• Title/Summary/Keyword: Confocal laser microscope

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Fabrication of PLGA/Dextran Double-Layered Microspheres by Oil-in-Water Solvent Evaporation Method (O/W 용매 증발법을 이용한 PLGA와 덱스트란의 이중층 미립구 제조)

  • Ko Jong Tae;Lee Jae-Ho;Lee Chang-Rae;Shin Hyung Sik;Yuk Soon Hong;Kim Moon Suk;Khang Gilson;Rhee John M.;Lee Hai Bang
    • Polymer(Korea)
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    • v.29 no.6
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    • pp.543-548
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    • 2005
  • Double-layered spheres play an important role in controlling drug delivery for pharmaceutical application, because of the low initial burst compared with single-layered spheres and targetable delivery to specific organ. But it has drawback in loading drug and controlling size. In this study, we developed double-layered spheres using relatively simple oil-in-water (O/W) solvent evaporation method witw/without ultrasonication and investigated the size variation of the double-layered microspheres on the contents of poly(lactide- co-glycolide) (PLGA). Double - layered spheres were char-acterized by scanning elecron microscope (SEM), camscope, and confocal fluorescence laser microscope (CFLM). Double-layered spheres showed smooth surfaces and obvious difference between core and corona by SEM observation and camscope. We observed the fluorescent core in the double-walled spheres composed of FlTC-dextran and PLGA using CFLM. It was found that the core of the microsphere was dextran and the corona of the fabricate microsphere was PLGA. Also, the more PLGA concentration, the more the size of the fabricating double-layered sphere observed.

Red-Colored Phenomena and Morphochemical Characteristics of Red-Colored Substances in Ginseng Roots (Panax ginseng C.A. Meyer) (인삼 적변현상과 적변물질의 형태-화학적 특성)

  • 윤길영;양덕조
    • Journal of Ginseng Research
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    • v.24 no.3
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    • pp.107-112
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    • 2000
  • One of the physiologically important ginseng diseases is red-colored phenomena (RCP) that is caused by accumulation of red-colored substances on the epidermis of ginseng roots. Although RCP severely deteriorates the quality of ginseng products, there has been little information on what red-colored substance is and how RCP occurs. Therefore, the heavy losses of cultivators and ginseng industry are suffering by RCP, For this reason, we have investigated with the morphochernical characteristics of RCP to find out main cause of it. The red-colored substances (RS) on the epidermis of red-colored ginseng (RCG) were examined using inverted light microscope, confocal laser scanning microscope (CLSM)and furier transform infrared (FT/IR) spectrometer. Red brown substances were accumulated in the cell wall of the epidermis from early stage to late stage of RCC. Especially, cell wall of the late stage of RCG was covered with the sub-stances with 80~ 130 fm thick. Therefore, the cell wall of RCG cannot protect the ginseng root cells from the mechanical damages, bacteria and fungi. To analyse red substances of roots, RS were isolated from epidermis of RCG and extracted using various solvents. RS is strongly insoluble but it was bleached by oxidizing agents including 12% (v/v) NaOCl. Therefore, RS was Presumed to make up of high chelation power. The proriles of FT/IR spectra or both healthy ginseng (HEG) and RCG showed a significant difference at two wavelength,2857 cm$\^$-1/(C-H) and 1032 cm$\^$-1/(S=O), respectively. Furthermore, absorption peak of 2857cm$\^$-l/ appears on the only epidermis of RCG. The other peak is shown lower absorption rate on the epidermis of RCG than that of healthy ginseng. Also, FT/IR spectra of the mixture of carboxym-ethylcellulose (CMC) and iron (Fe$\^$3+/) were very similar to RCG spectrum profiles. One of a interesting fact is that the contents of phenolic compounds at the epidermis of healthy ginseng were highest. The results of these experiments sup-port the RCP was closely related with the chemical interaction between inorganic elements (Fe) of rhizosphere and organic matters (cellulose, cellobiose, cell sap, etc.) of ginseng roots.

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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.

Development of Female Gametophyte of Six Species of Corydalis (Fumariaceae) (현호색속(현호색과) 6종의 자성배우자체 발달과정)

  • Oh, Byoung-Un;Jang, Chang-Gee
    • Korean Journal of Plant Taxonomy
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    • v.32 no.4
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    • pp.467-480
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    • 2002
  • The development of the female gametophyte of Corydalis albipetala, C. ambigua, C. filistipes, C. nobilis, C. solida, C. ophiocarpa have been comparatively investigated using laser scanning confocal microscope (LSCM) and light microscope. An archesporium was originated from one of the outmost parietal cells beneath the one-layered epidermis of protuberant nucellus, and acted directly as a megaspore mother cell (MMC). These species had linear tetrads after successional meiotic division during the megasprogenesis. A functional megasprore developed from one of the tetrad in the chalazal end, and the rest three being degenerated. The developmental type of the female gametophyte was monosporic in accordance with the Polygonum type. Prior to anthesis the female gametophyte was organized. So mature embryo sac was comprised a three-celled egg apparatus, three large antipodals were developed from the apex of each antipodal cell, and extended toward micropylar end to be contacted with egg apparatus. Two synergids were usually observed as degenerated condition, and in this time the apices of antipodal haustoria were connected with the degenerated synergids. The developmental characteristics of seven-nucleate female gametophytes were common in all the species investigated. But the shape of mature embryo sac was ovoidal in C. albipetala, C. filistipes, C. ophiocarpa and C. solida, reflexed in C. ambigua, and rather flattened ovoidal in C. nobilis. Also, the type of megasporangium was anatropous in all the species except C. ambigua with campylotropous ovule.

