• Title/Summary/Keyword: Cell viability test

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Inhibitory Effect of YBR on Hepatic Fibrogenesis (YBR의 간섬유화(肝纖維化)억제 효과(效果)에 관한 연구(硏究))

  • Seung, Hyun-Seok;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.31 no.2
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    • pp.314-330
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    • 2010
  • Objective : This study was performed to investigate the anti-fibrogenic effect and changes of inflammation-related genes by YBR I and YBR II (YBR I: Arteisiae Capillaris Herba, Atractylodis Rhizoma Alba, Hoelen/ YBR II: YBR I +Sanguisorbae Radix, Biotae Cacumen, Cirsii Japonici Herba) on HSC(hepatic stellate cells)-T6 and TAA-induced rat liver tissue. Materials and Methods : HSC-T6 were treated with various concentrations of distilled-water extract YBR I and YBR II extract for 24, 48 and 72 hours. After the treatment, cell viability, proliferation, procollagen levels and IL-6 levels were measured by using MTT Assay, BrdU Assay, Procollagen Type 1 C-peptide EIA kit, and Murine IL-6 ELISA Development kit. Rat liver fibrosis was induced by intraperitoneal TAA injection of 150mg/kg 3 times a week for 6 weeks. After the treatment, body weight, liver & spleen weights, liver function test, complete blood cell count and change of portal pressure were studied. In addition, gene expressions of ASMA, IL-6, MMP-2, TIMP-1 and TIMP-2, all of which are known to be associated with liver fibrosis, were analyzed by using Real-Time PCR. After YBR I and YBR IItreatment, percentages of collagen in TAA-induced rat liver tissue were measured. Results : The viability and proliferation of the HSC-T6 decreased as the concentration increased. The production of procollagen decreased as the concentration increased. The production of IL-6 was little influenced by YBR I and YBR II. There was no difference in rat body weight between the TAA-only group and the YBR groups. Compared with rat liver weight of TAA-only group, that of the YBR groups increased. In the YBR I group, the serum level of AST elevated by TAA injection significantly decreased and in the YBR I and II group, the serum level of ALP and ALT elevated by TAA injection decreased. In the YBR I group, white blood cell count elevated by TAA injection decreased but platelets increased. In the YBR I group, the portal pressure elevated by TAA injection significantly decreased. Decreases in the gene expression of ASMA and MMP-2 were observed in the YBR I group. The gene expression of IL-6 was little influenced by YBR I and YBR II -treated groups. In the histological finding, TAA injections caused severe fibrosis, but YBR I and YBR II treatment significantly reduced the amounts of hepatic collagens. Conclusions : These results suggest that YBR I and II have inhibitory effects on the hepatic fibrogenesis.

Physiological Activities using Root and Stem Extracts of Cymbidium (심비디움 뿌리 및 줄기 추출물의 생리 활성)

  • Kim, Hye-Ran;Park, Gyu-Nam;Jung, Bo-Kyoung;Shin, Yu-Su;Chang, Kyung-Soo
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.4
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    • pp.848-854
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    • 2016
  • Cymbidium is one of perennial herbs belonging to the Orchidaceae and is known as a medicinal plant. However, its scientific data are insufficient. The purpose of this study is to extract from root and stem of Cymbidium, to investigate the biological effects of them. Cymbidium antibacterial effects of the extracts were performed by antibacterial test against Staphylococcus aureus (S. aureus), Staphylococcus saphrophyticus (S. saprophyticus), Proteus vulgaris (P. vulgaris) and Klebsiella pneumonia (K. pneumonia). Antioxidant effects of the extracts were carried out by DPPH radical scavenging. Total phenolic contents were also determined. Moreover, Cell viability of extracts against MTT assay was cell viability against HepG2 cell and also measures Cholesterol adsorptivity of extracts. In this study, the extracts inhibited the growth of bacteria. Particularly Cymbidium root extracts by only ethanol extraction showed highest antimicrobial effect against S. aureus. The Cymbidium stem extracts by both ethanol extraction and sonication for 1 hour had higher antioxidant activites as well as total phenolic contents. Cell cytotoxicity showed higher than $50{\mu}g/mL$. Cholesterol adsorptivity showed lower than 20%. These results suggest that the Cymbidium might be a source of anti-bacterials and anti-oxidants.

