• Title/Summary/Keyword: MTT-test

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Antioxidant effect of Raphani Semen (Raphanus sativus L.) (나복자의 항산화 효과)

  • Seon Been, Bak;Seung-Ho, Kang;Kwang-Il, Park;Won-Yung, Lee
    • Herbal Formula Science
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    • v.31 no.1
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    • pp.41-51
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    • 2023
  • Objectives : Raphani Semen (Raphanus sativus L.) is known for the various beneficial effects in Korean medicine. This study aimed to investigate the effect of Raphani Semen extract (RSE) against arachidonic acid (AA)+iron-induced oxidative stress in cells. Methods : Ingredients, their target information, oxidative stress liver injury-related proteins was obtained from various network pharmacology databases and software. A hypergeometric test and enrichment analysis were conducted to evaluate associations between protein targets of RSE. The cell viability was assessed by MTT assay, and immunoblot analysis was used to confirm the molecular mechanisms. Results : A compound-target network of RSE was constructed, which consisted of 336 edges between 18 ingredients and 123 protein targets. PI3K-Akt signaling pathway, ErbB signaling pathway, HIF-1 signaling pathway, PPAR signaling pathway, and AMPK signaling pathway was significantly associated with protein targets of RSE. RSE protected HepG2 cells against AA+iron-induced oxidative stress as mediated with AMPK signaling. Conclusion : RSE was found to protect the cells against oxidative stress via the AMPK signaling pathway.

Study on Anti-allergic Effect and Safety of Bangpung-galgeun-tang (방풍갈근탕(防風葛根湯)의 항알레르기효과에 관한 연구)

  • Lee, Joo-Eun;Park, Seong-Ha;Kang, Kyung-Hwa;Lee, Yong-Tae
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.5
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    • pp.1118-1126
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    • 2007
  • The purpose of this study was to examine the anti allergic effect in vivo and in vitro, and to observe single and four weeks repeated toxicity in mice of Bangpung-galgeun-tang (BGT). We investigated anti DNP IgE-mediated passive cutaneous anaphylaxis in rodents and compound 48/80-induced active systemic anaphylatic shock in mice after oral administration with BGT of 0.4 g/kg and 0.8 g/kg for 8 days, and also examined MTT assay, ${\beta}-hexosaminidase$ activity, IL-4 and $TNF-{\alpha}$ from RBL-2H3 and $TNF-{\alpha}$ from Raw264.7 after pre-treatment with BGT of 0.25 mg/ml, 0.5 mg/ml, 1 mg/ml and 2 mg/ml. To ascertain safety and toxicity of BGT, we divided into single and four weeks repeated administration test. In single test, three groups were administrated different dosages and routes (2 g/kg/i.p., 4 g/kg/i.p. and 15 g/kg /p.o.) of BGT, and in four weeks repeated test, 0.8 g/kg BGT was administrated. Control groups were administrated with only saline according to on Korean Food and Drug Administration, respectively. We observed attentively motality, abnormal clinical sign, body weight change, organ weight, AST and ALT of mice after BGT administration. BGT inhibited passive cutaneous anaphylaxis and active systemic anaphylatic shock by oral administration. All the concentrations of BGT from 0.25 to 2 mg/ml didn't have an effect on cell viability and cytotoxicity. In RBL-2H3, ${\beta}-hexosaminidase$ release, IL-4 and $TNF-{\alpha}$, and in Raw264.7, $TNF-{\alpha}$ were significantly reduced by treated all concentrations of BGT. During toxicity experiment period, there was no difference in body weight change, organ weight, AST and ALT among different dose groups. Death were found 3 mice from day 2 to day 3 in single test i.p. group. (2 g/kg, 4 g/kg). Several individuals of single test i.p. group were observed that decreased locomotor activity, exophthalmos, bloodshot eyes, loss of eyesight and so on in early period after administration. But there was no difference in clinical signs among p.o. group. These results indicate that BGT have inhibition effects on allergy and suggest that no observable effect level of the test orally administration was considered to be more than 2 g/kg in mice under the conditions employed in this study.