Modification of surface pretreatment of white spot lesions to improve the safety and efficacy of resin infiltration

  • Yim, Hyun-Kyung;Min, Ji-Hyun;Kwon, Ho-Keun;Kim, Baek-Il
    • The korean journal of orthodontics
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    • v.44 no.4
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    • pp.195-202
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    • 2014
  • Objective: A low-viscosity resin (infiltrant) was used to inhibit the progression of white spot lesions (WSLs) and resolve associated esthetic issues. An alternative pretreatment was explored to increase the pore volume of the surface layer of the WSLs. Also, the penetration effects of the infiltrant were evaluated for various pretreatments. Methods: Sixty two artificial lesions were fabricated on bovine teeth. As a positive control, 15% HCl gel was applied for 120 seconds. Further, 37% $H_3PO_4$ gel was applied for 30 seconds using three methods. The samples were divided as follows: $H_3PO_4$ only group, $H_3PO_4$ sponge group, and $H_3PO_4$ brush group. The acid was gently rubbed with the applicators (i.e., a sponge or brush) throughout the application time. To compare the effects of resin infiltration, twenty paired halves of specimens were treated with an infiltrant (ICON$^{(R)}$). Results: Thicknesses of the removed surface layers and infiltrated areas were evaluated by confocal laser scanning microscope. The positive control and the 37% $H_3PO_4$ brush group failed to show significant differences in the removed thickness (p > 0.05); however, the mean percentage of the infiltrated area was higher in the 37% $H_3PO_4$ brush group ($84.13{\pm}7.58%$%) than the positive control ($63.51{\pm}7.62%$, p < 0.001). Scanning electron microscope observations indicate higher pore volumes for the 37% $H_3PO_4$ brush group than for the positive control. Conclusions: Application of 37% $H_3PO_4$ with a brush for 30 seconds increased the pore volume of WSL surface layers and the percentage of infiltrated areas in comparison to the use of 15% HCl for 120 seconds.

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.

Spectroscopic Comparison of Photo-oxidation of Outside and Inside of Hair by UVB Irradiation (자외선B 조사에 의한 모발 외부와 내부의 광산화에 관한 분광학적 비교)

  • Ha, Byung-Jo
    • Applied Chemistry for Engineering
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    • v.31 no.2
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    • pp.220-225
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    • 2020
  • Hair is made of proteins containing various amino acids. Ultraviolet (UV) radiation is believed to be responsible for the most damaging effects of sunlight, and also plays an important role in hair aging. The purpose of this study was to investigate the changes in morphological and chemical structures after ultraviolet B (UVB) irradiation of human hair. The UVB-irradiated hair showed characteristic morphological and structural changes, compared to those of the normal hair. The result from a scanning electron microscope (SEM) equipped with an energy dispersive X-ray diffractometer (EDX) showed that the scale of UV-irradiated hair appeared to be rough and the amount of oxygen element was higher than that of the normal hair. Fluorescence and three dimensional (3D) topographical images were obtained by a confocal laser scanning microscope (CLSM). In 3D images, the green emission intensity of normal hair was much higher than that of fluorescing UVB-irradiated hair. The intensity of green emission reflects the intrinsic fluorescence of hair protein. Also, a fluorescent imaging method using fluorescamine reagent was used to identify the free amino groups resulting from a peptide bond breakage in UVB-irradiated hair. Strong blue fluorescence of UVB-irradiated hair, which indicates a very high level of amino groups, was observed by CLSM. Therefore, the fluorescamine as an extrinsic fluorescence could provide a useful tool to identify the peptide bond breakage in UVB-irradiated hair. Infrared image mapping was also employed to assess the cross-sections of normal and UVB-irradiated specimens to examine the oxidation of disulfide bonds. The degree of peak areas with strong absorbance for the disulfide mono-oxide was spread from the outside to the inside of hair. The spectroscopic techniques used alone, or in combination, launch new possibilities in the field of hair cosmetics.

