• Restorative Dentistry and Endodontics
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• v.48 no.1
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• pp.2.1-2.12
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• 2023

Objectives: In this study, natural substances were introduced as primary dental pulp caps for use in pulp therapy, and the antimicrobial and cytotoxic properties of these substances were investigated. Materials and Methods: In this in vitro study, the antimicrobial properties of calcium-enriched mixture (CEM) cement, propolis, and propolis individually combined with the extracts of several medicinal plants were investigated against Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. Then, the cytotoxicity of each substance or mixture against pulp stem cells extracted from 30 primary healthy teeth was evaluated at 4 concentrations. Data were gathered via observation, and optical density values were obtained using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) test and recorded. SPSS software version 23 was used to analyze the data. Data were evaluated using 2-way analysis of variance and the Tukey test. Results: Regarding antimicrobial properties, thyme alone and thyme + propolis had the lowest minimum inhibitory concentrations (MICs) against the growth of S. aureus, E. coli, and P. aeruginosa bacteria. For E. faecalis, thyme + propolis had the lowest MIC, followed by thyme alone. At 24 and 72 hours, thyme + propolis, CEM cement, and propolis had the greatest bioviability in the primary dental pulp stem cells, and lavender + propolis had the lowest bioviability. Conclusions: Of the studied materials, thyme + propolis showed the best results in the measures of practical performance as a dental pulp cap.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.3
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• pp.893-899
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• 2004

To test the cytoprotective effect of sophorae radix (SR) against hydrogen peroxide (H₂O₂)-induced cytotoxicity, we investigated the cell viability using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay in the presence of ethyl acetate subfractions of SR water extracts In H9c2 cells. And to clarify the cytoprotective mechanism of SR extracts, we evaluated the cellular glutathione (GSH) contents in the presence of subfraction 1, 2, 3, and 4 of SR ethyl acetate soluble fractions. Among 1 -12 subfractions of SR ethyl acetate soluble fractions, 1, 2, 3 and 4 subfractions have an efficacy inhibiting the cytotoxicity induced by H₂O₂ in H9c2 cells. Also, the protective effects of 1, 2, 3 and 4 subfractions of SR ethyl acetate soluble fractions resulted from the anti-oxidant effects. These results suggest that ethyl acetate soluble fractions of SR water extracts is effective in the prevention of H₂O₂-induced cytotoxicity and 1, 2, 3 and 4 subfraction of ethyl acetate soluble fractions possess the anti-oxidant component.

• Bulletin of the Korean Chemical Society
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• v.31 no.12
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• pp.3567-3572
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• 2010

Black pine barks from the southern region of Korea were extracted using pressurized hot water and the water soluble extracts were then separated in a stepwise fashion using a variety of solvents, column chromatography (CC), thin layer chromatography (TLC), and high pressure liquid chromatography (HPLC). The antioxidant activities of each fraction and the active compounds were determined based on the radical scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH), reductive potential of ferric ion, and total phenol contents. A DPPH test showed that the half maximal effective concentration ($EC_{50}$ value : $6.59{\pm}0.31\;{\mu}g/mL$) of the ethyl acetate fraction (ca. 0.67%) was almost the same as that of the control compounds and inversely proportional to the value of the total phenol contents. The cell viability of the water extracts was confirmed by methyl thiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT) with enzyme linked immune sorbent assay (ELISA). Catechin, epicatechin, quercetin and ferulic acid were isolated from the ethyl acetate fraction as active compounds and identified by nuclear magnetic resonance. The antioxidant activity as value of DPPH of each of the separated compounds was lower than the ethyl acetate fraction, and ferulic acid was the lowest among these compounds.

