• Title/Summary/Keyword: Formazan

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Effect of (-)-epigallocatechin-3-gallate on maintaining the periodontal ligament cell viability of avulsed teeth: a preliminary study

  • Jung, Im-Hee;Yun, Jeong-Ho;Cho, Ah-Ran;Kim, Chang-Sung;Chung, Won-Gyun;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.41 no.1
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    • pp.10-16
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    • 2011
  • Purpose: Avulsed tooth can be completely recovered, if sound periodontal ligament (PDL) of tooth is maintained. Although a lot of storage solutions have been explored for the better storage of avulsed tooth, there is a shortcoming that the preservation time is much short. On the other hand, there has been studies that (-)-epigallocatechin-3-gallate (EGCG), the most abundant polyphenol in green tea, which is related to the anti inflammatory, antioxygenic, and antibacterial effects, allows the successful preservations of tissues and cells. This study evaluated the effect of EGCG on avulsed-teeth preservation of Beagle dogs for a period of time. Methods: The atraumatically extracted teeth of Beagle dogs were washed and preserved with 0/10/$100\;{\mu}M$ of EGCG at the time of immediate, period 1 (4 days in EGCG-contained media and additional 1 day in EGCG-free media), period 2 (8 days in EGCG-contained media and additional 2 days in EGCG-free media) and period 3 (12 days in EGCG-contained media and additional 2 days in EGCG-free media). Then, the cell viabilities of preserved teeth was calculated by dividing optical density (OD) of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay with OD of eosin assay to eliminate the measurement errors caused by the different tissue volumes. Results: From the results, the immediately analyzed group presented the highest cell viability, and the rate of living cells on teeth surface decreased dependent on the preservation period. However, the $100\;{\mu}M$ of EGCG-treated group showed statistically significant positive cell activity than EGCG-free groups throughout preservation periods. Conclusions: Our findings showed that $100\;{\mu}M$ EGCG could maintain PDL cell viability of extracted tooth. These results suggest that although EGCG could not be a perfect additive for tooth preservation, it is able to postpone the period of tooth storage. However, further in-depth studies are required for more plausible use of EGCG.

Effects of $TGF-{\beta}1$ on Cellular Activity of Minocycline-Pretreated Human Periodontal Ligament Cells (($TGF-{\beta}$)이 Minocycline을 전처리한 사람 치주인대세포의 활성에 미치는 영향)

  • Yang, Seung-Oh;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.469-490
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    • 1996
  • The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the effect on the cell activity than $TGF-{\beta}1$ only tested group and the tested group of minocycline inhibited cell activity. This results indicate that minocycline was effective on cell attachment in early stage, but it is harmful to cell activity, that inhibitory effect of minocycline was compensated with stimulatory effect of $TGF-{\beta}1$.

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Effect of Nutrient Solution Strength on pH of Drainage Solution and Root Activity of Strawberry 'Sulhyang' in Hydroponics (배양액의 농도가 배액의 pH와 딸기 '설향' 뿌리의 활성에 미치는 영향)

  • Jun, Ha-Joon;Byun, Mi-Soon;Liu, Shi Sheng;Jang, Mi-Soon
    • Horticultural Science & Technology
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    • v.29 no.1
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    • pp.23-28
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    • 2011
  • Experiments were conducted to investigate the optimum concentration of the nutrient solution in strawberry 'Sulhyang' with hydroponics in relationship between root activity and nutrient concentrations. Nutrient solutions for strawberry, made by Yamazaki, were supplied EC 0.5, 1.0, 2.0 $dS{\cdot}m^{-1}$ during experiment period. Growth of shoot and root of strawberries grown in visible plastic pot was observed during experiment. Petiole length was longest in plants grown in EC 1.0 $dS{\cdot}m^{-1}$, followed by 2.0 and 0.5 $dS{\cdot}m^{-1}$. Leaf width was longest in plants grown in EC 1.0 $dS{\cdot}m^{-1}$, followed by 0.5 and 2.0 $dS{\cdot}m^{-1}$. Fruit length, fruit diameter, fruit weight and yield were higher in EC 0.5 and 1.0 $dS{\cdot}m^{-1}$ than 2.0 $dS{\cdot}m^{-1}$ treatment but, soluble solids of the fruit did not show statistical differences among treatments. Shoot dry weight was heaviest in EC 1.0 $dS{\cdot}m^{-1}$, followed by 0.5 and 2.0 $dS{\cdot}m^{-1}$. Root dry weight was heavier in EC 0.5 and 1.0 $dS{\cdot}m^{-1}$ but significantly light in 2.0 $dS{\cdot}m^{-1}$. pH of the drainage solution was elevated in low nutrient concentration and lowered in high concentration. Also root activity was high in low nutrient concentration and low in high concentration. As a result, the optimum EC for strawberry 'Sulhyang' was EC 1.0 $dS{\cdot}m^{-1}$ in this experiment. It was confirmed that there was high relationship between root activity and pH of drainage solution. This result will be utilized as an indicator for strawberry hydroponics.