• 한국식품과학회지
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• 제54권2호
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• pp.134-140
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• 2022

다시마 추출물은 인체의 중금속 제거 효과 및 항암효과가 있다고 알려져 있다. 현재 국내에는 고령화, 야외활동의 증가, 오존층 파괴 등으로 인하여 흑색종 발병률이 증가하고 있다. 흑색종의 치료는 외과 수술적 제거를 원칙으로 하고 항암요법이나 방사선 치료, 면역요법 등이 보존적인 방법으로 사용되고 있다. 하지만 재발의 확률이 높고, 항암치료제에 대한 저항성을 나타내는 경우가 많아 치료에 어려움이 있다. 본 연구에서는 다시마 추출물이 B16-F0에서 항암효과를 확인하고 또한 어떠한 경로를 통하여 항암효과를 나타내는지 밝히기 위한 분자적 기전을 연구하였다. 다시마 추출물은 B16-F0의 증식을 억제하고 세포독성을 유도하였다. 그리고 세포의 군집형성을 억제하고, DNA 분절을 일으키며, 세포사멸을 일으키는 것으로 나타났다. 이러한 다시마 추출물의 흑색종 사멸효과를 유도하는 분자적 기전을 확인한 결과, 내인성 세포사멸 경로인 cytochrome c를 증가시켜 caspase-9 활성화하였고, 외인성 세포사멸 경로인 FADD를 증가시켜 caspase-8을 활성화하였다. Caspase-9의 활성화와 caspase-8의 활성화는 caspase-3를 활성화 시켜 결과적으로 PARP를 활성화하여 세포사멸을 유도하였다. 본 연구의 제한점으로는 단일물질이 아닌 추출물로 인한 다시마 추출물의 정확한 농도를 구하기 어려운 한계가 있으며 추후 연구에서 보완되어야 하는 부분이라 생각된다. 그럼에도 불구하고 본 실험에서 다시마 추출물은 내인성과 외인성 세포사멸 경로를 활성화시켜 흑색종에 독성을 가지며, 증식을 억제하고 세포사멸을 유도하는 것으로 나타났다. 다시마는 갈조류에 속해 있으며, 다양한 성분들이 포함되어 있는데 그중 fucoidan이 항암효과를 가지는 것으로 알려져 있다(Kang 등, 2006). Fucoidan과 관련한 흑색종의 연구는 미백 연구가 대부분이기 때문에, 이후 연구에서는 in vitro 및 in vivo에서 흑색종에 대한 fucoidan의 항암효과 검토 등 다양한 데이터들이 축적된다면 향후 다시마 추출물이 흑색종의 치료에 사용될 가능성이 있을 것으로 생각된다.

• 대한소아치과학회지
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• 제24권1호
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• pp.204-219
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• 1997

