• Title/Summary/Keyword: MTT-test

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In Vitro Cytotoxicity of Pt(II) Complexes Containing Ethylenediamine in Rabbit Kidney Proximal Tubular and Human Renal Cortical Cells (에틸렌디아민을 배위자로 한 백금(II)착체의 토끼 및 인체 신장세포에 대한 in vitro 독성)

  • Rho, Young-Soo;Lee, Kyung-Tae;Jung, Jee-Chang;Chang, Sung-Goo
    • YAKHAK HOEJI
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    • v.40 no.2
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    • pp.218-224
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    • 1996
  • This laboratory has recently reported the synthesis and in vitro antitumor activity of PT(II) complexes containing ethylenediamine and diphosphine. In view of the reports of others, cisplatin is toxic to the kidney since the kidney's vulnerability to PT(II) complexes may originate in its ability to accumulate and retain platinum to a greater degree than other organs. The in vitro cytotoxicity of these synthetic PT(II) complexes on the primary cultured proximal tubular cells of rabbit kidney and renal cortical cells of human kidney was investigated. Three endpoints for cytotoxicity tests were evaluated:3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), $^3H$-thymidine uptake and the glucose consumption tests. The rank order of sensitivity exhibited $^3H$-thymidine uptake>MTT>glucose consumption test. The agents with diphosphine leaving group were significantly less cytotoxic than cisplatin. Moreover, 1,2-bis(diphenylphosphino)ethane (DPPE) exhibited less cytotoxicity than 1.3-bis (diphenylphosphino)propane (DPPP) against on rabbit and human cultured kidney cells. Based on these results, the decreased nephrotoxicity of these new complexes over cisplatin appeared to be partially attributable to a leaving group of DPPP and DPPE. This novel class of platinum compound represents a valuable lead in the development of a "third-generation" agent.

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CHARACTERISTICS OF FIBROUS DYSPLASIA DERIVED CELLS (섬유성이형성증 유래세포의 특성연구)

  • Lee, Chan-Hee;Han, Ihn;Seo, Byoung-Moo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.35 no.5
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    • pp.304-309
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    • 2009
  • Purpose: Fibrous dysplasia (FD) is a fibro-osseous disease associated with activating missense mutations of the gene encoding the $\alpha$-subunit of stimulatory G protein. FD may affect a single bone (called monostotic form) or multiple bones (called polyostotic form). The extent of lesions reflects the onset time of mutation. In this study, cells from monostotic FD in maxilla of a patient were isolated and cultured in vitro for characterization. Materials and Methods: The single cells were released from FD lesion which was surgical specimen from 15 years-old boy. These isolated cells were cultured in vitro and tested their proliferation activity with MTT assay. In osteogenic media, these cells underwent differentiation process comparing with its normal counterpart i.e. bone marrow stromal cells. The proliferated FD cells were detached and transplanted into the dordsal pocket of nude mouse and harvested in 6 weeks and 12 weeks. Results and Summary: FD cells have an increased proliferation rate and poor differentiation. As a result, cells isolated from FD lesion decreased differentiation into osteoblast and increased proliferation capacity. MTT assay presented that proliferation rate of FD cells were higher than control. However, the mineral induction capacity of FD was lesser than that of control. Monostotic FD cells make fewer amounts of bone ossicles and most of them are woven bone rather than lamellar bone in vivo transplantation. In transplanted FD cells, hematopoietic marrow were not seen in the marrow space and filled with the organized fibrous tissue. Therefore, they were recapitulated to the original histological features of FD lesion. Collectively, these results indicated that the FD cells were shown that the increased proliferation and decreased differentiation potential. These in vitro and in vivo system can be useful to test FD cell's fate and possible.

Purification of Materials Produced by Amylocolatosis sp. and Anticancer Effect in Oral Cancer Model (Amylocolatosis sp.가 생산하는 항암물질의 정제 및 구강암 모델에 미치는 항암 효과)

  • Kim, Jung;Park, Young-Min;Lim, Do-Seon
    • Journal of dental hygiene science
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    • v.3 no.1
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    • pp.11-14
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    • 2003
  • A methylotrophic Actinomycetes strain, which produce the anti-oral cancer activity compound, was isolated from soil and estimated as Amylocolatosis sp. based on taxonomic studies. A methanol didn't have influence on the production of the anticancer compounds. These compound were isolated by ethylacetate extract, silica gel column chromatography, sephadex LH-20 column and reverse phase HPLC. The compounds were very stable under heat ($121^{\circ}C$), acid(pH 2.0) and alkali(pH 11.0) treatment. The cytotoxic effect of isolated anticancer compounds on various cancer cell lines such as A549, SNU-1, KB, L1210, and Sarcoma 180 was investigated by MTT assay method. And these produced compounds also showed the broad antimicrobial spectrum to test strains such as bacteria and yeast.

