) were observed in a part of the 9 sections of normal and 24 days after nerve axotomy groups. Whereas, red cell bodies were observed in all of the 9 sections of 14 days after nerve axotomy group. 5. The results indicates that number and fluorescence intensity of VIP immunoreactive cells were increased in the mandibular part of trigeminal ganglion following inferior alveolar nerve axotomy.

• 대한치과보철학회지
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• 제44권3호
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• pp.343-355
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• 2006

Statement of problem. The success of osseointegration can be enhanced with an implant that has improved surface characteristics. Anodic oxidation is one of the surface modifying method to achieve osseointegration. Voltage of anodic oxidation can change surface characteristics and cell activity Purpose. This study was performed to evaluate MG63 cell responses such as affinity, proliferation and to compare surface characteristics of anodic oxidized titanium in various voltage. Material and method. The disks for cell culture were fabricated from grade 3 commercially pure titanium,1 m in thickness and 12 mm in diameter. Surfaces of 4 different roughness were prepared. Group 1 had a machined surface, used as control. Group 2 was anodized under 220 V, group 3 was anodized under 300 V and group 4 was anodized under 320 V. The microtopography of specimens was observed by scanning electron microscope (JSM-840A, JEOL, Japan) and atomic force microscope(Autoprobe CP, Park Scientific Instrument, USA). The surface roughness was measured by confocal laser scanning microscope(Pascal, LSM5, Zeiss, Germany). The crystal structure of the titanium surface was analyzed with x-ray diffractometer(D8 advanced, Broker, Germany). MG63 osteoblast-like cells were cultured on these specimens. The cell morpholgy was observed by field emission electron microscope(Hitachi S-4700, Japan). The cell metabolic and proliferative activity was evaluated by MTT assay Results and conclusion. With in limitations of this in vitro study, the following conclusions were drawn. 1. In anodizing titanium surface, we could see pores which did not show in control group. In higher anodizing voltage, pore size was increased. 2. In anodizing titanium surface, we could see anatase. In higher anodizing voltage, thicker oxide layer increased crystallinity(anatase, anatase and rutile mixed). 3. MG63 cells showed more irregular, polarized and polygonal shape and developed more lamellipodi in anodizing group as voltage increased. 4. The activity of cells in MTT assay increased significantly in group 3 and 4 in comparison with group 1 and 2. However, there was no difference between group 3 and 4 at P<0.05. Proliferation of MG63 cells increased significantly in pore size($3-5.5{\mu}m$) of group 3 and 4 in comparison with in pore size($0.2-1{\mu}m$ ) of group 2.

• Restorative Dentistry and Endodontics
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• 제43권1호
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• pp.2.1-2.10
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• 2018

Objectives: To determine the effect of size and insertion depth of irrigation needle on the amount of apical extruded debris and the amount of penetration depth of sealer using a confocal laser scanning microscope (CLSM). Materials and Methods: Twenty maxillary premolars were assigned to 2 groups (n = 10), according to the size of needle tip, 28 G or 30 G. Buccal roots of samples were irrigated with respective needle type inserted 1 mm short of the working length (WL), while palatal roots were irrigated with respective needle type inserted 3 mm short of the WL. Prepared teeth were removed from the pre-weighed Eppendorf tubes. Canals were filled with F3 gutta-percha cone and rhodamine B dye-labeled AH 26 sealer. Teeth were transversally sectioned at 1 and 3 mm levels from the apex and observed under a CLSM. Eppendorf tubes were incubated to evaporate the irrigant and were weighed again. The difference between pre- and post-weights was calculated, and statistical evaluation was performed. Results: Inserting needles closer to the apex and using needles with wider diameters were associated with significantly more debris extrusion (p < 0.05). The position of needles and level of sections had statistically significant effects on sealer penetration depth (p < 0.05 for both). Conclusions: Following preparation, inserting narrower needles compatible with the final apical diameter of the prepared root canal at 3 mm short of WL during final irrigation might prevent debris extrusion and improve sealer penetration in the apical third.

• 생명과학회지
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• 제22권10호
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• pp.1415-1419
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• 2012

Thymosin ${\beta}4$ 는 대장암에서 암 줄기세포 마커인 CD133을 지닌 세포에서 지니지 않은 세포에 비해 강하게 발현된다고 보고되어 있다. 본 연구에서는 thymosin ${\beta}4$와 줄기세포 마커인 CD133의 상관관계를 정상 위 조직에서 관찰하였다. Thymosin ${\beta}4$와 CD133의 발현 양상은 tissue microarray 조직상에서 면역화학적 방법으로 관찰하였으며 thymosin ${\beta}4$와 CD133의 존재 위치는 면역형광 염색법 및 confocal microscope를 사용하여 조사하였다. Thymosin ${\beta}4$와 CD133은 동일한 양상으로 발현되었으며 모두 위의 선조직에서 강하게 발현되었다. 면역 형광 염색법으로 두가지 단백질을 동시에 염색한 결과 두 단백질이 동일한 위치에서 함께 존재하는 것으로 규명되었다. 이러한 결과는 thymosin ${\beta}4$와 CD133은 정상위의 선조직에서 발현되며 두 단백질의 발현 양상 및 위치가 동일하여 서로 긴밀한 상호작용을 할 가능성을 제시하고 있다.

