• Title/Summary/Keyword: Cell viability test

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Synergism of Cytotoxicity Effects of Triptolide and Artesunate Combination Treatment in Pancreatic Cancer Cell Lines

  • Liu, Yao;Cui, Yun-Fu
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.5243-5248
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    • 2013
  • Background: Triptolide, extracted from the herb Tripteryglum wilfordii Hook.f that has long been used as a natural medicine in China, has attracted much interest for its anti-cancer effects against some kinds of tumours in recent years. Artesunate, extracted from the Chinese herb Artemisia annua, has proven to be effective and safe as an anti-malarial drug that possesses anticancer potential. The present study attempted to clarify if triptolide enhances artesunate-induced cytotoxicity in pancreatic cancer cell lines in vitro and in vivo. Methods: In vitro, to test synergic actions, cell viability and apoptosis were analyzed after treatment of pancreatic cancer cell lines with the two agents singly or in combination. The molecular mechanisms of apoptotic effects were also explored using qRT-PCR and Western blotting. In vivo, a tumor xenograft model was established in nude mice, for assessment of inhibitory effects of triptolide and artesunate. Results: We could show that the combination of triptolide and artesunate could inhibit pancreatic cancer cell line growth, and induce apoptosis, accompanied by expression of HSP 20 and HSP 27, indicating important roles in the synergic effects. Moreover, tumor growth was decreased with triptolide and artesunate synergy. Conclusion: Our result indicated that triptolide and artesunate in combination at low concentrations can exert synergistic anti-tumor effects in pancreatic cancer cells with potential clinical applications.

An Anti-Tumor Effects of Dioscorea bulbifera L. on Stomach Cancer Cells (황약자(黃藥子)의 위암세포(胃癌細胞)에 대(對)한 항암(抗癌) 효능(效能) 검색(檢索))

  • Ryu, Bong-Ha;Ryu, Ki-Won;Yoon, Sang-Hyub;Kim, Jin-Seong;Kim, Sung-Hee
    • The Journal of Internal Korean Medicine
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    • v.23 no.1
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    • pp.99-106
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    • 2002
  • Background : Nowadays a lot of research is based on natural substances or materials world wide since many kinds of side effects are accompanied by anti tumor chemotherapy. In Chinese medicine, Dioscorea bulbifera L is widely used to treat many kinds of cancer, but in Korea it is rarely used. Therefore, we need to scientifically identify anti tumor effects of Dioscorea bulbifera L. Objective : We aimed to identify anti tumor effects of Dioscorea bulbifera L on the stomach cancer cells through molecular biological methods. Materials & Methods : We used AGS, a stomach cancer cell from American Type Culture Collection. We injected the boiled extract of Dioscorea bulbifera L 5 ul(sample 1), 10ul(sample 2) to cultured media(ml) for 0,6,12,18,24 hours. We measured the killing effect on stomach cancer cells through Tryphan blue exclusion test and suppressive effect on viability of stomach cancer cells via MTT assay. Results : Tryphan blue exclusion test showed that each test group killed more stomach cancer cells than the controlled group with a dosage-dependent, but not significantly. MTT assay showed that each test group had a more suppressive effect on viability of stomach cancer cells than the controlled group without a dosage-independent, but not significantly. The cell cycle analysis via flow cytometry showed that the test group extended cell cycle, and there was no peak in M phase, the number of sub G1, G0, G1 phase cells increased a little, but not significantly. Conclusion : This experiment showed that Dioscorea bulbifera L. has an anti-tumor effect, but not significantly. This is in vitro experiment and basic experiment on Dioscorea bulbifera L. We hope more progressive researches on Dioscorea bulbifera L. will be conducted and its anti tumor will be more accurately identified.

