• Title/Summary/Keyword: MTT-test

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Cellular Uptake and Transfection Efficiency of Plasmid DNA Using Low Molecular Weight Polyethylenimine (저분자량 폴리에틸렌이민을 이용한 유전자 송달 및 발현 유효성 연구)

  • Jeong, Gil-Jae;Park, Kui-Lye;Shin, Ji-Young;Choi, Han-Gon;Oh, Yu-Kyoung
    • Journal of Pharmaceutical Investigation
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    • v.34 no.4
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    • pp.263-267
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    • 2004
  • Branched and linear polyethylenimines (PEIs) have been studied as efficient and versatile agents for gene delivery in vitro and in vivo. PEIs exist in a linear or branched topology and are available in a wide range of molecular weight (Mw). Most studies have been done using PEIs with Mw higher than 10Kd. This study was aimed to test the transfection efficiency and the cell viability following gene delivery using PEI of Mw 2Kd, a relatively lower Mw cationic polymer. We used murine interleukin-2(mIL-2) plasmid DNA complexed with branched PEI 2Kd or 25Kd, and transfected them into a myoblast muscle cell line, C2C12. The cellular uptake of mIL-2 plasmid DNA was determined using quantitative polymerase chain reaction. RNA transcript levels were studied in the myoblast cells. Our results show that PEI 2Kd was as effective as PEI 25Kd in celluar gene delivery and transfection efficiency in C2C12 cells. Moreover, MTT assay indicated that PEI 2Kd/DNA complexes did not significantly reduce the cell viability regardless of N/P ratios. These results suggest that PEI of Mw 2Kd might play a role as effective and low toxic nonviral vector systems for muscular cell lines.

Anti-Proliferative Effect of Tetraphenylporphine (TPP) as an Iron Chelator on Vascular Smooth Muscle Cells and its Release Profiles from Polymer Coating Layer (철 킬레이터로서의 tetraphenylporphine의 혈관평활근세포의 성장억제효과와 고분자 코팅막으로부터의 방출 특성)

  • Park, Min-Hee;Kang, Soo-Yong;Park, Hyun-Jeong;Seo, Jin-Seon;Park, Young-A;Kim, Ji-Eun;Kim, Yang-Geun;Whang, Bae-Geon;Munkhjargal, Odonchimeg;Shim, Young-Key;Kho, Weon-Gyu;Lee, Woo-Kyoung
    • Journal of Pharmaceutical Investigation
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    • v.38 no.2
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    • pp.93-98
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    • 2008
  • The drug-eluting stent (DES) implantation is a widely acceptable treatment for coronary heart disease. It was reported that iron chelator had anti-proliferative effect on human vascular smooth muscle cells (HA-VSMCs). In this study, tetraphenylporphine (TPP) was selected as an iron chelator and drug for DES. MTT assay showed that TPP had antiproliferative effect on HA-VSMCs. TPP and polycaprolactone (PCL) were coated onto stainless steel plate using a spraycoating method. From the surface morphology examination of the coated plate by SEM, smooth polymer coating layer could be observed. The thickness of coating layer could be controlled by changing repeating time of coating. From in vitro release test, sustained release of TPP was observed from plate during two weeks. Thus, TPP as iron chelator can be used as drug for stent coating because of its antiproliferative effect and sustain release profile.

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.

Neurotoxicity Assessment of Methamphetamine and Cadmium Using Cultured Neuronal Cells of Long-Evans Rats (신경세포 배양법을 이용한 methamphetamine과 cadmium의 신경독성 평가)

