• Journal of The Korean Dental Society of Anesthesiology
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• v.14 no.3
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• pp.157-165
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• 2014

Background: Incisional site of surgical operation become transient ischemic state and then occur reoxygenation due to vasodilatation by inflammatory reaction, the productive reactive oxygen species (ROS) give rise to many physiologic results. Apoptosis have major role on elimination of inflammatory cell and formation of granulation tissue in normal wound healing process. Remifentanil can prevent the inflammatory response and can suppress inducible nitric oxide synthase expression in a septic mouse model. After cardiopulmonary bypass for coronary artery surgery, remifentanil can also inhibit the release of biomarkers of myocardial damage. Here we investigated whether remifentanil pretreatment has cellular protective effect against hypoxia-reoxygenation in HaCaT human keratinocytes, if so, the role of apoptosis and autophagy on this phenomenon. Methods: The HaCaT human keratinocytes were exposed to various concentrations of remifentanil (0.01, 0.05, 0.1, 0.5 and 1 ng/ml) for 2 h before hypoxia (RPC/HR group). These cells were cultured under 1% oxygen tension for 24h at $37^{\circ}C$. After hypoxia, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at $37^{\circ}C$. 3-MA/RPC/HR group was treated 3-methyladenine (3-MA), autophagy inhibitor for 1h before remifentanil treatment. Cell viability was measured using a quantitative colorimetric assay with thiazolyl blue tetrazoliumbromide (MTT, amresco), showing the mitochondrial activity of living cells. To investigate whether the occurrence of autophagy and apoptosis, we used fluorescence microscopy and Western blot analysis. Results: The viability against hypoxia-reoxygenation injury in remifentanil preconditioning keratinocytes were increased, and these cells were showed stimulated expression of autophagy 3-MA suppressed the induction of autophagy effectively and the protective effects on apoptosis. Atg5, Beclin-1, LC3-II and p62 were elevated in RPC/HR group. But they were decreased when autophagy was suppressed by 3-MA. Conclusions: Remifentanil preconditioning showed the protective effect in human keratinocytes, and we concluded that autophagy may take the major role in the recovery of wound from hypoxia-reoxygenation injury. We suggest that further research is needed about the cell protective effects of autophagy.

• Journal of Periodontal and Implant Science
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• v.31 no.1
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• pp.149-165
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• 2001

This study was performed to define the cytotoxicity and the anti-inflammatory action of Pulsatilla koreana extracts. To analyze cytotoxic effects, gingival and periodontal ligament fibroblasts were used, and anti-inflammatory actions related to reduction of $IL-1{\beta}$ and $PGE_2$ production were performed in vitro, for the suggestion of efficacy and safety on periodontal therapeutic use of Pulsatilla koreana extracts. We extracted ethylacetate and butylalcohol from well-dried and ground Pulsatilla koreana throughout multiple processing, then used different concentration solution(0.1 %, 0.2 %, 0.4 %, 0.01 %, 0.02 %, 0.04 %, 1 %, 2 %) of ethylacetate and butylalcohol extracts to examine eytotoxic effects and anti-inflammatory actions Cytotoxic effects were examined by ELISA reader using MTT(Methyl Thiazol-2-YL-2, 5-diphenyl Tetrazolium bromide)solution following culture of human gingival and periodontal ligament fibroblasts. Synthesis of $IL-1{\beta}$was examined by $IL-1{\beta}$ enzyme-immunoassay(EIA)system after separation and culture of monocyte, and $PGE_2$ was examined by $PGE_2EIA$ system after culture of gingival fibroblasts. The results were as follows: 1. In the MTT test of gingival fibroblasts, the change of optical density was decreased significantly at 2 % of butylalcohol extracts and 0.04 %, 0.1 %, 0.2 %, 0.4 %, 1 %, 2 % of ethylacetate extracts.(p<0.05) 2. In the MTT test of periodontal ligament cells, the change of optical density were not differ significantly. but butylalcohol and ethylacetate extracts except from butylalcohol 0.01 % showed high cell cytotoxity. 3. Both ethylacetate and butylalcohol extracts from Pulsatilla koreana inhibited the synthesis of $IL-1{\beta}$and inhibition effect of ethylacetate extracts were higher than butylalcohol extracts. 4. Both ethylacetate and butylalcohol extracts from Pulsatilla koreana inhibited the synthesis of $PGE_2$, and ethylacetate extracts were higher than butylalcohol extracts. In conclusion, ethylacetate and butylalcohol extracts from Pulsatilla koreana showed little cell cytotoxity for gingival and periodontal ligament fibroblasts, and the inhibition of $IL-1{\beta}$ and $PGE_2$ sysnthesis, therefore it is considered that these extracts can be developed as the therapeutics of the periodontal disease.

