• Title/Summary/Keyword: In vitro cytotoxicity

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Attachment of Human Gingival Fibroblast to Various Subgingival Restorations;A Comparative Study in Vitro (다양한 치은 연하 수복물에 대한 치은 섬유아 세포 부착 연구)

  • Lee, Eun-Suk;Song, In-Taeck;Lim, Jeong-Su;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.29 no.3
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    • pp.621-636
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    • 1999
  • When mucoperiosteal flaps are positioned and sutured to desirable position, the wound contains several interface between tissues which differ fundamentally in composition & biological reaction. Thus the C-T surface of the flap will, on one hand, oppose another vascularized surface, and on the other, the avascular dental material for example, when root resoptions, fractured root, endodontic perforation, deep root carious lesions were filled with amalgam, glass ionomer, resin etc. Recently, a number of case report described the successful treatment of a subgingival root lesion with restorative material & free gingival graft, open flap surgery, but more objective research was needed . Most of study on restorative materials were concerned for cytotoxicity not for actual healing event on that materials and its influencing factors such as biocompatibility, surface wettability, surface topography . The aim of this in vitro study was to evaluate the effect of amalgam, resin modified glass ionomer, composite resin per se, and their surface roughness on the growth of human gingival fibroblast. The cells were obtained and placed on culture flask and incubated for 3 days with the prepared test materials. Then count the attached cell number with hemocytometer,(n=12) and 2 samples were examined with SEM about attachment cell morphology . Another 4 samples were evaluated on their surface roughness with Talysurf and average surface roughness value(Ra) were obtained. Statistical difference in attached cell number, roughness value were analyzed using ANOVA. The number of attached cell was as follows, for root dentin specimen 16.7${\pm}$4.41, resin modified glass ionomer 14.0${\pm}$4.15, resin 8.13${\pm}$3.63, amalgam 0.72${\pm}$3.33(${\times}10^3$). Between root dentin and resin-modified glass ionomer, no significant difference was observed, but resin, amalgam showed a significant less cell numbers than for root dentin, resin modified glass ionomer cement. SEM examination expressed many cell surface attachment apparatus in root dentin and resin modified glass ionomer specimens. For resin specimen, cell attachment was observed but exposed less appratus. The average surface roughness value are following results. Dentin specimen 0.6972${\pm}$ 0.104, resin modified glass ionomer 0.0822${\pm}$0.009, resin 0.0875${\pm}$0.005, amalgam 4.2145${\pm}$0.985(${\mu}m$). Between root dentin, resin-modified glass ionomer, and resin, no significant difference was observed, but amalgam showed a significant more rough surface than other groups. When evlauated the interrelationship between cell attachment and surface roughness, therefore, there was weak reverse correlation.(pearson correlation : - 0.593) These results suggest that resin modified glass ionomer have the favorable healing potential when used for subgingival restoration. And for relationship between cell attachment and surface characteristics, further investigations were needed.

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Glucosylation of Resveratrol Improves its Immunomodulating Activity and the Viability of Murine Macrophage RAW 264.7 Cells (당화된 레스베라트롤의 대식세포 RAW 264.7세포의 생존능력과 레스베라트롤의 면역제어 활성을 증가)

  • Pandey, Ramesh Prasad;Lee, Jisun;Park, Yong Il;Sohng, Jae Kyung
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.19-26
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    • 2017
  • Effects of resveratrol glucosylation on the immunomodulation properties of resveratrol and on the viability of macrophage cells have been studied by using murine macrophage RAW 264.7 cells. Nitric oxide (NO) and interleukin 6 (IL-6) expression in macrophages in vitro were studied after treatment with different concentrations of (E)-resveratrol, (E)-resveratrol 3-O-${\beta}$-${\small{D}}$-glucoside (R-3-G), or (E)-resveratrol 4'-O-${\beta}$-${\small{D}}$-glucoside (R-4'-G). In vitro viability of RAW 264.7 cells after treatment with the aforementioned three compounds was also studied. As demonstrated by macrophage cell viability assays, two different resveratrol monoglucosides, R-3-G and R-4'-G, exhibited 50-80% reduced cytotoxicity in comparison to (E)-resveratrol in A549 and HepG2 cells. Compared to the resveratrol aglycon, both glucosylated resveratrol derivatives positively modulated NO and IL-6 production in macrophages positively via transcriptionally up-regulating IL-6 and iNOS expression. Conjugation of a glucose moiety on resveratrol was found to enhance the immunomodulating activity of resveratrol and the viability of RAW 264.7 cells.

