• Title/Summary/Keyword: MTT method.

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Bioactivity behavior of biphasic calcium phosphate powders prepared by co-precipitation method (공침법으로 합성된 biphasic calcium phosphate 분말의 생체활성 거동)

  • Kim, Tae-Wan;Kim, Dong-Hyun;Jin, Hyeong-Ho;Lee, Heon-Soo;Park, Hong-Chae;Yoon, Seog-Young
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.22 no.2
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    • pp.99-104
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    • 2012
  • The co-precipitation technique has been applied to synthesize biphasic calcium phosphate (BCP). $Ca(NO_3)_2{\cdot}4H_2O$ and $(NH_4)_2HPO_4$ as the starting materials was used. X-ray diffraction (XRD) and Fourier transformed infrared (FT-IR) spectroscopy were used to characterize the structure of as-synthesized and calcined BCP powders. After immersion in Hanks' Balanced Salt Solution (HBSS), for 1 week a precipitation started to be formed with individual small granules on the specimen surface. An MTT assay indicated that BCP powders have no cytotoxic effects on MG-63 cells, and that they have good biocompatibility.

Effects of Paeoniae Radix Aqua-Acupuncture Solution on Tert-Butyl Hydroperoxide Induced Lipid Peroxidation and Antioxidative Enzymes in Cultured Rat Liver Cells (작약 약침액이 tert-butyl hydroperoxide 로 유도된 흰쥐 배양 간세포의 지질과산화반응 및 항산화효소 활성에 미치는 영향)

  • Moon, Jin-Young
    • Journal of Acupuncture Research
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    • v.17 no.3
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    • pp.176-187
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    • 2000
  • Objectives : This study was purposed to investigate the antioxidative effects of Paeoniae radix aqua-acupuncture solution(PR) on culture liver cell system, lipid peroxidation and antioxidative enzyme activities in tert-butyl hydroperoxide(t-BHP) treatmented conditions. Methods : Cultured normal rat liver cell(Ac2F) were prepared and incubated with or without PR(at 2% volume in culture medium). After 16~18hr, cells placed in DMEM medium without serum, and then incubated with 1mM t-BHP for 2hr. Viable cells were detected by MTT assay, and the levels of lipid peroxide(LPO) were measured by TBA method. And catalase activity was measured as the decrease in hydrogen peroxide absorbance at 240nm on spectrophotometer using 30mM hydrogen peroxide. Superoxide dismutase(SOD) were assayed by recording the inhibition of nitro blue tetrazolium reduction with xanthine and xanthine oxidase. Glutathione peroxidase(GPX) activity was determined by the modified coupled assay developed by Paglia and Lawrence. The reaction was started by addition of 2.2mM hydrogen peroxide as substrate. The change in absorbance at 340nm was measured for 1min on spectrophotometer. Glutathione-S-transferase(GST) activity was assayed with CDNB as substrate and enzyme activity of GST towards the glutathione conjugation of CDNB. Results : Cell killing was significantly enhanced by addition of t-BHP compared to those of untreated group. PR pretreated cell resisted the toxic effects of t-BHP. LPO levels of t-BHP treatment group were significantly higher than other groups. This increased level was significandy reduced by PR pretreatment. The t-BHP treatment resulted in a decrease of catalase, GPX and GST activities. By contrast, PR pretreatment markedly increased compare to those of untreated groups. Conclusions : T-BHP which can produce intracellular free radical was used for inducer of the peroxidation of cellular lipids. PR protected the cell death induced by t-BHP and significantly increased cell viabiliry in the normal rat liver cell, and showed effective inhibition of lipid peroxidation, and elevations of catalase, GPX and GST activities. These results suggested that PR might play a protective role in lipid peroxidation by free radicals.