• The korean journal of orthodontics
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• v.23 no.4 s.43
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• pp.503-527
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• 1993

This paper describes a new method to create an animal model for TMJ internal derangement in the New Zealand white rabbits and the light and electron microscopical changes of posterior attachment of them. Twenty six rabbits(2.5-3.0kg), four normal and twenty two experimental, were used. The right disc of experimental animal was displaced anteriorly without sectioning the posterior attachment and tied to the zygomatic arch with nylon not to be reduced to the original position. The left TMJ was sham-operated to be compared with its right experimental one. Normal animals were sacrificed one day and eight weeks after experiment. Experimental animals were sacrificed one day, ten days, three weeks, five weeks and eight weeks after surgery respectively. They were fixed intravenously with $2\%$ glutaldehyde under general anesthesia and the samples of them were processed for light and electron microscopic examination. The purpose of this experiment is to make a suitable animal model of disc displacement without reduction for studying and understanding the cellular and morphologic events in posterior attachment of TMJ including early changes which were difficult to be observed in human TMJs. The results of this investigation suggest the following conclusions : 1. Authors induced anterior disc displacement surgically in rabbits with new method to examine histologic changes of posterior attachment. Tissue reactions of this model seem to be similar to those observed in human disc displacement. We think this animal model for anterior disc displacement may be used to explore and evaluate objectively the effects of many treatment modalities in disc displacements. 2. The animal disease model showed inflammation at early stage(one and ten days). At this stage there were mild-to-severe mononuclear inflammatory cell infiltration, numerous newly formed vessels, vessel dilatation and engormement and many fibroblasts. 3. At middle stage(three weeks), fibrosis occurred, where fibroblasts decreased in number, but their cytoplasm was profuse indicating high activity. Collagen fibers increased in number and the tissue looked more dense. 4. At late stage(five weeks and eight weeks) showed degenerative changes including perforation of posterior attachment, disintegration of collagen fiber bundles, degeneration of fibroblasts, metastatic ossification, and dystrophic calcification.

• KSBB Journal
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• v.10 no.2
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• pp.212-220
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• 1995

An obligatory type II methanotroph Methylosinus trichosporium OB3b was cultivated on methanol as a sole carbon and energy source. The effects of methanol concentration, pH, temperature, nitrogen source and phosphate concentration on cell growth were investigated and the results were compared with the growth on methane, which had been studied previously. When $(NH_4)_2SO_4$ was used as a nitrogen source, the maximal specific growth rate (${\mu}max$) on methanol was $0.20hr^{-1}$ and the carbon conversion efficiency(CCE) was 43%. In comparison, on methane, ${\mu}max$ and CCE were $0.08hr^{-1}$ and 32%, respectively. Ammonia was found to be a better nitrogen source for methanol-growing cells. Cell yield on nitrogen (YX/N) was the same regardless of nitrogen source as 7.14g dry cells/g N, but the yield on methanol(YX/N) was higher with ammonia(0.8g dry cells/g MeOH) than with nitrate(0.64g dry cells/g MeOH). Optimal pH and temperature were 7.0 and $30^{\circ}C$, respectively. Methanol inhibition on cell growth was observed at above 0.5%(v/v). Inhibition by phosphate was observed at above 60mM, although the inhibition on methanol dehydrogenase activity started at a much lower level of 20mM. Based on the experimental findings, the cellular physiology of M. trichosporium OB3b growing on the two closely-related carbon sources were discussed extensively.