• Journal of Korean Medicine for Obesity Research
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• v.13 no.1
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• pp.24-32
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• 2013

Objectives: The aim of this study is to investigate anti-imflammatory and protective effect for intestinal epithelial cells with Atractylodes macrocephae (AM), a traditional Korean Herbal medicine and fermented Atractylodes macrocephae (FAM) with Lactobacillus plantarum. Methods: HCT-116 and Raw 264.7 cells were used in this study. Using NO assay, we measured lipopolysaccharide (LPS)-induced anti-inflammatory effect. We measured permeability of intestinal epithelial cells with transepithelial electrical resistance and horseradish peroxide flux assay. Water soluble tetrazolium salt assay was used to see cell proliferation. All the results were presented in mean and standard deviation. We used Student's t-test for analyzing significance of results. Results: In Raw 264.7 cells NO production decreased 22.4% with pre-treatment of AM and FAM, especially with FAM in high concentration. In HCT-116 cells LPS-induced intestinal permeability had a protective effect with both AM and FAM, which was also tend to be proportional to the concentration. Cell viability increased up to 135.52% after treatment of high concentration of FAM in HCT-116, while there was no significant change in Raw 264.7 cells with herb treatments. Conclusions: These results show evidence that AM, especially fermented ones, significantly reduced intestinal membrane permeability. They also had a protective effect as well as an anti-inflammation effect for HCT-116 and Raw 264.7 cells. This suggest that FAM may be a therapeutic agent for Leaky gut syndrome by reducing intestinal permeability.

• Journal of dental hygiene science
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• v.20 no.4
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• pp.213-220
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• 2020

Background: The leaves of Perilla frutescens, commonly called perilla and used for food in Korea, contain components with a variety of biological effects and potential therapeutic applications. The purpose of this study was to identify the components of 70% ethanol extracted Perilla frutescens (EEPF) and determine its inhibitory effects on oral microbial activity and production of nitric oxide (NO) and prostaglandin E2 (PGE2) in lipopolysaccharides (LPS)-stimulated Raw264.7 macrophages, consequently, to confirm the possibility of using EEPF as a functional component for improving the oral environment and preventing inflammation. Methods: One kg of P. frutescens leaves was extracted with 70% ethanol and dried at -70℃. EEPF was analyzed using high-performance liquid chromatography analysis, and antimicrobial activity against oral microorganisms was revealed using the disk diffusion test. Cell viability was elucidated using a methylthiazolydiphenyl-tetrazolium bromide assay, and the effect of EEPF on LPS-induced morphological variation was confirmed through microscopic observation. The effect of EEPF on LPS-induced production of pro-inflammatory mediators, NO and PGE2 was confirmed by the NO assay and PGE2 enzyme-linked immunosorbent assay. Results: The main component of EEPF was rosemarinic acid, and EEPF showed weak anti-bacterial and anti-fungal effects against microorganisms living in the oral cavity. EEPF did not show toxicity to Raw264.7 macrophages and had inhibitory effects on the morphological variations and production of pro-inflammatory mediators, NO and PGE2 in LPS-stimulated Raw264.7 macrophages. Conclusion: EEPF can be used as a functional material for improving the oral environment through the control of oral microorganisms and for modulating inflammation by inhibiting the production of inflammatory mediators.

• The Korean Journal of Pain
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• v.35 no.2
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• pp.140-151
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• 2022

Background: Essential oils are of great interest for their analgesic and anti-inflammatory properties. We aimed to study the content of the essential oil of the Origanum vulgare of the Armenian highlands (OVA) in different periods of vegetation and to investigate its antinociceptive and anti-inflammatory effects in mice (in vivo) and cytotoxic action in cultured cells (in vitro). OVA essential oil was extracted from fresh plant material by hydro-distillation. Methods: For OVA essential oil contents determination the gas chromatography-mass spectrometry method was used. Formalin and hot plate tests and analysis of cell viability using the methyl-thiazolyl-tetrazolium (MTT) assay were used. Results: The maximal content of β-caryophyllene and β-caryophyllene oxide in OVA essential oil was revealed in the period of blossoming (8.18% and 13.36%, correspondently). In the formalin test, 4% OVA essential oil solution (3.5 mg/mouse) exerts significant antinociceptive and anti-inflammatory effects (P = 0.003). MTT assay shows approximately 60% cytotoxicity in HeLa and Vero cells for 2.0 µL/mL OVA essential oil in media. Conclusions: The wild oregano herb of Armenian highlands, harvested in the blossoming period, may be considered as a valuable source for developing pain-relieving preparations.

