• Title/Summary/Keyword: barrier membrane

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The Relationship between Intracellular Protein Kinase C Concentration and Invasiveness in U-87 Malignant Glioma Cells (교모세포종 세포주 U-87에서 세포내 PKC 농도와 종양침습성과의 상관 관계)

  • Ji, Cheol;Cho, Kyung-Keun;Lee, Kyung Jin;Park, Sung Chan;Cho, Jung Ki;Kang, Joon Ki;Choi, Chang Rak
    • Journal of Korean Neurosurgical Society
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    • v.30 no.3
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    • pp.263-271
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    • 2001
  • Objective : Glioblastomas, the most common type of primary brain tumors, are highly invasive and cause massive tissue destruction at both the tumor invading edges and in areas that are not in direct contact with glioma cells. As a result, patients with high-grade gliomas are faced with a poor prognosis. Such grim statistics emphasize the need to better understand the mechanisms that underlie glioma invasion, as these may lead to the identification of novel targets in the therapy of high grade gliomas. Protein kinase C(PKC) is a family of serine/threonine kinases and an important signal transduction enzyme that conveys signals generated by ligand-receptor interaction at the cell surface to the nucleus. PKC appears to be critical in regulating many aspects of glioma biology. The purpose of this study was to assess accurately the role of PKC in the invasion regulation of human gliomas based on hypothesis that protein kinase C(PKC) is functional in the process of glial tumor cell invasion. Method : To test this hypothesis, U-87 malignant glioma cell line intracellular PKC levels were up and down regulated and their invasiveness was tested. Intracellular PKC level was characterized using PKC activity assays. Invasion assays including barrier migration and spheroid confrontation were used to study the relationship between PKC concentration and invasiveness. Result : The cell line which were treated by PKC inhibitor tamoxifen and hypericin exhibited decreased PKC activity and decreased invasive abilities dose dependently both in matrigel invasion assay and tumor spheroid fetal rat brain aggregates(FRBA) confrontation assay. However, the cell line that was treated by PKC activator 12-O-tetradecanylphorbol-13acetate(TPA) did not exhibit increases in either PKC activity or invasive ability. Conclusion : These studies suggest that PKC may be a useful molecular target for the chemotherapy of glioblastoma and other malignancies and that a therapeutic approach based on the ability of PKC inhibitors may be helpful in preventing invasion.

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A study of the effects of PDGF-BB on the characteristics of bone stromal and periodontal ligament cells (혈소판유래성장인자-BB가 골간질세포와 치주인대세포의 성상에 미치는 영향)

  • Kwon, Young-Hyuk;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.949-965
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    • 1996
  • The main goal of periodontal therapy is to restore the lost periodontal tissue and establish the attachment appratus. Current acceptable therapeutic techniques are included : removal of diseased soft tissue, demineralization of exposed root surface, using the barrier membrane for preventing the downgrowth of gingival epithelial cell, insertion of graft materials as a scaffolding action, and biological mediators for promoting the cell activity. The latest concept one among them has been studied which based on the knowledge of cellular biology of destructed tissue. Platelet-derived growth factor(PDGF) is one of the polypeptide growth factor which have been reported as a biological mediator to regulate activities of wound healing progress including cell proliferation, migration, and metabolism. The purposes of this study is to evaluate the influences of the PDGF as biological mediator to periodontal ligament and bone marrow cell. Both right and left maxillary first molar were extracted from rat which had treated with 0.4% ${\beta}-Aminopropionitril$ for 5 days, and feeded until designed date to sacrifice under anesthesisa. Periodontal ligament were removed from the extracted socket of the rat, and cultured with Dulbecco's Modified Essential Medium(DMEM) contained with 10% Fetal Bovine Serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. Bone marrow cell were culture from bone marrow suspension with which washed out from femur with same medium. The study was performed to evaluate the effect of PDGF to periodontal ligament and bone cell, cell proliferation rate, total protein synthesis, and alkaline phosphatase activity of rat periodontal ligament(PDL) cell and bone stromal(RBS) cell in vitro. The effects of growth factors on both cells were measured at 3, 5th day after cell culture with (control group) or without growth factors(experimental group). The results were as follows: 1. The tendency of cell proliferation under the influence of PDGF showed more rapid proliferation pattern than control at 3 and 5 days after inoculation. 2. The activity of Alkaline phosphatase revealed 14, 80% increased respectively at 3, 5 days culture than control group. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(P<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P<0.05). 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group l(PDL 100%), 2(PDL 70%:GF 30%), and 3(PDL 50%:GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30%:GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05). From the above results, it is assumed that the PDGF on PDL cells and RMB cell culture. GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density. In conclusion, platelet-derived growth factor can promote rapid osteogenesis during early stage of periodontal tissue regeneration.

