• Archives of Pharmacal Research
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• v.18 no.4
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• pp.256-261
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• 1995

The elevation of intracellular $Ca^{2+}$ in various tissue through oxidative stress induced by menadione has been well documented. Increase of $Ca^{2+}$ level inplatelets results in aggreaction of patelets. To test the hypothesis that menadione-induced $Ca^{2+}$ elevations can play a role in platelet aggregation, we have studied the effect of menadione on aggragation of platelets isolated from female rats. Treatment with menadione to platelet rich plasma (PRP), which proved to be 60% as determined by aggregometry. however, exposure of PRP to menadione leads to a loss of cell viability, as measured by lactae dehydrogenase (LDH) leakage, suggesting that menadione might induce cell lysis rather than aggregation of platelets. Turbidty changes induced by menadione were unaffected by addition ofl dicoumarol, which is a quinone reducellular factions of patelets. These data, which indicate an absence of the QR detoxifying pathway, suggest that platelets may be more susceptible to menadione-induced cytotoxicity than certain other cell, as hepatocytes.

• KSBB Journal
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• v.30 no.4
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• pp.191-196
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• 2015

Blueberry juice possesses rich-procyanidins and - anthocyanidin, comprised a group of with numerous health benefits such as protection against coronary heart disease, detoxification, and obesity. Blueberry (Vaccinium virgatum) juice extracts were analyzed and separated by an HPLC method for the purpose of the separation and quantification in polyphenolic groups. In specific HPLC conditions, a binary mobile phase consisting of formic acid: water (10:90, v/v, solvent A) and formic acid: water: acetonitrile (10:60:30, v/v/v, solvent B) was utilized and it is detected at 546 nm wavelength. The phenolic contents of the extracts are determined using Folin-Ciocalteu phenol reagent. In order to test anti-inflammation activity assay, after producing nitric oxide (NO) in lipopolysaccharide activated RAW 264.7 cells, at concentration of $20-500{\mu}g/mL$ it reduced to NO production at a dose-dependent manner. Importantly, cytotoxicity assay with up to $500{\mu}g/mL$ of the extract from blueberry juice showed ~100% cell viability for RAW264.7 cell line. Therefore, Korean blueberry juice might have potential as anti-oxidant and antiinflammation agents.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.3
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• pp.247-257
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• 2014

Oil-in-water nanoemulsions of astaxanthin prepared by new vesicle, glyceryl citrate/ lactate/ linoleate/ oleate, were evaluated thoroughly in terms of cosmeceutical properties such as antioxidant effect, cell viability, influence of protein related enzyme, skin penetration, skin hydration and elasticity. Antioxidant effect and cell viability of nanoemulsion of astaxanthin were evaluated by DPPH and MTT assay. Also other properties of nanoemulsions of astaxanthin were measured by proteome analysis using 2D-PAGE, confocal laser scanning microscope and in-vivo test. We were able to find that the nanoemulsion of astaxanthin is good at scavenging of radical and inhibits the degradation of dermal extracellular matrix with the down-regulation of MMPs and other proteins related to MMP expression. CLSM was adopted for observing penetration of nanoemulsion of astaxanthin and showed high effective penetration rate compared to the nanoemulsion of astaxanthin prepared by conventional lecithin. In-vivo measurement of the nanoemulsions in hydration and elasticity were conducted to 11 Korean female adults for 28 days and showed better results.

• Microbiology and Biotechnology Letters
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• v.50 no.2
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• pp.277-282
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• 2022

Whitening effect of the extract of Phaseolus angularis (PA) shell was investigated for its potential application as a functional ingredient for cosmetic products. The tyrosinase inhibitory effect, which is related to whitening, was 76.4% at a concentration of 1,000 ㎍/ml. Cell viability of melanoma cells was measured to test toxicity of the PA shell extract at concentrations showing at least 90% viability. In addition, western blot and RT-PCR assays of the PA shell extract showed concentration-dependent decreases of MITF, TRP-1, TRP-2 and tyrosinase protein and inhibitory effect of mRNA expression. These findings suggested that extract from PA shell has great potential as a cosmeceutical ingredient with whitening effect.