The use of fluoride is one of the most effective methods for caries prevention. Fluoridation of public water supply has been recognized, for many years, as an effective way to reduce dental caries. The fluoride supplement has been recommended when the natural fluoride was unavailable or below the optimal range. However the mechanism of caries prevention by fluoride has not yet been clarified and it is well known that an overdose of fluoride results inacute and chronic toxicity, especially dental fluorosis. Fluoride mouthrinsing solution is widely used in dentistry due to its effectiveness in carrying anticariogenic action. Understanding the effects of fluoride mouthrinsing solution on human gingival fibroblasts will provide the safety rationale for its use during the caries preventive therapy. The purpose of this study was to evaluate the cytotoxic effect of fluoride mouthrinsing solution on the human gingival fibroblast in vitro. The human gingival fibroblasts were cultured from healthy gingiva on the extracted deciduous teeth of children. Cells were inoculated into a 24-well plate with $1{\times}10^4cells/well$ of medium at $37^{\circ}C$, 100% humidity, 5% $CO_2$ incubator for 24 hours. And the cells were counted by using the hemocytometer at each designed study. Human gingival fibroblasts were cultured in growth medium after one minute application range of 0.02%-0.2% NaF solution and 0.1% $SnF_2$ solution. The cells used in this study were between fifth to eighth passage number. The cell morphology was examined by inverted microscope and cell proliferation was measured by incorporating $[^3H]$-thymidine into DNA. DNA synthesis by human gingival fibroblasts was assessed by $[^3H]$-thymidine uptake assays while the cell activity was measured by MTT assay. Each concentrated fluoride mouthrinsing solution was estimated for its biocompatability with fibroblasts by the tissue culture technique. The results of this study were as follows : 1. It was observed that at 0.05%, 0.2% NaF mouthrinsing solution the cytoplasmic processes became globular. When 0.1% $SnF_2$ mouthrinsing solution was applied, the cytoplasmic process and cell morphology were disappeared. 2. DNA synthetic activity was reduced regardless of the concentration of the fluoride mouthrinsing solution. However, the result is statistically insignificant except 0.1% $SnF_2$ mouthrinsing solution(p<0.05). 3. Our results indicate that 0.02%, 0.05% concentrations of NaF mouthrinsing solution caused minimal cytotoxicity. But 0.2% NaF and 0.1% $SnF_2$ concentration were a significant difference between the cell activity in the experimental group and control group (p<0.05). 4. After appling 0.05% & 0.02% NaF fluoride mouthrinsing solution, cell activity was restored to the control groups level according to incubating time. The results suggest that direct exposure to fluoride solution inhibits gingival fibroblast activity. Therefore, for the most effective use of fluoride use, lowering the concentration of fluoride mouthrinsing is advisable because it maintains biocompatability and free ion in the oral fluid.

• 대한소아치과학회지
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• 제42권4호
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• pp.291-301
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• 2015

본 연구의 목적은 다양한 MTA 재료(ProRoot MTA, RetroMTA, Endocem Zr)에 대한 유치 치수 세포의 생활력 및 분화능을 비교 평가하는 것이다. 유치 치수세포는 각 재료별로 경화된 원형 디스크를 이용하여 직접법 및 간접법으로 세포 생활력을 관찰하였다. 또한 재료별 추출물을 이용하여 pH를 측정하였으며, alkaline phosphatase(ALP) 활성도 및 Alizarin Red S 염색법을 통하여 세포의 분화능을 관찰하였다. 직접법에서 유치 치수세포는 ProRoot MTA와 RetroMTA에서 Endocem Zr에 비해 높은 세포 생활력을 보였으나, 반면 간접법에서는 Endocem Zr에서 다른 재료에 비해 높은 세포 생활력이 관찰되었다. pH의 경우 Endocem Zr가 다른 두 재료에 비해 낮은 알칼리성을 나타냈다. 모든 재료에서 ALP 활성도는 대조군에 비해 증가하지 않았으며, Alizarin Red S 염색결과 유치 치수세포의 분화능이 대조군에 비해 낮았다. 본 실험에서 재료별 차이는 있었으나 모든 재료에서 어느 정도의 세포 독성이 관찰되었으며, 유치 치수세포의 생활력과 분화능을 증진시키지 못하였다. 하지만 Endocem Zr의 경우 ProRoot MTA나 RetroMTA에 비해 낮은 알칼리성과 높은 생활력을 보였다.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.756-766
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• 2005

The signaling mechanism underlying aburatubolactam C-induced FasL upregulation was investigated in human Jurkat T cells. After treatment with aburatubolactam C, the src-family PTKs $p56^{lck}\;and\;p59^{fyn}$, and MAP kinases ERK2 and JNK1, were activated prior to FasL upregulation; Both $p56^{lck}\;and\;p59^{fyn}$ were directly activated 2.4- and 2.2-fold, respectively, in vitro by aburatubolactam C. The aburatubolactam C-induced cellular changes, including the activation of ERK2 and INK1, and FasL upregulation, were completely prevented by the PTK inhibitor genistein. The activation of protein kinase C (PKC$\delta,\;\epsilon\;and\;\mu$ was also induced following aburatubolactam C treatment. Although the activation of $p56^{lck}$ and tyrosine phosphorylation of the cellular proteins were not blocked by the PKC inhibitor GFl09203X, the activation of ERK2 was completely abrogated, along with a detectably enhanced JNK1 activation; FasL upregulation, and apoptosis. However, the FasL upregulation and apoptosis were significantly inhibited by the PKC activator PMA, with a remarkable increase in the ERK2 activation. The cytotoxic effect of aburatubolactam C was reduced in the presence of the anti-Fas neutralizing antibody ZB-4. Although ectopic expression of Bcl-2 failed to completely block the cytotoxicity of aburatubolactam C, it was clearly suppressed. The c-Fos mRNA expression was upregulated in a biphasic manner, where the second phasic expression overlapped with the FasL upregulation. Accordingly, these results demonstrate that aburatubolactam C-induced apoptosis is exerted, at least in part, by FasL upregulation dictated by activation of the PTK ($p56^{lck}\;and\;p59^{fyn}$) /JNKI pathway, which is negatively affected by the concurrent activation of the PKC/ERK2 pathway proximal to PTK activation.