  • Cho, Dae-Hyun;Kim, Jun-Gyon;Jeong, Yong;Lee, Bong-Hun;Kim, Eun-Youb;Kim, Jeong-Goo;Cho, Tai-Soon;Kim, Jin-Suk;Moon, Hwa-Hwey
    • Toxicological Research
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    • v.12 no.1
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    • pp.69-79
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    • 1996
  • Primary culture of cerebellar neuronal cells derived from 8-day old Long-Evans rats was used. Pure granule cells, astrocytes or mixed cells culture systems were prepared. These cells were differentiated and developed synaptic connections. And the astrocytes were identified by immunostaining with glial fibrillary acidic protein (GFAP). Methamphetamine (MAP), which acts on dopaminergic system and cadmium (Cd), a toxic heavy metal, were applied and biochemical assays and electrophysiological studies were performed. $LC_50$ values estimated by MTT assay of MAP and Cd were 3 mM and 2$\mu M$ respectively. Cells were treated with 1 mM or 2 mM MAP and 1$\mu M$ $CdCl_2$ for 48 hour, and the incubation media were analyzed for the content of released LDH. MAP (2 mM) and Cd significantly increased the LDH release. Cell viability was decreased in both groups and some cytopathological changes like cell swelling or vacuolization were seen. The cerebellar granule cells were used for measuring membrane currents using whole-cell clamp technique. Sodium and potassium currents were not affected by MAP neither Cd, but calcium current was significantly reduced by Cd but not affected by MAP. Therefore, in vitro neurotoxicity test system using neuronaI cells and astrocytes cultures were established and can be used in screening of potential neurotoxic chemicals.

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The Effects of Gyejijakyakjimo-Tang on the Allergic Contact Dermatitis induced by DNCB (계지작약지모탕(桂枝芍藥知母湯)이 DNCB로 유발된 알레르기성 접촉피부염에 미치는 영향)

  • Kim, Sung-Ho;Kim, Hee-Taek
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.23 no.1
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    • pp.8-22
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    • 2010
  • Objectives : The purpose of this study is to investigate the effects of Gyejijakyakjimo-Tang on the Allergic Contact Dermatitis caused by 2,4-dinitro-chlorobezene(DNCB). Methods : Twenty eight mice were divided into four groups ; normal, control, experimental group A and B. Control and experimental groups were induced allergic contact dermatitis by DNCB. Experimental group A was orally administered the Gyejijakyakjimo-Tang and experimental group B was orally administered the prednisolone. In this study, ear thickness measurement, auricle microphotograph observation, MPO(Myeloperoxidase) activity measurement, Reverse transcription-polymerase chain reaction(RT-PCR) analysis of the mRNA level of TNF-$\alpha$, IL-$1{\beta}$ were performed on these four groups. In addition, the effect of Gyejijakyakjimo-Tang on cell viability and the effect of Gyejijakyakjimo-Tang on the compound 48/80-induced histamine release from HMC and RPMC were measured. Results: 1. Both experimental group A and B had decreased ear thickness compared with control group In contact hypersensitivity assay. 2. In experimental group A, inflammatory edema was similarly observed comparing to control group. Nevertheless, inflammatory edema was obviously reduced in experimental group B. In both experimental group A and B, pathological lesion of dermatitis were alleviated. In addition, the numbers of infiltrated inflammatory cells were decreased compared with control group. 3. Compared to the normal group, there was a noticeable increase in MPO activity in control group. However, in experimental group A and B, it showed remarkable inhibition of the increase in MPO activity comparing with control group. 4. The level of expression of TNF-$\alpha$, IL-$1{\beta}$ in experimental group A and B were meaningfully lower than those in control group. 5. In MTT assay, the concentrations of Gyejijakyakjimo-Tang that were used on the test had no cytotoxicity. 6. Gyejijakyakjimo-Tang dose-dependently inhibited the compound 48/80-induced histamine release from both HMC and RPMC. Conclusions : According to above experiments, Gyejijakyakjimo-Tang was effective on allergic contact dermatitis.