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The effect of 100KHz PWM LED light irradiation on RAT bone-marrow cells (100kHz PWM LED 광조사가 백서 골수세포에 미치는 영향)

  • Cheon, Min-Woo;Kim, Seong-Hwan;Kim, Young-Pyo;Lee, Ho-Sic;Park, Yong-Pil;Yu, Seong-Mi;Lee, Hee-Gap;Kim, Tae-Gon
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.06a
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    • pp.512-513
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    • 2008
  • The study examined what effects 100kHz PWM LED light irradiation causes to bone marrow cells of SD-Rat when LED characterized cheap and safe is used onto the light therapy by replacing the low 1evel laser. We developed the equipment palpating cell proliferation using a high brightness LED. This equipment was fabricated using a micro-controller and a high brightness LED, and designed to enable us to control light irradiation time, intensity, frequency and so on. Especially, to control the light irradiation frequency, FPGA was used, and to control the change of output value, TLC5941 was used. Consequent1y, the current value could be controlled by the change of 1eve1 in Continue Wave(CW) and Pulse Width Modulation(PWM), and the output of a high brightness LED could be controlled stage by stage. MTT assay method was chosen to verify the cell increase of two groups and the effect of irradiation on cell proliferation was examined by measuring 590nm transmittance of ELISA reader. As a result, the cell increase of Rat bone marrow cells was verified in 100kHz PWM LED light irradiation group as compared to non-irradiation group.

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Immuno-stimulating Effects of Oga-Power (OP) Containing Extract of Acanthopanax sessiliflorus on Immune Cells in Mice (오가피를 활용한 음료인 오가파워가 생쥐의 면역세포 활성화에 미치는 영향)

  • Kim, Hyung-Woo;Kim, Kyung-Yoon;Lee, Sang-Yeong;Kim, Gye-Yeop;Jeon, Byung-Gwan;Cho, Su-In;Jeong, Hyun-Woo
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.141-147
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    • 2008
  • Objectives : The cortex and root of Acanthopanax sessiliflorus (AR), a herbal medicine, have been used for several diseases including cancer in Oriental countries. Recently, we reported that AR has an immune-potentiating action. Oga-Power(OP) was made using extract from AR. For these reasons, we hypothesized that OP can potentiate the immune system in terms of accelerating proliferation rates of immune cells such as thymocytes and splenocytes. Methods : In this experiment, proliferation rates of thymocytes and splenocytes were measured using modified 3-[4,5-dimethy -lthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT). No production levels in macrophages isolated from normal mice were measured using Griess method. Results : In our results, treatment with OP accelerated proliferation rates of splenocytes, but did not affect those of thymocytes in vitro. On the other hand, proliferation rates of thymocytes was elevated in vivo. In addition, level of NO production from macrophage separated from abdominal cavity of normal mice was elevated by treatment with OP. Conclusions : In conclusion, OP has immune-potentiating action, by acceleration of splenocyte proliferation and elevation of NO production level from macrophages.

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In vitro Protective Effects of Glehnia Littoralis on Alpha-amanitin Induced Hepatotoxicity (알파 아마니틴에 의한 간독성에 대한 갯방풍의 보호 효과)

  • Kim, Bo Hyun;Sun, Kyung Hoon;Kim, Sun Pyo;Park, Yongjin
    • Journal of The Korean Society of Clinical Toxicology
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    • v.15 no.2
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    • pp.107-115
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    • 2017
  • Purpose: Glehnia littoralis has been used to treat ischemic stroke, phlegm, cough, systemic paralysis, antipyretics and neuralgia. The pharmacological mechanisms of Glehnia littoralis include calcium channel block, coumarin derivatives, anticoagulation, anti-convulsive effect, as well as anti-oxidant and anti-inflammatory effects. Alpha-amanitin (${\alpha}$-amanitin) is a major toxin from extremely poisonous Amanita fungi. Oxidative stress, which may contribute to severe hepatotoxicity was induced by ${\alpha}$-amanitin. The aim of this study was to investigate whether Glehnia littoralis ethyl acetate extract (GLEA) has the protective antioxidant effects on ${\alpha}$-amanitin -induced hepatotoxicity. Methods: Human hepatoma cell line HepG2 cells were pretreated in the presence or absence of GLEA (50, 100 and $200{\mu}g/ml$) for 4 hours, then exposed to $60{\mu}mol/L$ of${\alpha}$-amanitin for an additional 4 hours. Cell viability was evaluated using the MTT method. AST, ALT, and LDH production in a culture medium and intracellular MDA, GSH, and SOD levels were determined. Results: GLEA (50, 100 and $200{\mu}g/ml$) significantly increased the relative cell viability by 7.11, 9.87, and 14.39%, respectively, and reduced the level of ALT by 10.39%, 34.27%, and 52.14%, AST by 9.89%, 15.16%, and 32.84%, as well as LDH by 15.86%, 22.98%, and 24.32% in culture medium, respectively. GLEA could also remarkably decrease the level of MDA and increase the content of GSH and SOD in the HepG2 cells. Conclusion: In the in vitro model, Glehnia littoralis was effective in limiting hepatic injury after ${\alpha}$-amanitin poisoning. Its antioxidant effect is attenuated by antidotal therapy.