• The Journal of Korean Academy of Prosthodontics
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• v.54 no.2
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• pp.120-125
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• 2016

Purpose: The purpose of this study is to identify the effect of mesenchymal stem cell proliferation on recombinant human transforming growth factor-beta (rhTGF-${\beta}2$) / poly (D,L-lactide-co-glycolide) (PLGA) treated titanium discs by electrospray. Materials and methods: Anodized titanium surface coated with PLGA was used for a control group to compare anodized titanium surface coated with 125 ng/ml and 500 ng/ml rhTGF-${\beta}2$ as test groups. Atomic force microscope (AFM) test was utilized to determine the difference in coating surface roughness, and field-emission scanning electron microscopy (FE-SEM) was taken to visualize even distribution of coating particles on titanium discs. The mesenchymal stem cell proliferation was tested by using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl-tetrazolium bromide) assay on 1st, 4th, 7th days. Results: According to AFM results, there was no statistically significant difference in titanium discs treated with PLGA and with rhTGF-${\beta}2$/PLGA (P>.05). MTT assay test results showed that there was statistically significant difference in mesenchymal stem cell proliferation on test groups compared to control groups at 7th day, and cell viability on discs coated with rhTGF-${\beta}2$ was significantly higher than control groups (P<.05). Conclusion: Titanium surface coated with rhTGF-${\beta}2$/PLGA shows statistically significant higher cell proliferation and the titanium surface coated with the higher concentration of rhTGF-${\beta}2$ presents faster cell growth activity.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.28 no.1
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• pp.127-143
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• 1998

The author evaluated the effects of taxol, a microtubular inhibitor, as a possible radiation sensitizer and the production of prostaglandins on three human cancer cell lines(KB, RPMI-2650 and SW-13) and one murine cell line(L929). Each cell line was divided into four groups (control, taxol only, radiation only and combination of taxol and radiation). The treatment consisted of a single irradiation of 10Gy and graded doses (5, 50, 100, 200, 300, 500 nM) of taxol for a 24-h period. The cytotoxicity of taxol alone was measured at 1 day after(1-day group) and 4 days after(4-day group) the treatment. The survival ratio of cell was analyzed by MTT (3-(4,5-dimethylthiazol-2-yl) -2,5-dimethyl tetrazolium bromide) test. Prostaglandins(PGE2 and PGI2) were measured in the culture medium by a radioimmunoassay. The results obtained were as follows. 1. There was a significantly increased cytotoxicity of KB cells in 4-day group than those in I-day group. There was a high correlation between doses of taxol and cell viability in both groups(l-day group R=0.82741, 4-day group R=0.84655). 2. There was a significantly increased cytotoxicity of RPMI -2650 cells treated with high concentration of taxol in 4-day group than those in I-day group. Also there was a high correlation between doses of taxol and cell viability in 4-day group(R=0.93917). 3. There was a significantly increased cytotoxicity of SW-13 cells treated with high concentration of taxol in 4-day group than those in 1-day group. However no high correlation was observed between doses of taxol and cell viability in both groups(1-day group R=0.46362, 4-day group R=0.65425). 4. There was a significantly increased cytotoxicity of L929 cells treated with low concentration of taxol in 4-day group than those in 1-day group. At the same time, there was a low correlation between doses of taxol and cell viability in both groups(1-day group R=0.34237, 4-day group R=0.23381). 5. In I-day group of L929 cells, higher cytotoxicities were observed in the groups treated with 500 nM taxol than given 10 Gy radiation alone. L929 cells in I-day group alone showed a radiosensitizing effect by taxol.. 6. In addition to L929 cells, all cancer cells treated with a combination of taxol and radiation in 4-day group appeared to have some fragmented nuclei and to float on the medium. In addition, L929 cells appeared to be more confluent. 7. The level of PGE2 production was the highest in the contol KB cells. This appeared to increase in every experimental group of all three cancer cells except L929 cells. There was a significantly increased production of PGE2 in SW -13 cells treated with a combination taxol and radiation compared to the other experimental groups. 8. The level of PGE2 production in the control group of RPMI-Z650 cells was the highest. This appeared to increase in every experimental group of all cells except in SW-13 cells. This also increased significantly in RPMI-2650 cells treated with a combination of taxol and radiation compared to the other experimental groups.

