• 한국재료학회지
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• 제33권4호
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• pp.130-134
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• 2023

This study evaluated cell viability and cytokine release in immortalized human oral fibroblasts (hTERT-hNOFs) and keratinocytes (IHOK) exposed to a dental-impregnated gingival retraction cord. To prepare the extracts, dental gingival retraction cords impregnated with aluminum chloride hexahydrate were immersed in a cell culture medium for 24 h at 37 ℃. hTERT-hNOFs and IHOK were cultured for 24 h. The cell culture medium was removed and extracts of the dental gingival retraction cords were added. After incubation with the extract solution, cell viability was evaluated using an MTT assay. The levels of the cytokines IL-1α and IL-8 were measured in the supernatants of each cell type. The cell viability after exposure to the extract solution for 10 min exceeded 70 % in both cell types. The ET50 values for hTERT-hNOF and IHOK were 35.75 and 28.98 min, respectively. For IHOK, the IL-1α level was (5.35 ± 5.22) pg/mL at 10 min, (3.58 ± 5.38) pg/mL at 20 min, and (2.85 ± 4.28) pg/mL at 60 min of exposure (p > 0.05). The IL-8 level in IHOK was (67.16 ± 18.70) pg/mL at 10 min, (78.36 ± 7.50) pg/mL at 20 min, and (111.9 ± 26.10) pg/mL at 60 min of exposure (p > 0.05). Cytokine release was not observed from hTERT-hNOFs. Based on these results, cell viability and cytokine release were confirmed in cells exposed to the impregnated gingival retraction cord. In addition, the application of the extracts to hTERT-hNOF and IHOK during the actual contact time and determination of ET50 may be beneficial for evaluating the biocompatibility of dental-impregnated gingival retraction cords.

• Journal of Microbiology and Biotechnology
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• 제13권2호
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• pp.217-224
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• 2003

Previously, we developed a CHO cell line (CHO-OTC1-Al9) that expresses the first two enzymes in the urea cycle and exhibits a higher ammonia-removing ability and faster growth rate than a vector-controlled CHO cell line (CHO-neo-5). The current study was undertaken to develop a cell line with an ammonia-removing ability higher than the cell line developed previously. To accomplish this, CHO cell lines expressing the first three, first four, or all five enzymes of the urea cycle were constructed using a stable transfection method. Finally, the CHO-AS-16, CHO-AL-19, and CHO-Arg-11 cell lines expressing the first three, first four, and all five enzymes of the urea cycle, respectively, were selected and found to exhibit higher ammonia-removing ability than the CHO-OTC1-Al9 cell line. Among the three selected cell lines, CHO-AL-19 showed the highest ammonia-removing ability and highest cell viability at a higher cell density, with 40% and 15% lower ammonia concentration in the, culture media than that of CHO-neo-5 and CHO-OTC1-A19 cell lines, respectively. CHO-AL-19 also showed 44% and 10% higher cell viability than the CHO-neo-5 and CHO-OTC-Al9 cell lines, at a higher cell density, respectively. The ammonia concentrations in the culture media were expressed as the ammonia concentration/cell, and the CHO-AL-19 cells revealed 45-60% and 20% lower ammonia concentration/cell than the CHO-neo-5 and CHO-OTC1-Al9 cells, respectively.

• Journal of Nutrition and Health
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• 제32권3호
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• pp.318-326
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• 1999

Apoptotic cell death, characterized by DNA fragmentation and morphological changes, has previously been shown to occur in vascular endothelial cells cultured with hydrogen peroxide. The present study examined the induction of apoptosis by hydrogen peroxide and whether pyruvate, a key glycolytic intermediate and $\alpha$-keto-monocarboxylate, can inhibit the apoptotic effects in bovine pulmonary artery endothelial cells(BPAECs). Culture with 500uM hydrogen peroxide resulted in 30% cell death and induced morphological changes and DNA fragmentation. Cell injury was inhibited by the treatment with pyruvate. Pyruvate(0.1-5.0mM), and cell viability increased in a dose-dependent manner. In the presence of pyruvate(10~20mM), the viability was improved to over 95%. In contrast, treatment with lactate, a reduced form of phyuvate, did not protect against cell death oxidative stress-induced loss of viability and apoptosis was examined with $\alpha$-cyano-3-hydroxycinnarmate(COHC) as a selective mitochondrial monocarboxylate transport blocker. Incubation with COHC(500uM) did not significantly affect cell viability in the presence of hydrogen peroxide. The cytoprotection by pyruvate(3mM)against hydrogen peroxide stress was abolished by COHC. This indicates that the cytoprotection by pyruvate against oxidative stress in endothelial cells is mediated, at least in part, by mitochondrial pyruvate uptake and hence endothelial enerygetics. However, cytosolic mechanisms related, at least in part, by mitochondrial pyruvate uptake and hence endothelial energetics. However, cytosolic mechanisms related to the glutathione system may also contribute. The results suggest that pyruvate has therapeutic potential in the treatment of oxidative stress-induced cytotoxicity associated with increased apoptosis.