Antibacterial Effects and Cytotoxicity of Crassirhizomae Rhizoma (관중의 항균작용 및 세포독성에 관한 연구)

  • Kim, Seung-Nam;Ku, Young;Rhyu, In-Cheol;Hahm, Byung-Do;Bae, Ki-Hwan;Han, Soo-Boo;Chung, Chong-Pyoung;Choi, Sang-Mook
    • Journal of Periodontal and Implant Science
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    • v.30 no.1
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    • pp.65-77
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    • 2000
  • The purpose of this study was to evaluate the antimicrobial activity of Crassirhzimae rhizoma and its possible use as an oral antiseptics for prevention of periodontitis. Its antibacterial activity against periodontopathic microorganisms including Actinobacillus actiomycetem - comitans, Capnocytophaga ochracea, Streptococcus mutans, Porphyromonas gingivalis, Prevotella intermedia, Actinomyces viscosus, Fusobacterium nucleatumwas evaluated via modified stab culture method. The cytotoxicity against gingival fibroblasts and rat osteoblasts was investigated via [$^3H$]thymidine incorporation and cellular activity was investigated via MTT assay. Chlorhexidine was used as control group. Crassirhizomae rhizoma was prepared at concentrations of 0.2, 0.15, 0.1, 0.05%. Chlorhexidine was also prepared at the same concentration. Crassirhizomae rhizoma showed lower antimicrobial antivity against these microorganism than chlorhexidine, but this difference was not significant. And, Crassirhzomae rhizoma showed more cellular activity and less cytotoxicity than chlorhexidine on human gingival fibrablast and rat osteoblast. This study suggests that Crassirhzomae rhizoma might be a candidate for a safe oral antiseptic for the prevention and treatment of periodontal disease.

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Effects of Taeumjowetang on Lipid Peroxidation by Free Radicals and Oxidative Damage of Hepatocytes by tert-Butyl Hydroperoxide (태음조위탕(太陰調胃湯)의 항산화(抗酸化) 효능(效能)에 의한 간세포(肝細胞) 보호(保護) 효과(效果)에 관한 연구(硏究))

  • Kim, Man-woo;Park, Seong-sik
    • Journal of Sasang Constitutional Medicine
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    • v.13 no.1
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    • pp.51-60
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    • 2001
  • Effects of Taeumjowetang on Lipid Peroxidation by Free Radicals and Oxidative Damage of Hepatocytes by tert-Butyl Hydroperoxide. 1. Purpose The present study was carried out to evaluate the antioxidant effects of Taeumjowetang in vitro. 2. Methods In this study, antioxidant effects of TJT on lipid peroxidation were determined according to the method of TBA. (Abbreviation) TJT : Taeumjowetang, TBA : 2-thiobarbituric acid. 3. Results : 1) TJT inhibited markedly peroxidation of linoleic acid during the autoxidation. 2) TJT inhibited lipid peroxidation induced by hydroxyl radical derived from H2O2-Fe2+ in rat liver homogenate. 3) TJT showed 66% scavenging effect on DPPH radical. 4) TJT exhibited a 25% inhibitory effect on superoxide generation from xanthine-xan thine oxidase system. 5) To investigate the antioxidative effects of TJT on the hepatocytes, cultured normal rat liver cells(Ac2F) were prepared and incubated with or without TJT. After 16~18hr, cells placed in DMEM medium without serum, and then incubated with 1mM t-BHP for 2hr. Viable cells were detected by MTT assay. In this test, TJT protected the cell death induced by t-BHP and significantly increased cell viability in the normal rat liver cell. (Abbreviation) DPPH : ${\alpha},{\alpha}$-diphenyl-${\beta}$-picryl hydrazyl, DMEM : Dulbecco's Modified Eagle Medium, t-BHP : terr-butyl hydroperoxide, 4. Conclusion These results suggested that TJT might play a protective role in lipid peroxidation by free radicals.