Whitening Effects of Angelica dahurica Radix Ethanol Extract (백지 에탄올추출물의 미백효능 연구)

  • Kim, Pil-Sun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.12 no.9
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    • pp.4038-4045
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    • 2011
  • To investigate whitening Effects of Angelica dahurica Radix Ethanol Extract (ADEE), we used melan-a cell line, brown guinea pig, and HMB-45. We treated with ADEE of 6.25, 12.5, 25, and 50 ${\mu}g/m{\ell}$ concentration in order to evaluate the effect of ADEE on cell viability and on morphological observation of melan-a cells. Also we were induced the artificial tanning spots by 1,500 mJ/$cm^2$ of ultraviolet B radiation on the backs of brown guinea pigs (approximately 450~500g) and then the test agent of $30{\mu}{\ell}$ was applied on the spots twice a day, five days a week, for five weeks respectively. The visible whitening effect was evaluated once a week. At the end of the experiment, the animals were sacrificed under anesthetization. The artificial tanning spots were obtained by biopsy punch and stained with HMB-45 to observe the gp100 proteins which were melanosomes. Our results show that cell viability was not reduce at ADEE concentrations between 6.25 and 50 ${\mu}g/m{\ell}$, melanin synthesis and melanocyte dendricity were decreased in ADEE treated melan-a cells increasing ADEE concentration. In the gross observation, ADEE treated groups had lower pigmentation than the vehicle control groups. And in the histological observation, ADEE treated groups had lower melanocytes than the vehicle control groups. Also in the quantitative analysis of the gp100 proteins using image analysis software, ADEE treated groups had a significantly lower value (p<0.001) than the vehicle control group and this resultsagreed with the results of observation under microscope. From these results, weconcluded that ADEE had positive whitening effect.

Antimicrobial Activity and Safety Test of Natural extract including Phellodendro namurense, Eucommia ulmides Oliv extracts (황벽나무, 두충나무 추출물 등을 포함한 천연 추출물의 항균력 및 안전성)

  • Noh, Daeyoung;Joe, Suehyeon;Yang, Hyunguk;Han, Donggyun;Kim, Jinhong;Kim, Donguk
    • Korean Chemical Engineering Research
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    • v.54 no.6
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    • pp.762-766
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    • 2016
  • In this study, natural extract including Phellodendro namurense, Eucommia ulmides Oliv and Prunus padus were tested for antimicrobial activity and safety. Antimicrobial activity was measured by using opportunistic microbes such as Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans. As safety test, cell viability test, single dose oral toxicity test, single dose inhalation toxicity test, repeat dose inhalation toxicity test and eye irritation test were done. From antimicrobial test, natural extract showed execellent antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, Escherichia coli. From 5 kinds of safety tests, toxicity was not observed. From experimental results, natural extract including Phellodendro namurense, Eucommia ulmides Oliv and Prunus padus showed superb safety and antimicrobial effect.

Colorimetric Evaluation of the Time-Killing Assay for Citropin 1.1, Lipopeptide Palm-KK-$NH_2$, and Temporin A

  • Baranska-Rybak, Wioletta;Dawgul, Malgorzata;Bielinska, Sylwia;Kraska, Bartlomiej;Piechowicz, Lidia;Kamysz, Wojciech
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.536-539
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    • 2011
  • Nowadays, there are a number of colorimetric techniques available for the determination of a time killing assay in a manner much easier and faster than those previously more commonly used, which were much more time-consuming and laborious colony counting procedures. Here, an attempt has been made to test the antimicrobial peptides of Citropin 1.1, Palm-KK-$NH_2$, and Temporin A on a reference strain of Staphylococcus aureus using resazurin as the cell viability reagent. Staphylococcus aureus was exposed to the test compounds over varying periods of time and the metabolic activity measured, with a profile of antimicrobial activity then established. The results are in agreement with data from previous literature, thus confirming the relevance of the application of resazurin for the testing of antimicrobial agents.