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Development of Anti-Wrinkle Agent from Nelumbo nucifera Root Extract (연근 추출물에서 주름개선 소재의 개발)

  • Kim, Hee Jin;Kim, Tagon;Kang, Whan Yul;Baek, Hyun;Cheon, Hae Young;Kim, Bo Young;Kim, Donguk
    • Korean Chemical Engineering Research
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    • v.48 no.4
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    • pp.413-416
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    • 2010
  • In this research, root extracts of Nelumbo nucifera was tested to see the possibility for functional cosmetic agent. 70-100% ethanol was used as solvent and nuciferin was confirmed as active component. To test cosmetic effect of root extracts of Nelumbo nucifera, safety effect(MTT assay), anti-wrinkle effect(elastase inhibition assay) and antioxidation effect(DPPH free radical scavenging assay) were measured. When 100% ethanol was used as extracting solvent, cell viability was over 80% at $100{\mu}g/ml$, which indicated that root extract of Nelumbo nucifera was suitable for cosmetic agent. Root extract of Nelumbo nucifera showed 40~50% elastase inhibition at $100{\mu}g/ml$ so that it had good anti-wrinkle characteristics. 50% antioxidation capacity($FSC_{50}$) was $5.0{\sim}38{\mu}g/ml$ and root extract of Nelumbo nucifera showed excellent antioxidation effect. From the research, root extracts of Nelumbo nucifera showed strong possibility for anti-wrinkle functional cosmetic agent.

Cell attachment and proliferation of bone marrow-derived osteoblast on zirconia of various surface treatment

  • Pae, Ahran;Lee, Heesu;Noh, Kwantae;Woo, Yi-Hyung
    • The Journal of Advanced Prosthodontics
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    • v.6 no.2
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    • pp.96-102
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    • 2014
  • PURPOSE. This study was performed to characterize the effects of zirconia coated with calcium phosphate and hydroxyapatite compared to smooth zirconia after bone marrow-derived osteoblast culture. MATERIALS AND METHODS. Bone marrow-derived osteoblasts were cultured on (1) smooth zirconia, (2) zirconia coated with calcium phosphate (CaP), and (3) zirconia coated with hydroxyapatite (HA). The tetrazolium-based colorimetric assay (MTT test) was used for cell proliferation evaluation. Scanning electron microscopy (SEM) and alkaline phosphatase (ALP) activity was measured to evaluate the cellular morphology and differentiation rate. X-ray photoelectron spectroscopy (XPS) was employed for the analysis of surface chemistry. The genetic expression of the osteoblasts and dissolution behavior of the coatings were observed. Assessment of the significance level of the differences between the groups was done with analysis of variance (ANOVA). RESULTS. From the MTT assay, no significant difference between smooth and surface coated zirconia was found (P>.05). From the SEM image, cells on all three groups of discs were sporadically triangular or spread out in shape with formation of filopodia. From the ALP activity assay, the optical density of osteoblasts on smooth zirconia discs was higher than that on surface treated zirconia discs (P>.05). Most of the genes related to cell adhesion showed similar expression level between smooth and surface treated zirconia. The dissolution rate was higher with CaP than HA coating. CONCLUSION. The attachment and growth behavior of bone-marrow-derived osteoblasts cultured on smooth surface coated zirconia showed comparable results. However, the HA coating showed more time-dependent stability compared to the CaP coating.

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.

The Effects on Antimicrobial and Anticarcinogenic Activity of Momordica Charantia L. (메탄올로 추출한 여주 분획성분의 항균 및 항발암 효과)

  • 배송자
    • Journal of Nutrition and Health
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    • v.35 no.8
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    • pp.880-885
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    • 2002
  • This study was performed to determine the antimicrobial and anticarcinogenic activities of the Momordica charantia L. (MC) on several microorganisms and human cancer cell lines. In the paper disk test, its antimicrobial activity was increased in proportion to its concentration. Among the various solvent fractions of Momordica charantia L., the ethylether partition layer (MCMEE) showed the strongest antimicrobial activity. Also, the ethylacetate partition layer (MCMEA) and the butanol partition layer (MCMB) showed antimicrobial activity. We also determined the cytotoxicity and chemopreventive effect of Momordica charantia L. extract and fractions on human cancer cells. The experiment was conducted to determine the cytotoxicity of Momordica charantia L. partition layers on HepG2, HeLa and MCF-7 cells by MTT assay. Among the various partition layers of Momordica charantia L., MCMEE and MCMEA showed strong cytotoxic effects on all cancer cell lines. The chemopreventive effect of the quinone reductase induced activities of HepG2 cell, the hexane partition layer (MCMH) at a dose of 50 $\mu\textrm{g}$/mL was 3.62 times more effective compared with the control values of 1.0. Therefore, based on these studies, Momordica charantia L. may be developed into a potentially useful cancer chemopreventive agent.