• Restorative Dentistry and Endodontics
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• v.48 no.2
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• pp.18.1-18.10
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• 2023

Objectives: This study aimed to determine whether collagen triple helix repeat containing-1 (CTHRC1), which is involved in vascular remodeling and bone formation, can stimulate odontogenic differentiation and angiogenesis when administered to human dental pulp stem cells (hDPSCs). Materials and Methods: The viability of hDPSCs upon exposure to CTHRC1 was assessed with the WST-1 assay. CTHRC1 doses of 5, 10, and 20 ㎍/mL were administered to hDPSCs. Reverse-transcription polymerase reaction was used to detect dentin sialophosphoprotein, dentin matrix protein 1, vascular endothelial growth factor, and fibroblast growth factor 2. The formation of mineralization nodules was evaluated using Alizarin red. A scratch wound assay was conducted to evaluate the effect of CTHRC1 on cell migration. Data were analyzed using 1-way analysis of variance followed by the Tukey post hoc test. The threshold for statistical significance was set at p < 0.05. Results: CTHRC1 doses of 5, 10, and 20 ㎍/mL had no significant effect on the viability of hDPSCs. Mineralized nodules were formed and odontogenic markers were upregulated, indicating that CTHRC1 promoted odontogenic differentiation. Scratch wound assays demonstrated that CTHRC1 significantly enhanced the migration of hDPSCs. Conclusions: CTHRC1 promoted odontogenic differentiation and mineralization in hDPSCs.

• Journal of Applied Biological Chemistry
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• v.62 no.4
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• pp.347-353
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• 2019

Stings of jellyfish, which frequently occur in a warm season, cause severe pain, inflammation and sometimes irreversible results such as the death. Harmful venoms from jellyfish, therefore, have been studied for finding the therapeutic agents to relieve pain or to neutralize toxic components. However, it is still unclear if and how jellyfish venom reveal neuronal toxicity even though pain induction seems to result from the activation of nociceptors such as nerve endings. In this study, using HT-22 cell line, we investigated neurotoxic effects of the venom of Chrysaora pacifica (CpV) which appears in South-East ocean of Korea. In 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, CpV significantly reduced the viability of HT-22 cells in a dose-dependent manner. Additionally, in 2',7'-Dichlorofluorescin diacetate fluorescence test under the culture condition lacking dominant inflammatory factors, CpV remarkably increased the production of intracellular reactive oxygen species (ROS). Reduced responsive fluorescence to Rhodamine123 and increased expression of intracellular cytochrome c were also observed in HT-22 cells treated with CpV. These indicate that CpV-reduced viability of HT-22 cells may be due to the activation of apoptotic signalings mediated with oxidative stress and mitochondrial dysfunction. Furthermore, removing Ca²⁺ ion or adding N-acetyl-Lcystein remarkably blocked the CpV effect to reduce the viability of HT-22 cells. The findings in this study clearly demonstrate that CpV may activate Ca²⁺-mediated ROS signalings and mitochondrial dysfunction resulting in neuronal damage or death, and suggest that blocking Ca²⁺ pathway is a therapeutic approach to possibly block toxic effects of jellyfish venoms.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.4105-4105
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• 2001