• 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.

• Restorative Dentistry and Endodontics
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• 제24권2호
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• pp.310-321
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• 1999

In this in vitro study, confocal laser scanning microscopic morphology of dentin-resin interface and its relationship to shear bond strength were investigated after the exposed dentin surfaces were treated with 3 different kinds of dentin adhesive systems[three-step; Scotchbond Multi-Purpose Plus(SMPP), self-priming bonding resin; Single Bond(SB), self-etching primer; Clearfil Liner Bond 2(LB2)]. 52 extracted human molar teeth without caries and/or restorations. The experimental teeth were randomly divided into three groups of seventeen teeth each. In five teeth of each group, class V cavities(depth: 1.5mm) with 900 cavosurface angles were prepared at the cementoenamel junction on buccal and lingual surfaces. Bonding resins of each dentin adhesive system were mixed with rhodamine B. Primer of SMPP was mixed with fluorescein. In group 1. the exposed dentin was conditioned with etchant, applied with above primer and bonding resin of SMPP. In group 2, with etchant and self-priming bonding agent of SB. In group 3, with self-etching primer and bonding agent of LB2. After treatment with dentin adhesive systems, composite resin were applied and photocured. The experimental teeth were cut longitudinally through the center line of restoration and grounded so that about $90{\mu}m$-thick wafers of buccolingually orientated dentin were obtained. And, $70{\sim}80{\mu}m$-thick wafers sectioned horizontally, thus presenting a dentinal tubules at 900 to the cut surface of a remaining tooth, were obtained. Primer of SMPP mixed with rhodamine B was applied to these wafers. Confocal laser scanning microscopic investigations of these wafers were done within of 24 hours after treatment. To measure shear bond strength, the remaining twelve teeth of each group were grounded horizontally below the dentinoenamel junction, so that no enamel remained. After applying dentin adhesive systems on the dentin surface, composite was applied in the shape of cylinder. The cylinder was 5mm in diameter, and 2mm in thickness. Shear bond strength was measured using Instron with a cross-head speed of 0.5mm/min. It was concluded as follows ; 1. Hybrid layer of SMPP(mean: $4.56{\mu}m$) was thicker than that of any other groups. This value was not statistically significant thicker than that of SB(mean: $3.41{\mu}m$, p>0.05), and significant thicker than that of LB2(mean: $1.56{\mu}m$, p<0.05). There was a statistical difference between SB and LB2(p<0.05). 2. Although there were variations in the length of resin tag even in a sample, and in a group, most samples in SMPP and SB showed resin tags extending above $20{\mu}m$. But samples in LB2 showed resin tags of $10{\mu}m$ at best. 3. Besides primer's infiltration into demineralized peritubular dentin and dentinal tubules, fluorophore of primer was detected in the lateral branches of dentinal tubules. 4. All groups demonstrated statistically significant differences from one another(p<0.05), with shear bond strengths given in descending order as follows: SMPP(18.3MPa), SB(16.0MPa) and LB2(12.4MPa). 5. LB2 having thinnest hybrid layer($1.56{\mu}m$) showed the lowest shear bond strength(12.4MPa).

• 한국지하수토양환경학회지:지하수토양환경
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• 제9권1호
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• pp.79-86
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• 2004