Characterization of Immuno-stimulating Polysaccharides Isolated from Korean Persimmon Vinegar (감식초에서 분리한 면역활성다당의 특성)

  • Hwang, Yong-Chul;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.40 no.2
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    • pp.220-227
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    • 2008
  • In this study, polysaccharides were isolated from Korean persimmon vinegar to characterize the polysaccharides existing as soluble forms within traditional Korean fermented beverages, and their immuno-stimulating activities were examined. Three successive chromatographies were used to purify the main polysaccharide in the persimmon vinegar, PV-1b-I, to homogeneity from the crude polysaccharide (PV-0). The molecular mass of PV-1b-I was estimated as 110 kDa and it contained significant proportions of mannose (46.8%), galactose (28.5%) and arabinose (19.1%). PV-1b-I strongly reacted with ${\beta}$-glucosyl Yariv reagent, suggesting the presence of an arabino-3,6-galactan moiety. PV-1b-I also induced high levels of macrophage activation and mitogenicity on murine splenocytes in vitro. The intravenous administration of PV-1b-I significantly augmented NK cytotoxicity against YAC-1 tumor cells. PV-1b-I also showed potent anticomplementary activity in a dose-dependent manner. Finally, C3 activation products were identified by crossed immunoelectrophoresis using anti-human C3 and the anti-complementary activity of PV-1b-I under $Ca^{2+}$-free conditions, suggesting that this PV-1b-I causes complementary activations via both alternative and classical pathways. From these results, one can conclude that Korean persimmon vinegar contains select polysaccharides in addition to healthy components, and these polysaccharides appear to provide immuno-stimulating activities beneficial to human health.

Concentration-dependent in vitro Anti-osteoarthritis Effects of Mixed Formula - Pomegranate Concentrate Powder: Eucommiae Cortex: Achyranthis Radix 5:4:1 (g/g) on the Primary Cultured Rat Articular Chondrocytes

  • Choi, Beom Rak;Ku, Sae Kwang;Kang, Su Jin;Park, Hye Rim;Sung, Mi Sun;Lee, Young Joon;Park, Ki Moon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.33 no.2
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    • pp.131-140
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    • 2019
  • The objective of present study is to evaluate concentration-dependent in vitro anti-osteoarthritic (OA) effects of synergic mixed formula consisted of dried pomegranate juice concentrate powder, Eucommiae Cortex aqueous extract and Achyranthis Radix aqueous extract 5:4:1 (g/g) mixture on the primary cultured rat articular chondrocytes. First, any cytotoxic effect of mixture was observed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay. Next, cyto-protective effect of test substances was evaluated by using the recombinant human interleukin $(rhIL)-1{\alpha}$ induced chondrocytes. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin $E_2(PGE_2)$ productions and 5-lipoxygenase (LPO) activities, and inhibitory effects on matrix metalloproteinase (MMP)-2 and MMP-9 activities were observed on $rhIL-1{\alpha}$ treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions. No obvious cytotoxic effects of mixture were demonstrated. Inflammatory damages of chondrocytes and related ECM degradations induced by treatment of LPS or $rhIL-1{\alpha}$ were significantly and concentration-dependently inhibited by pretreatment of mixture from a concentration level of 0.001 mg/ml to 1 mg/ml. In addition, mixture showed $IC_{50}$ for $rhIL-1{\alpha}-induced$ MMP-2 and MMP-9 activities as 44.01 and $162.47{\mu}g/ml$, and also showed $EC_{50}$ for $rhIL-1{\alpha}-induced$ inhibition of collagen type II, SOX9 and aggrecan mRNA expression as 8.61, 10.79 and $4.47{\mu}g/ml$, respectively. It is observed that mixture showed concentration-dependent anti-inflammatory and cytoprotective ECM preserved effects on the primary cultured rat articular chondrocytes without cytotoxicity.