Near infrared spectroscopy (NIRS) was employed to qualify and quantify on survival, the injury rate and apoptosis of the human breast cancer cell line MCF-7 cells. MCF-7 cells were cultured in RPMI medium supplemented with 10% FCS in a 95% air and 5% CO2 atmosphere at 37$^{\circ}C$. For the viable cells preparation, cells were de-touched by 0.1% of trypsin treatment and washed with RPMI supplemented with 10% FCS medium by centrifugation at 1000 rpm for 3min. For the dead cells preparation, cells were de-touched by a cell scraper. The cells were counted by a hemacytometer, and the viability was estimated by the exclusion method with frypan blue dye. Each viable and dead cells were suspended in PBS (phosphate bufferred saline) or milk at the cell density desired. For the quantitative determination of cell death by measuring the LDH (lactate dehydrogenase) activity liberated from cells with cell membrane injuries, LDH-Cytotoxic Test Wako (Wako, Pure Pharmaceutical Co. Ltd., Japan) was used. We found that NIRS measurement of MCF-7 cells at the density range could evaluate and monitor the different characteristics of living cells and dead cells. The spectral analysis was performed in two wavelength ranges and with 1,4, 10 mm pathlength. Different spectral data pretreatment and chemometrics methods were used. We applied SIMCA classificator on spectral data of living and dead cells and obtained good accuracy when identifying each class. Bigger variation in the spectra of living cells with different concentrations was observed when compared to the same concentrations of dead cells. PLS was used to measure the number of cells in PBS. The best model for measurement of dead cells, as well as living cells, was developed when raw spectra in the 600-1098 nm region and 4 mm pathlength were used. Smoothing and second derivative spectral data pretreatment gave worst results. The analysis of PLS loading explained this result with the scatter effect found in the raw spectra and increased with the number of cells. Calibration for cell count in the 1100-2500 nm region showed to be very inaccurate.

• Journal of Preventive Medicine and Public Health
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• v.39 no.3
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• pp.199-204
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• 2006

Objectives: Mercury is a hazardous organ-specific environmental contaminant. It exists in a wide variety of physical and chemical states, each of which has unique characteristics for the target organ specificity. Exposure to mercury vapor and to organic mercury compounds specifically affects the CNS, while the kidney is the target organ for inorganic Hg compounds. Methods: In this study, mercury chloride $(HgCl_2)$ was studied in a renal derived cell system, i.e., the tubular epithelial Madin-Darby canine kidney (MDCK) cell line, which has specific sensitivity to the toxic effect of mercury. MDCK cells were cultured for 6-24 hr in vitro in various concentrations (0.1-100 M) of $HgCl_2$, and the markers of apoptosis or cell death were assayed, including DNA fragmentation, caspase-3 activity andwestern blotting of cytochrome c. The influence of the metal on cell proliferation and viability were evaluated by the conventional MTT test. Results: The cell viability was decreased in a time and concentration dependent fashion: decreases were noted at 6, 12 and 24 hr after $HgCl_2$, exposure. The increases of DNA fragmentation were also observed in the concentrations from 0.1 to 10 M of $HgCl_2$ at 6 hr after exposure. However, we could not observe DNA fragmentation in the concentrations more than 25 M because the cells rapidly proceeded to necrotic cell death. The activation of caspase-3 was also observed at 6 hr exposure in the $HgCl_2$ concentrations from 0.1 to 10 M. The release of cytochrome c from the mitochondria into the cytosol, which is an initiator of the activation of the caspase cascade, was also observed in the $HgCl_2-treated$ MDCK cells. Conclusions: These results suggest that the activation of caspase-3 was involved in $HgCl_2-induced$ apoptosis. The release of cytochrome c from the mitochondria into the cytosol was also observed in the $HgCl_2-treated$ MDCK cells. These findings indicate that in MDCK cells, $HgCl_2$ is a potent inducer of apoptosis via cytochrome c release from the mitochondria.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.6
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• pp.461-467
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• 2005