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SECOND-PHASE VALIDATION STUDY OF AN ALTERNATIVE DEVELOPMENTAL TOXICITY TEST USING MOUSE EMBRYONIC STEM CELL-DERIVED EMBRYOID BODIES

  • J.-H. LEE;S.Y. PARK;C. AHN;Y.-M. YOO;C.-W. KIM;J.-E. KIM;N.R. JO;H.Y. KANG;E.-M. JUNG;K.-S. KIM;K.-C. CHOI;S.D. LEE;E.-B. JEUNG
    • The Korean Journal of Physiology and Pharmacology
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    • v.71 no.2
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    • pp.223-233
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    • 2020
  • The embryoid body test (EBT) is a developmental toxicity test method that measures the size of embryoid bodies (EBs) and the viability of mouse embryonic stem cells (mESCs) and fibroblasts (3T3 cells). The previous pre-validation study confirmed the high accuracy (above 80%) of EBT using 26 coded test chemicals. This second-phase validation study assessed the inter-laboratory reproducibility (5 chemicals in common) and predictive capacity (10 chemicals in each laboratory) test using the coded test chemicals at three laboratories. For the prediction model, the accuracy is increased when more data is accumulated. Therefore, we updated the prediction model and analyzed the results of the second year with the newly created-prediction model. Statistical analysis of the inter-laboratory reproducibility test results indicated that accuracy, sensitivity, and specificity were 87%, 78%, and 100%, respectively. The results of the statistical analysis of the predictive capacity test showed an accuracy of 80%, sensitivity of 78%, and specificity of 81%. In conclusion, the EBT can accurately classify various embryotoxicants within a short period and with relatively little effort. Therefore, EBT can be used as a good way to test developmental toxicity.

Cytotoxicity(MTT) evaluation of dental instruments made of polymers (치과용 폴리머 기구의 세포독성(MTT) 평가)

  • Choi, Eun-Mi
    • Journal of the Korea Convergence Society
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    • v.12 no.8
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    • pp.187-195
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    • 2021
  • In order to assess the cell toxicity of 10 instruments made of polymers, the MTT assay which utilizes the L-929 cell was selected. Specimens were eluted at a temperature of 37℃ for 24 hours at a rate of 4g per 20mL, RPMI 1640, and then was positively and negatively contrasted with a control test solution, in accordance with the Notification No. 2020-12 Protocols of Medical Apparatus Biological Safety from the Ministry of Drug and Food Safety. As a result of 24 hours of incubation in 37℃, 5% CO2 Incubator and assessment using an ELISA reader, the results of Intraoral camera indiciated a cellular viability of more than 70% at a 50% eluate. But, the Plastic impression tray, 3D printing tweezer, Impression disposable syringe, Dental floss holder, Hand implant scaler, Surgical retractor, Oral scanner tip, Dental mirror, and the Water pick tip all reported a cellular viability of more than 70% at a 100% eluate, which indicates that do not exhibit cytotoxicity, thus allowing it to be used in contact with the mucous membrane of the oral cavity.

A Review on Experimental Studies of Parkinson's Disease in Korean Medical Journals (파킨슨병에 대한 국내 한의학 관련 실험 연구 동향)

  • Joo, Hwan Soo;Yu, Ok Cheol;Han, Chang;Yang, Kyu Jin;Lee, Sung Yub;Moon, Hye Yeon
    • Journal of Oriental Neuropsychiatry
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    • v.28 no.3
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    • pp.145-156
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    • 2017
  • Objectives: The purpose of this study is to review experimental studies of Parkinson's Disease in Korean medical journals, which have been published since 1990. Methods: Experimental studies related to Parkinson's disease published in Korean medical journals, registered with Korea research foundation, were searched. Articles were grouped into journal of publication, subject, and publication year. Experimental study subjects and models were analysed. Results and Conclusions: Twenty-two articles about experimental studies of Parkinson's Disease in Korean medical journals were included in our review. There were 13 articles in which herbs were used for treatment, 4 articles in which bee venom was used for treatment, 4 articles in which acupuncture was used for treatment, and 1 article in which pharmacopuncture was used for treatment. Most of the studies used experimental models of MPTP-induced Parkinson's disease and MTT assay was used to analyse cell toxicity and cell viability. Pole test, rotarod test, Morris water maze test, and forced swimming test were commonly used to analyse MPTP-induced behavioral deficits in animals. Analysis on experimental studies of Parkinson's disease was categorized into the subjects of dopaminergic cell protective effect, cell-apoptosis protectiveness, anti-inflammatory effect, and anti-oxidative effect.

