• Title/Summary/Keyword: In vitro cytotoxicity

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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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Antitumor Effect of an Adenoviral Cytosine Deaminase/Thymidine Kinase Fusion Gene in C6 Glioma Cells (아데노 바이러스 Cytosine Deaminase/Thymidine Kinase 융합 유전자의 항 종양효과)

  • Kim, Young Woo;Choi, Jae Young;Chang, Jin Woo;Park, Yong Gou;Chung, Sang Sup
    • Journal of Korean Neurosurgical Society
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    • v.30 no.sup1
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    • pp.13-19
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    • 2001
  • Objective : We investigated the feasibility of a double suicide gene/prodrug therapy, involving direct introduction of the herpes simplex virus Type 1 thymidine kinase(TK) gene and the Escherichia coli cytosine deaminase(CD) gene, via a recombinant adenoviral vector and ganciclovir(GCV) and/or 5-fluorocytosine(5-FC) treatment, in C6 glioma cells. Methods : Efficient gene transfer and transduction of C6 glioma cells via a recombinant adenovirus were evaluated by infecting cells with adenovirus bearing the ${\beta}$-galactosidase gene and then staining cells with 5-bromo-4-chloro-3-indolyl-13-D-galactoside. CD/TK expression in cells infected with adenovirus bearing the CD/TK gene(ad-CD/TK) was examined by immunoblotting analysis. For in vitro cytotoxicity experiments, the cells were infected with ad-CD/TK or ad-${\Delta}E1$(as a control). After addition of a variety of concentrations of GCV and 5-FU, either separately or in combination, cell viability was determined by staining the cells with crystal violet solution 6 days after infection. Result : C6 glioma cells were efficiently transduced with recombinant adenoviral vector at multiplicities of infection of 200 or more. In vitro cytotoxicity of GCV and/or 5-FC, either alone or in combination, was exclusively observed in the cells transduced with ad-CD/TK. Obvious cytotoxicity(>50% inhibition) was observed in the presence of 5-FC at concentrations greater than 30ug/ml or GCV at concentrations greater than 0.3ug/ml at a multiplicity of infection of 100. Additionally, cytotoxicity in the presence of both GCV and 5-FC was greater than that after sinlge-prodrug treatments, indicating additive effects of the prodrug treatments. Conclusion : The administration of a double-suicide gene/prodrug therapy might have great potential in the treatment of brain tumors.

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Cytopathic Change and Inflammatory Response of Human Corneal Epithelial Cells Induced by Acanthamoeba castellanii Trophozoites and Cysts

  • Sohn, Hae-Jin;Seo, Ga-Eun;Lee, Jae-Ho;Ham, A-Jeong;Oh, Young-Hwan;Kang, Heekyoung;Shin, Ho-Joon
    • Parasites, Hosts and Diseases
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    • v.57 no.3
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    • pp.217-223
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    • 2019
  • Acanthamoeba castellanii has ubiquitous distribution and causes primary acanthamoebic keratitis (AK). AK is a common disease in contact lens wearers and results in permanent visual impairment or blindness. In this study, we observed the cytopathic effect, in vitro cytotoxicity, and secretion pattern of cytokines in human corneal epithelial cells (HCECs) induced by A. castellanii trophozoites and/or cysts. Morphological observation revealed that panked dendritic HCECs co-cultured with amoeba cysts had changed into round shape and gradually died. Such changes were more severe in co-culture with cyst than those of co-cultivation with trophozoites. In vitro cytotoxicity assay revealed the highest cytotoxicity to HCECs in the co-culture system with amoeba cysts. A. castellanii induced the expression of $IL-1{\alpha}$, IL-6, IL-8, and CXCL1 in HCECs. Secreted levels of $IL-1{\alpha}$, IL-6, and IL-8 in HCECs co-cultured with both trophozoites and cysts were increased at an early incubation time (3 and 6 hr). These results suggested that cytopathic changes and pro-inflammatory cytokines release of HCECs in response to A. castellanii, especially amoebic cysts, are an important mechanism for AK development.

CHANGE OF THE INVASIVENESS WITH SELECTIVE COX-2 INHIBITION IN AN ORAL SQUAMOUS CELL CARCINOMA CELL LINE, KB ; PRELIMINARY IN VITRO STUDY (선택적 COX-2 저해를 통한 구강암세포주 KB의 침습성 변화에 관한 예비연구)

  • Lee, Eun-Jin;Kim, Myung-Jin;Myoung, Hoon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.2
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    • pp.103-108
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    • 2007
  • Cyclooxygenase-2 (Cox-2) is known as one of the critical factor in carcinomas of various organs. However, the importance of Cox-2 in oral squamous cell carcinoma has not been fully described yet. The purpose of this study is to evaluate the anti-cancer effect of selective cox-2 inhibitor, celecoxib in an oral squamous cell carcinoma cell line, KB with respect to cytotoxicity test, in vitro invasion and MMP-2 expression. In cytotoxicity test, celecoxib treated group showed definitely concentration dependent cytotoxicity. In addition, administration of celecoxib reduced the invasive potential of KB cell line significantly in invasion assay. However, there was no remarkable difference of the MMP-2 expression between the celecoxib treated group and the control group. Considering these data, celecoxib had a potential cytotoxic agent to oral squamous cell carcinoma cells. Also, it had anti-invasive property without acting on the MMP-2 expression mechanism. Therefore, it was postulated that celecoxib had the possibility of anti-cancer agent in treatment strategies of oral squamous cell carcinoma.