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The effects of a combination of calcium sulfate and platelet-derived growth factor on periodontal ligament cells in vitro (Calcium sulfate와 혈소판 유래성장인자의 혼합사용이 치주인대세포에 미치는 영향)

  • Kim, Jun-Seong;Choi, Seong-Ho;Yu, Yun-Jung;Chai, Jung-Kiu;Kim, Chong-Kwan;Cho, Kyoo-Sung
    • Journal of Periodontal and Implant Science
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    • v.27 no.4
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    • pp.785-804
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    • 1997
  • It was well known that calcium sulfate was biocompatible, resorbed rapidly in the body, had potential as a good barrier membrane. Platelet-derived growth factor(PDGF) was one of polypeptide growth factor that had been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purpose of this study was to evaluate the effects of a combination of calcium sulfate and PDGF on periodontal ligament cells in vitro to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the premolar tooth extracted for the orthodontic treatment. Cells were cultured in ${\alpha}-MEM$ contained with 20% FBS, at the $37^{\circ}C$, 100% of humidity, 5% $Co_2$ incubator. Cells were inoculated and cultured into 96 well culture plate with $1{\times}10^4cells/well$ of ${\alpha}-MEM$ for 1 day. After discarding the medium, those cells were cultured in ${\alpha}-MEM$ contained with 10% FBS alone(control group), in calcium sulfate(calcium sulfate group), in calcium sulfate treated with 15ng/ml of PDGF-BB(calcium sulfate+PDGF group), in ${\alpha}-MEM$ contained with 10% FBS treated with 15ng/ml of PDGF-BB(PDGF group) for 1, 2, 3 day respectively. And then each group was characterized by examining of the cell counting, MTT assay, collagen synthesis. The results were as follows. 1. In the analysis of cell proliferation by cell counting, both calcium sulfate group and calcium sulfate plus PDGF group showed no stastically significant difference compared to control group, but there was stastically significant difference between PDGF group and calcium sulfate group at 1, 2 day(P<0.05). 2. In the analysis of cell proliferation by MTT assay in calcium sulfate extracts, both calcium sulfate group and calcium sulfate plus PDGF group showed no stastically significant difference compared to control group, but there was stastically significant difference between PDGF group and calcium sulfate group at 2, 3 day, and between calcium sulfate plus PDGF group and calcium sulfate group at 2 day(P

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Characteristics of Percutaneous Absorption of Glycol ethers (Glycol ethers에 대한 피부 투과 특성)

  • Lee, Han-Seob;Choi, Sung-Boo;Kim, Nac-Joo;Keun, Jang-Hyoun;Hwang, Hyun-Suk;Baek, Jung-Hun;Choi, Jin-Ho;Lee, Ho-Joon
    • Journal of the Korean Applied Science and Technology
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    • v.30 no.1
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    • pp.116-126
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    • 2013
  • Glycol ethers are a group of solvents based on alkyl ethers of ethylene glycol commonly used in paints. These solvents typically have a higher boiling point, together with the favorable solvent properties of lower-molecular weight ethers and alcohols. The word "Glycol ethers" was registered as a United States trademark by Union Carbide Corp. Typically, glycol ethers are found in pharmaceuticals, sunscreens, cosmetics, inks, dyes and water based paints. On the other hand, glycol ethers are used in degreasers, cleaners, aerosol paints and adhesives. Most glycol ethers are relatively water soluble, biodegradable and only a few are considered toxic. Therefore, they are unlikely to pose an adverse risk to the environment. Recent study suggests that occupational exposure to glycol ethers is related to low motile sperm count in men, but the finding has been disputed by others. In this study, skin permeation of 3 types glycol ethers were studied in vitro using matrix such as solvent and detergent. The absorption of glycol ethers[methyl glycol ethers(MC), ethyl glycol ethers(EC) and butyl glycol ethers(BC)] has been measured in vitro through rat skin. Epidermal membranes were set up in Franz diffusion cells and their permeability to PBS measured to establish the integrity of the skin before the glycol ethers were applied to the epidermal surface. Absorption rates for each glycol ethers were determined and permeability assessment made to quantify any irreversible alterations in barrier function due to contact with the esters. Types of glycol ethers in vitro experimental results on MC> EC> BC quickly appeared in the following order: skin permeation was beneficial to the skin permeation small molecular weight, the difference in chemical structure, such as hydrophilic, because with the partition coefficient and solubility mechanisms and passive diffusion to increase the speed at which transmission is considered.

The Effect of Seasoning on the Intestinal Absorption -Absorption by Passive Transport and the Effect of Red Pepper- (조미료가 창자 운동과 흡수기능에 미치는 영향 -소장의 피동적 흡수에 대한 고추의 영향-)