Mineralization-inducing potentials of calcium silicate-based pulp capping materials in human dental pulp cells

  • Kang, Sohee
    • Journal of Yeungnam Medical Science
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    • v.37 no.3
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    • pp.217-225
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    • 2020
  • Background: This study was performed to provide a long-term bacterial seal through the formation of reparative dentin bridge, calcium silicate-based pulp capping materials have been used at sites of pulpal exposure. The aim of this study was to evaluate the mineralization-inducing potentials of calcium silicate-based pulp capping materials (ProRoot MTA [PR], Biodentine [BD], and TheraCal LC [TC]) in human dental pulp cells (HDPCs). Methods: Specimens of test materials were placed in deionized water for various incubation times to measure the pH variation and the concentration of calcium released. The morphology of HDPCs cultured on the specimens was examined using a confocal laser scanning microscope (CLSM). Alizarin red S staining and alkaline phosphatase assays were used to evaluate mineralization-inducing potentials of the capping materials. Results: BD showed the highest calcium release in all test periods, followed by PR and TC. (p<0.05). All experimental groups showed high alkalinity after 1 day, except at 14 days. BD showed the highest cell viability compared with PR and TC after 1 and 3 days, while TC showed the lowest value (p<0.05). The CLSM analysis showed that cells were well adhered and expressed actin filaments for all pulp capping materials. Mineralization by PR and BD groups was higher than that by TC group based on alizarin red S staining. BD showed significantly higher alkaline phosphatase activity than PR and TC, while TC showed the lowest value (p<0.05). Conclusion: Within the limitations of the in vitro study, BD had higher mineralization-inducing potential than PR and TC.

Processed Panax ginseng, Sun Ginseng, Decreases Oxidative Damage Induced by tert-butyl Hydroperoxide via Regulation of Antioxidant Enzyme and Anti-apoptotic Molecules in HepG2 Cells

  • Lee, Hye-Jin;Kim, Jin-Hee;Lee, Seo-Young;Park, Jeong-Hill;Hwang, Gwi-Seo
    • Journal of Ginseng Research
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    • v.36 no.3
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    • pp.248-255
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    • 2012
  • Potential antioxidant effect of processed ginseng (sun ginseng, SG) on oxidative stress generated by tert-butyl hydroperoxide (t-BHP) was investigated in HepG2 cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and lactate dehydrogenase (LDH) leakage test demonstrated that SG dose-dependently prevents a loss of cell viability against t-BHP-induced oxidative stress. Also, SG treatment dose-dependently relieved the increment of activities of hepatic enzymes, such as aspartate aminotrasferase and alanine aminotransferase, and lipid peroxidation mediated by t-BHP treatment in HepG2 cells. SG increased the gene expression of antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase. However, high dose of SG treatment caused decrease in mRNA level of glutathione peroxidase as compared to low dosage of SG-treated cells. The gene expression of glutathione reductase was found to be slightly increased by SG treatment. In addition, SG extract attributed its hepaprotective effect by inducing the mRNA level of bcl-2 and bcl-xL but reducing that of bax. But, the gene expression of bad showed no significant change in SG-treated HepG2 cells. These findings suggest that SG has hepatoprotective effect by showing reduction of LDH release, activities of hepatic enzymes and lipid peroxidation and regulating the gene expression of antioxidant enzymes and apoptosis-related molecules against oxdative stress caused by t-BHP in HepG2 cells.

