• Biomedical Science Letters
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• v.13 no.4
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• pp.355-360
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• 2007

In order to evaluate on the cytotoxicity of methyl mercury (MM) and antioxidant effect of phenolic compound, poncirin against MM-induced cytotoxicity, XTT assay was performed to determine the cell viability after human skin fibroblasts (Detroit 51) were grown in the media containing various concentrations of methylmercuric chloride (MMC). And also, the antioxidant effect of poncirin on the cytotoxicity induced by MMC was examined by cell viability and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in these cultures. MMC decreased cell viability in dose-dependent manner in these cultures and the midcytotoxicity value was determined at concentration of 30 ${\mu}M$ MMC after human skin fibroblasts were treated with $10\sim50{\mu}M$ MMC for 72 hours, respectively. MMC was highly toxic on cultured human skin fibroblasts by toxic criteria. MMC-mediated cytotoxicity was related with oxidative stress by the diminution of toxic effect according to the treatment of vitamin E. In the antioxidant effect of poncirin, it showed vitamin E-like DPPH radical scavenging activity at 90 ${\mu}g/ml$ poncirin and also, remarkably increased cell viability compared with MMC-treated group. From these results, it is suggested that MMC-mediated cytoxicity was highly toxic and was related with oxidative stress in cultured human skin fibroblasts, and also phenolic compound such as poncirin showed the protection on MMC-induced cytotoxicity by antioxidant effect in these cultures.

• Journal of Physiology & Pathology in Korean Medicine
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• v.26 no.1
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• pp.74-80
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• 2012

Candida albicans is a common opportunistic pathogen and is frequently associated with biofilm formation occurring on the surfaces of host tissues and medical devices. On account of the distinct resistance of C. albicans biofilms to the conventional antifungal agents, new strategies are required to cope with these infections. The root of Polygonum cuspidatum has been used for medicinal purposes in East Asia. The aim of this study was to assess the anticandidal potential of the P. cuspidatum ethanol extract by evaluating biofilm formation, integrity of the cell membranes of C. albicans and adhesion of C. albicans cells to polystyrene surfaces. The growth and development of the biofilm was assessed using an XTT reduction assay, and the extract (0.39 mg/ml) significantly reduced ($41.1{\pm}17.8%$) biofilm formation of 11 C. albicans strains. The extract damaged the cell membranes of C. albicans and remarkably inhibited cell adhesion to polystyrene surfaces. The plant extract displayed fungistatic activity without significant hemolytic activity. Based on the results of this study, the P. cuspidatum extract has promising potential for use in treating biofilm-associated Candida infection.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.5
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• pp.989-1000
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• 2002

The purpose of this study is to examine the toxic effects caused by xanthine oxidase/hypoxanthine(XO/HX) and the effects of herbal extracts such as JHYJ and GJHYJ on the treatment of the toxic effects. For this purpose, experiments with the cultured hippocampal cells from new born mice were done. The results of these experiments were as follows. 1. XO/HX, a oxygen radical-generating system, decreased the survival rates of the cultured cells on XTT assay and INT assay, the amount of DNA syntheses, and the amount of neurofilaments, and increased the lipid peroxidation. 2. JHYJ and GJHYJ have the efficacy of increasing the survival rates of the cultured cells. 3. JHYJ and GJHYJ have the efficacy of increasing the amount of neurofilaments and of decreasing the lipid peroxidation. 4. JHYJ and GJHYJ have the efficacy of increasing the amount of DNA syntheses. From the above results, it is suggested that Jihwangyeumja and Gamijihwangyeumja have marked efficacy as a treatment for the damages caused by the XO/HX-mediated oxidative stress. And Jihwangyeumja and Gamijihwangyeumja are thought to have certain pharmacological effects. Further dinical study of this pharmacological effects of Jihwangyeumja and Gamijihwangyeumja should be complemented.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.2
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• pp.699-704
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• 2012

Natural products have been the target for cancer therapy for several years but there is still a dearth of information on potent compounds that may protect normal cells and selectively destroy cancerous cells. The present study was aimed to evaluate the cytotoxic potential of n-butanolic leaf extract of $Annona$ $muricata$ L. on WRL-68 (normal human hepatic cells), MDA-MB-435S (human breast carcinoma cells) and HaCaT (human immortalized keratinocyte cells) lines by XTT assay. Prior to cytotoxicity testing, the extract was subjected to phytochemical screening for detecting the presence of compounds with therapeutic potential. Their relative antioxidant properties were evaluated using the reducing power and $DPPH^*$radical scavenging assay. Since most of the observed chemo-preventive potential invariably correlated with the amount of total phenolics present in the extract, their levels were quantified and identified by HPLC analysis. Correlation studies indicated a strong and significant (P<0.05) positive correlation of phenolic compounds with free radical scavenging potential. The results revealed that the extract was moderately cytotoxic to normal cells with a mean IC50 value of 52.4 ${\mu}g$ when compared with those obtained for cancerous cells (IC50 values of 29.2 ${\mu}g$ for MDA-MB-435S and 30.1 ${\mu}g$ for HaCaT respectively). The study confirms the presence of therapeutically active antineoplastic compounds in the n-butanolic leaf extract of $Annona$ $muricata$. Isolation of the active metabolites from the extract is in prospect.

