• Title/Summary/Keyword: MTT-test

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Fabrication of Cross-linked Nano-Fibrous Chitosan Membranes and Their Biocompatibility Evaluation

  • Nguyen, Thi-Hiep;Lee, Seong-Jin;Min, Young-Ki;Lee, Byong-Taek
    • Korean Journal of Materials Research
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    • v.21 no.2
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    • pp.125-132
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    • 2011
  • Fibrous chitosan membranes were fabricated as a substrate for skin applications using an electro-spinning process with different solvents and varying concentrations. Scanning electron microscopy (SEM) images confirmed that the formation of the chitosan fibrous membrane in trifluoroacetic acid was better than that in acetic acid. Fourier transform infrared spectroscopy showed that the chitosan fibers were cross-linked with glutaraldehyde, and that the cytotoxicity of the aldehyde groups was reduced by glycine and washing by NaOH and DI water. Chitosan cross-linked fibrous membranes were insoluble in water and could be washed thoroughly to wash away glycine and excess NaOH and prevent the infiltration of other water soluble bio-toxic agents using DI water. MTT assay method was employed to test the cytotoxicity of chitosan membranes during fabricating, treating and washing processes. After the dehydration of cell cultured chitosan membranes, cell attachment behavior on the material was evaluated using SEM method. Effect of the treatment processes on the biocompatibility of the chitosan membranes was shown by comparing of filopodium and lamellipodium of fibroblast cells on grown washed and unwashed chitosan fibrous membrane. The MTT assay and SEM morphology confirmed that the washed chitosan fibrous membrane increased cell attachment and cell growth, and decreased toxicity compared to results for the unwashed chitosan fibrous membrane.

Neuroprotective effects of modified Bo-Yang-Hwan-Oh-Tang in N2a neuroblastoma cells (가감보양환오탕(加減補陽還五湯)의 N2a 뇌신경세포에 대한 보호효과)

  • Lim, Kyu;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.77-84
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    • 2006
  • Objectives : To evaluate the neuroprotective effects of modified Bo-Yang-Hwan-O-Tang (BHT), we investigated the neuronal death protection effects to oxidative damages in N2a neuroblastoma cells. Methods : To study the cytotoxic effect of BHT on N2a neuronal cells, the cell viability was determined by MTT assay. To investigate the neuronal death protection of BHT, N2a neuronal cells were induced oxidative damages by H2O2, and assayed the cell viability and DNA fragmentation. We also investigated DPPH free radical scavenging effects of BHT by tube test. Results : In MTT assay, $500{\mu}g/ml$ of BHT was not showed cytotoxic effect on N2a neuronal cells. BHT protected N2a neuronal cells from H2O2-induced cell death in a dose-dependent manner. BHT also protected N2a neuronal cells from H2O2-induced DNA fragmentation. BHT scavenged DPPH free radicals in a dose-dependent manner. Conclusion : These data suggest that BHT may have strong anti-oxidant effects through the free radical scavenging and neuroprotective effects in neuronal cells.

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Protective effects of added Bo-Yang-Hwan-Oh-Tang on $H_2O_2-induced$ neurotoxicity in SH-SY5Y neuronal cells (가매보양환오탕(加昧補陽還五湯)의 SH-SY5Y 뇌신경세포에서 산화적 손상에 의한 세포사멸에 대한 보호효과)

  • Han, Hyung-Soo;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.85-92
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    • 2006
  • Objectives : To evaluate the neuroprotective effects of added Bo-Yang-Hwan-Oh-Tang (BHT), we investigated the neuronal death protection effects to oxidative damages in SH-SY5Y neuronal cells. Methods : To study the cytotoxic effects of BHT on SH-SY5Y cells, the cell viability was determined by MTT assay. To investigate the neuronal death protection of BHT, SH-SY5Y cells were induced oxidative damages by $H_2O_2$ and then assayed the cell viability and DNA fragmentation. We also investigated DPPH free radical scavenging effect of BHT by tube test. Results : In MTT assay, $1000{\mu}g/ml$ of BHT was not showed the cytotoxic effect on SH-SY5Y cells. BHT protected SHSY5Y cells from $H_2O_2-induced $ neuronal cell death in a dose-dependent manner. BHT also protected SH-SY5Y cells from $H_2O_2-induced$ DNA fragmentation. BHT effectively scavenged DPPH free radicals in a dose-dependent manner. Conclusion : These data suggest that BHT may have strong antioxidant effects through the free radical scavenging and neuroprotective effects in human neuronal cells.