• Korean journal of applied entomology
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• v.42 no.4
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• pp.353-360
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• 2003

Immunodepression can be required for entomopathogenic bacteria to induce their potent pathogenicities to the target insects. Here, we raise a hypothesis that the capacity of a pathogenic bacterium to induce the target insect immunodepression has positive relationship with the degree of pathogenicity. X. nematophilus had 1,200 times as potent as another entomopathogenic bacterium, Staphylococcus gallinarum against the fifth instar larvae of silkworm, Bombyx mori, when they were Injected into the hemocoel. Although both bacteria had significant cytotokic effect on the hemocytes of B. mori, X. nematophilus gave faster and greater cytotoxicity than did S. gallinarum. In cellular immune reactions, B. mori could form 20 hemocyte nodules against the bacterial injection with 5${\times}$10$\^$5/ cells. The number of the hemocyte nodules was significantly depressed when live X. nematophilus was inject-ed, but not in S. gallinarum. Activation of prophenoloxidase (proPO) was depressed in the bacterial injection. The depression of PO activation was significantly greater in X. nematophilus infection than in S. gallinarum injection. Lysozyme activity was induced by the injection of S. gallinarum at 4 h after the treatment, but not induced in X. nematophilus at all the time. These results showed that X. nemato-philus induced greater immunodepression against B. mori and resulted in higher pathogenicity than did S. gallinarum. Therefore, this study suggests that the immunodepression induced by entomopathogenic bacteria has positive relationship with their pathogenicity.

• Journal of the korean academy of Pediatric Dentistry
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• v.33 no.4
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• pp.587-596
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• 2006

Most of oral streptococci express the Antigen I/II (AgI/II) proteins, cell wall anchored adhesions. AgI/II protein binds to salivary agglutinin glycoprotein, a component of tooth pellicle and to ligands in other bacteria. These associations play important roles in bacterial colonization. Recently, it was reported that diverse host molecules also interact with AgI/II protein and that these interactions induce inflammatory responses from host cells. Among mutans streptococci containing -type hemolytic activity, Streptococcus mutans is a causative agent for dental caries. Compared with many other strains of S. mutans, GS-5 strain is unique in that this bacterium expresses truncated secretory AgI/II protein due to the nonsense mutation in the agI/II gene. This indicates that S. mutans GS-5 has a different clinical role and a recent report supported this idea based on the results from clinically isolated S. mutans strains. Previously, we had cloned agI/II gene from S. mutans GS-5 and generated recombinant N-terminal AgI/II protein. In this study, we further produced a hybridoma line expressing anti-AgI/II monoclonal antibodies named as 1C11A. This antibody showed high sensitivity to AgI/II protein in Western blot and ELISA. This new reagent will provide a basis for investigating the mechanisms of AgI/II-related diseases.

• Environmental Mutagens and Carcinogens
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• v.22 no.2
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• pp.65-69
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• 2002

To elucidate the effect of microsphere coinjection on the administration of oligodeoxynucleotides (ODN), we have investigated biodistribution of ［S-35］-labeled antisense ODN targeted to cAMP-dependent protein kinase (PKA) RI-$\alpha$ subunit in nude mice xenografted with WiDr (human colon cancer, ATCC CCL218). The strategy of using microsphere has been proposed for cancer treatment as a carrier of therapeutic ODN so that it could offer an advantage with respect to maintaining constant ODN levels in blood and obtaining higher therapeutic ODN concentration at tumor sites. Comparative biodistribution studies were performed in nude mice (female, 20 g of body weight, n = 4-6) xenografted with WiDr cancer cells, when 0.1 $\mu$Ci (specific activity, 2.94 mCi/$\mu$mole) of ［S-35］-labeled RI-$\alpha$ antisense ODN was injected alone or with microsphere (PLG-18, polylactic copolymer with cationic surfactant DDAB18). Peak tumor uptake of ［S-35］-labeled ODN was significantly increased from 17.7% (at 6 h) of injected dose per gram of tissue (ID/g) to 42.5% (at 24 h) ID/g when microsphere was coinjected with ODN. The different biodistribution in the kidney accumulation (e.g., 100.2% ID/g for ODN alone and 54.9%/ID/g for microshpere coinjection) may contribute to higher blood concentration (e.g., 21.5%ID/$m\ell$ for ODN alone and 37.5%ID/$m\ell$ for microsphere coinjection) of radiolabeled ODN. Of importance is the fact that the whole body retention of radioactivity increased with microsphere coinjection from 50.8%ID/g to 68.0%ID/g after 24-h of injection. This decreased kidney accumulation and increased whole body retention of ［S-35］-labeled ODN resulted in a significant improvement of ODN targeting to the tumor site. In conclusion, the coinjection of microsphere appears to be an important carrier system in vehiculation of antisense oligonucleotide to the tumor tissue in vivo.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.12
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• pp.1815-1819
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• 2009