CHANGE OF VASOACTIVE INTESTINAL POLYPEPTIDE(VIP) IMMUNOREACTIVE CELLS FOLLOWING PULP EXTIRPATION IN RAT TRIGEMINAL GANGLION: A CONFOCAL LASER SCANNING MICROSCOPIC STUDY (치수제거 후 흰쥐 삼차신경절에서 VIP 면역반응세포의 변화: 공초점레이저주사현미경적 연구)

  • Kim, Heung-Joong;Kim, Seung-Jae;Park, Joo-Cheol;Lee, Chang-Seop;Lee, Sang-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.1
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    • pp.25-31
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    • 2001
  • The purpose of this study was to investigate the distribution and fluorescence intensity of vasoactive intestinal polypeptide immunoreactive (VIP-IR) cells in rat trigeminal ganglion following pulp extirpation of rat mandibular molar. The animals were divided into control group(n=6) and experimental group(n=6). The experimental animals were sacrificed at 14 days after pulp extirpation. The trigeminal ganglion was removed and immersed in the 4% paraformaldehyde in 0.1M phosphate buffer. Serial frozen sections about $20{\mu}m$ 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 (CLSM). Unprocessed optical sections were obtained and stored on a optical disk. Color pictures were printed by a video copy processor. The results were as follows; 1. The positive ratio of VIP-IR cells in mandibular part of trigeminal ganglion were 7.40% in control group and 28.42% in experimental group(14 days after pulp extirpation). 2. The relative fluorescence intensity of VIP-IR cells in mandibular part of trigeminal ganglion were 87.78 in control group and 138.65 in experimental group. The relative fluorescence intensity of experimental group was 58% higher than that of control group. 3. In optical serial section analysis of VIP-IR cells of experimental group, most of the 9 sections showed high fluorescence intensity. At high magnification, axons of the experimental group displayed greater VIP-IR than in the control group, and the positive cells were mainly of medium size. The result indicate that number and fluorescence intensity of VIP-IR cells were increased in the mandibular part of trigeminal ganglion following pulp extirpation of mandibular molar, and it suggests that VIP could play a role in processing of nociception.

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The Changes of Occludin in Tight Junction of Blood-Brain Barrier by ROS (치밀이음부 구조단백질인 Occludin에 대한 활성산소종의 영향)

  • Lee, Hee-Sang;Kim, Dae-Jin;Sohn, Dong-Suep;Jeong, Bong-Su;Choi, Hyung-Taek;Sim, Kyu-Min;Lee, Keum-Jeong;Cho, Hye-Jin;Kim, Suk-Joong;Lee, Jong-Chan;Jeong, Yoon-Hee;Kim, Sung-Su;Lee, Won-Bok
    • Applied Microscopy
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    • v.34 no.4
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    • pp.231-239
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    • 2004
  • Cerebral microvessel endothelial cells that form blood-brain barrier (BBB) have tight junction for maintaining brain homeostasis. Occludin, one of tight junction protein, is crucial for BBB function. $H_2O_2$ induced occludin changes and effects in bovine brain BBB endothelial cells were examined in this study. The decrease of transendothelial electrical resistance (TEER) by $H_2O_2$ was due to disruption of occludin localization. Cytotoxicity test revealed that $H_2O_2$ did not cause cell death below 1 mM $H_2O_2$ within 4 hr. $H_2O_2$ caused intermittent disruption and loss of occludin at tight junctions and occludin disappeared with dose dependent manner from tight junction in confocal laser microscopy. But Western blot revealed that the total amounts of occludin increased by $H_2O_2$ administration. Transmission electron microscopy revealed that the ultrastructure of tight junction was not changed by $H_2O_2$. These data suggest that functional disruption of BBB by $H_2O_2$ was due to the localized loss of occludin in tight junction, but the expression of occludin increased in order to compensate the disrupted function in BBB.

Influence of finish line design on the marginal fit of nonprecious metal alloy coping fabricated by 3D printing, milling and casting using CAD-CAM (CAD-CAM을 이용한 3D printing, milling, casting 방법의 비귀금속 코핑의 지대치 변연 적합도 연구)

  • Seo-Rahng Kim;Myung-Joo Kim;Ji-Man Park;Seong-Kyun Kim;Seong-Joo Heo;Jai-Young Koak
    • The Journal of Korean Academy of Prosthodontics
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    • v.61 no.1
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    • pp.1-17
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    • 2023
  • Purpose. The purpose of this study was to examine the correlation between the finish line designs and the marginal adaptation of nonprecious metal alloy coping produced by different digital manufacturing methods. Materials and methods. Nonprecious metal alloy copings were made respectively from each master model with three different methods; SLS, milling and casting by computer aided design and computer aided manufacturing (CAD-CAM). Twelve copings were made by each method resulting in 72 copings in total. The measurement was conducted at 40 determined reference points along the circumferential margin with the confocal laser scanning microscope at magnification ×150. Results. Mean values of marginal gap of laser sintered copings were 11.8 ± 7.4 ㎛ for deep chamfer margin and 6.3 ± 3.5 ㎛ for rounded shoulder margin and the difference between them was statistically significant (P < .0001). Mean values of marginal gap of casted copings were 18.8 ± 20.2 ㎛ for deep chamfer margin and 33 ± 20.5 ㎛ for rounded shoulder margin and the difference between them was significant (P = .0004). Conclusion. Within the limitation of this study, the following conclusions were drawn. 1. The variation of finish line design influences the marginal adaptation of laser sintered metal coping and casted metal coping. 2. Laser sintered copings with rounded shoulder margin had better marginal fit than deep chamfer margin. 3. Casted copings with deep chamfer margin had better marginal fit than rounded shoulder margin. 4. According to the manufacturing method, SLS system showed the best marginal fit among three different methods. Casting and milling method followed that in order.