Cytocompatibility and cell proliferation evaluation of calcium phosphate-based root canal sealers

  • Mestieri, Leticia Boldrin;Zaccara, Ivana Maria;Pinheiro, Lucas Siqueira;Barletta, Fernando Branco;Kopper, Patricia Maria Polli;Grecca, Fabiana Soares
    • Restorative Dentistry and Endodontics
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    • v.45 no.1
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    • pp.2.1-2.7
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    • 2020
  • Objectives: This study aimed to evaluate the cell viability and migration of Endosequence Bioceramic Root Canal Sealer (BC Sealer) compared to MTA Fillapex and AH Plus. Materials and Methods: BC Sealer, MTA Fillapex, and AH Plus were placed in contact with culture medium to obtain sealers extracts in dilution 1:1, 1:2 and 1:4. 3T3 cells were plated and exposed to the extracts. Cell viability and migration were assessed by 3-(4,5-dimethylthiazoyl)-2,5-diphenyl-tetrazolium bromide (MTT) and Scratch assay, respectively. Data were analyzed by Kruskal-Wallis and Dunn's test (p < 0.05). Results: The MTT assay revealed greater cytotoxicity for AH Plus and MTA Fillapex at 1:1 dilution when compared to control (p < 0.05). At 1:2 and 1:4 dilutions, all sealers were similar to control (p > 0.05) and MTA Fillapex was more cytotoxic than BC Sealer (p < 0.05). Scratch assay demonstrated the continuous closure of the wound according to time. At 30 hours, the control group presented closure of the wound (p < 0.05). At 36 hours, only BC Sealer presented the closure when compared to AH Plus and MTA Fillapex (p < 0.05). At 42 hours, AH Plus and MTA Fillapex showed a wound healing (p > 0.05). Conclusions: All tested sealers demonstrated cell viability highlighting BC Sealer, which showed increased cell migration capacity suggesting that this sealer may achieve better tissue repair when compared to other tested sealers.

Chemical Composition and Biological Activities of Essential Oils Extracted from Korean Endemic Citrus Species

  • Baik, Jong-Seok;Kim, Sang-Suk;Lee, Jung-A;Oh, Tae-Heon;Kim, Ji-Young;Lee, Nam-Ho;Hyun, Chang-Gu
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.74-79
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    • 2008
  • The aim of this study was to analyze the chemical composition of 14 kinds of citrus oils and to test their biological activities. Citrus essential oils were obtained by steam distillation from immature fruits collected from Jeju Island and were analyzed using gas chromatograph (GC)-flame ionization detectors (FID) and GC-MS. Limonene (55.4% to 91.7%), myrcene (2.1% to 32.1%), ${\alpha}$-pinene (0.6% to 1.6%) and linalool (0.4% to 6.9%) were the major components in most citrus species. To evaluate in vitro antibacterial activity, all essential oils were tested against Propionibacterium acnes and Staphylococcus epidermidis. Nine out of fourteen citrus oils exhibited antibacterial activity against P. acnes, but not against S. epidermidis. The effects of the citrus oils on DPPH radical scavenging, superoxide radical anion scavenging, nitric oxide radical, and cytotoxicity were also assessed. Three essential citrus oils, Joadeung, Dongjunggyul, and Bujiwha, exhibited potent inhibitory effects on nitric oxide production. Two essential oils, Dongjunggyul and Joadeung, showed potent free radical scavenging activities in the DPPH assay. For future applications in cosmetic products, we also performed MTT assays in a human dermal fibroblast cell line. The majority of the essential oils showed no cytotoxicity. The results indicate that citrus essential oils can be useful natural agents for cosmetic application.

Analysis of Antibacterial Activity against Food Spoilage and Food-borne Pathogens and Cytotoxicity on Human Cancer Cell Lines of Extracts from Pericarp and Seed of Vitis coignetiea (머루 과피와 종자 추출물의 식품 위해성 세균에 대한 항균성 및 인체 암세포주에 대한 cytotoxicity 분석)

  • Won, Ji-Hye;Kim, Mee-Ra
    • Korean journal of food and cookery science
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    • v.28 no.2
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    • pp.175-182
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    • 2012
  • In this study, antibacterial activity and cytotoxicity of the extracts from pericarp and seed of $Vitis$ $coignetiea$, which were extracted with 0.1% HCl-60% ethanol, were analyzed. The antibacterial activity of the extracts was determined by paper disc diffusion method against food spoilage and food-borne pathogens. The pericarp extract showed high antibacterial activity against $Bacillus$ $cereus$, $Escherichia$ $coli$ O157:H7, and $Pseudomonas$ $aeruginosa$, and the seed extract represented the high antibacterial activity against $B.$ $cereus$, $E.$ $coli$ O157:H7, and $Staphylococcus$ $aureus$. The cytotoxicity of the $Vitis$ $coignetiea$ extract against human cancer cells was determined using the MTT assay and SRB assay. The pericarp extract represented strong growth-inhibition activity against G361 and Hep3B cells and the seed extract greatly inhibited the growth of HeLa and G361 cells in the MTT assay. In addition, the pericarp extract displayed a high inhibition activity against the growth of AGS cells and the seed extract greatly inhibited the growth of HeLa, Hep3B, and MCF7 cells in the SRB assay. Especially, the cytotoxicities of the seed extract against HeLa were significantly higher than those of the extract against other cancer cells at all test concentrations. This study demonstrates that the extract from pericarp and seed of $Vitis$ $coignetiea$ possess high antibacterial activity and cytotoxicity.