The Effects of Anti-Oxidant and Phototoxic Inhibitory Activity of Rubus coreanus Fruits (복분자의 항산화 및 광독성 억제활성에 미치는 영향)

  • Lee, In-Su;Kim, Yong-Min;Kim, Hee-Taek
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.28 no.1
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    • pp.1-10
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    • 2015
  • Objectives : Oxidation by active oxygen in the body, phototoxicity and photoallergic interest has grown, antioxidant and phototoxicity inhibiting substances for research progress. The purpose of this study was to examine the effects on antioxidant and phototoxicity of Rubus coreanus Fruits. Meterial and Methods : Hs68 cell lines using the DPPH radical scavenging activity, cytotoxic, phototoxic inhibitory activity and apoptosis were measured. Results : 1. In MTT assay, the concentrations of Rubus coreanus Fruits that were used on the test had no cytotoxicity. 2. In DPPH radical scavenging activity, the concentration of $50{\mu}g/ml$, $100{\mu}g/ml$ anti-oxidant effect of Rubus coreanus Fruits was statistically significant increased than control group in dose-dependantly. 3. In phototoxic inhibitory activity, Rubus coreanus Fruits dose-dependently increased the cell viability of Hs68 cell lines. 4. The concentration of $50{\mu}g/ml$ Rubus coreanus Fruits inhibited the enrichment of nucleus in the Hs68 cell stimulated with UVB. Conclusions : These results indicate that Rubus coreanus Fruits has anti-oxidant effects and Phototoxic Inhibitory Activity. If further study is performed, the use of Rubus coreanus Fruits will be valuable and beneficial in the therapy of skin aging and damage.

Safety evaluation of bacteriophages for application as sanitizers (박테리오파지의 살균소독제 응용을 위한 안전성 평가)

  • Park, Do-Won;Lee, Young-Duck;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.52 no.1
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    • pp.109-112
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    • 2020
  • To evaluate the safety of bacteriophages for application of sanitizer, endotoxin content and cell cytotoxicity of two Escherichia coli and four Staphylococcus aureus phages were determined. Endotoxin ratio was determined by the Limulus amebocyte lysate (LAL) assay as a test for representative biological endotoxin content. The average endotoxin average content of the 9 log PFU/mL lysate was 18.6 EU/mL and that of the 10 log PFU/mL lysate was 5.9 EU/mL, suggesting that the phage lysate was not suitable for clinical applications, but suitable for food pathogen control applications. To confirm the cell cytotoxicity of the phage lysates, MTT assay was performed using Raw 264.7 cells treated with 9 log PFU/mL phages. Results of the assay indicated that the phage lysates did not significantly decrease the cell viability (p>0.05). These results indicated that bacteriophages would be suitable as a food safety sanitizer.

Fabrication of Endothelial Cell-Specific Polyurethane Surfaces co-Immobilized with GRGDS and YIGSR Peptides

  • Choi, Won-Sup;Bae, Jin-Woo;Joung, Yoon-Ki;Park, Ki-Dong;Lee, Mi-Hee;Park, Jong-Chul;Kwon, Il-Keun
    • Macromolecular Research
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    • v.17 no.7
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    • pp.458-463
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    • 2009
  • Polyurethane (PU) is widely used as a cardiovascular biomaterial due to its good mechanical properties and hemocompatibility, but it is not adhesive to endothelial cells (ECs). Cell adhesive peptides, GRGDS and YIGSR, were found to promote adhesion and spreading of ECs and showed a synergistic effect when both of them were used. In this study, a surface modification was designed to fabricate an EC-active PU surface capable of promoting endothelialization using the peptides and poly(ethylene glycol) (PEG) spacer, The modified PU surfaces were characterized in vitro. The density of the grafted PEG on the PU surface was measured by acid-base back titration to the terminal-free isocyanate groups. The successful immobilization of pep tides was confirmed by amino acid analysis, following hydrolysis, and contact angle measurement. The uniform distribution of peptides on the surface was observed by scanning electron microscopy (SEM) and atomic force microscopy (AFM). To evaluate the EC adhesive property, cell viability test using human umbilical vein EC (HUVEC) was investigated in vitro and enhanced endothelialization was characterized by the introduction of cell adhesive peptides, GRGDS and YIGSR, and PEG spacer. Therefore, GRGDS and YIGSR co-immobilized PU surfaces can be applied to an EC-specific vascular graft with long-term patency by endothelialization.