• Title/Summary/Keyword: in vitro cytotoxicity test

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A STUDY OF THE CYTOTOXICITY OF ROOT CANAL SEALER IN VITRO (생체외 실험을 이용한 근관충전용 Sealer의 세포독성에 관한 연구)

  • Lee, Sang-Tag;Lee, Chung-Sik
    • Restorative Dentistry and Endodontics
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    • v.16 no.2
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    • pp.62-84
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    • 1991
  • The purpose of this study was to evaluate the cytotoxicity of four root canal sealers(Tubliseal, AH26, Apatite Root Canal Sealer I, Apatite Root Canal Sealer II) in Vitro. The root canal sealers were mixed and filled in molds which were $14{\times}1.25mm$ in diameter, in height to use for cell counting and agar overlary method, and $7{\times}1.25mm$ for millipore filter method and set for 7 days to use for experiment. Silicone and copper plate were used for negative and positive control respectively. Using the culture of L929 fibroblast, total cell number and vital cell number were counted and the ratio of vital cell number to total cell number was calculated on 2 nd, 4 th, 6 th experimental day, and the change of cell membrane permeability was tested by agar overlay method, and the succinate dehydrogenase activity was tested by millipore filter method. The obtained results were as follows. 1. In ail experimental groups, the mitotic activity of fibroblast was reduced when compared with that of negative control group, so ail experimental groups showed cytotoxicity. Apatite Root Canal Sealer I group exhibited mild cytotoxicity, and Tubliseal, AH26, Apatite Root Cenal Sealer II groups exhibited severe cytotoxicity. 2. In the test of the change of cell membrane permeability by agar overlay method, all experimental groups showed cytotoxicity. AH26 group exhibited mild cytotoxicity, and Apatite Root Canal Sealer I group exhibited moderate cytotoxicity, and Tubliseal and Apatite Root Canal Sealer II group exhibited severe cytotoxicity. 3. In the test of SDH activity by millipore filter method, there was no cytotoxicity in Apatite Root Canal Sealer I and Apatite Root Canal Sealer II group, but Tubliseal and AH26 group showed mild cytotoxicity.

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Effect of aging on tear strength and cytotoxicity of soft denture lining materials; in vitro

  • Landayan, Jordi Izzard Andaya;Manaloto, Adrian Carlos Francisco;Lee, Jeong-Yol;Shin, Sang-Wan
    • The Journal of Advanced Prosthodontics
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    • v.6 no.2
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    • pp.115-120
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    • 2014
  • PURPOSE. The aim of this in vitro study was to evaluate the effect of aging on the tear strength and cytotoxicity of four soft denture lining materials. MATERIALS AND METHODS. Four commonly used soft denture lining materials, (Coe-Comfort$^{TM}$ GC America Inc., Alsip, IL, USA; Coe-SOFT$^{TM}$ GC America Inc., Alsip, IL, USA; Visco-gel Dentsply Caulk Milford, DE, USA; and Sofreliner Tough M Tokuyama Dental Corporation Tokyo, Japan) were selected. Sixty trouser-leg designed specimens per lining material were fabricated using a stainless steel mold for tear strength testing. The specimens were divided into non-thermocycling and 1000-, and 3000-thermocycling groups. For the cytotoxicity test, twenty-four disk shaped specimens per material were fabricated using a stainless steel mold. The specimens were soaked in normal saline solution for 24 h, 48 h and 72 h. Cytotoxicity was measured by XTT assay in L929 mouse fibroblasts. Data were analyzed by two way analysis of variance and Dunnett's test (P<.05). RESULTS. Before thermocycling, Sofreliner Tough M ($10.36{\pm}1.00N$) had the highest tear strength value while Coe-Comfort$^{TM}$ ($0.46{\pm}0.10N$) had the lowest. After 3000 cycles, Sofreliner Tough M ($9.65{\pm}1.66N$) presented the highest value and Coe-Comfort$^{TM}$ ($0.42{\pm}0.08N$) the lowest. Sofreliner Tough M, in all incubation periods was the least toxic with significant differences compared to all other materials (P<.05). Coe-Comfort$^{TM}$, Coe-$SOFT^{TM}$, and Sofreliner Tough M did not show any significant differences within their material group for all incubation periods. CONCLUSION. This in vitro study revealed that aging can affect both the tear strength and cytotoxicity of soft denture materials depending on the composition.

