• Title/Summary/Keyword: MTT-test

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Cytotoxic effects of different self-adhesive resin cements: Cell viability and induction of apoptosis

  • Sismanoglu, Soner;Demirci, Mustafa;Schweikl, Helmut;Ozen-Eroglu, Gunes;Cetin-Aktas, Esin;Kuruca, Serap;Tuncer, Safa;Tekce, Neslihan
    • The Journal of Advanced Prosthodontics
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    • v.12 no.2
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    • pp.89-99
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    • 2020
  • PURPOSE. The effects of four different self-adhesive resin cement materials on cell viability and apoptosis after direct and indirect exposure were evaluated using different cell culture techniques. MATERIALS AND METHODS. Self-adhesive cements were applied to NIH/3T3 mouse fibroblasts by the extract test method, cell culture inserts, and dentin barrier test method. After exposure periods of 24 h and 72 h, the cytotoxicity of these self-adhesive materials was evaluated using the MTT assay (viability) and the Annexin-V-FITC/PI staining (apoptosis). RESULTS. The lowest cell viability was found in cells exposed to BeautiCem SA for 24 h in the extract test method. Cell viability was reduced to 70.6% compared to negative controls. After the 72 h exposure period, viability rate of cell cultures exposed to BeautiCem SA decreased more than 2- fold (29.5%) while cells exposed to RelyX U200 showed the highest viability rate of 71.4%. In the dentin barrier test method, BeautiCem SA induced the highest number of cells in apoptosis after a 24 h exposure (4.1%). Panavia SA Cement Plus was the material that caused the lowest number of cells in apoptosis (1.5%). CONCLUSION. The used self-adhesive cements have showed different cytotoxic effects based on the evaluation method. As exposure time increased, the materials showed more cytotoxic and apoptotic effects. BeautiCem SA caused significantly more severe cytotoxic and apoptotic effects than other cements tested. Moreover, cements other than BeautiCem SA have caused necrotic cell death rather than apoptotic cell death.

Studies on the Biological Behaviors of Taxol Derivatives (Taxol 유도체들의 생물학적 거동에 관한 연구)

  • Awh, Ok-Doo;Yoo, Dae-Wung;Im, Sang-Moo
    • The Korean Journal of Nuclear Medicine
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    • v.31 no.4
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    • pp.440-451
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    • 1997
  • This study was designed to prospect the $^{111}In$-labelled paclitaxel as tumor imaging agent. In order to provide a taxol molecule with a functional group which is able to chelate In-111, taxol-DTPA conjugate and 2'-hemisuccinyltaxol were synthesized by esterification of taxol at C-2'on C-13 carbon with DTPA anhydride and succinic anhydride, respectively. Synthesis yield of the taxol derivatives was 34% for taxol-DTPA and 80% for 2'-hemisuccinyltaxol. Cytotoxicity of the taxol derivatives were measured by MTT method toward cell lines HT29, B16, P388, and CT26. The cytotoxic activities of the taxol derivatives were maintained, although less active than taxol. Radiolabelling of the taxol derivatives were proceeded directly with $^{111}InCl_3$ or indirectly with $^{111}In$-citrate(ligand-exchange method). The ligand-exchange method was not suitable because some precipitates appeared during the reaction. On the contrary, by direct radiolabelling method, we were able to obtain taxol-DTPA-$^{111}In$ in 100% radiochemical yield. However, 2'-hemisuccinyltaxol was not labelled by both methods. Yield and radiochemical purity of the radiolabelled com-pound were determined by HPLC, paper chromatography and instant thin layer chromatography. Taxol-DTPA-$^{111}In$ was characterized to be hydrophilic by lipophilicity test, and nearly non-adhesive to HT29, B16, P388, and CT26 by cell binding affinity test. Binding affinity of the taxol-DTPA-$^{111}In$ complex to serum proteins was also examined by protein precipitation with 30% trichloroacetic acid. The results showed that 30% of the taxol-DTPA-$^{111}In$ complex binds with serum proteins.