• 한국수산과학회지
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• 제42권5호
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• pp.434-441
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• 2009

This study was conducted to compare the biological activities of 7 melania snails from the family Pleuroceridae (Semisulcospira coreana, Koreanomelania nodifila, Semisulcospira forticosta, Koreoleptoxis globus ovalis, Semisulcospira libertina, Semisulcospira tegulata and Semisulcospira gottschei) in Korea. Among the 7 species, S. coreana, Korean. nodifila, S. forticosta and S. gottschei showed over 80% cytotoxicities on three cancer cell lines (SNU-1, A549 and Hep 3B) compared to the non-treatment, whereas S. libertina and S. tegulata showed almost no growth inhibition activities on the same cancer cell lines. In relation to ACE inhibition activity, only S. coreana, Korean. nodifila, and S. forticosta showed over 60% ACE inhibition activities, whereas other melania snails exhibited inhibition activities of lower than 25%. DPPH radical scavenging activities were also determined, and used to categories melania snails into three groups based on Duncan's multiple range test at P<0.05. The amount of TNF-${\alpha}$ produced by in vitro mouse peritoneal macrophage was determined according to bioactivity on L-929 cells. Three melania snails, S. coreana, Korean. nodifila and S. gottschei, exhibited 95.2%, 89.7% and 93.7% cell death(%) on L-929 cells, respectively. Glucose-6-phosphate dehydrogenase inhibitory activity was also obtained in the extract of S. coreana (31.9%) and Korean. nodifila (28.1%), showing that these extracts can be used as supplemental dietary health foods. In conclusion, we believe that the extracts of melania snails should be given due consideration in functional health food development.

• 혜화의학회지
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• 제22권1호
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• pp.145-170
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• 2013

Objectives : This study was carried out to investigate the anti inflammatory and anti allergy effects of Vitex rotundifolia Linne fil. extract(VRE). Results : 1. In vitro test, VRE was used to determine the modulation of cytokine secretion, the activation of inflammatory and allergic factor and the inhibition of gene expression. The cell survival rate of Raw 264.7 and Jurkat T cells didn't decrease and accordingly cytotoxicity wasn't observed. In anti-allergic assay, the secretion of IL-2, TNF-${\alpha}$, IL-4, IL-5 and IFN-${\gamma}$ were suppressed on Jurkat T cells induced by dust mites. And the gene expression of COX-2 was suppressed in HMC-1 stimulated by calcium ionophore A23187. In anti-inflammatory assay, the gene expression of TNF-${\alpha}$, COX-2 were suppressed on LPS-activated Raw 264.7 cells. And the secretion of IL-6 and IL-8 were suppressed on EoL-1 cells induced by dust mites. P38 and ERK activation of MAPK decreased generally. VRE showed potent inhibitory activity of NO production. 2. In vivo test, we used NC/Nga mouse induced by atopic dermatitis to observe the effects of VRE on the weight, water and feed, blood test, weight of organs, total IgE and histological change of main organs. Quantity of water and feed were not changed, therefore it didn't affect the weight directly, and no change was observed in related main organs, thus maybe there is no organ toxicity by test substances. And the symptoms were decreased significantly, and the thickness of epithelial cell layer and the number of mast cells were inhibited significantly by the difference of dosage. The number of total complete blood cells and IgE in serum were not changed significantly. Conclusion : These results suggest that VRE has anti-inflammatory and anti-allergic effects. Therefore VRE could be used effectively on improvement or treatment of atopic dermatitis. However, further study is needed to prove which component of VRE indicates effective pharmacological action.

