• Title/Summary/Keyword: MTT method.

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A simple chemical method for conversion of Turritella terebra sea snail into nanobioceramics

  • Sahin, Yesim Muge;Orman, Zeynep;Yucel, Sevil
    • Journal of Ceramic Processing Research
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    • v.19 no.6
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    • pp.492-498
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    • 2018
  • In this study, a sea shell was converted into bioceramic phases at three different sintering temperatures ($450^{\circ}C$, $850^{\circ}C$, $1000^{\circ}C$). Among the obtained bioceramic phases, a valuable ${\beta}-TCP$ was produced via mechanochemical conversion method from sea snail Turritella terebra at $1000^{\circ}C$ sintering temperature. For this reason, only the bioceramic sintered at $1000^{\circ}C$ was concentrated on and FT-IR, SEM/EDX, BET, XRD, ICP-OES analyses were carried out for the complete characterization of ${\beta}-TCP$ phase. Biodegradation test in Tris-buffer solution, bioactivity tests in simulated body fluid (SBF) and cell studies were conducted. Bioactivity test results were promising and high rate of cell viability was observed in MTT assay after 24 hours and 7 days incubation. Results demonstrated that the produced ${\beta}-TCP$ bioceramic is qualified for further consideration and experimentation with its features of pore size and ability to support bone tissue growth and cell proliferation. This study suggests an easy, economic method of nanobioceramic production.

Preparation and Characterization of Hyaluronic Acid Loaded PLGA Scaffold by Emulsion Freeze-Drying Method (히알루론산을 함유한 PLGA 지지체의 제조 및 특성결정)

  • Ko, Youn-Kyung;Kim, Soon-Hee;Jeong, Jae-Soo;Park, Jung-Soo;Lim, Ji-Ye;Kim, Moon-Suk;Lee, Hae-Bang;Khang, Gil-Son
    • Polymer(Korea)
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    • v.31 no.6
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    • pp.505-511
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    • 2007
  • Poly(lactide-co-glycolide)(PLGA) and hyaluronic acid (HA) has been widely used as biocompatible scaffold materials to regenerate tissue. In this present study, we fabricated microporous PLGA and HA loaded PLGA scaffolds by a emusion freeze-drying method. In order to confirm that the release profile of cytokine or water-soluble drugs, we manufactured the granulocyte macrophage colony stimulating factor(GM-CSF) loaded PLGA and HA-PLGA scaffold. All scaffolds were characterized using scanning electron microscope(SEM), mercury porosimeter and wettability measurement. Cell proliferation and viability were assessed by a 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) test. The porosity of HA-PLGA scaffold was greater than 95% with the total pore area of $261\;m^2/g$. The HA-FLGA scaffold exhibited well interconnected pores to allow greater cell adhesion and prolixferation. It was proven by higher cell viability in the HA-PLGA scaffold than PLGA alone. This may be due to the enhanced natural properties and higher water retention capacity of HA.

Studies on Biological Activities of the Polysaccharides and Oligosaccharides of Orostachys japonicus (와송 다당체 및 올리고당류의 생리 활성)

  • Kim Ki Hoon;Kim Eun Young;Kim Yea Oon;Baek Geum Ok;Kim Han Bok;Lee Dong Seok
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.334-341
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    • 2004
  • Polysaccharides were prepared from Orostachys japonicus by extration with hot steam water (OJPl). The OIPl fraction was further purified by Sephadex G-50 gel filtration chromatography to produce FI (polysaccharides) and FII (oligosaccharides) fraction. The average molecular masses o fFI and FII fraction were determined to be 3050 kDa and 13 kDa, respectively. The antimicrobial activity of OIPl was tested against 8 strains of bacteria and one strain of yeast by the disc diffusion method, fluorescein diacetate (FDA) method and broth dilution method. The OIPl exhibited a very strong growth inhibition to Candida albicans. The OIPl remarkably sup­pressed the growth of Salmonella typhimurium and Staphylococcus aureus. The OIPl showed higher growth inhibition to Escherichia coli and Pseudomonas aeruginosa than propolis, positive control. When the anticancer activity of the OIPl, FI or FII was examined against human cancer cell lines and the Sarcoma 180 cells, these widely suppressed the proliferation of cell lines in the MTT assay and morphology study. Especially, they remarkably inhibited the growth of A549, HeLa and AGS cells. Also treatment of cancer cells with OJPl, FI or FII induced apoptotic cell death characterized by DNA fragmentation. The OJPl, FI or FII exhibiting various biological activities such as antimicrobial activity and anticancer activity is expected to be developed as new biohealth products.