Synthesis of New Semisynthetic Analogs of Epi-xanthatin by Modification of the Side Chain and Their Cytotoxic Activity (Epi-xanthatin의 Side Chain 변환을 통한 새로운 반합성 유도체들의 합성 및 세포독성)

  • Baek Du-Jong;Ahn Jong-Woong;Lee Chongock
    • YAKHAK HOEJI
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    • v.49 no.1
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    • pp.68-73
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    • 2005
  • Epi-xanthatin analogs containing hydrophilic substituents such as carboxylic acid, alcohol, morpholine, amino acid, and glucose derivatives were synthesized and their in vitro cytotoxicity and in vivo antitumor activity were evaluated. The target compounds were generally cytotoxic against tumor cell lines of human origin with $ED_{50}$ values of $0.1{\sim}30{\mu}g/ml$, except the highly hydrophilic analog 6 containing aspartic acid. Contrary to the potent cytotoxicity weakly hydrophilic analogs 2 and 8 were not active in vivo, or even toxic to the test animals. As a result, hydrophilic analog of epi-xanthatin did not show in vitro cytotoxicity and hydrophobic analogs did not show in vivo antitumor activity, thus it is presumed that amphiphilic analogs or those with medium hydrophilicity would exhibit the antitumor potency in vivo.

In-vitro Antimicrobial Activity Phytochemical and Cytotoxicity of Methanolic Fruits Extract of Capsicum frutescent

  • Elbashir, Habiballah A.;Mubarak, Elnaeim E.;Kabbashi, Ahmed S.;Garbi, Mohamed I.;Elshikh, Ahmed A.
    • The Korean Journal of Food & Health Convergence
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    • v.4 no.4
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    • pp.10-17
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    • 2018
  • Capsicum frutescen is known in Sudan, is one of the most commonly used pepper species in cooking and in Sudanese folk medicine. The present study was conducted to investigate antimicrobial (bacteria and fungi) and cytotoxicity (Brine Shrimp Lethality Test) of methanolic extract of Capsicum frutescen (fruits). The extract have been tested in the present study to investigate the in vitro potential effects against Gram positive, Gram negative bacteria and fungi. The selected organisms were Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia and Candida albicans using the cup plate agar diffusion method. The methanol extract of Capsicum frutescen (fruits) exhibited inhibitory effects against Escherichia coli with zone of inhibition (23 mm) and Klebsiella pneumonia with zone of inhibition (17 mm). The phytochemical screening revealed the presence of Tannins, Saponin, Alkaloids, Anthroquinoles and Terpenoids. The Cytotoxicity of methanolic extract of Capsicum frutescens was $LD_{50}$ $64.68{\mu}g/ml$. The activity and presence of compounds known to be biologically active are a validation for the use of Capsicum as a food ingredient and as a therapeutic element of traditional medicine.

Toxicological Evaluation of Chitosan Cross-linked Collagen-GAG Matrix (CCGM) In vitro and In vivo (Chitosan Cross-linked Collagen-GAG Matrix(CCGM)의 독성학적 고찰)

  • Lee, Hae-Yul;Kim, Dong-Hwan;Cho, Hyun;Ahn, Byoung-Ok;Kang, Soo-Hyung;Kim, Won-Bae
    • Toxicological Research
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    • v.16 no.1
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    • pp.17-25
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    • 2000
  • Chitosan cross-linked collagen-glycosaminoglyan (CCGM) is an artificial skin substitute made to form a sponge like dimensional matrix. It can be used to facilitate reconstruction of dermal tissue when applied on large wounds such as severe burns. In order to study the toxicological effects of CCGM the cytotoxicity, local irritation and skin sensitization test were carried out according to the standards of ISO 10993. In the cytotoxicity test utilizing LDH and MTT test, both the CCGM and its extract had no toxicity of Balb/c 3T3 cells. The local irritatioin test on rabbit skin demonstrated that CCGM did not promote any harmful when directly applied on skin. In addition, it did not elicit any allergic reaction in the guinea pig maximization test. Based on these results, it is suggested that CCGM is a material without cytotoxicity, local irritation and allergenicity.