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Protective Effect of Wheat Bran Extract against β-Amyloid-induced Cell Death and Memory Impairment (베타아밀로이드로 유도된 신경세포 사멸과 기억력 손상에 대한 밀기울추출물의 보호효과)

  • Lee, Chan;Park, Gyu-Hwan;Lee, Jong-Won;Jang, Jung-Hee
    • The Korea Journal of Herbology
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    • v.30 no.1
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    • pp.67-75
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    • 2015
  • Objectives : The aim of this study is to examine the neuroprotective effect of wheat bran extract (WBE) against ${\beta}$-amyloid ($A{\beta}$)-induced apoptotic cell death in SH-SY5Y human neuroblastoma cells and memory impairment in triple transgenic animal model's of Alzheimer's disease (3xTg AD mice). Methods : In SH-SY5Y cells, MTT assay and TUNEL staining were conducted to evaluate the protective effect of WBE against $A{\beta}_{25-35}$-induced neurotoxicity and apoptosis. Alterations in mitochondrial transmembrane potential (MMP), expression of proapoptotic Bax and antiapoptotic Bcl-2 proteins, cleavage of PARP, and brain-derived neurotrophic factor (BDNF) levels were analyzed to elucidate the neuroprotective mechanism of WBE. To further investigate the memory enhancing effect of WBE, Morris water maze test was performed in 3xTg AD mice. Results : In SH-SY5Y cells, WBE protected against $A{\beta}_{25-35}$-caused cytotoxicity and apoptosis as shown by the restoration of cell viability in MTT assay and inhibition of DNA fragmentation in TUNEL staining. $A{\beta}_{25-35}$-induced apoptotic signals such as dissipation of MMP, decreased Bcl-2/Bax ratio, and cleavage of PARP were suppressed by WBE. Moreover, WBE up-regulated the protein levels of BDNF, which seemed to be mediated by activation of cAMP response element-binding protein (CREB). In 3xTg AD mice, oral administration of WBE attenuated learning and memory deficit as verified by reduced mean escape latency in water maze test. Conclusions : WBE protects neuronal cells from $A{\beta}_{25-35}$-induced apoptotic cell death and restores learning and memory impairments in 3xTg AD mice. These findings suggest that WBE exhibit neuroprotective potential for the management of AD.

Induction of Apoptosis by Curcuma aromatica on Lung Cancer Cells(A549), Cervical Cancer Cells(HeLa), Glioma Cancer Cells(A172) and Prostate Cancer Cells(PC3) (울금(鬱金)이 폐암(肺癌), 자궁암(子宮癌), 신경교종(神經膠腫) 및 전립선암(前立腺癌)에 대한 세포자살유도(細胞自殺誘導)에 미치는 영향(影響))

  • Park, Sang-Hyun;Kim, Jin-Sung;Yoon, Sang-Hyub;Ryu, Bong-Ha
    • The Journal of Internal Korean Medicine
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    • v.27 no.2
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    • pp.379-393
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    • 2006
  • Objectives: We are aimed to identify anti-tumor effects of Curcuma aromatics on some kinds of cancer cells through molecular biologic methods. Materials & Methods: We used 4 kinds of cancer cell lines such as lung cancer cells(AS49), cervical cancer cells(HeLa), glioma cancer cells(A172) and prostate cancer cells(PC3). We treated the boiled extract of Curcuma aromatica $5{\mu}g,\;10{\mu}g$ to cultural media(ml) for 24 hours. We measured the cytotoxicitv on 4 kinds of cancer cells through tryphan blue exclusion test and the suppressive effect on viability of 4 kinds of cancer cells via MTT assay. We measured change of mitochondria membrane potential via flow cytometry. The quantitative RT-PCR was used to examine the effect on the revelation of Bcl-2 and Bax which are genes related to apoptosis. We examined the effect on the revelation of Bcl-2 Protein and Bar protein by western blot analysis. Results : In the experiment of tryphan blue exclusion test, the extract of Curcuma aromatica showed more significant killing effect on AS49, HeLa than the control group with density dependent manner, which was statistically significant. In the experiment of MTT assay the extract of Curcuma aromatica showed more suppressive effect on viability of A549, HeLa than the control group with density dependent manner, which was statistically significant. Curcuma aromatica induced apoptosis by decreasing the membrane potential of mitochondria in A549, HeLa. In the experiment of the revelation of genes related to apoptosis, the revelation of Bcl-2 decreased and the revelation of Bax increased in A549, HeLa treated with Curcuma aromatica with dose dependent manner. In the experiment of the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in AS49, HeLa treated with Curcuma aromatica with dose dependent manner. Conclusions: From this study, we can infer that Curcuma aromatica has anti-tumor effect on lung cancer cells and uterine carcinoma cells but not on glioma cells and prostate cancer cells.