• Journal of Microbiology and Biotechnology
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• v.24 no.9
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• pp.1216-1225
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• 2014

Biofilm-related infections of Candida albicans are a frequent cause of morbidity and mortality in hospitalized patients, especially those with immunocompromised status. Options of the antifungal drugs available for successful treatment of drug-resistant biofilms are very few, and as such, new strategies need to be explored against them. The aim of this study was to evaluate the efficacy of phenylpropanoids of plant origin against planktonic cells, important virulence factors, and biofilm forms of C. albicans. Standard susceptibility testing protocol was used to evaluate the activities of 13 phenylpropanoids against planktonic growth. Their effects on adhesion and yeast-to-hyphae morphogenesis were studied in microplate-based methodologies. An in vitro biofilm model analyzed the phenylpropanoid-mediated prevention of biofilm development and mature biofilms using XTT-metabolic assay, crystal violet assay, and light microscopy. Six molecules exhibited fungistatic activity at ${\leq}0.5mg/ml$, of which four were fungicidal at low concentrations. Seven phenylpropanoids inhibited yeast-to-hyphae transition at low concentrations (0.031-0.5 mg/ml), whereas adhesion to the solid substrate was prevented in the range of 0.5-2 mg/ml. Treatment with ${\leq}0.5mg/ml$ concentrations of at least six small molecules resulted in significant (p < 0.05) inhibition of biofilm formation by C. albicans. Mature biofilms that are highly resistant to antifungal drugs were susceptible to low concentrations of 4 of the 13 molecules. This study revealed phenylpropanoids of plant origin as promising candidates to devise preventive strategies against drug-resistant biofilms of C. albicans.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.2
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• pp.631-637
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• 2013

Luteolin is a naturally occurring flavonoid present in many plants with diverse applications in pharmacology. Despite several studies elucidating its significant anti-cancer activity against various cancer cells, the mechanism of action in skin cancer is not well addressed. Hence, we investigated the effects of luteolin in HaCaT (human immortalized keratinocytes) and A375 (human melanoma) cells. The radical scavenging abilities of luteolin were determined spectrophotometrically, prior to a cytotoxic study (XTT assay). Inhibitory effects were assessed by colony formation assay. Further, the capability of luteolin to induce cell cycle arrest and apoptosis were demonstrated by flow cytometry and cellular DNA fragmentation ELISA, respectively. The results revealed that luteolin possesses considerable cytotoxicity against both HaCaT and A375 cells with $IC_{50}$ values of 37.1 ${\mu}M$ and 115.1 ${\mu}M$, respectively. Luteolin also inhibited colony formation and induced apoptosis in a dose and time-dependent manner by disturbing cellular integrity as evident from morphological evaluation by Wright-Giemsa staining. Accumulation of cells in G2/M (0.83-8.14%) phase for HaCaT cells and G0/G1 (60.4-72.6%) phase for A375 cells after 24 h treatment indicated cell cycle arresting potential of this flavonoid. These data suggest that luteolin inhibits cell proliferation and promotes cell cycle arrest and apoptosis in skin cancer cells with possible involvement of programmed cell death, providing a substantial basis for it to be developed into a potent chemopreventive template for skin cancer.

• The Journal of Advanced Prosthodontics
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• v.7 no.2
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• pp.166-171
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• 2015

PURPOSE. The aim of this study was to evaluate antibacterial activity and osteoblast-like cell viability according to the ratio of titanium nitride and zirconium nitride coating on commercially pure titanium using an arc ion plating system. MATERIALS AND METHODS. Polished titanium surfaces were used as controls. Surface topography was observed by scanning electron microscopy, and surface roughness was measured using a two-dimensional contact stylus profilometer. Antibacterial activity was evaluated against Streptococcus mutans and Porphyromonas gingivalis with the colony-forming unit assay. Cell compatibility, mRNA expression, and morphology related to human osteoblast-like cells (MG-63) on the coated specimens were determined by the XTT assay and reverse transcriptase-polymerase chain reaction. RESULTS. The number of S. mutans colonies on the TiN, ZrN and $(Ti_{1-x}Zr_x)N$ coated surface decreased significantly compared to those on the non-coated titanium surface (P<0.05). CONCLUSION. The number of P. gingivalis colonies on all surfaces showed no significant differences. TiN, ZrN and $(Ti_{1-x}Zr_x)N$ coated titanium showed antibacterial activity against S. mutans related to initial biofilm formation but not P. gingivalis associated with advanced periimplantitis, and did not influence osteoblast-like cell viability.

