• Title/Summary/Keyword: in vitro cytotoxicity test

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Effects of Bujasasim-tang Ethanol Extract on Oxidative Stress, Inflammation and Osteoarthritic Rat Model (부자사심탕(附子瀉心湯)이 산화적 손상, 염증 및 골관절염 병태모델에 미치는 영향)

  • Woo, Chang-Hoon;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.25 no.2
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    • pp.15-35
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    • 2015
  • Objectives This study was performed to investigate the effects of Bujasasim-tang ethanol extract (BST) on oxidative stress, inflammation and osteoarthritic rat model. Methods To ensure safety of BST, heavy metal levels were measured and cytotoxicity test was done. In vitro, To evaluate antioxidative effects of BST, total phenolic contents, 1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging activity, reactive oxygen species (ROS) levels were measured. Also, to evaluate anti-inflammatory effects of BST treated group, total nitric oxide (NO) and pro-inflammatory cytokines (IL-$1{\beta}$, IL-6, TNF-${\alpha}$) levels were measured in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. In vivo, We injected MIA $50{\mu}l$ (60 mg/ml) into knee joints of rats to induce osteoarthritis. Rats were divided into total 3 groups (normal, control, BST treated group, each n=7). Normal group was not treated at all without inducing osteoarthritis and taken normal diet. Control group was induced osteoarthritis by MIA and taken with 2 ml of distilled water once a day for 4 weeks. BST treated group was induced osteoarthritis by MIA and taken BST 2 ml (200 mg/kg/mouse) once a day for 4 weeks. We evaluated dynamic weight bearing with the Incapacitance Test Meter. At the end of experiment, the rats were sacrificed to observe the functions of liver and kidney, changes of WBC, neutrophil, lymphocyte, monocyte levels in blood, to evaluate the levels of pro-inflammatory cytokines, tissue inhibitor of metallopreteinases-1 (TIMP-1), matrix metalloproteinase-9 (MMP-9), prostaglandin $E_2$ ($PGE_2$), leukotriene $B_4$ ($LTB_4$) within serum. We observed change of articular structures by Hematoxylin & Eosin (H&E), safranin-O staining method and measured amount of cartilage by micro CT-arthrography. Statistical analysis was done by unpaired student's t-test with significance level at p<0.05 in SPSS 11.0 for windows. Results 1. Safety of the BST was identified. 2. AST, ALT, BUN, creatinine levels of BST treated group were within normal limit. In vitro, 1. DPPH and ABTS free radical scavenging activities of BST showed dose-dependent increase. 2. ROS production were significantly decreased. 3. Total nitric oxide (NO) and IL-$1{\beta}$ production were decreased. 4. IL-6 and TNF-${\alpha}$ production were significantly decreased. In vivo, 1. Weight bearing ability was significantly increased. 2. WBC, neutrophil, lymphocyte, monocyte levels in blood were decreased. 3. IL-$1{\beta}$ and TNF-${\alpha}$ levels in serum were significantly decreased. and the IL-6 level was decreased. 4. TIMP-1, MMP-9, $LTB_4$, $PGE_2$ levels in serum were significantly decreased. 5. Cartilage volume of BST treated group was significantly increased. Also changes of cartilage, synovial membrane, fibrous tissue were suppressed. Conclusions The results obtained in this study Bujasasim-tang have effects of antioxidative, anti-inflammatory, relieve pain and protection of cartilage. Therefore we expect that Bujasasim-tang is effective treatment for osteoarthritis.

Biocompatibility of experimental mixture of mineral trioxide aggregate and glass ionomer cement (Glass ionomer cement와 혼합한 mineral trioxide aggregate의 생체친화성)