• Animal cells and systems
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• 제11권1호
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• pp.17-22
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• 2007

The effects of antimetabolite 6-AN (6-amino-nicotinamide) on viability and morphology of L6 myoblast cells have been investigated. 6-AN ($100{\mu}M$) induced a time-dependent decrease in cell viability with respect to the untreated control cells. Following 6-AN administration the viability rate started to decline sharply, reaching about 23% of the untreated control cells at 48 h. Inverted phase-contrast microscopy revealed that 6-AN caused characteristic morphological changes such as irregularly elongated and stellate shape of cells, round-shaped nucleus, cytoplasmic vacuolization, irregular cell arrangements and formation of large spaces among cell clusters. The concentrations of ATP and $NAD^{+}$ in the 6-AN treated cells were significantly lower (p < 0.01) than those of the untreated control cells. In contrast, the concentration of AMP was significantly increased by the 6-AN treatment. Activities of catalase, superoxide dismutase and glutathione peroxidase in 6-AN treated cells were significantly higher (p < 0.01) than those of the untreated control cells. The activities of glyceraldehyde-3-phosphate dehydrogenase in 6-AN treated cells were significantly lower (p < 0.01) than those of the untreated control cells. The results suggest that 6-AN caused marked reduction of cell viability and alterations of some important metabolites and enzymes.

• Environmental Analysis Health and Toxicology
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• 제18권4호
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• pp.295-303
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• 2003

Physiological cellular activities responses to cadmium (Cd) exposure in green algae with several reductases activities and viability of the cell were examined. The cell division of green algae, Selenastrum capricornutum treated with 5ppm was significantly decreased than that of normal algae. The mean cell number of normal algal culture was as twice much as than that of algae at 6 days after Cd treatment. The cellular viability of algae was analysed by flow-cytometry with fluorescent dye after esterase reaction on cell membrane. The 85.35% of cellular viability of normal culture was decreased to 34.35% when algae was treated with 5 ppm of Cd at 6 days after treatment. It was considered that those method of flow-cytometry is useful tool for toxicity test on micro-organisms in the respect of identifying cellular viability. Also, the activities of both glutathione peroxidase (GPX) and ascorbate peroxidase (APX), which are indirectly react against oxidative stress through reduction of glutathione by Cd were significantly increased with 25%. It is considered that both GPX and APX are involved in the metabolic pathway of Cd -detoxification with similar portion in Selenasturm capricornutum.

• Journal of Yeungnam Medical Science
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• 제36권2호
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• pp.105-108
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• 2019

Background: Although kidney transplantation outcomes have improved dramatically after using calcineurin inhibitors (CNIs), CNI toxicity continues to be reported and the mechanism remains uncertain. Here, we investigated the neurotoxicity of CNIs by focusing on the viability of glioma cells. Methods: Glioma cells were treated with several concentrations of CNIs for 24 hours at $37^{\circ}C$ and their cell viability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results: Exposure to 0, 0.25, 0.5, 2.5, 5.0, and 10.0 mM concentrations respectively showed 100%, 64.3%, 61.3%, 68.1%, 62.4%, and 68.6% cell viability for cyclosporine and 100%, 38.6%, 40.8%, 43.7%, 37.8%, and 43.0% for tacrolimus. The direct toxic effect of tacrolimus on glioma cell viability was stronger than that of cyclosporine at the same concentration. Conclusion: CNIs can cause neurological side effects by directly exerting cytotoxic effects on brain cells. Therefore, we should carefully monitor the neurologic symptoms and level of CNIs in kidney transplant patients.

• 센서학회지
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• 제32권6호
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• pp.412-417
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• 2023

There is increasing interest in the rapid and highly sensitive monitoring of cell viability in biological and toxicological research. Conventional methods depend on optical assays using Water Soluble Tetrazolium-8 (WST-8) or 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay, which requires a large volume of samples and special instruments, necessitating shipment of clinical samples to laboratories. This paper reports on the development of a rapid and sensitive electrochemical (EC) sensor using screen printed electrode (SPE) and surface modification using 4'-mercapto-N-phenylquinone diamine (4'-NPQD), as double electron mediators, for monitoring cell viability via the measurement of nicotinamide adenine dinucleotide (NADH). We used the sensor to observe the viability of MCF-7 and doxorubicin (Dox)-treated cells. The oxidation current of NADH was measured via chronoamperometry (CA), and the EC results showed a good linear relationship when compared with NADH quantification using WST-8 assay. The analysis time was only 10 s and limit of detection (LOD) of NADH was 1.78 µM. Our EC method has the potential to replace conventional WST assays for cell viability and cytotoxicity experiments.