• Korean Journal of Acupuncture
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• v.29 no.1
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• pp.37-46
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• 2012

Objectives : The gnarl of Pinus densiflora, called Songjeol in Korea, has been used as a medicinal herb for the treatment of inflammatory-related diseases such as arthralgia, myalgia and bruise. However, the molecular actions and mechanisms have not been clearly investigated. The aim of this study was to clarify the anti-inflammatory activity of Pinus densiflora gnarl pharmacopuncture (PDGP) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Methods : Cytotoxicity was assessed by XTT assay. The amount of nitric oxide (NO) production was determined by nitrite assay. The mRNA expressions of interleukin-$1{\beta}$ (IL-$1{\beta}$), interleukin-6 (IL-6), cyclooxygenase-2 (COX-2) and heme oxygenase-1 (HO-1) were analyzed by RT-PCR. Reactive oxidative species (ROS) generation was measured using the fluorescence microscopy. In addition, inducible nitric oxide synthase (iNOS) and redox factor-1 (Ref-1) protein expressions were detected by Western blotting. Results : PDGP inhibited NO production and ROS generation in LPS-stimulated RAW264.7 cells. At the mRNA level, PDGP suppressed IL-$1{\beta}$, IL-6 and COX-2 expression. On the other hand, PDGP induced HO-1 mRNA expression. Furthermore, PDGP suppressed iNOS and Ref-1 protein expression. Conclusions : This result suggests that PDGP can act as a suppressor agent on NO and iNOS through induction of HO-1, and play an useful role in blocking inflammatory responses.

• The Journal of Korean Academy of Prosthodontics
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• v.53 no.1
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• pp.9-18
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• 2015

Purpose: The purpose of this study is to examine characteristics of implant surface with RBM and anodizing treatments, and to evaluate the responses of osteoblast-like cell (MG-63 cell). Materials and methods: Grade IV titanium disks were fabricated (Diameter 10 mm, thickness 3 mm). Anodizing treatment (ASD) group, RBM and anodizing treatment (RBM/ASD) group, control (machined surface) group were divided. In this study, osteoblast-like cell was used for experiments. The experiments consist of surface characteristics evaluation by FE-SEM images, energy dispersive spectroscopy and stereo-SEM. In order to evaluate cell adhesion evaluation by crystal violet assay and observe cells form by confocal laser microscopy. To assess cell proliferation by XTT assay, cell differentiation by RT-PCR and mineralization by Alizarin red S stain assay. ELISA analyzer was used for Quantitative evaluation. Comparative analysis was run by one-way ANOVA (SPSS version 18.0). Differences were considered statistically significant at P<.05. Results: In ASD group and RBM/ASD group, the surface shape of the crater was observed and components of oxygen and phosphate ions in comparison with the control group were detected. The surface average roughness was obtained $0.08{\pm}0.04{\mu}m$ in the control group, $0.52{\pm}0.14{\mu}m$ in ASD group and $1.45{\pm}0.25{\mu}m$ in RBM/ASD group. In cell response experiments, ASD group and RBM/ASD group were significantly higher values than control group in cell adhesion and mineralization phase, control group was the highest values in the proliferative phase. In RT-PCR experiments, RBM/ASD group was showed higher ALP activity than other groups. RBM/ASD group in comparison with ASD group was significantly higher value for cell adhesion and proliferation phase. Conclusion: In the limitation of this study, we are concluded that the surface treatment with RBM/ASD seems more effective than ASD alone or machined surface on cellular response.

• Korean Journal of Microbiology
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• v.49 no.1
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• pp.17-23
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• 2013

Candida biofilms are organized microbial communities growing on the surfaces of host tissues or indwelling medical devices, and the biofilms show enhanced resistance against the conventional antifungal agents. The roots of Coptidis chinensis have been widely used for medicinal purposes in East Asia. The present study was aimed to assess the effect of C. chinensis aqueous extract upon preformed biofilms of 10 clinical Candida albicans isolates and the antifungal activities which contribute to inhibit the C. albicans biofilm formation. Its effect on preformed biofilms was judged using XTT [2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide)] reduction assay, and metabolic activity of all tested strains was reduced significantly ($57.3{\pm}14.7%$) at $98{\mu}g/ml$ of the C. chinensis extract. The extract damaged the cell membrane of C. albicans which was analyzed by fluorescein diacetate and propidium iodide staining. The anticandidal activity was fungicidal, and the extract obstructed the adhesion of C. albicans biofilms to polystyrene surfaces, arrested C. albicans cells at $G_o/G_1$ as well, and reduced the growth of biofilms or budding yeasts finally. The data suggest that C. chinensis has multiple antifungal effects on target fungi resulting in preventing the formation of biofilms. Therefore, C. chinensis holds great promise for exploring antifungal agents from natural products in treating and eliminating biofilm-associated Candida infection.