Effect of Kaempferol on the Cytotoxicity Induced Oxygen Free Radicals in Skin Fibroblast Derived from Human In Vitro

  • Lee, Jai-Kyoo;Ha, Dae-Ho
    • Biomedical Science Letters
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    • v.14 no.3
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    • pp.193-198
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    • 2008
  • In order to evaluate on the effect of kaempferol on the cytotoxicity of oxygen tree radicals, XTT assay was performed to determine the cell viability after skin fibroblasts derived from human (Detroit 51) that were treated with various concentrations of hydrogen peroxide $(H_2O_2)$. And also, the effect of kaempferol on the cytotoxicity induced by H202 that was examined by cell viability, lactate dehydrogenase (LDH) activity and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in these cultures. $H_2O_2$ decreased cell viability in dose-dependent manner in these cultures and the $XTT_{90}\;and\;XTT_{50}$ values were determined at concentration of $35{\mu}M\;and\;90{\mu}M$ of $H_2O_2$ after skin fibroblasts derived from human were treated with $15{\sim}90{\mu}M$ of $H_2O_2$ for 6 hours, respectively. $H_2O_2$ was highly toxic on cultured skin fibroblasts derived from human by toxic criteria of Brenfreund and Puerner (1984). In the protective effect of kaempferol on $H_2O_2$-induced cytotoxicity, kaempferol increased DPPH radical scavenging activity and significantly decreased LDH activity. From these results, it is suggested that oxygen tree radical, $H_2O_2$, was highly toxic on cultured skin fibroblasts derived from human, and also kaempferol of flavonoid showed the protection on $H_2O_2$-induced cytotoxicity.

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In Vitro Antineoplastic Effects of Chitosan Hydrolysates on Various Tumor Cell Lines (키토산 가수분해물의 In Vitro 항종양성)

  • Park, Heon-Kuk
    • The Korean Journal of Food And Nutrition
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    • v.22 no.4
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    • pp.639-643
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    • 2009
  • In this study, the antineoplastic effects of chitosan hydrolysates were assessed. The chitosan hydrolysates showed no cytotoxicity in in vitro trials using the normal cell line, Vero E6(Africa green monkey kidney cells). The $IC_{50}$ value of the chitosan hydrolysates on Vero E6 was 1,107.95 ${\mu}g/m{\ell}$. The hydrolysates exhibited in vitro antineoplastic activity in five human tumor (lung carcinoma, bladder carcinoma, colon carcinoma, stomach carcinoma, breast carcinoma) cell lines. The $IC_{50}$ values of the hydrolysates on A549, J82, SNU-C4, SNU-1, and ZR75-1 cells were 421.06, 417.99, 445.54, 380.65 and 460.49 ${\mu}g/m{\ell}$, respectively.

A comparative evaluation of cytotoxicity of root canal sealers: an in vitro study

  • Badole, Gautam Pyarelal;Warhadpande, Manjusha Madhukar;Meshram, Ganesh Kothiramji;Bahadure, Rakesh Namdeoraoji;Tawani, Shubha Gopal;Tawani, Gopal;Badole, Shital Gautam
    • Restorative Dentistry and Endodontics
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    • v.38 no.4
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    • pp.204-209
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    • 2013
  • Objectives: The objective of this in vitro study was to evaluate and compare the cytotoxicity of four different root canal sealers i.e. Apexit Plus (Ivoclar Vivadent), Endomethasone N (Septodont), AH-26 (Dentsply) and Pulpdent Root Canal Sealer (Pulpdent), on a mouse fibroblast cell line (L929). Materials and Methods: Thirty two discs for each sealer (5 mm in diameter and 2 mm in height) were fabricated in Teflon mould. The sealer extraction was made in cell culture medium (Dulbecco's Modified Eagle's Medium, DMEM) using the ratio 1.25 $cm^2/mL$ between the surface of the sealer samples and the volume of medium in a shaker incubator. Extraction of each sealer was obtained at 24 hr, 7th day, 14th day, and one month of interval. These extracts were incubated with L929 cell line and 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay was done. Two-way ANOVA for interaction effects between sealer and time and Post-hoc multiple comparison using Tukey's test across all the 16 different groups were used for statistical analysis. Results: Apexit Plus root canal sealer was significantly less toxic than other sealers (p < 0.05) and showed higher cellular growth than control. Endomethasone N showed mild cytotoxicity. AH-26 showed severe toxicity which became mild after one month while Pulpdent Root Canal Sealer showed severe to moderate toxicity. Conclusions: Apexit Plus was relatively biocompatible sealer as compared to other three sealers which were cytotoxic at their initial stages, however, they became biocompatible with time.