  • Shin, Dong-Hoon;Kim, Joong-Soo;Koh, Jae-Pyong;Ahn, Seung-Woon
    • The Korean Journal of Physiology
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    • v.7 no.1
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    • pp.23-31
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    • 1973
  • Numerous factors concern with the absorption of substances through the membrane of the gastrointestinal tract. To simplify the experimental condition, present work has been restricted to observe the disappearance rate of substance from the intestinal loop which was made in the jejunum, 70 cm apart from the pylorus of the adult rabbit. The purpose of the study is to clarify the absorption of urea through the jejunal wall is solely attributable to the concentration difference between the luminal fluid and plasma, and to observe the effect of adding red pepper upon the rate of absorption. The rabbits were anesthetized with nembutal, 35mg/kg I.V. Jejunal loop was made by ligating at 2 spots, 70 cm and 80cm apart from the pylorus. After rinsing with normal saline solution through the polyethylene tubing inserted from the end of the loop, 8 ml of test solution was placed through the same tubing. The test solution contained 200 mg% of urea and 150mg% of polyethylene glycol(M.W. 4,000) in normal saline solution. Right after placing the test solution the first specimen was taken through the tubing, and successive samplings were performed at 5, 10, 20, and 30 minutes. Logarithm of the difference of urea concentration between the luminal fluid and plasma was plotted against time elapsed after the onset of the experiment. If straight line is revealed, it would verify the nature of transport mechanism as diffusion, obeying the Fick's principle. The concentration of polyethylene glycol (PEG) was also measured in order to examine the change in the volume. PEG was used as the marker substance because it is not absorbable in the intestinal tract. Consequently the concentration of PEG relates inversely to the volume of the loop. Instantaneous concentration of urea in the loop times the volume will give the amount of urea remaining in the luminal fluid. The change in the amount of any substance is directly relate to the volume of the compartment and differs from the change in the concentration which is independent of the volume. After completion of the experiment without red pepper, it was added in the test solution and was centrifuged after thorough mixing. Supernatant of the mixture was placed in the loop and similar sampling were performed with the same time intervals that of previous run in order to observe the effects of the red pepper on the passive transport of the water soluble small substance, urea. The results obtained were as follows: 1. Logarithm of the concentration difference of urea between the luminal fluid and plasma was diminished exponentially as time elapsed. The decay constant in the experiment without red pepper was 0.0563/min. By adding red pepper in the test solution as much as the concentration rose to 4,000 mg% and 8,000 mg%, the decay constants were lowered to 0.0493/min and to 0.0506/min, respectively. The time interval by which the concentration difference dropped to one half of the initial value was prolonged. Without red pepper the half concentration time was 13.30 minutes, and by adding extract of red pepper, 15.31 minutes and 15.71 minutes were revealed. 2. The profile of the diminishing rate of tile amount of urea was quite different from that of the concentration because of the change in the volume of the loop during the observed period. 3. By adding the extract of red pepper, it slowed down the rate of absorption of urea in the intestinal loop, suggesting an increase in the diffusional barrier. 4. Larger dosage of red pepper brought an increase in the secretion of intestinal fluid with concomitant expansion of the luminal volume, and the retardation of the absorption of urea was noticed. This effect was largely dependent on the sensitivity of the individual animal to the red pepper, extract. The amount of urea remained after 10 minutes interval was 55.5% of the initial amount in the experiment without red pepper. On the other hand it was not consistent after administration of red pepper, showing 50.6% and 66.5% of the initial figures by adding 400 mg and 800 mg of red pepper in the test solution, respectively. It was postulated that symptom of diarrhea often encountered by taking a hot (red pepper) food might be attributable to the increase of secretion and the retardation of absorption in the intestinal tract.

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Development of Dermal Transduction Epidermal Growth Factor (EGF) Using A Skin Penetrating Functional Peptide (피부투과 기능성 펩타이드를 이용한 경피투과성 상피세포성장인자의 개발)

  • Kang, Jin Sun;La, Ha Na;Bak, Sun Uk;Eom, Hyo Jung;Lee, Byung Kyu;Shin, Hee Je
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.2
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    • pp.175-184
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    • 2019
  • The epidermal growth factor (EGF) has a intrinsic function of inducing growth and proliferation of cells through interacting with cell membrane receptors in human epidermis and dermis layer. These functions of EGF are used as a main ingredient for wound healing medicines and anti-aging cosmetics. As a cosmetic ingredient, the EGF has a problem in exhibiting its natural efficacy due to the lack of the ability to penetrate through the stratum corneum, which is known as the skin barrier. In this study, a recombinant human epidermal growth factor ($MTD_{151}-EGF$) fused with the macromolecule transduction domain $(MTD)_{151}$ with the skin penetration ability was developed to improve the skin penetration efficiency of the EGF. Expression of $MTD_{151}-EGF$ was performed in E. coli transformed with a vector encoding the $MTD_{151}-EGF$ gene and then purified. The purified $MTD_{151}-EGF$ was evaluated using cell proliferation assay, cytotoxicity test and skin penetration test by franz diffusion cell assay and artificial skin. Cell proliferation activity of $MTD_{151}-EGF$ purified to high purity of 99% or above was equivalent to the EGF or better, and cytotoxicity was not observed. In addition, the $MTD_{151}-EGF$ showed an excellent penetration efficiency compared to the EGF in the skin penetration test with EGF and $MTD_{151}-EGF$ labeled by FITC in an artificial skin penetration model. Based on the quantitative analysis of the penetrating substance using franz diffusion cell assay, the amount of penetration was about 16 times more than that of EGF. These results can be regarded as an effective alternative to improve the existing physical transdermal penetration method related to the use of various active ingredients for cosmetics.