Fabrication of Luminescent Hydroxyapatite Microspheres for Drug Loading and Release

  • Park, So Yeon;Kwon, Seung Lee;Baek, Mi Yeon;Yoo, Jin Sun;Kim, Min-Cheol;Jung, Hyun Suk
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.360.1-360.1
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    • 2016
  • Hydroxyapatite (Ca10(PO4)6(OH)2, HAP) particles have attracted a great deal of attention in biomedical fields due to their good biocompatibility, bioactivity and fairly broad applications as drug delivery, dental implant, bone cement, and etc. Thus, many researchers have made an effort to add new functionalities such as luminescence, drug delivery, and bone regeneration properties up to HAP powders by controlling their nanostructure as well as composition. In this research, the mesoporous strontium substituted HAP (Sr-HAP) microspheres were synthesized using a hydrothermal method. In this synthesis, aspartic acid monomers were utilized to form microsphere by controlling surface energy of HAP particles and Sr ions were substituted into Ca ion sites, which induced luminescence property in HAP powders. Moreover, the change in the amount of Sr substitution was found to influence the particle size, morphology, and concurrently surface area, which led to changing drug loading as well as drug release property. The amount of Sr influences the morphology, luminescent properties, particle size, surface area cell viability and drug loading property, which are investigated by SEM, TEM, XRD, FTIR, BET, XPS and in vitro test such as MTT assay and drug release test. In particular, the multifunctional Sr-HAP with molar ratios of 0.25 (Sr/(Ca+Sr)) possessed the strongest luminescent property as well as the superior drug loading and sustained release properties that were correspondent with large surface area and pore size. Our study indicates that the fabricated multifunctional Sr-HAP microspheres are quite useful for bone regeneration and drug delivery.

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In vitro evaluation of a newly produced resin-based endodontic sealer

  • Song, Yoo-Seok;Choi, Yoorina;Lim, Myung-Jin;Yu, Mi-Kyung;Hong, Chan-Ui;Lee, Kwang-Won;Min, Kyung-San
    • Restorative Dentistry and Endodontics
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    • v.41 no.3
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    • pp.189-195
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    • 2016
  • Objectives: A variety of root canal sealers were recently launched to the market. This study evaluated physicochemical properties, biocompatibility, and sealing ability of a newly launched resin-based sealer (Dia-Proseal, Diadent) compared to the existing root canal sealers (AHplus, Dentsply DeTrey and ADseal, Metabiomed). Materials and Methods: The physicochemical properties of the tested sealers including pH, solubility, dimensional change, and radiopacity were evaluated. Biocompatibility was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. For microleakage test, single-rooted teeth were instrumented, and obturated with gutta-percha and one of the sealers (n = 10). After immersion in 1% methylene blue solution for 2 weeks, the specimens were split longitudinally. Then, the maximum length of staining was measured. Statistical analysis was performed by one-way analysis of variance followed by Tukey test (p = 0.05). Results: Dia-Proseal showed the highest pH value among the tested sealers (p < 0.05). ADseal showed higher dimensional change compared to AHplus and Dia-Proseal (p < 0.05). The solubility values of AHplus and Dia-Proseal were similar, whereas ADseal had the lowest solubility value (p < 0.05). The flow values of sealer in increasing order were AHplus, DiaProseal, and ADseal (p < 0.05). The radiopacity of AHplus was higher than those of ADseal and Dia-Proseal (p < 0.05). The cell viability of the tested materials was statistically similar throughout the experimental period. There were no significant differences in microleakage values among the tested samples. Conclusions: The present study indicates that Dia-Proseal has acceptable physicochemical properties, biocompatibility, and sealing ability.

Antimicrobial, Antioxidant and Cytotoxic Activities of Dendropanax morbifera Léveille extract for mouthwash and denture cleaning solution

  • Kim, Ryeo-Woon;Lee, Sook-Young;Kim, Su-Gwan;Heo, Yu-Ri;Son, Mee-Kyoung
    • The Journal of Advanced Prosthodontics
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    • v.8 no.3
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    • pp.172-180
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    • 2016
  • PURPOSE. The purpose of this study was to analyze the antimicrobial, antioxidant activity and cytotoxicity of Dendropanax morbifera $L{\acute{e}}veille$ extract for assessing whether Dendropanax morbifera $L{\acute{e}}veille$ can be used for the development of natural mouthwash and denture cleaning solution. MATERIALS AND METHODS. The extract was obtained from branches of Dendropanax morbifera $L{\acute{e}}veille$. The solvent fractions were acquired by fractionating Dendropanax morbifera $L{\acute{e}}veille$ extract using n-hexane, ethyl acetate, chloroform and butanol solvent. Paper disc test was used to evaluate the antimicrobial and antifungal activity of Dendropanax morbifera $L{\acute{e}}veille$ extract and solvent fractions against Streptococcus mutans and Candida albicans. The analysis of antioxidant activity was carried out through DPPH radical scavenging assay. The cytotoxicity of Dendropanax morbifera $L{\acute{e}}veille$ extract was analyzed through MTT assay using normal human oral keratinocytes. RESULTS. Dendropanax morbifera $L{\acute{e}}veille$ extract showed antimicrobial activity against Streptococcus mutans and especially Candida albicans. The solvent fractions of Dendropanax morbifera $L{\acute{e}}veille$ showed strong antimicrobial activity against Streptococcus mutans and Candida albicans in n-hexane and butanol solvent fraction, respectively. Dendropanax morbifera $L{\acute{e}}veille$ extract also showed outstanding antioxidant activity. Butanol, ethyl acetate, and chloroform solvent fraction of Dendropanax morbifera $L{\acute{e}}veille$ tended to have increased antioxidant activity as the concentration increased. Dendropanax morbifera $L{\acute{e}}veille$ extract showed high cell survival rate in cytotoxicity test. CONCLUSION. Dendropanax morbifera $L{\acute{e}}veille$ extract turned out to have antimicrobial, antioxidant activity and cytophilicity. Based on these results, it is expected that Dendropanax morbifera $L{\acute{e}}veille$ is applicable as an ingredient for natural mouthwash and denture cleanser.