Surface Immobilization of Amphiphilic Comb-like Polymer on Polydimethylsiloxane and in vitro Cytotoxicity Assay (양친성 빗 모양 고분자의 PDMS 표면 고정화 및 세포독성 평가)

  • Choi, Jaeyoo;Jung, Jaeyeon;Cheng, Jie;Lee, Jonghwan;Hyun, Jinho;Kim, Hyunjoong
    • Journal of Adhesion and Interface
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    • v.11 no.2
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    • pp.70-75
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    • 2010
  • It described the modification of polydimethylsiloxane (PDMS) with amphiphilic methyl methacrylate-based polyethylene glycol (PMMA-b-PEG) to enhance the hydrophilicity of a PDMS surface and cytotoxicity of it. PMMA-b-PEG solutions in water/ethanol mixture was spun-cast on the PDMS surface and the surface was characterized by long-term measurement of water contact angle. The morphology of PDMS surfaces coated with PMMA-b-PEG was characterized by field emission scanning electron microscopy and atomic force microscope. Cytotoxicity of the modified surfaces was investigated by MTT assay which would be necessary for the evaluation of tissue compatibility after implantation of the materials. Based on the MTT assay, PDMS coated with PMMA-b-PEG didn't show any significant cytotoxcity.

Synthesis and In Vitro Cytotoxicity of Cinnamaldehydes to Hyman Solid Tumor Cells

  • Kwon, Byoung-Mog;Lee, Seung-Ho;Choi, Sang-Un;Park, Sung-Hee;Lee, Chong-Ock;Cho, Young-Kwon;Sung, Nack-Do;Bok, Song-Hae
    • Archives of Pharmacal Research
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    • v.21 no.2
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    • pp.147-152
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    • 1998
  • Cinnamaldehydes and related compounds were synthesized from various cinnamic acids based on the $2^{I}$-hydroxycinnamaidehyde isolated from the bark of Cinnamomum cassia Blume. The cytotoxicity to human solid tumor cells such as A549, SK-OV-3, SK-MEL-2, XF498 and HCT15 were measured. Cinnamic acid, cinnamates and cinnamyl alcohols did not show any cytotoxicity against the human tumor cells. Cinnamaldehydes and realted compounds were resistant to A549 cell line up to 15 .mu.g/ml. In contrast, HCT15 and SK-MEL-2 cells were much sensitive to these cinnamaidehyde analogues which showed $ED{50} values 0.63-8.1{\mu}g/ml.$Cytotoxicity of the saturated aldehydes was much weak compared to their unsaturated aldehydes. From these studies, it was found that the key functional group of the cinnamaldehyde-related compounds in the antitumor activity is the propenal group.p.

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Synthesis and in vitro Antitumor Activity of lsoazamitosene and lsoiminoazamitosene Derivatives

  • Ahn, Chan-Mug;Kim, Soo-Kie
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.535-542
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    • 1996
  • Seven isoazamitosene derivatives, mitomycin analogues, were synthesized and tested for cytotoxicities against leukemia and gastric cancer cell lines. Preparation of a pyrrolo[1, 2-a]benzimidazole (3) (azamitosene ring system) was completed by utilizing the Lewis acid-catalized cyclization, with .omicron.-chloronitrotoluene as the starting material. Nitration of 3 produced a mixtue of two isomers (5-nitro isomer (4) and 7-nitro isomer (5)) in product ratio of 36 : 52. 4 was directly converted into quinone (7) by reduction and Fremy oxidaton. Finally, quinone derivatives (8, 9, 10, and 11) were synthesized by 1, 4-addition of 7 with cyclic secondary amines. From above-mentioned 5, 8-nitro compound (15) was prepared in 4 steps. At pH 3, Fremy oxidation of 15 produced quinone (16), whereas iminoquinone derivatives (17a and 17b) at pH 7. Isoazamitosene derivatives (8, 9, 10, and 11), containing cyclic amino groups at the 7-position, showed potent cytotoxicity on P388, SNU-1, and KHH tumor cell lines. Among them, 8 had stronger cytotoxicity against SNU-1 cell line than mitomycin and adriamycin. Considering these results, isoazamitosene derivatives may had unique cytotoxicity profiles. However, isoiminoazamitosene derivatives (17a and 17b) revealed very weak cytotoxicity.

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