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Anti-oxidation and Anti-atopic Dermatitis Effect of Herbal Wood Vinegar (한방목초액의 항산화 및 항 아토피 효과)

  • Kim, Tagon;Nho, Hwa Jung;Jun, Sang Hui;Kim, Kang Bae;Kim, Donguk
    • Korean Chemical Engineering Research
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    • v.48 no.6
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    • pp.690-694
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    • 2010
  • In this study, herbal wood vinegar including Bambusoideae, Cinnamomi Cortex, Zingiberis Rhizoma was tested to see possibility for cosmetic or skin related medicine. Anti-oxidation effect of herbal wood vinegar was tested by DPPH free radical scavenging activity, and showed 97% inhibition rate at $50{\mu}g/ml$. Anti-bacterial effect was tested by disc diffusion method, and it indicated strong anti-bacterial activity against normal skin flora Staphylococcus aureus. Whitening effect was measured by tyrosinase inhibition assay, and it was lower compared with vitamin C. Stability test was done by MTT assay, and cell toxicity was relatively high. Stability was also checked, and there was not significant change in color, aroma, appearance and pH during storage. Anti-atopic dermatitis test was done by hairless mouse and herbal wood vinegar recovered damaged skin to almost normal condition after 9 days of application. IgE concentration in herbal wood vinegar treated mouse was also reduced 30% compared with control. From the study, herbal wood vinegar showed good anti-oxidation, anti-bacterial and anti-atopic dermatitis effect, and had promising application in cosmetic or skin related medicine.

Antioxidative, Antimutagenic, and Cytotoxic Activities of Ethanol Extracts from Cornus officianalis (산수유(Cornus officianalis) 에탄올 추출물의 항산화, 항돌연변이 활성 및 암세포 성장 억제 효과)

  • Jeon, Yeon-Hee;Kim, Mi-Hyun;Kim, Mee-Ra
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.1
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    • pp.1-7
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    • 2008
  • The antioxidative, antimutagenic and cytotoxic activities of ethanol extract from Cornus officianalis have been studied. The antioxidant activity of the ethanol extract was measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. The inhibition effects on the mutagenicity in Salmonella Typhimurium TA100 were evaluated by Ames test and cancer cell inhibitory effects in Hep3B cell and HeLa cell were tested by MTT assay. Cornus officianalis had an important free radical-scavenging activity towards the DPPH radical. At a concentration of 500 ppm, the DPPH radical-scavenging activity of Cornus officianalis was similar to that of L-ascorbic acid. None of the extracts produced a mutagenic effect on S. Typhimurium TA100. The ethanol extract from Cornus officianalis showed about 77% of inhibition at 500 ppm on the mutagenicity induced by 4-nitroquinoline-1-oxide. The extract from Cornus officianalis showed strong cytotoxicity against Hep3B and HeLa cells, with inhibition of 83 and 78% at a dose of $700{\mu}g$/plate, respectively. Moreover, the ethanol extracts had 34.33 mg H.E/g of polyphenols and 5.67 mg Q.E/g of flavonoids, respectively. Therefore, the present study showed antioxidative, antimutagenic and anticancer potential of the ethanol extract from Cornus officianalis.

Cognitive Enhancing Activity of the Steamed and Fermented Extracts of Codonopsis lanceolata Radix (양유(洋乳)의 증숙 및 발효 추출물의 인지능 개선 활성)

  • Weon, Jin Bae;Yun, Bo-Ra;Lee, Jiwoo;Eom, Min Rye;Lee, Hyeon Yong;Park, Dong-Sik;Chung, Hee-Chul;Chung, Jae Youn;Ma, Choong Je
    • YAKHAK HOEJI
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    • v.57 no.5
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    • pp.323-329
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    • 2013
  • This study was designed to determine and compare the cognitive enhancing effect of various Codonopsis lanceolata extracts by steaming, fermentation and high hydrostatic pressure process. We prepared water extract of C. lanceolata, steamed C. lanceolata, steamed and fermented C. lanceolata and C.lanceolata by high hydrostatic pressure process and fermentation. Cognitive enhancing effect of extracts was evaluated in scopolamine-induced memory impairment mice using by passive avoidance test and Morris water maze tests. MTT assay was conducted to investigate neuroprotective effect on glutamate induced cell death in HT22 cells. Steamed and fermented C. lanceolata water extract decreased escape latency in Morris water maze test and increased the latency time of the passive avoidance test compared to other extracts. Furthermore, the steamed and fermented C. lanceolata water extract showed neuroprotective effect. These results suggest that steaming and fermentation process more improve cognitive enhancing effect of C. lanceolata than other process.