Based on the soft and rigid extents, tissues are mainly divided into two groups in mammals, soft tissues including heart, lung, kidney and brain, and hard tissues including tendon, cartilage, teeth and bone. Among various tissues, bone, a dynamic rigid organ, is continuously remodeled by the opposing functional activity between bone formation by osteoblasts and bone destruction by osteoclasts. Bone protects the soft tissues and provides mineral reservoirs, which can supply the mineral needs of other soft tissues to normally maintain cellular function. While calcification in bone is an important action to fundamentally support the body and protect the soft tissues, calcification in soft tissues, including the heart, aorta, kidney, lung and spleen, results in severe organ damages, eventually causing sudden death. A growing body of evidence indicates that the osteoporotic patient who are aging, post-menopausal, diabetes and chronic kidney disease simultaneously represent a high clinical incidence of soft tissue calcification, illustrating a link between soft tissue calcification and bone decalcification (osteoporosis). This study will review what is currently known about the connection between bone decalcification and soft tissue calcification.

• Biomedical Science Letters
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• v.5 no.1
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• pp.85-93
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• 1999

Nitric oxide (NO) plays an important role in immunologic defense, and influences upon the functioning of secretory tissues and cells. It also exhibits cytotoxic/cytostatic activity as one of major operating effectors of the cellular immunity system. We investigated the effects of serum on the cell damages and NO production in the mouse liver cell line BNL CL.2 to establish the role of NO. We observed that, when BNL CL.2 cells were cultured in serum-free medium, they were induced to cell damage by the stimulation of IFN-$\gamma$ alone or IFN-$\gamma$ plus LPS. Serum-starved cells showed large amount of nitrite accumulation and NO synthase (NOS) expression in response to IFN-$\gamma$ alone in dose- and time- dependent manners, but serum-supplied cells did not The production of NO was blocked by protein tyrosine kinase (PTK) inhibitors, genistein and herbimycin. These results suggest that the deprivation of serum in the BNL CL.2 cell culture medium might primed with the cells to produce NO when the cells are triggered by IFN-$\gamma$ and the involvement of PTK signal transduction pathway in the expression of NOS gene in murine hepatocytes.

• Molecules and Cells
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• v.37 no.7
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• pp.547-553
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• 2014

Glioblastoma multiforme (GBM) is one of the most common brain malignancies and has a very poor prognosis. Recent evidence suggests that the presence of cancer stem cells (CSC) in GBM and the rare CSC subpopulation that is resistant to chemotherapy may be responsible for the treatment failure and unfavorable prognosis of GBM. A garlic-derived compound, Z-ajoene, has shown a range of biological activities, including anti-proliferative effects on several cancers. Here, we demonstrated for the first time that Z-ajoene specifically inhibits the growth of the GBM CSC population. CSC sphere-forming inhibition was achieved at a concentration that did not exhibit a cytotoxic effect in regular cell culture conditions. The specificity of this inhibitory effect on the CSC population was confirmed by detecting CSC cell surface marker CD133 expression and biochemical marker ALDH activity. In addition, stem cell-related mRNA profiling and real-time PCR revealed the differential expression of CSC-specific genes, including Notch, Wnt, and Hedgehog, upon treatment with Z-ajoene. A proteomic approach, i.e., reverse-phase protein array (RPPA) and Western blot analysis, showed decreased SMAD4, p-AKT, 14.3.3 and FOXO3A expression. The protein interaction map (http://string-db.org/) of the identified molecules suggested that the AKT, ERK/p38 and $TGF{\beta}$ signaling pathways are key mediators of Z-ajoene's action, which affects the transcriptional network that includes FOXO3A. These biological and bioinformatic analyses collectively demonstrate that Z-ajoene is a potential candidate for the treatment of GBM by specifically targeting GBM CSCs. We also show how this systemic approach strengthens the identification of new therapeutic agents that target CSCs.