• Journal of Periodontal and Implant Science
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• 제26권4호
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• pp.889-905
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• 1996

It is known that growth factors function as potent biologic mediators regulating numerous activities of wound healing via cell proliferation, migration and extracellular matrix formation and they also promote periodontal regeneration. But, method of growth factor application is controversial yet. So purpose of this study is to evaluate the effect of demineralized root surface as one of method of growth factor application. The ginigival fibroblasts were primary cultured and fifth or sixth subpassages were used in these experiments. In first experiment, root surface blocks demineralized with 100mg/ml tetracycline for 5 minutes and pH 1 citric acid for 3 minutes(experimental groups) and nonteminerilized root surface blocks (control groups) were placed in 100ng/ml PDGF-BB for 5 minutes. Then the cells were seeded on each root surface blocks and cultured for 6, 24, 48, 72 hours. In second experiment, root surface blocks deminerilized with tetracycline and citric acid and nondemineralized root surface blocks were placed in 200ng/ml PDGF-BB for 5 minutes and another non-demineralized root surfcae blocks were placed in DMEM without PDGF-BB. At 1, 2, 4, 6, 8 days, the cells were seeded in 24-well plate and using of each eluent, cultured for 72 hours. The results of the four determinants were presented as mean and S.D.. The results were as follows : The attachment and proliferation of human gingival fibroblast on root surface were more increased when PDGF-BB was applicated on root surfrace demineralized with tetracycline or citric acid than non-demineralized root surface. And, in comparision tetracycline with citric acid, there were more attachment and proliferation of human gingival fibroblast on root surface demineralized with tetracycline than citric acid, and proliferation of human gingival fibroblast on demineralized root surface was increased time dependently 1 day to 3 days. In second experiment using eluent, proliferation of human gingival fibroblast was more increased to 6 days when human gingival fibroblast was cultured in eluent that PDGF-BB was applicated on demineralized root surface than two control groups, and degree of proliferation was decreased time dependently 1 day to 6 days. Proliferation of human gingival fibroblast cultured in eluent without PDGF-BB was constant 1 day to 6 days. After 6 days, degree of proliferation of human gingival fibroblast was similar in four groups. This means that release duration of PDGF-BB from demineralized root surface is 6 days. And in comparision tetracycline with citric acid, there was more proliferation of human gingival fibroblast in tetracycline-treated group than citric acid. In conclusion, demineralized root surface as primary site for PDGF-BB application, especially demineralized with tetracycline has important roles in attachment and proliferation of human gingival fibroblast, and may be useful clinical applications in periodontal regenerative procedures.

• Journal of Periodontal and Implant Science
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• 제33권2호
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• pp.301-310
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• 2003

The purpose of this study is to evaluate the biocompatibility and the biorsorbability of several types of calcium polyphosphate made through change of manufacturing process for 12 month. To solve limitation of calcium phosphate, we developed a new ceramic, Calcium Polyphosphate(CPP), and report the biologic response to CPP in extraction sites of beagle dog. Porous CPP blocks were prepared by condensation of anhydrous $Ca(H_2PO_4)_2$ to form non-crystalline $Ca(PO_3)_2$ and then milled to produce CPP powder. CPP powder, CPP block, and CPP granules added with $Na_2O$ were implanted in extraction sockets and histologic observation were performed at 12 months later. Like 3 months results, histologic observation at 12 months revealed that CPP matrix were mingled with and directly apposed to new bone without any adverse tissue reaction, CPP powder show direct bony contact, but new bone formation and fibrous tissue encapsulation showed in CPP block. 10% $Na_2O$ CPP granules show more inflammatory cells infiltration around graft materials compared at 3 month, but 15% $Na_2O$ CPP granules show less. This result revealed that regardless of addition of $Na_2O$, CPP had a high affinity for bone and had been resorbed slowly. From this results, it was suggested that CPP is promising ceramic as a bone substitute and addition of $Na_2O$ help biodegradation but optimal concentration of $Na_2O$ and other additive component to increase degradation rate should be determined in further study.