Preparation and Functional Properties of Dendropanax morbiferus Kombucha

  • Jie Rong;Ki-Bae Hong;Yun Jae Cho;Sung-Soo Park
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.442-448
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    • 2023
  • This study aimed to prepare kombucha, a fermented tea beverage, containing Dendropanax morbiferus (DM) leaves and roots, and analyze its antioxidant and intracellular activities. We compared the pH change, total acidity, radical scavenging activity, and oxygen radical absorbance capacity (ORAC) of kombucha fermented with black tea alone and that with added DM leaves or roots during fermentation. Using RAW 264.7, we evaluated the effects of kombucha containing different DM parts on nitric oxide (NO) production and inflammation-related cytokine content in cells. Kombucha containing ethanol extracts of DM leaves (BTK-E-DML) and roots (BTK-E-DMR) showed higher radical scavenging activity and ORAC 3 d after fermentation than that prepared from black tea alone (BTK-Ori). In an in vitro experiment using RAW 264.7, samples were treated with 8 mg/mL kombucha considering cytotoxicity; the lipopolysaccharide (LPS)-induced NO content significantly reduced after BTK-E-DML and BTK-EDMR treatments compared with that after BTK-Ori treatment. Additionally, the levels of interleukin-6 and tumor necrosis factor-alpha, which were LPS-stimulated inflammatory cytokines, significantly decreased in cells treated with BTK-E-DML and BTK-E-DMR 15 d after fermentation compared with those treated with BTK-Ori. In conclusion, these results demonstrate that kombucha fermented with the leaves and roots of DM increases antioxidant activity and can significantly regulate inflammatory responses at the cellular level.

Cytotoxicity Evaluation of Essential Oil and its Component from Zingiber officinale Roscoe

  • Lee, Yongkyu
    • Toxicological Research
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    • v.32 no.3
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    • pp.225-230
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    • 2016
  • Zingiber officinale Roscoe has been widely used as a folk medicine to treat various diseases, including cancer. This study aims to re-examine the therapeutic potential of co-administration of natural products and cancer chemotherapeutics. Candidate material for this project, ${\alpha}$-zingiberene, was extracted from Zingiber officinale Roscoe, and ${\alpha}$-zingiberene makes up $35.02{\pm}0.30%$ of its total essential oil. ${\alpha}$-Zingiberene showed low $IC_{50}$ values, $60.6{\pm}3.6$, $46.2{\pm}0.6$, $172.0{\pm}6.6$, $80.3{\pm}6.6$ (${\mu}g/mL$) in HeLa, SiHa, MCF-7 and HL-60 cells each. These values are a little bit higher than $IC_{50}$ values of general essential oil in those cells. The treatment of ${\alpha}$-zingiberene produced nucleosomal DNA fragmentation in SiHa cells, and the percentage of sub-diploid cells increased in a concentration-dependent manner in SiHa cells, hallmark features of apoptosis. Mitochondrial cytochrome c activation and an in vitro caspase-3 activity assay demonstrated that the activation of caspases accompanies the apoptotic effect of ${\alpha}$-zingiberene, which mediates cell death. These results suggest that the apoptotic effect of ${\alpha}$-zingiberene on SiHa cells may converge caspase-3 activation through the release of mitochondrial cytochrome c into cytoplasm. It is considered that anti-proliferative effect of ${\alpha}$-zingiberene is a result of apoptotic effects, and ${\alpha}$-zingiberene is worth furthermore study to develop it as cancer chemotherapeutics.

Isolation and Identification of Cytotoxic and Biological Active Toxin from the Puffer Fish Arothron stellatus

  • Veeruraj, Anguchamy;Pugazhvendan, Sampath Renuga;Ajithkumar, Thipramalai Thankappan;Arumugam, Muthuvel
    • Toxicological Research
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    • v.32 no.3
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    • pp.215-223
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    • 2016
  • This study is to investigate the biological, biochemical and cytotoxic effects of puffer fish (Arothron stellatus) toxin extracts under in-vitro condition. Extracted toxins from various organs of puffer fish were purified by using active charcoal column, and Bio-gel-P2 column chromatography. The lethality of toxin was tested in crabs, which consists of neurotoxic compounds. The degree of the brine shrimp lethality assay was found directly proportional to the concentration of the toxin extracts, which was well supported by hemolytic assay. The experimental results suggested that the gonad was found higher toxins than the liver and muscles. The mortality rate of brine shrimp nauplii was increased with the raise of concentrations of toxin level. Among the different doses and time dependent cytotoxic effect of human cervical carcinoma (HeLa) cells were showed $4.0{\mu}g/mL$ of toxin, which was effectively inhibited cancer cell proliferation. HPLC and TLC analysis was revealed that the A. stellatus toxin contains tetrodotoxin (TTX), related compounds 4-epi TTX and anhydro-TTX. The present results suggested that the A. stellatus contain TTX as a major and anh-TTX as a minor toxin. It could be the potential candidate in the field of anticancer drug discovery against human cervical cancer cells. The present data is confirming that the puffer fish toxin as an interesting source of novel bioactive natural compounds with potent applications in pharmacology.