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Antimutagenic and Cytotoxic Activity of Ethanol and Water Extracts from Rubus coreanum (복분자 딸기(Rubus coreanum) 에탄올 및 열수추출물의 항돌연변이 활성과 암세포 성장 억제 효과)

  • Jeon, Yeon-Hee;Choi, Sang-Won;Kim, Mee-Ra
    • Korean journal of food and cookery science
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    • v.25 no.3
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    • pp.379-386
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    • 2009
  • The antimutagenic and cytotoxic activities of ethanol and water extracts from Rubus coreanum were investigated in this study. Their antimutagenic activities were measured by the Ames test and their cytotoxic activities were evaluated by the growth inhibition of cancer cells via the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay and the sulforhodamine B (SRB) assay. In the results, the inhibition rates of the ethanol and water extracts toward mutagenicity induced by 4-NQO were 95.0% and 93.6% at 5 mg/plate, respectively, while their inhibition rates against mutagenicity induced by sodium azide were 27.2% and 40.8%, respectively. According to MTT assay, the cytotoxicity values of the ethanol extract against Hep3B and HeLa cells were 67.2% and 68.5%, respectively, and the values for the water extract were 65.8% and 66.4%, respectively. In the SRB assay, the ethanol and water extracts inhibited over 60% of cancer cell growth. In conclusion, both the ethanol and water extracts of Rubus coreanum offer potentially good antimutagenic and anticancer effects.

Ferutinin, an Apoptosis Inducing Terpenoid from Ferula ovina

  • Matin, Maryam Moghaddam;Nakhaeizadeh, Hossein;Bahrami, Ahamd Reza;Iranshahi, Mehrdad;Arghiani, Nahid;Rassouli, Fatemeh Behnam
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.5
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    • pp.2123-2128
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    • 2014
  • A current hurdle in cancer management is the intrinsic or acquired resistance of cancer cells to chemical agents that restricts the efficacy of therapeutic strategies. Accordingly, there is an increasing desire to discover new natural compounds with selective toxicity to combat malignancies. In present study, the cytotoxic and apoptosis-inducing activities of ferutinin, a terpenoid derivative from Ferula ovina, were investigated on human breast (MCF7) and bladder (TCC) cancer cells as well as normal fibroblasts (HFF3).The toxicity and DNA damage inducing effects of ferutinin were studied by MTT and comet assays, DAPI and PI staining and DNA laddering. The $IC_{50}$ values of ferutinin were identified and compared with routine prescribed drugs, doxorubicin and vincristine, by MTT test. Alkaline comet assay and DAPI staining revealed DNA damage due to ferutinin, which was significantly (p<0.001) higher in MCF7 and TCC than HFF3 cells. Apoptosis induction was evidenced by PI staining and DNA laddering. Our results suggest that ferutinin could be considered as an effective anticancer agent for future in vivo and clinical experiments.

Screening of Cytotoxicity and Antimicrobial Effects of Hexane Extracts from Cornis fructus (산수유 헥산 추출물의 항균효과 및 세포독성)

  • Chun Hyun Ja;Choi Won Hyung;Lee Jeong Ho;Yang Hyun Ok;Baek Seung Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.476-480
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    • 2003
  • Cornis fructus was extracted by successive extraction and then fractionated with hexane extract to get active fractions. This study was performed to determine the cytotoxic effect of hexane extract from Cornis fructus on NIH 3T3 fibroblasts and cancer cell lines using MTT assay. Hexane extract showed cytotoxic effect against A549, B16 melanoma and MDA-MB-231. Futher fractionation with hexane extract was performed to obtain effective fraction, fraction 3 showed the cytotoxic effect against A549 and MDA-MB-231. In antimicrobial test of each fraction of hexane extract, fraction 5 showed antimicrobial activities against P. putida and P. aeruginosa.

Correlation Between Skin Irritation and Cytotoxicity of Anti-wrinkle Agents (화장품 원료의 피부자극성과 세포독성의 관련성)

  • 이은희;이종권;김용규;박기숙;안광수
    • YAKHAK HOEJI
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    • v.45 no.3
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    • pp.310-319
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    • 2001
  • To compare skin irritation and cytotoxicity of anti-wrinkle agents, we examined skin irritation of six anti-wrinkle agents (ascorbic acid, glycolic acid, all trans-retinoic acid, ginseng extract, retinol, EB) in New Zealand white rabbit. Cytotoxicity of these agents was determined by MTT [tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] at multi-time points in cultured HaCaT cell, a human immortalized keratinocyte cell. We then analyzed correlation between skin irritation and cytotoxicity by spearman's rank correlation analysis. All trans-retinoic acid showed the highest primary irritation index (0.92) in skin irritation test. Being all the six agents not irritant, retinal showed the most cytotoxic agents. The correlation between skin irritation and cytotoxicity ($IC_{50}$/ at different time point was 0.814, 0.757, 0.814 and 0.7 at 3, 24, 48 and 72 h, respectively. We also fecund that IC$_{20}$ and IC$_{80}$ of these agents showed similar correlation with skin irritation. These results therefore demonstrated that there is close correlation between skin irritation and cytotoxicity $IC_{50}$/ value by MTT in HaCaT cell at early time points by anti-wrinkle agents or IC$_{20}$ value. $IC_{50}$/ at earily time point or IC$_{20}$ values may be reliable alternative determinant of skin irritation.n.