• 대한한방내과학회지
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• 제37권1호
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• pp.65-76
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• 2016

Objectives: The purpose of this study was to identify the inhibitory effects of Hwangheuk-san (HHS), a Korean multi-herb formula comprising four medicinal herbs, on cell migration and invasion, two critical cellular processes that are often deregulated during metastasis, using the human bladder cancer 5637 cell line.Methods: Cell viability, motility, and invasion were assessed by 3-(4,5-dimethyl-2 thiazolyl)-2,5-diphnyl-2H-tetrazolium bromide (MTT), wound healing migration, and Transwell assays, respectively. Gene expression was detected by Western blot analysis. In addition, the activities of matrix metalloproteinases (MMPs) and the values for transepithelial electrical resistance (TER) were analyzed using a Gelatinase Activity Assay Kit and an Epithelial Tissue Voltohmmeter, respectively.Results: Our data indicated that within the concentration range that was not cytotoxic, HHS effectively inhibited the cell motility and invasiveness of 5637 cells. HHS markedly decreased the expression and activity of MMP-2 and MMP-9, which was associated with unregulation of tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2. Further investigation revealed that phosphorylation of phosphatidylinositol 3-kinase (PI3K) and AKT was decreased in HHS-treated 5637 cells, and a PI3K/AKT inhibitor synergistically reduced the inhibition of migration and invasion and also inactivated MMP-2 and MMP-9. Moreover, HHS increased the tightening of tight junctions (TJs), which was demonstrated by an increase in the TER, and reduced the expression the levels of claudin family members (claudin-3 and -4), which are major components involved in the tightening of TJs.Conclusions: The present findings demonstrated that HHS attenuated the migration and invasion of bladder cancer 5637 cells by modulating the activity of the PI3K/Akt signaling pathway and also through TJ tightening.

• Molecular & Cellular Toxicology
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• 제1권2호
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• pp.116-123
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• 2005

The application of high pressure on cellular morphology, proliferation and protein expression of Jurkat cells (human T lymphocyte cell line) has been extensively investigated. In the present study, we manufactured a novel pressure chamber that modulates 5% $CO_{2}$, temperature and pressure (up to 3 ATA). Jurkat cells was incubated 2 ATA pressure and analyzed cellular morphology and growth using an electron microscopy and MTT assay. The cells showed the morphological changes in the cell surface, which appeared to cause a severe damage in cell membrane. The growth rate of the cells under 2 ATA pressure decreased as cultured time got increased. Furthermore, a long term exposure of high pressure on Jurkat cells may act as one of the important cellular stresses that leads to inducing cell death. Cellular proteomes were separated by 2-dimensional electrophoresis with pH 3-10 ranges of IPG Dry strips. And many proteins showed significant up-and-down expressions with hyperbaric pressure. Out of all, 10 spots were identified significantly using matrix-assisted laser desorption/ionization-time of fight (MALDI-TOF) mass spectrometry. We and found that 9 protein expressions were decreased and one protein, heat shock protein HSP 60, was increased in Jurkat cells under 2 ATA. Identified proteins were related to lipid metabolism and signal transduction.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2005년도 제22차 정기총회 및 학술발표회
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• pp.66-67
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• 2005

배자 생식세포 발달에 관련된 메카니즘을 밝혀내기 위해서, 닭 배자 생식기에서 추출한 원시 생식세포의 단백질체 지도를 만들었다. 총 500 배자를 6일간 배양하여 배자 생식기를 획득했고, 7-10일 배양 후, 배양된 원시생식세포는 2차원 젤 전기 영동법에 의해 분할되어 졌다. 유의적 발현 수준을 나타낸 많은 단백질 스팟 들은 MALDI-TOP 와 LC-MS/MS에 의해 확인되었으며, 89개의 단백질 스팟 중에 50개의 mass spectra 들이 데이터베이스에서 조류 단백질과 일치함을 확인하였다. 본 실험에서 행한 단백질체 지도는 형질전환 연구와 생식세포 생물학 분야에 중요한 참고 문헌으로 가치를 가질수 있을 것이다.