Effect of Retrovirus Mediated TNF-$\alpha$ Gene Transfer to Tumor Necrosis Factor(TNF) Sensitive Tumor Cell Lines on Sensitivity to TNF (Retroviral Vector를 이용한 TNF-$\alpha$ 유전자의 이입이 암세포의 종양괴사인자(TNF) 감수성에 미치는 효과)

  • Oh, Yeon-Mok;Park, Kyeo-Yeong;Jung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Goo;Sim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.2
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    • pp.87-96
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    • 1994
  • Background : Since tumor necrosis factor was discovered in 1975, TNF has been well known about its cytotoxic effect on tumor cells in vivo and in vitro. According to the recent improvement of molecular biological techinques, it is possible that exogenous TNF gene is transferred to tumor cells and is expressed in theirs. By virtue of TNF gene transfer, we have expected that TNF expressed in TNF-gene-transferred tumor cells would kill tumor cells in vivo without systemic side effect. The expected mechanisms in which antitumor effects of TNF expressed in TNF-gene-transferred tumor cells are working would be as followings. In the first mechanism, TNF expressed in TNF-gene-transferred tumor cells would kill tumor cells around(like homicide). In the second mechanism, TNF expressed in TNF-gene-transferred tumor cells would kill themselves(like suicide). In the third mechanism, TNF expressed in TNF-gene-transferred tumor cells would recruit immune effector cells and kill tumor cells indirectly. In the last mechanism, TNF expressed in TNF-gene-transferred tumor cells would augment cytokine such as interferon-$\gamma$ to kill tumor cells. Among these four mechanisms of antitumor effect, only the second mechanism has not been established yet. Therefore, to elucidate the second mechanism, We performed this study. Method : We transferred TNF-$\alpha$ gene to NCI-H2058, a human mesothelioma cell line and WEHI164, a murine fibrosarcoma cell line by using retroviral vector(pLT12SNTNF). And, We determined by using MTT assay whether TNF expressed in TNF-gene-transferred tumor cell lines would kill themselves like suicide or not. Then, if TNF-gene-transferred tumor cell lines would not suicide themselves, I would know more about the TNF sensitivity of TNF-gene-transferred tumor cell lines to exogenous TNF also by MTT assay. Result : NCI-H2058 and WEHI164 which were sensitive to TNF, became far less sensitive to endogenous and exogenous TNF after being transferred TNF-$\alpha$ gene to. Conclusion : TNF-gene-transfer to NCI-H2058 and WEHI164 gave them resistance to TNF.

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Preparation of CdSe QDs-carbohydrate Conjugation and its Application for HepG2 Cells Labeling

  • Jiang, Mingxing;Chen, Yan;Kai, Guiqing;Wang, Ruijun;Cui, Huali;Hu, Meili
    • Bulletin of the Korean Chemical Society
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    • v.33 no.2
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    • pp.571-574
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    • 2012
  • In present study, CdSe quantum dots (QDs) were prepared with a novel but simple, effective and exercisable method. Nine different types of carbohydrate molecules were used to modify CdSe QDs. D-mannose (Man)-coated quantum dots were prepared for labeling human hepatoma (HepG2) cells, because of the high expression of mannose receptor (MR) on HepG2 cells. The uptake characteristics of CdSe QDs-Man were investigated in HepG2 cells. The absorption rate result of MTT assay in 48 h suggested the extremely low cytotoxicity of CdSe QDs-Man. The presence of quantum dots was confirmed with fluorescence microscopy. These results were encouraging regarding the application of QDs molecules for early detection of HepG2 cells.

Fabrication and characterization of PCL/TCP-coated PHBV composite multilayer as a bone plate

  • Kim, Yang-Hee;Song, Ho-Yeon;Lee, Byong-Taek
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2009.11a
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    • pp.39.2-39.2
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    • 2009
  • In this work, Poly($\varepsilon$-Caprolactone)(PCL) andpoly(3-hydroxybutyrate-co-3-hydroxyvalerate)(PHBV) mats were fabricated usingelectrospinning process. The electrospinning process is a simple and efficient method to fabricate the nanofibrous mats. PCL and PHBV is a kind of biodegradable polymer but their mechanical properties aren't good. For improving mechanical properties, PHBV mats were coated by TCP. Using PCL mats and TCP-coated PHBV composite mats, a bio-resorbablebone plate were made by pressing. Detailed micro-structural characterization was done by SEM techniques. Tensile strength and bending strength were also evaluated for mechanical properties. The cytotoxicity evaluation ofPCL/TCP-coated PHBV composite multilayer was done by MTT assay. The evidence obtained in this work implies the potential for use as a biodegradable boneplate.