Evaluation of vitrification for cryopreservation of teeth

  • Dissanayake, Surangi C.;Che, Zhong-Min;Choi, Seong-Ho;Lee, Seung-Jong;Kim, Jin
    • Journal of Periodontal and Implant Science
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    • v.40 no.3
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    • pp.111-118
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    • 2010
  • Purpose: The aim of this study was to investigate whether vitrification in the cryopreservation of periodontal ligament (PDL) cells could be useful for tooth banking. Methods: In step 1, primary cultured human PDL cells were cryopreserved in 100% conventional cryopreservation media and 100% vitrification media (ESF40 media) in different temperatures for 2 weeks. In step 2, a series of modified vitrification formulae named T1 (75% vitrification media + 25% F-media), T2 (50% vitrification media + 50% F-media) and T3 (25% vitrification media + 75% F-media) were used to store PDL cells for 2 weeks and 4 weeks in liquid nitrogen. MTT assay was performed to examine the viability of PDL cells. Results: Maximum cell viability was achieved in cells stored in 100% conventional cryopreservation media at $-196^{\circ}C$ (positive control group) in step 1. Compared to the positive control group, viability of the cells stored in 100% vitrification media was very low as 10% in all test conditions. In step 2, as the percentage of vitrification media decreased, the cell viability increased in cells stored for 2 weeks. In 4-week storage of cells in step 2, higher cell viability was observed in the T2 group than the other vitrification formulae while the positive control group had the highest viability. There was no statistically significant difference in the cell viability of 2-week and 4-week stored cells in the T2 group. Conclusions: These observations indicate 100% vitrification media is not successful in PDL cell cryopreservation. Conventional cryopreservation media is currently the most appropriate media type for this purpose while T2 media would be interesting to test for long-term storage of PDL cells.

Antioxidative and Antimutagenic Effects of Ganoderma lucidum Krast Extracts (영지버섯(Ganoderma lucidum Krast) 추출물의 항산화 및 항돌연변이 효과)

  • Oh Se-In;Lee Mee-Sook
    • The Korean Journal of Food And Nutrition
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    • v.18 no.1
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    • pp.54-62
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    • 2005
  • The antioxidative effect, antimutagenic capacity and inhibitory effect of cancer cell proliferation in ethanol extracts of Ganoderma lucidum were studied for suggestion of prevention and dietetic treatment of chronic diseases and development of antioxidative, antimutagenic and anticancer functional food by employing biological and biochemical assay. The IC/sub 50/ of MDA with BSA conjugation reaction, lipid peroxidation and scavenging effect on DPPH radical in ethanol extracts of Ganoderma lucidum showed 1026.21 mg/mL, 0.152 mg/mL and 0.412 mg/mL, respectively. So, the most effective antioxidative capacity in ethanol extracts of Ganoderma lucidum was the lipid peroxidation, among the method used this study. The indirect and direct antimutagenic effects of ethanol extracts of Ganoderma lucidum were examined by Ames test using Salmonella typimurium TA98 and TA100. The inhibition rates on indirect mutagenicity mediated by 2-Anthramine showed 100% in the Salmonella typimurium TA98 and 98.74% in the Salmonella typimurium TA100 and the direct mutagenicity mediated by sodium azide in Salmonella typimurium TA100 was 82.96%. But, the inhibitory effect on indirect mutagenicity mediated by 2-Nitrofluorene in Salmonella typimurium TA98 was low(7.81%). The inhibitory Effect of Ganoderma lucidum ethanol extracts on cell proliferation in Hela and MCF-7 by MTT test were 74.36% in HeLa and 73.90% in MCF-7 at the 0.50 mg/assay concentration and IC/sub 50/ were 0.163 mg/mL and 0.196 mg/mL respectively. From this result, it is suggested that Ganoderma lucidum is believed to have a possible antioxidative, antimutagenic and anticancer capacities.