• Journal of Periodontal and Implant Science
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• 제33권1호
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• pp.1-11
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• 2003

Hydroxyapatite(HA) has been extensively used as bone graft materials and tooth implant surface coating materials because of its biocompatibility and osteoconductive properties. However, as HA is intrinsically poor in mechanical properties, zirconia($ZrO_2$) was incorporated with HA as reinforcing phases for improvement of mechanical properties. The purpose of this study was to investigate the biological activities of HA-coated zirconia through the cell proliferation test, measurements of alkaline phosphatase activity, and histologic examination. Four kinds of tested blocks were prepared according to the pore size (300-500${\mu}m$/500-700${\mu}m$) and the porosity (70%/90%). Cell proliferation and alkaline phosphatase activity was measured at 1, 7, 14 days. The number of cells proliferate after 7, 14 days were significantly increased in all groups when compared with that of the first day, but there was no significant difference between the 4 groups at each time period. At the 7 day, alkaline phosphatase activities of cells cultured in 4 groups were higher than that of the first day, but there was no significant difference between the 4 groups at each time period. The human gingival fibroblast and MG 63 cell was used to evaluate the cell cytotoxicity using MTT test. The materials tested in the current study turned out to be non-cytotoxic. In histologic examination(SEM), at 1 day there were many cells attached on the surfaces of all kinds of tested blocks. The number of cells were increased over time. At the 14 day, there were more cells proliferated than 1 day and some of the pores of blocks were partially filled with the proliferated cells. The in vitro response of osteoblast-like cells to the HA-coated zirconia showed comparable effect on transformation comparable to hydroxyapatite.

• 한국세라믹학회지
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• 제45권10호
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• pp.618-624
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• 2008

Various calcium phosphate bioceramics are distinguished by their excellent biocompatibility and osteoconductivity. Especially, the exceptional biodegradability of $\beta$-TCP makes it a bone graft substitute of choice in many clinical applications. The activation of osteoclasts, differentiated from macrophage precursor cells, trigger a cell-mediated resorption mechanism that renders $\beta$-TCP biodegradable. Based on this evidence, we studied the biodegradation process of granular-type $\beta$-TCP bone graft substitute through in vitro and in vivo studies. Raw 264.7 cells treated with RANKL and M-CSF differentiated into osteoclasts with macrophage-like properties, as observed with TRAP stain. These osteoclasts were cultured with $\beta$-TCP nano powders synthesized by microwave-assisted process. We confirmed the phagocytosis of osteoclasts by observing $\beta$-TCP particles in their phagosomes via electron microscopy. No damage to the osteoclasts during phagocytosis was observed, nor did the $\beta$-TCP powders show any sign of cytotoxicity. We also observed the histological changes in subcutaneous tissues of rats implanted with granule-type $\beta$-TCP synthesized by fibrous monolithic process. The $\beta$-TCP bone graft substitute was well surrounded with fibrous tissue, and 4 months after implantation, 60% of its mass had been biodegraded. Also, histological findings via H&E stain showed a higher level of infiltration of lymphocytes as well as macrophages around the granule-type $\beta$-TCP. From the results, we have concluded that macrophages play an important role in the biodegradation process of $\beta$-TCP bone graft substitutes.

• Asian Pacific Journal of Cancer Prevention
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• 제14권10호
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• pp.5825-5831
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• 2013