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Anti-tumor Effects of Soybeans and Fermented Soybean Paste

  • Lee, Sung-Lim;Kim, Jong-Gyu
    • Proceedings of the Korean Environmental Health Society Conference
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    • 2004.06a
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    • pp.182-184
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    • 2004
  • Oral cancer is the sixth most common cancer globally. The effects of several extracts from soybeans and Korean soybean paste (doen-jang) on the growth of human oral carcinoma cells in vitro were assessed. We prepared petroleum ether extract, ethyl acetate extract, chloroform extract, methanol extract, and water extract from soybeans and soybean paste. We used KB cell, which is an oral epidermoid carcinoma cell, and investigated proliferation of the tumor cells using MTT method. Each extract of soybean paste suppressed the KB cell proliferation. A dose-response relationship was observed between the level of ethyl acetate extract of soybean paste and its suppression of the cell proliferation. The effects of soybean extracts were lower than those of soybean paste extracts. The effects might be enhanced by the fermentation of soybeans. The results of this work indicate that extracts from soybeans and Korean soybean paste could have potential as anti-tumor substances.

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BIOCOMPATIBISITY OF ION BEAM PROCESSED FILMS DEPOSITED ON SURGICAL TI-6AI-4V

  • Lee, I-S;Song and I-j Yu
    • Journal of the Korean Vacuum Society
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    • v.6 no.S1
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    • pp.16-22
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    • 1997
  • ion beam processing of materials for medical application has gained increasing interest in the last decade and the implantation of nitrogen into TI-6AI-4V to improve corrosive-wear performance is currently used for processing of total hip and knee joints. Oxides and nitrides of Ti, Zr, Al, Cr were deposited on TI-6AI-4V substrates by DC magnetron sputtering dual ion beam sputtering and ion beam assisted deposition. The cytotoxicity of these films were investigated by MTT method and showed comparable to untreated TI-6AI-4V Plasm-sprayed hydroxyapatite(HAp) coatings showed excellent cytotoxicity regardless of heat treatment. intermediate layer coatings of nitrides and oxides increased the bond strength of HAp to substrate by intrdducing chemical bond at interface. Heat treatment of HAp coatings also improved the chemical bond at interfaces and increased the bond strength of untreated TI-6AI-4V to 16.4 kg/$\textrm{cm}^2$ but still lower than 33.1 kg./$\textrm{cm}^2$ of ir oxide as a imtermediate layer caoting.

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Expression of Human Stem Cell Factor with Recombinant Baculovirus in BmN Cell Line and Silkworm

  • Xijie, Guo;Yongfeng, Jin;Mingguan, Yang;Yaozhou, Zhang
    • International Journal of Industrial Entomology and Biomaterials
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    • v.4 no.1
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    • pp.51-56
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    • 2002
  • A recombinant transfer vector pBacSCF was constructed by inserting huamn stem cell factor (hSCF) cDNA into plasmid pBacPAK8. BmN cells were co-transfected with modified Bombyx mori, nuclear polyhedrosis virus (BmBacPAK) DNA and the recmbinant transfer vector to construct a recombinant baculovirus containing hSCE gene. DNA dot blotting and RNA dot blotting demonstrated that the hSCE gene was contained in the recombinant virus and transcribed. The recombinant baculovirus was infectious to BmN cells and to silkworm. SDS-PAGE analysis showed a specific band of expressed product in the extract of infected cells and in the heamolymph of infected larvae. Bioactivity of the recombinant hSCE was determined with W-1 cell line and MTT colorimetric method in synergy with interlukin-3 (IL-3). These results revealed that the hSCF gene was over-expressed in cultured cells and lavae of silkworm.

Effect of Codonopsis lanceolatae Radix Water Extract on Immunocytes (더덕(Codonopsis lanceolatae Radix) 추출물이 면역세포에 미치는 영향)

  • 서정숙
    • The Korean Journal of Food And Nutrition
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    • v.9 no.4
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    • pp.379-384
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    • 1996
  • The purpose of this research was to investigate the effect of Codonopsis lanceolatae Radix water extract(CLE) on immunocytes. The effect of CLE on the proliferation of thymocytes was estimated by MTT colorimetric assay, sub-population of thymocytes was estimated by laser flow cytometry, production of nitric oxide from macrophages was estimated by Griess method, and phagocytosis of human polymorphonuclear cells was estimated by lucigenin chemiluminescence. CLE increased the proliferation of thymocytes in vivo, but did not affect the proliferation of thymocytes in vitro. CLE accelerated the activation of heper T cells in thymocytes. CLE inhibited the production of nitric oxide from peritoneal macrophages, and increased the phagocytosis of human polymorphonuclear cells. These results suggest that CLE have immuno-regulatory action in vivo.

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