Chemical compositions and biological activities of marine invertebrates from the East Sea of South Korea

  • Kim, Geun-Hyeong;Park, Hyeon-Ho;Chandika, Pathum;Ko, Seok-Chun;Jung, Kyung-Mi;Yoon, Sang Chul;Oh, Taeg-Yun;Kim, Young-Mog;Jung, Won-Kyo
    • Fisheries and Aquatic Sciences
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    • v.22 no.6
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    • pp.13.1-13.9
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    • 2019
  • Background: Marine invertebrates are well known as pivotal bioresources with bioactive substances such as anti-inflammatory sterols, antitumor terpenes, and antimicrobial peptides. However, there are few scientific reports on chemical compositions and bioactivities of marine invertebrates from the East Sea of South Korea. Methods: In this study, chemical compositions and biological activities were evaluated on both 70% EtOH and hot water extracts of 5 species of marine invertebrates (Crossaster papposus japonicus, Actinostola carlgreni, Stomphia coccinea, Actinostola sp., and Heliometra glacialis) collected from the East Sea of South Korea. The antioxidant activities were measured by ABTS radical scavenging assay. The cytotoxicity and anti-inflammatory activity were evaluated using MTT and Griess reagents. Moreover, the antibacterial effect was evaluated using paper disc assay and minimum inhibitory concentration (MIC) assay. Results: In the results of antioxidant activities, 70% EtOH extract of A. carlgreni showed the highest activity ($IC_{50}\;0.19{\pm}0.03mg/ml$) compared to other extracts. Moreover, 70% EtOH extract of A. carlgreni could significantly suppress the nitric oxide (NO) production in lipopolysaccharide-induced RAW 264.7. All extracts treated under $400{\mu}g/ml$ have no cytotoxic effects on RAW 264.7 macrophages. In the antibacterial test, both 70% EtOH extracts of C. papposus japonicus and H. glacialis showed a significant antibacterial effect on Staphylococcus aureus. The MIC values were evaluated at 256 and $512{\mu}g/ml$, respectively. Conclusions: These results suggested the bioactive potentials of marine invertebrates from the East Sea of South Korea in pharmaceutical and nutraceutical applications.

Evaluation of the Cosmeceutical Activity of Apocynum lancifolium Russanov Extracts (개정향풀 추출물의 향장학적 활성 평가)

  • Park, Ji Hyo;Lee, Ji-An
    • Journal of Convergence for Information Technology
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    • v.11 no.1
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    • pp.236-243
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    • 2021
  • In this study, the functional cosmetic activity of hot-water (AW) and methanol(AM) extracts from Apocynum lancifolium Russanov are examined. In the DPPH antioxidant activity test, the AW extract showed the highest inhibition rate of 90.5% (IC50 37.717 ± 8.209 ㎍/mL) and the antioxidative activity of ABTS showed a high activity in the AW extract with an IC50 of 185.244 ± 12.602 ㎍/mL and 96.2%, respectively. In RAW264.7 macrophages, the two extracts significantly suppressed the LPS-induced nitric oxide (NO), TNF-α, IL-6 production and iNOS expression level. The MTT assay measured by the cell survival rate showed that AW and AM extracts have no toxicity. The astringent activity of the AW extract exhibited the highest astringent activity with 74.336±2.487 mg/mL. Therefore, these results suggest that A. lancifolium extracts have a health promotion potential which can be further developed as food additive, natural products for cosmetic purpose.

Physicochemical, Antibacterial Properties, and Compatibility of ZnO-NP/Chitosan/β-Glycerophosphate Composite Hydrogels