Platycarya strobilacea S. et Z. Extract Has a High Antioxidant Capacity and Exhibits Hair Growth-promoting Effects in Male C57BL/6 Mice

  • Kim, Eun Jin;Choi, Joo Yeon;Park, Byung Cheol;Lee, Bog-Hieu
    • Preventive Nutrition and Food Science
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    • v.19 no.3
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    • pp.136-144
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    • 2014
  • This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This study was conducted to evaluate the effects of Platycarya strobilacea S. et Z. (PSE) extract on mouse hair growth and to determine the mechanism of action of PSE. PSE was purchased and its antioxidant activities, such as electron donating ability, total polyphenol content, and flavonoid content were tested. Toxicity during topical treatment was determined by the CCK-8 assay, a cell viability test. Fifteen 4-week-old male C57BL/6 mice were assigned to receive one of three treatments: dimethyl sulfoxide (negative control), minoxidil (positive control) or PSE. Test materials were topically applied to the shaved dorsal skin of each mouse daily for 3 weeks. After 21 days, we observed skin tissue hair follicle morphology and length, mast cell number, and stem cell factor (SCF) expression using hematoxylin and eosin (H&E), toluidine blue, and immunohistochemical staining, respectively. Furthermore, the expression of cytokines involved in hair growth [i.e., insulin-like growth factor (IGF)-1, keratinocyte growth factor (KGF), and transforming growth factor (TGF)-${\beta}1$] was determined by PCR. PSE was found to have very high antioxidant activity. The cell viability rate of PSE-treated mice was markedly higher than that of mice in the control group. We also observed an increase in hair follicle length, strong SCF staining, and a decrease in mast cell number in the PSE group. In addition, PSE-treated mice had higher IGF-1 and KGF expression and lower TGF-${\beta}1$ expression than mice in the minoxidil-treated group. These results suggest that topical application of PSE promotes hair growth by intensifying SCF, suppressing mast cell production, and increasing hair growth-promoting cytokine expression.

In vitro Alternative Study of Phototoxitiy by Neutral Red Uptake Assay Using Human Fibroblast (사람유래의 섬유아세포 배양을 이용하여 NRU(neutral red uptake) 시험법에 의한 in vitro 광독성 대체시험연구)

  • 이종권;이은희;김형수;홍진태;류승렬;박기숙;김대병;김부영;조대현
    • Toxicological Research
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    • v.16 no.1
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    • pp.77-82
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    • 2000
  • This study was conducted to assess a possible alternative method as replacements for in vivo test. The human fibroblasts were exposed to several photoxic chemicals (promethazine, neutral red, chlortetracyclone, amiodatone, bithional, 8-methyooxypsorale) and non-phototoxi substance, ammonium laureth sulfate and irradiatied with 5 J/$cm^2$ of UVA (3320~420nm). The cell viability was measured by NRU (neutral red uptake) assay. The photoxic potential of test chemicals in the NRU PT (phototoxicity test) was assessed by determining the PIF (photoirritancy Factor) by using a cut-off value of 5. The NRU PT responses of most chemicals showed a close agreement with in vivo response except bithinol. There was a relatively good agreement between in vitro NRU assay and in vivo data. These results suggest that NRU assay using fibroblast could be used to predict the phototoxicity.

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Immunostimulatory effects of BCG-CWS on the proliferation and viability of mouse spleen cells (마우스 비장세포의 증식과 생존율에 대한 BCG-CWS의 면역자극 효과)