  • Oh, Min-Jae;Jeong, Yu-Na;Bae, In-Ho;Yang, So-Young;Park, Bum-Jun;Koh, Jeong-Tae;Hwang, Yun-Chan;Hwang, In-Nam;Oh, Won-Mann
    • Restorative Dentistry and Endodontics
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    • v.35 no.5
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    • pp.359-367
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    • 2010
  • Objectives: The purpose of the present in vitro study was to evaluate the biocompatibility of mineral trioxide aggregate (MTA) mixed with glass ionomer cement (GIC), and to compare it with that of MTA, GIC, IRM and SuperEBA. Materials and Methods: Experimental groups were divided into 3 groups such as 1 : 1, 2 : 1, and 1 : 2 groups depending on the mixing ratios of MTA powder and GIC powder. Instead of distilled water, GIC liquid was mixed with the powder. This study was carried out using MG-63 cells derived from human osteosarcoma. They were incubated for 1 day on the surfaces of disc samples and examined by scanning electron microscopy. To evaluate the cytotoxicity of test materials quantitatively, XTT assay was used. The cells were exposed to the extracts and incubated. Cell viability was recorded by measuring the optical density of each test well in reference to controls. Results: The SEM revealed that elongated, dense, and almost confluent cells were observed in the cultures of MTA mixed with GIC, MTA and GIC. On the contrary, cells on the surface of IRM or SuperEBA were round in shape. In XTT assay, cell viability of MTA mixed with GIC group was similar to that of MTA or GIC at all time points. IRM and SuperEBA showed significantly lower cell viability than other groups at all time points (p < 0.05). Conclusions: In this research MTA mixed with GIC showed similar cellular responses as MTA and GIC. It suggests that MTA mixed with GIC has good biocompatibility like MTA and GIC.

Antimicrobial and cytotoxic properties of calcium-enriched mixture cement, Iranian propolis, and propolis with herbal extracts in primary dental pulp stem cells

  • Mohammad Esmaeilzadeh;Shirin Moradkhani;Fahimeh Daneshyar;Mohammad Reza Arabestani;Sara Soleimani Asl;Soudeh Tayebi;Maryam Farhadian
    • 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.

Evaluation of biodegradability and tissue regenerative potential of synthetic biodegradable membranes (수종의 성분해성 차폐막의 생체분해도 및 조직 재생유도 능력에 관한 연구)

  • Kim, Dong-Kyun;Ku, Young;Lee, Young-Moo;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.27 no.1
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    • pp.151-163
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    • 1997
  • The purpose of this study was to evaluate on the biodegradability, biocompatibility and tissue regenerative capacity of synthetic biodegradable $mernbranes-Resolut^{(R)}$, $Guidor^{(R)}$ and $Biomesh^{(R)}$. To evaluate the cell attachment on the membranes, in vitro, the number of gingival fibroblasts attached to each membrane was counted by hemocytometer. Cytotoxicity test for the membranes was performed by MTT test with gingival fibroblast For evaluation of guided- bone regenerative potential, the amount of new bone formation in the rat calvarial defects(5mm in diameter) beneath the membranes was observed for two weeks and examined of the specimens by Massons trichrome staining. Biodegradability was observed for 2, 4, 8 and 12 weeks after implantation of each materials under the skin of rats and examined the specimens with H & E staining. The number of cell attachment were the greatest in $Biomesh^{(R)}$ and followed by $Resolut^{(R)}$. Cell viability of three membranes was almost similar levels. Biodegradability of $Resolut^{(R)}$ was the highest among three membrane and the potential of guided bone regeneration was the greatest in the $Biomesh^{(R)}$ and $Resolut^{(R)}$ was followed. These results suggested that commercially available biodegradable membranes were non-toxic and highly potential to guided bone regeneration.

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Sex determination of bovine embryos with hamster H-Y antibody and by polymerase chain reaction (햄스터 H-Y항체와 중합효소연쇄반응을 이용한 소 수정란의 성감별)

  • Yu, Il-jeong;Kim, Yong-jun;Lee, Kyung-kwang
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.189-203
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    • 1999
  • To determine sex of bovine embryos using hamster histocompatibility Y(H-Y) antibodies, bovine compact morulae were incubated for 6 hours in TCM199 supplemented with 10% hamster H-Y antiserum and the embryos with developmental arrest were diagnosed as male embryos, while the embryos showing development during the incubation as female embryos. This presumptive embryo sexing was confirmed by polymerase chain reaction(PCR)method. 1. In the result of hamster sperm cytotoxicity test to measure H-Y antibody titer, the rate of dead sperm was considerably lower in H-Y antiserum absorbed with hamster male splenocytes than in H-Y antiserum absorbed with hamster female splenocytes or H-Y antiserum unabsorbed with splenocytes(p<0.01). 2. The rate of oocytes fertilized in vitro and the rate of blastocysts of the fertilized oocytes were 58.5% and 32.4%, respectively. The rate of blastocysts on day 8 was 15.9%, denoting the highest rate during whole culture period posterior to in vitro fertilization (IVF). 3. The bovine 16 cell and compact morulae embryos incubated in the medium supplemented with hamster H-Y antibodies showed 37.1% and 48.9% of developmental arrest which were diagnosed as male, respectively, and rates of redeveloped embryos from the arrested were 24.1% in 16 cell and 44.3% in compact morulae embryos, respectively, denoting higher rate of sex determination and rate of redevelopment in compact morulae than 16 cell embryos. 4. Bovine compact morulae of Korean cattle and Holstein were treated with hamster H-Y antibodies for sex determination and the rates of developmental arrest(diagnosed as male) were 48.4% for Korean cattle and 47.9% for Holstein, respectively. The rates of redeveloped embryos to blastocyst after treatment were 42.6% for Korean cattle and 41.8% for Holstein, respectively, showing no significant differences of sex determination and redevelopment between both breed. 5. The sex determination of bovine embryos(Korean cattle and Holstein) using hamster H-Y antibodies was diagnosed by PCR for confirmation, denoting the rates of 86.1% for Korean cattle and 85.9% for Holstein male embryos, respectively, and the rates of 91.9% for Korean cattle and 90.1% for Holstein female embryos, respectively, with no significant differences of sex determination between both breed. These results indicated that hamster H-Y antibodies can be usable for sex determination of bovine embryos of Korean cattle and Holstein, the viability of bovine embryos was sustained while being cultured in the medium supplemented with hamster H-Y antibodies of appropriate titer and sex determination of bovine embryos by PCR can be feasible for confirmation.