Background. 5-(and-6)-carboxy-2',7'-dichlorofluorescein diacetate, succinimidyl ester mixed (CFSE) is the fluorescent labelling agent of living cells and used to trace the cells in vivo after transplatnation of various cells. The CFSE labelled cells can maintain fluorescence for up to 7 days after labelling. The MC3T3-E1 cell line (MC3T3) has been used for many studies about osteoblast, which is well known as a mouse preosteoblast. So the CFSE would be used to trace the transplanted MC3T3. However there are few reports about CFSE labelling of MC3T3. This study is aimed to know about adequate concenturation and incubation time of CFSE to MC3T3. Materials and methods. The MC3T3 was incubated in a humidified atmosphere of 95% air with 5% $CO_2$ at $37^{\circ}C$ using ${\alpha}$-minimal essential medium (${alpha}$-MEM) containing10% FBS and gentamycin. Ten mM CFSE solution in dimethylsulphoxide (DMSO: 1%) was diluted with phosphate buffered saline (PBS) and final concentration of culture medium was, respectively, 5, 10, 15, 20, 25 and 30 ${{\mu}M$. Then the MC3T3 was incubated with CFSE in a humidified atmosphere of 95% air with 5% $CO_2$ at $37^{\circ}C$ for 5, 10, 15, 20, 25, 30, 35, 40 and 45 minutes in each concentration. The fluorescence of CFSE labelled cells was analysed with a inverted fluorescence microscope. The duration of cell labelling was also studied. Trypan blue dye exclusion test was done for cell viability. Results. For concentration between 5 and 10 ${\mu}M$, CFSE did not significantly label the MC3T3 in vitro. The destruction of MC3T3 was observed at the concentration of 20 ${\mu}M$. In the concentration of 15 ${\mu}M$, the best labelling was obtained at an incubation period between 15 and 30 minutes. The MC3T3 labelled with an incubation period of 15 minutes at 15 ${\mu}M$ was still fluorescent 7 days after CFSE labelling. The mean cell viability was 95.93%. Conclusion. These results suggests an incubation period of 15 minutes at 15 ${\mu}M$ of CFSE provides best labelling of MC3T3 in vitro.

• Journal of the korean academy of Pediatric Dentistry
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• v.51 no.2
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• pp.149-164
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• 2024

This study compared the solubility, water absorption, dimensional stability, release of various ions (hydroxyl, calcium, sulfur, strontium, and silicon), and cytotoxicity of light-cured resin-modified pulp-capping materials. Resin-modified calcium hydroxide (Ultra-blend^TM plus, UBP), light-cured resin-modified calcium silicate (TheraCal LC^TM, TLC), and dual-cure resin-modified calcium silicate (TheraCal PT^TM, TPT) were used. Each material was polymerized; solubility, 24-hour water absorption, and 30- day dimensional stability experiments were conducted to test its physical properties. Solubility was assessed according to the ISO 6876 standard, and 24 hours of water absorption, 30 days of dimensional stability were assessed by referring to the previous protocol respectively. Eluates at 3 and 24 hours and on 7, 14, and 28 days were analyzed according to the ISO 10993-12 standard. And the pH, Ion-releasing ability, cell proliferation rate, and cell viability were assessed using the eluates to evaluate biochemical characteristics. pH was measured with a pH meter and Ion-releasing ability was assessed using inductively coupled plasma atomic emission spectrometry (ICP-AES). Cell proliferation rate and cell viability were assessed using human dental pulp cells (hDPCs). The former was assessed by an absorbance assay using the CCK-8 solution, and the latter was assessed by Live and Dead staining. TPT exhibited lower solubility and water absorption than TLC. UBP and TPT demonstrated higher stability than TLC. The release of sulfur, strontium, calcium, and hydroxyl ions was higher for TLC and TPT than for UBP. The 28-day release of hydroxyl and silicon ions was similar for TLC and TPT. TLC alone exhibited a lower cell proliferation rate compared to the control group at a dilution ratio of 1 : 2 in cell proliferation and dead cells from Live and Dead assay evaluation. Thus, when using light-cure resin-modified pulp-capping materials, calcium silicate-based materials can be considered alternatives to calcium hydroxide-based materials. Moreover, when comparing physical and biochemical properties, TPT could be prioritized over TLC as the first choice.