  • Lee, Che-Wook;Ko, Eun-Ju;Joo, Hong-Gu
    • Korean Journal of Veterinary Research
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    • v.52 no.2
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    • pp.89-97
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    • 2012
  • Mycobacterial cell-wall skeleton (CWS) is an immunoactive and biodegradable particulate adjuvant and has been tried to use for immunotherapy. The CWS of Mycobacterium bovis bacillus Calmette-Guerin (BCG-CWS) was studied as an universal vaccine vehicle for antigen conjugation, to develop potentially effective and safe vaccine. Although a variety of biological activities of BCG-CWS have been studied, the effects of BCG-CWS on spleen cells are not fully elucidated. Using MTT assay and trypan blue exclusion test, we found that BCG-CWS significantly enhanced the viability and proliferation of cells. Multiple clusters, indicating proliferation, were observed in BCG-CWS-treated spleen cells and surface marker staining assay revealed that BCG-CWS promoted the proliferation of $CD19^+$ B lymphocyte rather than $CD4^+$ or $CD8^+$ T lymphocyte. In addition, BCG-CWS up-regulated the expression of anti-apoptotic molecules such as bcl-2, bcl-xL. BCG-CWS increased the surface expression of CD25 and CD69 as well as IL-2 production of spleen cells, suggesting increased activation. Furthermore, BCG-CWS enhanced the antigen-specific cell proliferation and interferon-gamma production of spleen cells. Taken together, these results demonstrate the immunostimulatory effects of BCG-CWS on spleen cells via multiple mechanisms, providing valuable information to broaden the use of BCG-CWS in clinical and research settings.

Protective Effects of Sanyakbojungbangam-tang Ethanol Extracts on Cisplatin-induced Apoptosis in ECV304 Cells (혈관내피세포에서 cisplatin에 의한 세포고사에 대한 산약보정방암탕 에탄을 추출물의 방어효과)

  • Kwon Kang-Beom;Kim Eun-Kyung;Lee Young-Rae;Ju Sung-Min;Ryu Do-Gon;Kim Sung-Hoon;Jeon Byung-Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.1
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    • pp.20-24
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    • 2006
  • This study was designed to investigate the protective effect of Sanyakbojungbangam-tang Ethanol Extracts (SB Et-OH) on the cisplatin-induced apoptosis of human endothelial cell line ECV304 cells. After cells were treated with cisplatin, MTT assay was performed for cell viability test. To explore the mechanism of cytotoxicity, we used the several measures of apoptosis to determine whether this processes was involved in cisplatin-induced cell damage in ECV304 cells. Also, cells were treated with SB Et-OH and then, followed by the addition of cisplatin. Cisplatin decreased the viability of ECV304 cells in a dose-dependent manner and increased the caspase-3 enzyme activity ECV304 cells treated cisplatin were revealed as apoptosis characterized by nuclear staining. SB Et-OH protected ECV304 cells from cisplatin-induced nuclear fragmentation and chromatin condensation. Also, SB Et-OH inhibited the activation of caspase-3 pretense and the cleavage of poly(ADP-ribose) polymerase (PARP) in cisplatin-treated ECV304 cells. According to above results, SB Et-OH may protect ECV304 cells from the apoptosis induced by cisplatin.

Bone Regeneration Using Block-type Deproteinized Porcine Bone Mineral with Collagen Membrane Using 3,4-Dihydroxyphenylalanine as Bone Adhesive

  • Kang, Joo Hyun;Pae, Hyoung-Chul;Cha, Jae-Kook;Lee, Jung-Seok;Paik, Jeong-Won;Jung, Ui-Won;Choi, Seong-Ho
    • Journal of Korean Dental Science
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    • v.11 no.2
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    • pp.43-56
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    • 2018
  • Purpose: The purpose of this study was to assess the adhesiveness and cytotoxicity of 3, 4-dihydroxyphenylalanine (DOPA), and to evaluate the role of collagen membrane with DOPA in the guided bone regeneration. Materials and Methods: Peel resistance and cell cytotoxicity test were performed. Four defect types in nine rabbit calvaria were randomly allocated: i) control, ii) membrane, iii) deproteinized porcine bone mineral (DPBM) covered by membrane with DOPA, and iv) DPBM covered by membrane with cyanoacrylate. Animals were sacrificed at 2 (n=4) and 8 weeks (n=5) for microcomputed tomography and histomorphometric analysis. DOPA showed low peel resistance but high cell viability. Result: Cyanoacrylate and DOPA groups showed significantly higher mineralized tissue volume (MTV) compared to control and membrane groups at 2 weeks (P<0.05). At 8 weeks, DOPA group showed the highest MTV. Significantly higher new bone area was found in DOPA group at 8 weeks (P<0.05). Bone formation increased from 2 to 8 weeks in DOPA group (P<0.05). Conclusion: DOPA showed high cell viability and in vivo study revealed predictable performance in bone regeneration.