• Journal of Pharmacopuncture
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• v.20 no.2
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• pp.127-131
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• 2017

Objectives: Polymorphonuclear neutrophils (PMNs) constitute the first line of defense against invading microbial pathogens. Early events in inflammation involve the recruitment of neutrophils to the site of injury or damage where changes in intracellular calcium can cause the activation of pro-inflammatory mediators from neutrophils including superoxide generation, degranulation and release of myeloperoxidase (MPO), productions of interleukin (IL)-8 and tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$), and adhesion to the vascular endothelium. To address the anti-inflammatory role of flavonoids, in the present study, we investigated the effects of the flavonoids quercetin and vitexin on the stimulus-induced nitric oxide (NO), $TNF-{\alpha}$, and MPO productions in human neutrophils. Methods: Human peripheral blood neutrophils were isolated, and their viabilities were determined by using the Trypan Blue exclusion test. The polymorphonuclear leukocyte (PMNL) preparations contained more than 98% neutrophils as determined by morphological examination with Giemsa staining. The viabilities of cultured neutrophils with various concentrations of quercetin and vitexin ($1-100{\mu}M$) were studied using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without quercetin and vitexin ($25{\mu}M$) for 45 min, and stimulated with phorbol 12-myristate 13-acetate (PMA) ($10^{-7}M$). NO production was carried out through nitrite determination by using the Griess method. Also, the $TNF-{\alpha}$ and the MPO productions were measured using enzyme-linked immunosorbent assay (ELISA) kits and MPO assay kits. Results: Neutrophil viability was not affected up to a concentration of $100{\mu}M$ of quercetin or vitexin. Both quercetin and vitexin significantly inhibited $TNF-{\alpha}$, NO, and MPO productions in human neutrophils (P < 0.001). Conclusion:The present study showed that both quercetin and vitexin had significant anti-inflammatory effects. Thus, treatment with either quercetin or vitexin may be considered as a therapeutic strategy for treating patients with neutrophil-mediated inflammatory diseases.

• Journal of Korean Medicine for Obesity Research
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• v.19 no.2
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• pp.106-112
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• 2019

Objectives: The aim of this study was to observe the reduction of lipid accumulation by treatment with Akkermansia muciniphila extract on 3T3-L1 adipocytes. Methods: After treating pasteurized Akk. muciniphila strains in HT-29 colorectal cancer cell, the relative expression of interleukin (IL)-8, tumor necrosis factor-α, IL-6, and IL-1β mRNA was analyzed by real time polymerase chain reaction, respectively. 27 strains of Akk. muciniphila which have anti-inflammatory effects were selected. 3T3-L1 pre-adipocytes were treated with Akk. muciniphila for 24 hr and then measured the toxicity using water soluble tetrazolium salt assay. The cells were incubated for 4 days and then differentiated into adipocytes using the medium including adipogenic reagents for 10 days. The Akk. muciniphila was treated when the medium was exchanged for differentiation medium at 4^th day and insulin medium at 6^th day. To observe the lipid accumulation, the cells were stained with Oil red O dye and were measured using a spectrophotometer. Results: In the cytotoxicity test, the cell viability of 3T3-L1 pre-adipocytes was significantly increased compared to the control group which untreated with Akk. muciniphila, and there was no cytotoxicity of Akk. muciniphila at 1×10⁷ CFU/mL. The results on Oil red O staining and absorbance measurements were showed a significant decrease in lipid accumulation in the group which was treated with Akk. muciniphila compared to the control group. Conclusions: In our results, Akk. muciniphila has the inhibitory effect of lipid accumulation in 3T3-L1 adipocytes. This suggests that Akk. muciniphila could be help to improve obesity.