Evaluation of Biocompatibility of Anodized and Hydrothermally Treated Pure Niobium Metal (양극산화와 열수처리한 순수 니오비움 금속의 생체활성 평가)

  • Won, Dae-Hee;Choi, Un-Jae;Lee, Min-Ho;Bae, Tae-Sung
    • Journal of Technologic Dentistry
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    • v.27 no.1
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    • pp.79-88
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    • 2005
  • This study was performed to investigate the surface properties of electrochemically oxidized pure niobium by anodic oxide and hydrothermal treatment technique. Niobium specimens of $10\times10mm$ in dimension were polished sequentially from #600, #800, #1000 emery paper. The surface pure niobium specimens were anodized in an electrolytic solution that was dissolved calcium and phosphate in water. The electrolytic voltage was set in the range of 250 V and the current density was 10 $mA/cm^2$. The specimen was hydrothermal treated in high-pressure steam at 300$^{\circ}C$ for 2 hours using an autoclave. Then, specimens were immersed in the Hanks' solution with pH 7.4 at 37$^{\circ}C$ for 30 days. The surface of specimen was characterized by scanning electron microscope(SEM), energy dispersive X-ray microanalysis(EDX), potentiostat/galvanostat test, and cytotoxicity test. The results obtained was summarized as follows; According to the result of measuring corrosion behavior at 0.9% NaCl, corrosion resistance was improved more specimens treated with anodic oxide than in hydrothermal treated ones. The multi-porous oxide layer on surface treated through anodic oxidation showed a structure that fine pores overlap one another, and the early precipitation of apatite was observed on the surface of hydrothermal treated samples. According to the result of EDX after 30 days deposition in Hanks' solution, Ca/P was 1.69 in hydrothermal treated specimens. In MTT test, specimens treated through anodic oxidation and hydrothermal treated ones showed spectrophotometer similar to that of the control group. Thus no significant difference in cytotoxicity was observed (P>0.05).

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Antibacterial Effect of Ipyo-san against Methicillin-Resistant Staphylococcus aureus (입효산(立效散)의 Methicillin-Resistant Staphylococcus aureus에 대한 항균활성에 관한 연구)

  • Yoon, Jae-Hong;Choi, Yeun-Ju;Jeong, Seung-Hyun;Shin, Gil-Cho
    • The Journal of Internal Korean Medicine
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    • v.34 no.3
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    • pp.278-288
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    • 2013
  • Objectives : Methicillin-resistant Staphylococcus aureus (MRSA) has a cephalosporin and beta-lactam antibiotic-resistant strains. MRSA is one of the major pathogens causing hospital infection and the isolation ratio of MRSA has gradually increased. Consequently, increased resistance to antibiotics is causing serious problems in the world. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infectious diseases. Methods : The antibacterial activities of Ipyo-san were evaluated against 2 strains of MRSA and 1 standard Methicillin-susceptible staphylococcus aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MIC) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay performed under dark. Results : The MIC of Ipyo-san water extract against S. aureus strains ranged from 1000 to $2,000{\mu}g/ml$, so we confirmed that it had a strong antibacterial effect. Also, the combinations of Ipyo-san water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. We suggest that Ipyo-san water extract against MRSA has antibacterial activity so it has potential as alternatives to antibiotic agents. For the combination test, we used Triton X-100 (TX) and DCCD for measurement of membrane permeability and inhibitor of ATPase. As a result, antimicrobial activity of Ipyo-san water extract was affected by the cell membrane. Conclusions : We suggest that the Ipyo-san water extract lead the treatment of bacterial infection to solve the resistance and remaining side-effect problems that are the major weak points of traditional antibiotics.

Mutagenic and Antimutagenic Effects of Hemp Seed Oil Evaluated by Ames Salmonella Testing (삼종실유의 항돌연변이 효과)

  • Jeun, Jung-Ae;Cho, Hee-Joon;Jun, Hee-Jin;Lee, Ji-Hae;Jia, Yao-Yao;Cho, Kyoung-Sang;Kim, Eun-Soo;Lee, Sung-Joon
    • Korean Journal of Food Science and Technology
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    • v.43 no.3
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    • pp.396-400
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    • 2011
  • We examined the in vitro mutagenic and antimutagenic effects of hexane-extracted hemp (Cannabis sativa L. subsp. sativa var. sativa) seed oil (HO) with and without S9-mediated metabolic activation, using the TA98 and TA100 Salmonella Typhimurium strains. The MTT assay revealed no cytotoxicity in HepG2 cells for HO quantities $\leq400g$/mL. In the mutagenicity test, revertant colonies did not exceed spontaneous colonies in number. Colony numbers did not increase in either strain after HO treatment, with or without metabolic activation. HO showed no mutagenic effects and did not induce a mutation in either strain. In the antimutagenicity test, HO reduced the number of mutated colonies induced by 4NQO in both strains. The inhibition rates of HO (TA98, 21-91%; TA100, 21-85%) indicated a potent reduction in mutagenicity induced by 4NQO. HO showed no significant mutagenicity and may have antimutagenic effects, as assessed by Ames testing.