Synthesis and Optimization of Cholesterol-Based Diquaternary Ammonium Gemini Surfactant (Chol-GS) as a New Gene Delivery Vector

  • Kim, Bieong-Kil;Doh, Kyung-Oh;Bae, Yun-Ui;Seu, Young-Bae
    • Journal of Microbiology and Biotechnology
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    • v.21 no.1
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    • pp.93-99
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    • 2011
  • Amongst a number of potential nonviral vectors, cationic liposomes have been actively researched, with both gemini surfactants and bola amphiphiles reported as being in possession of good structures in terms of cell viability and in vitro transfection. In this study, a cholesterol-based diquaternary ammonium gemini surfactant (Chol-GS) was synthesized and assessed as a novel nonviral gene vector. Chol-GS was synthesized from cholesterol by way of four reaction steps. The optimal efficiency was found to be at a weight ratio of 1:4 of lipid:DOPE (1,2-dioleoyl-L-${\alpha}$- glycero-3-phosphatidylethanolamine), and at a ratio of between 10:1~15:1 of liposome:DNA. The transfection efficiency was compared with commercial liposomes and with Lipofectamine, 1,2-dimyristyloxypropyl-3-dimethylhydroxyethylammonium bromide (DMRIE-C), and N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTAP). The results indicate that the efficiency of Chol-GS is greater than that of all the tested commercial liposomes in COS7 and Huh7 cells, and higher than DOTAP and Lipofectamine in A549 cells. Confirmation of these findings was observed through the use of green fluorescent protein expression. Chol-GS exhibited a moderate level of cytotoxicity, at optimum concentrations for efficient transfection, indicating cell viability. Hence, the newly synthesized Chol-GS liposome has the potential of being an excellent nonviral vector for gene delivery.

Luteolin Induced-growth Inhibition and Apoptosis of Human Esophageal Squamous Carcinoma Cell Line Eca109 Cells in vitro

  • Wang, Ting-Ting;Wang, Shao-Kang;Huang, Gui-Ling;Sun, Gui-Ju
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5455-5461
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    • 2012
  • Luteolin is a plant flavonoid which exhibits anti-oxidative, anti-inflammatory and anti-tumor effects. However, the antiproliferative potential of luteolin is not fully understood. In this study, we investigated the effect of luteolin on cell cycling and apoptosis in human esophageal squamous carcinoma cell line Eca109 cells. MTT assays showed that luteolin had obvious cytotoxicity on Eca109 with an $IC_{50}$ of $70.7{\pm}1.72{\mu}M$ at 24h. Luteolin arrested cell cycle progression in the G0/G1 phase and prevented entry into S phase in a dose- and time-dependent manner. as assessed by FCM. Luteolin induced apoptosis of Eca109 cells was demonstrated by AO/EB staining assay and annexin V-FITC/PI staining. Moreover, luteolin downregulated the expression of cyclin D1, survivin and c-myc, and it also upregulated the expression of p53, in line with the fact that luteolin was able to inhibit Eca109 cell proliferation.

EFFECTS OF CHLORHEXIDINE AND L1STERINE ON CELL ACTIVITY OF HUMAN GINGIVAL FIBROBLAST IN VITRO (Chlorhexidine과 Listerine이 인체 치은 섬유모세포의 활성화에 미치는 영향)

  • Kang, Jung-Koo;Yoo, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.25 no.1
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    • pp.1-13
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    • 1995
  • Chlorhexidine and Listerine are widely used in dentistry due to its effectiveness on plaque control and bactericidal action. The effects of these agent on chronic gingivitis and wound healing following surgical periodontal therapy in human has been favorable. Understanding the effects of chlorhexidine and Listerine on human gingival fibroblast will provide the rationale for its use during the healing process of periodontal surgery. The purpose of this study was to compare the effects of chlorhexidine and Listerine on human gingival fibroblast. Human gingival fibroblasts were cultured from the healthy gingiva on the extracted premolar of orthodontic patients. Human gingival fibroblast were trypsinized and cultured in growth medium added range of 0.0012-0.12% chlorhexidine and 1-100% Listerine mouth wash solution. The cell used in this study were between fifth to eighth passage number. The cell morphology were examined by inverted microscope and the cell activity were measured by MIT assay. The Morphology of gingival fibroblast added Chlorhexidine and Listerine at the concentration of all range were became globular and lost their cytoplasmic process. Our results indicate that a 0.0012 concentration of chlorhexidine and 1% concentration of Listerine were shows minimal cytotoxicity, but above these concentraion, there was a significant difference between the cell activity in the experimental group and control group(p

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