  • Huang, Pingping;Su, Wen;Han, Rui;Lin, Hao;Yang, Jing;Xu, Libin;Ma, Lei
    • Journal of Microbiology and Biotechnology
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    • v.32 no.4
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    • pp.522-530
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    • 2022
  • In this study we aimed to develop novel ZnO-NP/chitosan/β-glycerophosphate (ZnO-NP/CS/β-GP) antibacterial hydrogels for biomedical applications. According to the mass fraction ratio of ZnO-NPs to chitosan, mixtures of 1, 3, and 5% ZnO-NPs/CS/β-GP were prepared. Using the test-tube inversion method, scanning electron microscopy and Fourier-transform infrared spectroscopy, the influence of ZnO-NPs on gelation time, chemical composition, and cross-sectional microstructures were evaluated. Adding ZnO-NPs significantly improved the hydrogel's antibacterial activity as determined by bacteriostatic zone and colony counting. The hydrogel's bacteriostatic mechanism was investigated using live/dead fluorescent staining and scanning electron microscopy. In addition, crystal violet staining and MTT assay demonstrated that ZnO-NPs/CS/β-GP exhibited good antibacterial activity in inhibiting the formation of biofilms and eradicating existing biofilms. CCK-8 and live/dead cell staining methods revealed that the cell viability of gingival fibroblasts (L929) cocultured with hydrogel in each group was above 90% after 24, 48, and 72 h. These results suggest that ZnO-NPs improve the temperature sensitivity and bacteriostatic performance of chitosan/β-glycerophosphate (CS/β-GP), which could be injected into the periodontal pocket in solution form and quickly transformed into hydrogel adhesion on the gingiva, allowing for a straightforward and convenient procedure. In conclusion, ZnO-NP/CS/β-GP thermosensitive hydrogels could be expected to be utilized as adjuvant drugs for clinical prevention and treatment of peri-implant inflammation.

Anti-Melanogenic Effect of Thymol, a Major Odorant in Essential Oils of Family Lamiaceae (꿀풀과 식물 정유의 주성분인 Thymol의 미백활성에 관한 연구)

  • Choi, Deok-Gyun;Park, Chan Ik;Lee, Sun-Mi;Baek, Jeong-In
    • The Korea Journal of Herbology
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    • v.34 no.4
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    • pp.19-25
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    • 2019
  • Objectives : Thymol (2-isopropyl-5-methylphenol), a natural monoterpenoid phenol, is one of the major odorant constituents found in natural essential oils of various herbal plants, such as Thymus quinquecostatus and Thymus vulgaris. Multiple biological activities of thymol, including antioxidative, antimicrobial, and anti-inflammatory effects, have been reported in numerous in vitro studies, and recently it was suggested that thymol may could inhibit oxidization of L-dihydroxyphenylalanine (L-DOPA) to dopaquinone required in melanogenesis pathway, as an antioxidant. Methods : MTT assay was performed to test the cytotoxic effect of thymol in B16F10 cells. Inhibitory effect of thymol to tyrosinase activities were examined using both mushroom tyrosinase and intracellular tyrosinase. Expression level of tyrosinase in B16F10 cells were investigated by western blot analysis. Results : The cell viability was decreased by thymol treatment in dose-dependant manner, leading significant cytotoxicity in 500 and $1000{\mu}M$ thymol-treated groups. In the alpha-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis, administration of thymol significantly decreased extracellular (secreted) melanin content in dose-dependent manner. Cellular tyrosinase activity assay and western blot analysis of intracellular tyrosinase showed that thymol has a strong anti-melanogenic effect by inhibition of tyrosinase activity and by decreasing expression of tyrosinase that contribute to melanin synthesis in the B1610 cells. Conclusions : As the first functional study that prove anti-melanogenic effect of thymol and its underlying mechanism in the living cells, our study suggests the applicability of fragrance as the functional materials of cosmetics or health supplement, not as just an additive.

Evaluation on the Whitening Effect of Nypa fruticans wurmb Extracts (해죽순 추출물의 미백활성 평가)

  • Kim, Il-Chool
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.3
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    • pp.462-470
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    • 2022
  • The purpose of this study was to investigate the role of the Nypa fruticans extracts as a cosmetic additive. The tyrosinase inhibitory effects showed 52.0% at 1,000 ㎍/mL concentration. A cell viability test, measured on melanoma cell (B16F10) hot water extract of nypa fruticans showed 84.8% at 100 ㎍/mL concentration. The protein expression inhibitory effects of nypa fruticans extracts were measured by western blot at 25, 50, 100 ㎍/mL concentration and the β-actin. Results showed that the expression inhibition rates of the MITF, TRP-1, TRP-2, tyrosinase protein were decreased by 70.7%, 83.3%, 45.7%, 45.9% at 100 ㎍/mL concentration, respectively. It was concluded that nypa fruticans extracts had the whitening effects and thus could be applied for cosmetics as a natural ingredient.