Effects of Duchesnea indica on Human Stomach Cancer Cells(KATOIII) (사매의 KATOIII 위암세포(胃癌細胞)에 대(對)한 항암효능(抗癌效能))

  • Lim, Joong-Hwa;Kim, Jin-Sung;Yoon, Sang-Hyub;Ryu, Ki-Won;Ryu, Bong-Ha
    • The Journal of Internal Korean Medicine
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    • v.26 no.2
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    • pp.302-309
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    • 2005
  • Background : Death by cancer, malignant tumor, is on the increase around the world. In South Korea, cancer is the number one cause of death, and, in South Korea, stomach cancer is the most common form of cancer. This may be due to genetic and enviornmental factors. Because stomach cancer is so prevalent here, this study was undertaken. Objectives: This study was conducted to investigate the anti-cancer effects of Duchesnea indica(Andr.) Focke. in human stomach cancer cells(KATOIII). Materials & Methods: For human stomach cancer cells KATO III cells were obtained from Korea Cell Line Bank. The water decoction of Duchesnea indica(Andr.) Focke was added to cultures. First, morphological changes were examined under an inverted microscope. The destruction of stomach cancer cells was measured through Trypan blue exclusion testing, and suppression of viability of stomach cancer cells was measured via MTT assay. Anti-cancer mechanisms were assessed by analyzing the cell cycle. Results & Conclusions : In morphologic change, stomach cancer cells showed the withdrawn and floating appearance that is typical in cellular impairment. In each test group more stomach cancer cells were killed than in the control group to a statistically significant degree. Each test group showed more suppression of viability of stomach cancer cells than that of the control group to a statistically significant degree. Analysis of the cell cycle showed that Duchesnea indica(Andr.) Focke inhibited division of stomach cancer cells. This experiment yielded empirical evidence for Duchesnea indica(Andr.) Focke as a statistically significant anticarcinogen, specifically against stomach cancer.

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Comparison of Contact Lenses and Storage Solution for Cytotoxic Potential using a Cell Growth Inhibition Assay (소프트 콘택트렌즈 용출액과 보존 용액의 세포 독성 비교)

  • Kim, Jai-Min;Seong, Jeong-Sub;Yoo, Keun-Chang;Yoon, Young;Ra, Myung-Suk;Lee, Jong-Bin
    • Journal of Korean Ophthalmic Optics Society
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    • v.4 no.2
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    • pp.81-86
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    • 1999
  • Human corneal epithelial cells and mouse fibroblast L929 cells were grown to 60-70% confluency in 96 well plates. 24 hours after the well plates are inoculated, the medium on the test plates is aspirated and replaced with an extract supplemented medium prepared from the materials to be tested. The contact lenses manufactured by 7 companies were collected from optical shops and used for this study. The exactracts having been prepared by autoclaving 8 lenses in 2.5ml saline. The cell monolayer is then cultured for a further 48 hour period. MTT and SRB assys were performed for cytotoxic effect on cultured cells An inhibition of 30% is considered clear indication of cytotoxic potential in the test material. All the materials were not cytotoxic, but 3 storage solutions of them inhibited growing L929 cells.

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Biocompatibility of Ti-8wt.%Ta-3wt.%Nb alloy with Surface Modification (표면 개질에 따른 Ti-8wt.%Ta-3wt.%Nb 합금의 생체적합성)

  • Lee, Doh-Jae;Lee, Kyung-Ku;Park, Bum-Su;Lee, Kwang-Min;Park, Sang-Won
    • Korean Journal of Materials Research
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    • v.16 no.5
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    • pp.277-284
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    • 2006
  • The alloys were prepared by a non-consumable vacuum arc melting and homogenized at $1050^{\circ}C$ for 24 hrs. Two kind of surface modifications were performed alkali treatment in 5.0M NaOH solution subsequent and heat treatment in vacuum furnace at $600^{\circ}C$, and were oxidizing treatment at the temperature range of 550 to $750^{\circ}C$ for 30 minutes. After surface modification, these samples were soaked in SBF which consists of nearly the same ion concentration as human blood plasma. Cytotoxicity tests were performed in MTT assay treated L929 fibroblast cell culture, using indirect methods. A porous and thin activated layer was formed on Titanium and Ti-8Ta-3Nb alloy by the alkali treatment. A bone-like hydroxyapatite was nucleated on the activated porous surfaces during the in vitro test. However, Ti-8Ta-3Nb alloys showed better bioactive properties than Titanium. According to XRD results, oxide layers composed of mostly $TiO_2$(rutile) phases. Cytotoxicity test also revealed that moderate oxidation treatment lowers cell toxicity and Ti-8Ta-3Nb alloy showed better results compared with Titanium.