암석 내 균열을 따른 수리전도도는 균열의 기하학적 요소, 즉 방향, 간극, 거칠기 그리고 상호 연결도에 주로 좌우된다. 따라서, 균열 내 투수계수를 정확하게 계산하기 위해서는 이와 같은 기하 요소들을 최대한 계산모델에 반영할 필요가 있다. 이 연구에서는 균열 기하양상을 최대한 정확히 반영한 균열모델에서 기존 수치해석과는 다른 새로운 방법인 균질화 해석법(homogenization analysis method)을 이용하여 균열을 따른 투수계수를 구하기 위해 수치해석을 수행하였다. 먼저, 공초점 레이저 스캔 현미경(Confocal Laser Scanning Microscope)을 이용하여 암석시료의 균열 조도와 균열에 가한 수직압축력의 변화에 따른 간극 변화량을 직접 측정하고, 이와 같이 획득한 자료는 균열모델 재현을 위한 입력자료로 사용되었다. 재현된 균열모델을 토대로 한 균질화 해석법은 미시규모(microscale) 매질특성과 거시규모(macroscale) 매질특성을 동시에 고려하여 투수계수를 계산할 수 있는 것이다. 즉, 균질화 해석법은 주기적 미세구조(microstructure)를 갖는 미소 불균질 물질의 거동특성을 구명하기 위해 개발된 새로운 형태의 섭동(perturbation) 이론이다. 이는 균질한 미시규모에서 미시 투수특성을 계산한 후, 거시규모에서의 균질화 투수계수를 계산하게 된다. 그러므로, 이 방법은 균열 기하양상의 국부적 영향을 고려한 투수특성을 정확히 해석할 수 있다. 균질화법을 이용한 투수계수 산정결과를 기존 연구에서 제안한 경험식과 비교하여 그 타당성을 검증하기 위해 전술한 2차원 균열모델을 이용한 투수계수 계산을 수행하였다. 균열모델은 거칠기(roughness)를 반영하고 동일한 간극을 할당한 평행판 모델을 가정하였다. 계산결과에 의하면, 균질화 해석법에 의해 계산한 C-투수계수는 실내투수시험에 의해 구한 투수계수와 같은 범위의 값을 가지거나 $10^1$ 정도의 차이를 보여, 그 계산결과는 타당하다고 볼 수 있다. 그러나, 균질화 해석법은 국부적으로 불균질한 균열 기하양상과 물질특성이 미시규모와 거시규모에서 모두 고려되므로, 이들 특성을 정확히 알고 있을 경우 기존에 제안된 경험식들에 의한 계산결과 보다 균질화 해석법의 결과가 훨씬 정확함을 주목하여야 한다.

• 지질공학
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• 제25권1호
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• pp.1-8
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• 2015

Deep geological disposal is the preferred storage method for high-level radioactive waste, because it ensures stable long-term storage with minimal potential for human disruption. Because of the risk of groundwater contamination, a buffer of steel and bentonite layers has been proposed to prevent the leaching of radionuclides into groundwater. Quartz is one of the most common minerals in earth's crust. To understand how deformation and dissolution phenomena affect waste disposal, here we study quartz samples at pressure, temperature, and pH conditions typical of deep geological disposal sites. We perform a dissolution experiment for single quartz crystals under different pressure and temperature conditions. Solution samples are collected and the dissolution rate is calculated by analyzing Si concentrations in a solution excited by inductively coupled plasma-atomic emission spectroscopy (ICP-AES). After completing the dissolution experiment, deformation of the quartz sample surfaces is investigated with a confocal laser scanning microscope (CLSM). An empirical formula is introduced that describes the relationship between dissolution rate, pressure, and temperature. These results suggest that bentonite layers in engineering barrier systems may be vulnerable to thermal deformation, even when exposed to higher temperatures on relatively short timescales.

• Advances in environmental research
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• 제7권1호
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• pp.39-52
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• 2018

Direct membrane filtration (DMF) of wastewater has many advantages over conventional biological wastewater treatment processes. DMF is not only compact, but potentially energy efficient due to the lack of biological aeration. It also produces more biosolids that can be used to produce methane gas through anaerobic digestion. Most of ammoniacal nitrogen in wastewater is preserved in effluent and is used as fertilizer when effluent is recycled for irrigation. In this study, a technical feasibility of DMF was explored. Organic and nitrogen removal efficiencies were compared between DMF and membrane bioreactor (MBR). Despite the extremely high F/V ratio, e.g., $14.4kg\;COD/m^3/d$, DMF provided very high COD removal efficiencies at ~93%. Soluble microbial products (SMP) and extracellular polymeric substances (EPS) were less in DMF sludge, but membrane fouling rate was far greater than in MBR. The diversity of microbial community in DMF appeared very narrow based on the morphological observation using optical microscope. On the contrary, highly diverse microbial community was observed in the MBR. Microorganisms tended to form jelly globs and attach on reactor wall in DMF. FT-IR study revealed that the biological globs were structurally supported by feather-like materials made of secondary amines. Confocal laser scanning microscopy (CLSM) study showed microorganisms mainly resided on the external surface of microbial globs rather than the internal spaces.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.12-12
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• 2001

To get insight into the nature of foreign mitochondria and syngamy during mammalian fertilization we compared fertilization processes in porcine oocytes following microinjection of porcine or mouse spermatozoa. Pronuclear movement, sperm mitochondria, and DNA synthesis were imaged with propidium iodide, mitotracker, and BrdU under confocal laser scanning microscope. Intracytoplasmic injection of either porcine or mouse spermatzoon activated porcine oocytes without additional parthengenetic stimulation. Foreign mitochondria in either mouse or porcine sperm midpiece were introduced into porcine oocytes following sperm injection, but rapidly disappeared from the actively developing porcine oocytes. BrdU experiment showed new DNA synthesis in porcine oocytes following injection of mouse spermatozoon or sperm head. At 24 h after injection of mouse isolated sperm head or a spermatozoon, mitoic metaphase was seen in oocyte, but they did not go to normal cell division (Table). These results suggest that pronuclear formation, foreign mitochondria disruption, DNA synthesis and syngamy formation during fertilization are not species specific processes.(Table Omitted).