• 한국재료학회지
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• 제8권3호
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• pp.268-273
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• 1998

Zircaloy-4 합금판재에 230-250ppm의 수소를 장입시키고 $400^{\circ}C$에서 72시간동안 균질하게 수소화물을 형성시킨후 $350^{\circ}C$의 static autoclave를 이용하여 여러 가지 농도의 LiOH 부식용액조건에서 부식시험을 수행하였다. 부식평가는 시간에 따른 무게증가의 변화로서 측정하였고, 시편의 미세구조는 광학현미경과 주사전자현미경을 사용하여 관찰하였다. 부식시험 후 형성된 산화막에서의 H와 Li의 분포를 확인하기 위해 secondary ion mass spectrometry(SIMS)를 이용해 각 원소의 분포를 두께방향에서 측정하였다. 여러 가지 농도의 LiOH 수용액조건에서 Li+ 이온의 용액농도가 30ppm 이상으로 증가하면 합금의 부식은 급격히 가속화되었다. 이것은 $Li^{+}$가 산화막 내의 $Zr^{4+}$ 자리를 치환함에 따라 산소공공이 증가로 산화반응이 증가되고, 이로 인해 형성되는 수소화물의 양이 증가하기 때문이다. LiOH용액조건에서 부식시험 전에 시편 내에 수소를 장입시켜 수소화물을 형성시키면 수소를 장입하지 않은 시편보다 부식이 더 빨리 가속되지만, 시험기간이 길어지면 오히려 수소를 미리 장입시키지 않은 시편의 부식속도가 더 빨라진다. 이것은 부식시험 전에 수소를 시편에 미리 장입을 시키면 이때 형성된 수소화물에 의해 초기에 부식이 빨리 가속되지만 이미 고용도 이상의 수소가 금속 내부에 존재하므로 부식과정 중에 생기는 수소가 금속의 내부로 확산되어 들어오는 것이 억제되어 부식속도가 둔화되는 것으로 생각된다.

• 대한방사성의약품학회지
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• 제1권1호
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• pp.38-45
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• 2015

Most tumors are believed to overexpress several receptors, and small peptides targeting these receptors were developed for diagnosis and tumor therapy during past decade. Here we report that fibroadenoma can be visualized by bladder cancer specific peptide. A 9-mer bladder cancer specific peptide, which was discovered from the phage display method, was synthesized by peptide synthesizer, and additional tyrosine was conjugated at the N-terminal for radioiodination (Y-BP). Y-BP was radiolabeled with $^{131/124}I$ using Iodogen tube. The rat treated with N-butyl-N-(4-hydroxybutyl)nitrosamine for 8 weeks was allowed to grow until large size tumor was developed under axilla. The tumor model was microPET imaged sequentially using [$^{18}F$]FDG and radioiodinated $^{124}I-Y-BP$. The tumor was excised and examined by immunostaining studies. Radioiodinated $^{124}I-Y-BP$ was purified using fast protein liquid chromatography (FPLC) in > 90% radiochemical purity. The whole tumor was well visualized by [$^{18}F$]FDG with several intense focal uptake within tumor. The tumor was also clearly seen with $^{124}I-Y-BP$ at 4 h post-injection, and to our surprise the tumor uptake of $^{124}I-Y-BP$ lasted up to three days. The tumor was diagnosed histologically as a fibroadenoma derived from mammary gland. In conclusion, the bladder cancer specific peptide showed the good potential as a new radiotracer for the detection of breast fibroadenoma.

• 동의신경정신과학회지
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• 제19권2호
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• pp.111-122
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• 2008

Objective : Herb medicines are potential sources of useful edible and medicinal plants. They are used as a drug because of their various biological activities such as immunomodulatory, antiviral, and antitumor functions. Nelumbo nucifera have been applied in Chinese herbal prescriptions to improve tissue inflammation. However, it has not been elucidated on the effect of the flower of Nelumbo nucifera in cells. Method : In the present study, to examine the effect of that on glioma cells, U87, the essential oil was extracted from the flower of Nelumbo nucifera (NN essential oil). U87 cells were exposed to different concentrations of 2-40 ug/ml of NN essential oil in ethanol. Cell viability was measured by MTT assay at 24 h. To find out the intracellular target signal molecule(s) for this antiproliferative activity of NN essential oil, phosphorylation of Akt, ERM, MAPK or p38 proteins were examined by Western blot analysis. To study long term effect of NN essential oil in U87 cells, the image of cells treated with NN essential oil for 4 days were obtained. Results and Conclusion : NN essential oil was shown to exhibit antitumor activity in glioma cells, at a broad range of concentrations of 10-40 ug/ml. The phosphorylation of Akt and Endoplasmic Reticulum Matrix (ERM) proteins which known to be involved in the cell death, were gradually decreased to 2 hours after addition 20 ug/ml of NN essential oil. However, the phosphorylation of mitogen-activated protein (MAPK) and p38 was found to increase in NN essential oil treated cells. NN essential oil treated cells showed decreased glioma cell number. These results provide a possible NN essential oil-induced inhibitory signal for tumor cell proliferation that is initiated by the decrease in Akt activity. Moreover, it is likely that the activation of p38 is required for the NN essential oil-induced inhibition of tumor proliferation.