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Biological Activities and Cell Proliferation effects of Red Ginseng Ethanol Extracts (홍삼 에탄올 추출물의 생리활성과 세포증식 효과)

  • Hwang, Sung-Yeoun;Ahn, Seong-Hun
    • Journal of Pharmacopuncture
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    • v.14 no.3
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    • pp.55-61
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    • 2011
  • Objectives: Reactive Oxygen Species(ROS) are continuously produced at a high rate as a by-product of aerobic metabolism. Since tissue damage by free radical, ROS such as hydrogen peroxide($H_2O_2$), nitric oxide(NO) increases with age. Several lines of evidence provided that ROS appears to cause to develop aging-related various diseases such as cancer, arthritis, cardiovascular disease. In this study, we have conducted to investigate the pharmacological effects of red ginseng for the development possibility to pharmacopuncture drug sources or healthy aid foods. Methods: For our aims, it was investigated the biological activities of Red Ginseng ethanol extracts (RGEE) by measuring total polyphenol contents, total flavonoid contents, DPPH radical scavenging activity, ABTS radical scavenging activity and cell viability of MCF 10A and SK-MEL-2 in vitro with MTT assay method. Results: The total polyphenol contents of RGEE was 3.06${\pm}$0.11mg/g in 10mg/ml, the total flavonoid contents of RGEE was 1.35${\pm}$0.01mg/g in same concentration. The ABTS radical scavenging activity was about 80% and that of DPPH activity was 65% in 50mg/ml of RGEE. The cell viability of SKMEL-2, skin cancer cell line was decreased and that of MCF 10A, skin normal cell line was increased. Conclusions: We conclude that RGEE may be useful as potential functional foods or pharmacopuncture drug sources on the diseases induced by oxidant stress.

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.

Effect of Progesterone on Cultured Human Dermal Fibroblast (배양된 인체진피섬유모세포의 증식에 대한 황체호르몬의 영향)

  • Kwon, Soon Sung;Oh, Myung June;Lee, Jin Hee;Park, Jong Lim;Chang, Hak;Minn, Kyung Won
    • Archives of Plastic Surgery
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    • v.34 no.4
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    • pp.420-425
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    • 2007
  • Purpose:The mechanism of scar formation is not fully understood. Fibroblast is an important cell in wound healing process. We experienced a patient who was taking progesterone orally. Upper blepharoplasty was performed on her but, wound healing was delayed. We hypothesized that progesterone was the cause of delayed wound healing and fibroblast proliferation inhibition. We investigated the effect of progesterone in vitro on human dermal fibroblasts to study the effects on fibroblast proliferation. Methods: Human dermal fibroblasts from four persons were cultured initially. Progesterone is mixed to them at various concentrations, and fibroblast cell count was measured by MTT assay method at 570 nm. We confirmed that progesterone has some inhibitory effect on fibroblast proliferation and maximal inhibitory concentration of progesterone was determined. Then fibroblasts from a total of nineteen persons were cultured and the effects of progesterone were studied. Results: The initial study showed the maximal inhibitory concentration of progesterone to be $50{\mu}g/ml$. The main study showed that progesterone had 70.9% inhibitory effect on human dermal fibroblast in vitro. Conclusion: Progesterone has inhibitory effect on cultured human dermal fibroblast proliferation in vitro.