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Inhibitory Effect of Saengangeonbitang-gasamchilgn on Liver Fibrosis in Rat (생간건비탕가삼칠근(生肝健脾湯加三七根)이 흰쥐의 간섬유화 억제에 미치는 영향)

  • Lee, Eun;Kim, Young-Chul;Ko, Heung
    • The Journal of Internal Korean Medicine
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    • v.29 no.2
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    • pp.500-511
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    • 2008
  • The aim of this study was to investigate the inhibitory effect of Saengangeonbitang-gasamchilgn(SGGBTGSCG) on collagen production in rat hepatic stellate cells(HSC) and on the TAA-induced chronic liver injury model in rats. Methods : 1) HSCs were treated with SGGBTGSCG extract powder(50% EtOH SGGBTGSCG, dw SGGBTGSCG). After the treatment, MTT assay, BrdU assay and procollagen assay were done. In addition, gene expressions of collagen type $1{\alpha}2$, ASMA, TIMP1, and TIMP2, all of which are known to be associated with liver fibrosis, were analyzed by RT-PCR. 2) Liver fibrosis was developed in rats by injection of TAA 3 times a week for 5 weeks. After the SGGBTGSCG-treatment, body weight, liver & spleen weight, liver function test, the complete blood cell count and the change of portal pressure were studied. Results : In MTT assay, SGGBTGSCG significantly decreased the viability of HSCs in a dose-dependent manner. In BrdU assay, SGGBTGSCG significantly inhibited the HSC proliferation in a dose-dependant manner. In procollagen assay, SGGBTGSCG decreased procollagen production by HSC. In the change of rats' liver and spleen weight, TAA+SGGBTGSCG groups showed little difference compared with TAA-only group. In the liver function test, SGGBTGSCG decreased the serum level of ALT, AST, and Alp elevated by TAA. In the complete blood cell count, SGGBTGSCG significantly decreased WBC elevated by TAA and increased RBC and Hct lowered by TAA. In the change of portal pressure, SGGBTGSCG decreased portal pressure elevated by TAA. Conclusions : These results suggest that SGGBTGSCG is beneficial in the treatment of cirrhotic patients as well as for patients with chronic hepatitis.

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Efficacy of Safflower on the Acne Skin and Its Application for Facial Cleansing Biomedical Material (홍화잇꽃의 여드름피부 개선효과 및 세안용 생약식물소재 응용)

  • Park, Young-Ho;Lee, Chang-Seop
    • Journal of the Korean Chemical Society
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    • v.55 no.3
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    • pp.400-404
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    • 2011
  • Safflower is called as the 'beneficial flower' because 'it helps human health', and it was introduced as red flower in Tonguibogam due to the red color of floral leaf. From old times, it has been used for the material of cloth and rouge. Recently, polyphenol compound, the main ingredient of safflower, known as anti-aging and anti-oxidizing material in the healthy food industry becomes the emerging hot topic. This study aims to confirm by DDT (Disk Diffusion Test) assay, MTT assay, and NF-${\kappa}$B Luciferase activity inhibition assay in vitro that polyphenol compound, which is the main ingredient of safflower, has the anti-microbial efficacy to inhibit the growth of acne germs that make troubles for the teenagers or middle aged. Also it aims to evaluate its clinical efficacy on the acne skin, utilizing the facial cleansing cosmetic form of soap sample. This study can contribute to take a major step forward to the development of cosmetic soap for acne in the cosmeceutical industry.

Gene expression of MC3T3-E1 osteoblastic cells on titanium and zirconia surface

  • Gong, Soon-Hyun;Lee, Heesu;Pae, Ahran;Noh, Kwantae;Shin, Yong-Moon;Lee, Jung-Haeng;Woo, Yi-Hyung
    • The Journal of Advanced Prosthodontics
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    • v.5 no.4
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    • pp.416-422
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    • 2013
  • PURPOSE. This study was performed to define attachment and growth behavior of osteoblast-like cells and evaluate the gene expression on zirconia compared to titanium. MATERIALS AND METHODS. MC3T3-E1 cells were cultured on (1) titanium and (2) zirconia discs. The tetrazolium-based colorimetric assay (MTT test) was used for examining the attachment of cells. Cellular morphology was examined by scanning electron microscopy (SEM) and alkaline phosphatase (ALP) activity was measured to evaluate the cell differentiation rate. Mann-Whitney test was used to assess the significance level of the differences between the experimental groups. cDNA microarray was used for comparing the 20215 gene expressions on titanium and zirconia. RESULTS. From the MTT assay, there was no significant difference between titanium and zirconia (P>.05). From the SEM image, after 4 hours of culture, cells on both discs were triangular or elongated in shape with formation of filopodia. After 24 hours of culture, cells on both discs were more flattened and well spread compared to 4 hours of culture. From the ALP activity assay, the optical density of E1 cells on titanium was slightly higher than that of E1 cells on zirconia but there was no significant difference (P>.05). Most of the genes related to cell adhesion showed similar expression level between titanium and zirconia. CONCLUSION. Zirconia showed comparable biological responses of osteoblast-like cells to titanium for a short time during cell culture period. Most of the genes related to cell adhesion and signal showed similar expression level between titanium and zirconia.