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Alternatives to In Vivo Draize Rabbit Eye and Skin Irritation Tests with a Focus on 3D Reconstructed Human Cornea-Like Epithelium and Epidermis Models

  • Lee, Miri;Hwang, Jee-Hyun;Lim, Kyung-Min
    • Toxicological Research
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    • v.33 no.3
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    • pp.191-203
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    • 2017
  • Human eyes and skin are frequently exposed to chemicals accidentally or on purpose due to their external location. Therefore, chemicals are required to undergo the evaluation of the ocular and dermal irritancy for their safe handling and use before release into the market. Draize rabbit eye and skin irritation test developed in 1944, has been a gold standard test which was enlisted as OECD TG 404 and OECD TG 405 but it has been criticized with respect to animal welfare due to invasive and cruel procedure. To replace it, diverse alternatives have been developed: (i) For Draize eye irritation test, organotypic assay, in vitro cytotoxicity-based method, in chemico tests, in silico prediction model, and 3D reconstructed human cornealike epithelium (RhCE); (ii) For Draize skin irritation test, in vitro cytotoxicity-based cell model, and 3D reconstructed human epidermis models (RhE). Of these, RhCE and RhE models are getting spotlight as a promising alternative with a wide applicability domain covering cosmetics and personal care products. In this review, we overviewed the current alternatives to Draize test with a focus on 3D human epithelium models to provide an insight into advancing and widening their utility.

In Vitro Cytotoxicity of Pt(II) Complexes Containing Ethylenediamine in Rabbit Kidney Proximal Tubular and Human Renal Cortical Cells (에틸렌디아민을 배위자로 한 백금(II)착체의 토끼 및 인체 신장세포에 대한 in vitro 독성)

  • Rho, Young-Soo;Lee, Kyung-Tae;Jung, Jee-Chang;Chang, Sung-Goo
    • YAKHAK HOEJI
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    • v.40 no.2
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    • pp.218-224
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    • 1996
  • This laboratory has recently reported the synthesis and in vitro antitumor activity of PT(II) complexes containing ethylenediamine and diphosphine. In view of the reports of others, cisplatin is toxic to the kidney since the kidney's vulnerability to PT(II) complexes may originate in its ability to accumulate and retain platinum to a greater degree than other organs. The in vitro cytotoxicity of these synthetic PT(II) complexes on the primary cultured proximal tubular cells of rabbit kidney and renal cortical cells of human kidney was investigated. Three endpoints for cytotoxicity tests were evaluated:3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), $^3H$-thymidine uptake and the glucose consumption tests. The rank order of sensitivity exhibited $^3H$-thymidine uptake>MTT>glucose consumption test. The agents with diphosphine leaving group were significantly less cytotoxic than cisplatin. Moreover, 1,2-bis(diphenylphosphino)ethane (DPPE) exhibited less cytotoxicity than 1.3-bis (diphenylphosphino)propane (DPPP) against on rabbit and human cultured kidney cells. Based on these results, the decreased nephrotoxicity of these new complexes over cisplatin appeared to be partially attributable to a leaving group of DPPP and DPPE. This novel class of platinum compound represents a valuable lead in the development of a "third-generation" agent.

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Characterization and Preparation of Low Molecular Weight Water Soluble Chitosan Nanoparticle Modified with Cell Targeting Ligand for Efficient Gene Delivery (효과적인 유전자전달을 위한 표적성 리간드가 도입된 저분자량 수용성 키토산 나노입자의 제조 및 특성)

  • Heo, Sun-Heang;Jang, Min-Ja;Kim, Dong-Gon;Jeong, Young-Il;Jang, Mi-Kyeong;Nah, Jae-Woon
    • Polymer(Korea)
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    • v.31 no.5
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    • pp.454-459
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    • 2007
  • Gene therapy using low molecular weight water soluble chitosan (LMWSC) as polycationic polymer shows good biocompatibility, but low transfection efficiency. The mechanism of folic acid (FA) uptake in the cells to promote targeting and internalization could improve transfection rates. The objective of this study was to synthesize and characterize the WSCFA-DNA complex and evaluate their cytotoxicity, in vitro. In $^1H-NMR$ spectra, specific peaks appeared both of FA and LMWSC in $D_2O$. WSCFA nanoparticles have spherical shapes with particle size show below 110 nm. In the cell cytotoxicity test, the WSCFA-DNA complex showed high cell viability, in vitro. Gel electrophoresis showed condensed DNA within the carriers. hi vitro transfection efficiency was assayed by fluorescence spectroscopy WSCFA nanoparticles have less cytotoxicity, good DNA condensation and particle size around 110 nm, which makes them a promising candidate as a non-viral gene vector.