• 열처리공학회지
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• 제20권2호
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• pp.84-93
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• 2007

Microstructural changes during tempering at the temperature range of $300^{\circ}C{\sim}700^{\circ}C$ for the nitrogen-permeated STS 410 and 410L martensitic stainless steels has been investigated. After nitrogen permeation at temperature between 1050 and $1150^{\circ}C$, the surface layer appeared fine $Cr_2N$ of square and rod types in the martensite matrices. Hardness of the nitrogen-permeated surface layer represented 680Hv and 625Hv, respectively, for 410 and 410L steels. It is considered that the fine homogeneously dispersive effect of precipitates by nitrogen caused the increased hardness. Due to the counter current effect of carbon from interior to surface during nitrogen diffusion from surface to interior, the 0.1%C alloyed 410 steel showed the low nitrogen content of 0.025% compared with 0.045% of 410L steel at the distance of $100{\mu}m$ from the surface. Tempering of nitrogen-alloyed 410 and 410L showed the maximum hardness at $450^{\circ}C$. This maximum hardness was considered to be the secondary hardening effect of very fine carbide and nitride. The decrease in hardness at $700^{\circ}C$ was the softening effect of the matrix due to the precipitation of many needle-shaped $Cr_2N$ for 410 steel and the precipitation of coarse nitride of $Cr_2N$ in line with the spherical precipitates with directionality for 410L steel. For 410 steel, the corrosion resistance of nitrogen permeated surface in the solution of 1 N $H_2SO_4$ were nearly unchanged, however the superior corrosion resistance was obtained for nitrogen permeated 410L steel compared to the solution annealed condition.

• Natural Product Sciences
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• 제26권1호
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• pp.83-89
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• 2020

Osteoporosis is a worldwide disease leading to significant economic and societal burdens globally. Osteoporosis is caused by unbalanced bone remodeling between the rate of osteoclast bone resorption and osteoblast bone formation. Acer tegmentosum Maxim (AT) is a traditional herbal medicine containing multiple biological activities such as anti-oxidant and anti-inflammatory purposes. However, its role in osteoporosis has not been fully studied. Therefore, we investigated whether AT has a potent inhibitory effect on osteoporosis and its mechanism through a systemic evaluation in ovariectomized (OVX) mice. OVX mice were orally administrated with the AT at doses of 50, 100, and 200 mg/kg for 10 weeks. Histological images and histomorphometry analyses were performed by H&E and Toluidine blue satin, and the expression levels of receptor activator for nuclear factor-kB ligand (RANKL), nuclear factor of activated T cells cytoplasm 1 (NFATc1), c-Fos, and matrix metalloproteinase 9 (MMP9) related to the osteoclast differentiation were investigated using immunohistochemical analysis. Administration of AT prevented bone loss and the alternations of osteoporotic bone parameters at the distinct regions of the distal femur and spongiosa region in OVX mice. Further, administration of AT increased periosteal bone formation in a dose-dependent manner. Meanwhile, AT inhibited not only the expression of NFATc1 and c-Fos, which are two major regulators of osteoclastogenesis but also reduced bone resorbed encoding expression of MMP9 and RANKL. Our results indicated that administration of AT prevented bone loss and the alternations of osteoporotic bone parameters at the distinct regions of the distal femur and spongiosa region in OVX mice. Also AT has the bone protective effect through the suppression of osteoclast and promotion of osteoblast, suggesting that it could be a preventive and therapeutic candidate for anti-osteoporosis.