• 대한한의학회지
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• 제23권4호
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• pp.45-54
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• 2002

Objectives: In order to investigate the relationship of Radix Trichosanthis components and the melanin synthesis, the author has analyzed the cell viability and tyrosinase activity, melanin content and morphologic changes in n-hexane, EtOAc, n-BuOH, and H2O fraction. Methods: At first, in order to determine the concentration of the Radix Trichosanthis component, the author investigated the viability of B16 melanoma cell. To measure the effects of Trichosanthes kirilowii extracts (n-BuOH, n-Hexane, EtOAc, H2O fractions) on the viability of A549 cells, A549 cells were treated with various concentrations (from 0.5 to $25{\;}\mu\textrm{g}/ml$) of components of Trichosanthes kirilowii. After 24hrs, the cell viability was measured by MTT assay. The EtOAc components of Trichosanthes kirilowii decreased the viability of A549 cells in a dose-dependent manner. H2O and n-BuOH components had no cell toxicity till $25{\;}\mu\textrm{g}/ml$, the n-hexane component showed minor cell toxicity at $25{\;}\mu\textrm{g}/ml$ and the EtOAc component cell toxicity was revealed at $5{\;}\mu\textrm{g}/ml$ concentration. Results: 1. The results of tyrosinase activity and the Radix Trichosanthis component; n-hexane and EtOAc components controlled it effectively; the n-BuOH components were less effective. 2. The results of melanin content analysis showed that the n-hexane and EtOAc components effectively inhibited, the n-BuOH fraction inhibited less, and H2O component didn't inhibit the terminal melanin formation. 3. In the n-BuOH and H2O component there were no changes, but in the n-hexane component the melanin content was effectively inhibited. 4. In the EtOAc fraction, although the melanin content was inhibited, the cell count was evidently suppressed, Of all of the Radix Trichosanthis components, the n-Hexane and EtOAc fractions inhibited the melanin synthesis best, but owing to its toxicity, the EtOAc components inhibited the cell count. Conclusion: The above results demonstrated that Radix Trichosanthis n-hexane fraction efficiently inhibited the tyrosinase activity and melanin synthesis.

• Journal of Ginseng Research
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• 제28권1호
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• pp.39-44
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• 2004

홍삼추출액의 신경성체줄기세포 성장과 생존에 미치는 영향을 분석하고자 human neuronal stem cell line인 F3 cell을 배양한 후 홍삼추출액을 여러 농도로 희석하여 MTT assay로 cell viability를 측정하였고 FACS analysis로 cell Cycle변화를 측정하였다. 특정한 농도에서는 세포가 증식되는 경향을 보였으며 농도가 증가되면서 viability가 감소되는 현상을 확인할 수 있었다. Cell cycle 분석상 세포증식시에는 5 phase 및 G2/M phase가 증가되는 경향을 보였고, viability가 감소되면서 S phase가 감소되고 G0/G1 phase 가 증가 되었다. 한편 DNA fragmentation이 cell viability감소에 따라 증가되었으나, Caspase 3 activation 또는 Bax expression과는 관련성이 적었다.

• Journal of Korean Dental Science
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• 제11권2호
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• pp.62-70
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• 2018

Purpose: Bone marrow has long been a source of primary cells. This study was performed to evaluate the effects of age and sex on the cellular viability and expression of stem cell markers of mRNA and on the protein expression of bone marrow stem cells (BMSCs) derived from healthy donors. Materials and Methods: Stem cells were isolated from human bone marrow and plated in culture plates. The shape of the BMSCs was observed under inverted microscope. Quantitative cellular viability was evaluated using a Cell-Counting Kit-8 assay. The expression of stem cell surface markers was tested and a series of quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot was performed to evaluate the expression in each group. Result: The shapes of the cells at 20s, 30s, and 50s were similar to each other. No significant changes in cellular viability were noted among different age groups or sex groups. The BMSCs expressed CD44, CD73, and CD90 surface markers but did not express CD14 and CD34. There were no noticeable differences in CD surface markers among the different age groups. The expressions of CD surface markers were similar between men and women. No significant differences in the secretion of vascular endothelial growth factors (VEGFs) were noted at Day 3 between different age groups. qRT-PCR regarding the expression showed differences between the age groups. However, Western blot analysis showed a decrease in expression but did not reach statistical significance (P>0.05). Conclusion: This study clearly showed no significant differences in shape, cell viability, expression of stem cell surface markers, or secretion of human VEGF among different age groups. However, western blot analysis showed a tendency of age-related decrease which did not reach statistical significance. Collectively, autologous or allogeneic BMSCs should be meticulously applied to obtain optimal results regarding age and sex.