• Journal of Periodontal and Implant Science
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• 제41권1호
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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.

• Journal of Periodontal and Implant Science
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• 제26권2호
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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 $TGF-{\beta}1$.

• 원예과학기술지
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• 제29권1호
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• pp.23-28
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• 2011

딸기에 적합한 수경재배기술을 개발하기 위하여, 배양액 농도와 뿌리의 활성과의 상관관계를 조사함으로써 딸기에 적정한 배양액의 농도를 구명하고자 하였다. '설향(雪香)' 딸기를 재료로 하여 야마자키 조성 딸기 전용배양액을 0.5, 1.0, $2.0dS{\cdot}m^{-1}$ 농도로 처리하였다. 그리고 투명 플라스틱 포트에 코코피트를 충진하고 처리별로 5주씩 정식하여 딸기의 지상부 및 뿌리의 발달을 관찰하였다. 뿌리의 활력은 TTC(Triphenyl-tetrazoliumchloride)법으로 조사하였다. 엽병장은 EC $1.0dS{\cdot}m^{-1}$ 처리구에서 가장 길었으며, 그 다음 EC 2.0, $0.5dS{\cdot}m^{-1}$ 순으로 나타났다. 엽폭은 EC $1.0dS{\cdot}m^{-1}$ 처리구에서 가장 넓었으며, EC 0.5, $2.0dS{\cdot}m^{-1}$ 순으로 나타났다. 과장, 과경, 과중 및 수량은 EC 0.5와 $1.0dS{\cdot}m^{-1}$ 처리구에서 EC $2.0dS{\cdot}m^{-1}$ 처리구 보다 통계적으로 유의하게 높게 나타났다. 그러나, 당도는 처리 간에 유의한 차이를 나타내지 않았다. 지상부 건물중은 EC $1.0dS{\cdot}m^{-1}$ 처리구에서 가장 높았으며 다음이 $0.5dS{\cdot}m^{-1}$ 처리구, $2.0dS{\cdot}m^{-1}$ 처리구 순으로 나타났다. 지하부의 건물중은 배양액의 농도가 0.5와 $1.0dS{\cdot}m^{-1}$에서 높게 나타났으며, $2.0dS{\cdot}m^{-1}$에서는 현저하게 낮았다. 배액의 산도 변화는 EC $1.0dS{\cdot}m^{-1}$ 처리구는 공급배양액의 pH와 비슷한 경향을 나타내었으나, EC 0.5 처리구에서는 상승하는 경향을 보였고 EC 2.0 처리구에서는 현저하게 낮아지는 경향을 보였다. Formazan의 농도는 EC $1.0dS{\cdot}m^{-1}$ 처리구에서 전 생육기간 동안 가장 높게 나타났으며, EC $2.0dS{\cdot}m^{-1}$ 처리구에서는 현저하게 낮은 경향을 보였다. 이상의 결과에서 '설향'딸기에 가장 적합한 배양액 농도는 EC $1.0dS{\cdot}m^{-1}$에 가까운 것으로 생각되었다. 또한, 배액의 pH는 뿌리의 활성과 직접적인 상관이 있어서, 배액의 pH가 높은 것은 뿌리의 활성이 양호한 것을 시사하고, 배액의 pH가 낮은 것은 뿌리의 활성이 낮은 것을 나타내는 지표가 될 수 있다는 것을 확인하였다. 이러한 결과는 딸기 수경재배에서 생육진단의 지표로 유용하게 활용될 수 있을 것으로 생각된다.