Background/Objectives: Maintenance of cellular function in culture is vital for transfer and development following adoptive immunotherapy. Dual properties of IL-21 in activating T cells and reducing activation induced cell death led us to explore the mechanism of action of IL-21 enhanced proliferation and cytotoxic potential of CIK cells. Method: CIK cells cultured from PBMCs of healthy subjects were stimulated with IL-21 and cellular viability and cytotoxicity to K562 cells were measured. To elucidate the mechanism of action of IL-21, mRNA expression of cytotoxic factors was assessed by RT-PCR and protein expression of significantly important cytotoxic factors and cytokine secretion were determined through flow cytometry and ELISA. Western blotting was performed to check the involvement of the JAK/STAT pathway following stimulation. Results: We found that IL-21 did not enhance in vitro proliferation of CIK cells, but did increase the number of cells expressing the CD3+/CD56+ phenotype. Cytotoxic potential was increased with corresponding increase in perforin ($0.9831{\pm}0.1265$ to $0.7592{\pm}0.1457$), granzyme B ($0.4084{\pm}0.1589$ to $0.7319{\pm}0.1639$) and FasL ($0.4015{\pm}0.2842$ to $0.7381{\pm}0.2568$). Interferon gamma and TNF-alpha were noted to increase ($25.8{\pm}6.1ng/L$ to $56.0{\pm}2.3ng/L$; and $5.64{\pm}0.61{\mu}g/L$ to $15.14{\pm}0.93{\mu}g/L$, respectively) while no significant differences were observed in the expression of granzyme A, TNF-alpha and NKG2D, and NKG2D. We further affirmed that IL-21 signals through the STAT-3 and STAT-5b signaling pathway in the CIK cell pool. Conclusion: IL-21 enhances cytotoxic potential of CIK cells through increasing expression of perforin, granzyme B, IFN-gamma and TNF-alpha. The effect is brought about by the activation of STAT-3 and STAT-5b proteins.

• 한국응용과학기술학회지
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• 제40권5호
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• pp.988-1000
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• 2023

Hesperetin은 Hesperidin에서 유도되는 강한 항산화 기능의 플라보노이드 비당체이다. 본 연구에서는 Hesperetin과 이의 Cyclodextrin 포접 복합체에 대하여 항산화, 항염증 및 항균 활성을 비교하였다. Hesperetin은 Hesperidin에 효소처리하여 제조되었으며, Hesperetin/Cyclodextrin 포접체는 용매 증류법에 의해 𝛽-Cyclodextrin 및 Hydroxypropyl-𝛽-Cyclodextrin을 사용하여 제조되었다. Hesperetin에 비해 Hesperetin/Hydroxypropyl-𝛽-Cyclodextrin 포접체의 용해도는 93.5배 증가하였고, Hesperetin/𝛽-Cyclodextrin 포접체의 용해도는 22.5배 증가하였다. 항산화 분석에서 Hydroxypropyl-𝛽-Cyclodextrin 포접체는 Hesperetin과 유사한 라디칼 소거 활성능을 보인 반면, 𝛽-Cyclodextrin 포접체는 Hesperetin 보다 약간 낮은 활성을 나타내었다. RAW 264.7 세포에 대한 세포독성은 Hydroxypropyl-𝛽-Cyclodextrin 포접체, 𝛽-Cyclodextrin 포접체, Hesperetin의 순으로 세포독성이 낮았다. Hesperetin과 Cyclodextrin 포접체는 모두 세포내 산화질소(NO), 종양괴사인자-𝛼(TNF-𝛼) 및 인터루킨-6(IL-6)과 같은 염증 매개체를 감소시켰다. Hesperetin 및 Hydroxypropyl-𝛽-Cyclodextrin 포접체는 상대적으로 𝛽-Cyclodextrin 포접체 보다 더 효과적이었다. 피부 유해성 세균인 황색 포도상구균과 녹농균에 대해 억제 효과를 시험한 결과, 황색 포도상구균에 대해서는 Hesperetin = Hydroxypropyl-𝛽-Cyclodextrin 포접체 > 𝛽-Cyclodextrin 포접체의 순서로 항균 효과를 나타내었으나, 녹농균에 대해서는 뚜렷한 억제효과를 나타내지 않았다. 결론적으로, Hesperidin의 비당체 형태인 Hesperetin과 이의 Cyclodextrin 포접체는 다양한 생물학적 활성을 보여주었으며, 용해도가 높은 Hydroxypropyl-𝛽-Cyclodextrin 포접체가 𝛽-Cyclodextrin 포접체에 비해 상대적으로 더 높은 활성을 나타내었다.