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Recent Studies of Breast Cancer in Traditional Chinese Medicine Journals (중의학 논문에 나타난 유방암의 연구 동향에 대한 고찰 - 중의학 논문을 중심으로 -)

  • Jerng, Ui-Min;Jeong, Jong-Soo;Park, Jae-Woo;Jung, Hyun-Sik;Yoon, Seong-Woo
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.1
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    • pp.263-278
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    • 2009
  • Purpose: The purpose of this study is to research trends in the study of breast cancer in Traditional Chinese Medicine (TCM) and to establish the further direction for its study. Methods: We reviewed TCM papers published in the last 29 years (1979-2008). Results: 1. We researched 49 papers and the patterns of study were as follows: in vitro studies were 27 papers (55.1%), in vivo studies were 9 papers (18.4%) and clinical studies were 19 papers (38.8%). 2. In vitro studies on breast cancer research in TCM were focused on cytotoxicity (17 papers) and apoptosis (8 papers). Most of in vivo studies (6 papers) were done for the purpose of inducing growth suppression of tumor cell after administration of the test drug. Each drug acted on this effect through various types of mechanism. 3. Unlike in vitro and in vivo studies, clinical studies on growth suppression of tumor cell were rare (4 papers). Most of the studies were focused on reduction of side effect of chemotherapy or synergistic effect with chemotherapy (7 papers), immune regulation (7 papers), and improvement of quality of life (6 papers). 4. Among the treatment method we reviewed, 'Runing Ⅱ(Ⅱ號方)' was the only medication that further studied as clinical trial after experimental study. 5. Since almost all studies have defects like poorly designed model or insufficient data description, it was difficult to make any definite conclusion about these studies. Conclusion: More subsequent clinical studies based on experimental study will be needed afterwards. Strict and high-level study design with detailed description will be needed in further study.

Development of Implantable Blood Pressure Sensor Using Quartz Wafer Direct Bonding and Ultrafast Laser Cutting (Quatrz 웨이퍼의 직접접합과 극초단 레이저 가공을 이용한 체내 이식형 혈압센서 개발)

  • Kim, Sung-Il;Kim, Eung-Bo;So, Sang-kyun;Choi, Jiyeon;Joung, Yeun-Ho
    • Journal of Biomedical Engineering Research
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    • v.37 no.5
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    • pp.168-177
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    • 2016
  • In this paper we present an implantable pressure sensor to measure real-time blood pressure by monitoring mechanical movement of artery. Sensor is composed of inductors (L) and capacitors (C) which are formed by microfabrication and direct bonding on two biocompatible substrates (quartz). When electrical potential is applied to the sensor, the inductors and capacitors generates a LC resonance circuit and produce characteristic resonant frequencies. Real-time variation of the resonant frequency is monitored by an external measurement system using inductive coupling. Structural and electrical simulation was performed by Computer Aided Engineering (CAE) programs, ANSYS and HFSS, to optimize geometry of sensor. Ultrafast laser (femto-second) cutting and MEMS process were executed as sensor fabrication methods with consideration of brittleness of the substrate and small radial artery size. After whole fabrication processes, we got sensors of $3mm{\times}15mm{\times}0.5mm$. Resonant frequency of the sensor was around 90 MHz at atmosphere (760 mmHg), and the sensor has good linearity without any hysteresis. Longterm (5 years) stability of the sensor was verified by thermal acceleration testing with Arrhenius model. Moreover, in-vitro cytotoxicity test was done to show biocompatiblity of the sensor and validation of real-time blood pressure measurement was verified with animal test by implant of the sensor. By integration with development of external interrogation system, the proposed sensor system will be a promising method to measure real-time blood pressure.