CYTOTOXICITY OF RETROGRADE FILLING MATERIALS TESTED BY 51Cr RELEASE, MIT AND LD ACTIVITY (51Cr방출과 MTT 및 LD활성도를 이용한 역충전재의 세포독성에 관한 연구)

  • Choi, La-Young;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.19 no.2
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    • pp.409-428
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    • 1994
  • Endodontic surgery is performed when conventional endodontic therapy fails or is contraindicated. In such cases, retrograde filling materials including amalgam, composite resin, and various cements have been used. Biocompatibilty and margin sealing ability of retrograde filling materials are important for the long term success of endodontic surgery. In vitro cell culture is frequently used as the method of measuring the biocompatibilty of dental materials. The purpose of this study was to evaluate the cytotoxicity of six kinds of retrograde filling materials including newly developed light curing glass ionomer cements. Each material was mixed according to. the manufacture's instruction and evaluated as : freshly mixed, 24-hour after mixing, and 168-hour after mixing respectively. The elution solution was extracted after 24-hour contact with materials using media. Cytotoxicity was evaluated by direct contact, or elution contact. Test results of radiochromium($^{51}Cr$) release, cell viability using tetrazolium dye (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl dimethyltetrazolium bromide(MTT) test and lactate dehydrogenase(LD) of damaged L929 cells were analyzed. In the $^{51}Cr$ release of direct contact, all experimental retrograde filling materials except amalgam and glass ionomer cement showed increased cytotoxicity compared to control. In the $^{51}Cr$ release of elution solution, the released $^{51}Cr$ was so minimal that it was impossible. to evlauate the cytotoxicity exactly. The elution solutions of glass ionomer cement and IRM showed marked cytotoxicity in MTT test. LD enzyme activity was highest in tests of direct contact with composite, light curing composite, and light curing glass ionomer cement and IRM. Amalgam revealed least cytotoxicity while IRM showed cytotoxicity using all three methods. Composite, light curing composite and light curing glass iomomer cement were cytotoxic in the tests of $^{51}Cr$ release and LD activity. Glass ionomer cement showed cytotoxic effect only in the MTT method. From these results it is suggested that the standardization and optimization of cytotoxicity testing, especially using elution solutions, should be strongly advised.

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In Vitro Skin Irritation Test of Anti-Inflammatory Drugs (소염진통제 약물에 대한 In vitro 피부자극 시험연구)

  • Lee, Jong-Kwon;Kim, Dai-Byung;Lee, Eun-Hee;Lee, Sun-Hee;Ryu, Seung-Rel;Choi, Ki-Hwan;Kim, Yoon-Jeong;Kim, Pu-Young
    • Toxicological Research
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    • v.14 no.3
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    • pp.315-320
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    • 1998
  • In vitro skin iritation of anti-inflammatory drugs was investigated in terms of the cytotoxicity method to human skin fibroblast cells. Five anti-inflammatory drugs (Diclofenac, Naproxen, Meclofenamic acid, Ibuprofen and Fnoprofen) which are commercially available as oral preparations or injections were tested. The cytotoxicity of 5 chemicals was evaluated by using MTT[tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. NRU (neutral red uptake) assay and Alamar Blue assay after fibroblast cells had been exposed to the chemicals for 24 hours or 489 hours. The $IC_{50}$ values of the chemicals showed the comparative strength of cytotoxicity as following order of Meclofenamic acid>Diclofenac>Fenoprofen>Ibuprofen>Naproxen. The values of $IC_{50}$ determined by Alamar Blue assay were lower than those of MTT and NRU assay. These data suggest Alamar Blue assay can be useful method for assessing in vitro skin irritation potential of anti-inflammatory drugs.

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