Response of Odontoblast to the Bio-Calcium Phosphate Cement

  • Kim, Jin-Woo;Kim, Sung-Won;Kim, Gyoo-Cheon;Kim, Yong-Deok;Kim, Cheol-Hun;Kim, Bok-Joo;Kim, Uk-Kyu
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.33 no.4
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    • pp.301-307
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    • 2011
  • Purpose: If the tooth structure is damaged, then it is impossible to regenerate the tooth. The materials used to restore the tooth structure are not related to the composition of the tooth. The materials used to restore the structure can't replace the natural tooth because they just fill the defective structure. Calcium phosphate cement remineralizes the dentin and almost replaces the natural tooth, but there are some disadvantages. We conducted basic tests with Biomimetic CPC (Bio-CPC) to make sure of the possibility of the biomaterial to remineralize the defective tooth structure. Methods: In this study, the bioactivity and biocompatibility of Bio-CPC were evaluated for its potential value as the bio-material for regeneration of damaged tooth structure by conducting a cell toxicity assay (WST-1 assay), a cytokinesis-block micronucleus assay, a chromosomal aberration test, total RNA extraction and RT-PCR on MDPC-23 mouse odontoblast-like cells. Results: The in vitro cytotoxicity test showed that the Bio-CPC was fairly cytocompatible for the MDPC-23 mouse odontoblast-like cells. Conclusion: Bio-CPC has a possibility to be a new biomaterial and further study of Bio-CPC is needed.

In vitro Studies on the Superoxide Scavenging Activities, the Cytotoxic and the Immunomodulating Effects of Thirteen Kinds of Herbal Extracts (13종 허브추출물의 수퍼옥사이드 소거능과 세포독성 및 면역증강 효과의 in vitro계 검색)

  • Chung, Ha-Yull;Kim, Hyun-Bae
    • Korean Journal of Food Science and Technology
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    • v.32 no.3
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    • pp.699-705
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    • 2000
  • The superoxide scavenging activities of thirteen kinds of herbal extracts were examined together with their cytotoxic and immunomodulating effects. The extracts of 6 kinds of herbs such as eucalyptus, mate, peppermint, sage, thyme and yarrow, so called medicinal herbs, showed above 70% of the superoxide scavenging activities. They also showed cytotoxicities against the cancer cell lines of Hepa-1c1c7 and KB-3-1 as well as the normal fibroblast cell line of mouse. The $IC_{50}$ values of above 6 herb extracts on the cancer cell lines were above or similar to the $IC_{50}$ values on the normal cell line, so it was unable to observe the herb extract which showed cytotoxicity only to the cancer cell lines. Considering the results of nitric oxide test that the sage extract was the only one having the immunomodulating effect(37% of positive control), there was no significant relationship between the superoxide scavenging activities of herbs and their immunomodulating effects.

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Studies on Genetic Toxicity of Epoxidized Soy Bean Oil (에폭시화 대두유의 유전독성 연구)

  • 한의식;정해관;김종원;박미선;엄미옥;강혁준;민수진;오혜영
    • Journal of Food Hygiene and Safety
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    • v.16 no.2
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    • pp.145-151
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    • 2001
  • EpoxidiBed soy bean oil (ESBO) is a plasticizer of PVC which is being widely used as a gaskets for the lid of glass jars including baby food. Using reverse mutation assay, chromosome aberration test and micronucleus test, ESBO were evaluated the mutagenicity. In the reverse mutation test, ESBO did not induced mutagenicity in Salmonella typhimurium TA98, TA100, TA1535, TA1537, TA102 with and without metabolic activation. In the chromosome aberration test using CHL cells, the results showed no increased structural and numerical aberrations in the concentration of sample producing cytotoxicity with and without metabolic activation. The in vivo induction of micronuclei was measured in polychromatic erythrocytes of bone marrow of young (3weeks old) and adult (6 weeks old) ddY mice of both sex. At 24 hours after treatment with ESBO 20, 10, 5, 2.5 g/B.W. kg/corn oil 10 ml by oral route animals were sacrificed and bone marrow cells were prepared for smear slides. The results showed no increased micronucleated polychromatic erythrocytes regardless of sex and age. It was concluded that water soluble ESBO did not show certain genotoxicity within our studies conducted.

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