• Title/Summary/Keyword: MTT analysis

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DNA microarray analysis of gene expression of MC3T3-E1 osteoblast cell cultured on anodized- or machined titanium surface

  • Park, Ju-Mi;Jeon, Hye-Ran;Pang, Eun-Kyoung;Kim, Myung-Rae;Kang, Na-Ra
    • Journal of Periodontal and Implant Science
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    • v.38 no.sup2
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    • pp.299-308
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    • 2008
  • Purpose: The aim of this study was to evaluate adhesion and gene expression of the MC3T3-E1 cells cultured on machined titanium surface (MS) and anodized titanium surface (AS) using MTT test, Scanning electron micrograph and cDNA microarray. Materials and Methods: The MTT test assay was used for examining the proliferation of MC3T3-E1 cells, osteoblast like cells from Rat calvaria, on MS and AS for 24 hours and 48 hours. Cell cultures were incubated for 24 hours to evaluate the influence of the substrate geometry on both surfaces using a Scanning Electron Micrograph (SEM). The cDNA microarray Agilent Rat 22K chip was used to monitor expressions of genes. Results: After 24 hours of adhesion, the cell density on AS was higher than MS (p < 0.05). After 48 hours the cell density on both titanium surfaces were similar (p > 0.05). AS had the irregular, rough and porous surface texture. After 48 hours incubation of the MC3T3-E1 cells, connective tissue growth factor (CTGF) was up-regulated on AS than MS (more than 2 fold) and the insulin-like growth factor 1 receptor was down-regulated (more than 2 fold) on AS than MS. Conclusion: Microarray assay at 48 hours after culturing the cells on both surfaces revealed that osteoinductive molecules appeared more prominent on AS, whereas the adhesion molecules on the biomaterial were higher on MS than AS, which will affect the phenotype of the plated cells depending on the surface morphology.

Antiproliferative Effect of Chungjogupae-tang Treatment was Associated with the Inhibition of Prostaglandin E2 Release in Human Lung Carcinoma Cells (인체폐암세포의 증식 및 prostaglandin E2 생성에 미치는 청조구폐탕의 영향에 관한 연구)

  • Im, Jae-Hyung;Kim, Hoon;Byun, Mi-Kyeon;Kam, Chul-Woo;Park, Dong-Il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.4
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    • pp.966-972
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    • 2006
  • The effect of water extract of Chungjogupae-tang (CJGPT) was investigated _on the growth of human lung carcinoma A549 cells. Methods: MTT assay and fluorescent microscope peformed to compare and examine the efficacy of CJGPT treatment on the cytostaticity of lung cancer cells in proportion to time and doses, and DAPI staining and Western blot analysis were used to examine their effect on apoptosis. In addition, the quantitative RT-PCR was used to examine to lung cancer cells growth, and Prostaglandin E2 activity were measured. Results: Exposure of A549 cells to CJGPT respited in the growth inhibition and apoptosis in a dose-dependent manner as measured by MTT assay and fluorescent microscope. The antiproliferative effect by CJGPT treatment in A549 cells was associated with morphological changes such as membrane shrinking and cell rounding up. CJGPT treatment resulted in an up-regulation of cyclin-dependent kinase inhibitor p21 (WAFl/CIPl) in a p53-independent fashion. We found that CJGPT treatment decreased the levels of cyclooxygenase (COX)-2 and inducible nitric oxide synthease (iNOS) expression without significant changes in the expression of COX-1 , which was correlated with a decrease in prostaglandin E2 (PGE2) synthesis. Conclusion: These findings suggested that CJGPT-induced inhibition of human lung carcinoma A549 cell growth was connected with the induction of apoptotic cell death and the results provided important new insights into the possible molecular mechanisms of the anti-cancer activity of CJGPT.

Preparation and Characterization of Tissue Engineered Scaffold Using Porcine Small Intestinal Submucosa and Hyaluronic Acid (돼지의 소장점막하 조직과 히알루론산을 이용한 조직공학적 담체의 제조 및 특성분석)

  • Lim, Ji-Ye;Kim, Soon-Hee;Kang, Gil-Son;Rhee, John M.
    • Polymer(Korea)
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    • v.32 no.5
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    • pp.415-420
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    • 2008
  • The porcine small intestinal submucosa (SIS) without immunorejection responses and hyalunonic acid (HA) can be used as biomaterials. In this study, we tried to design and characterize novel sponge. SIS- HA sponge was prepared by freeze-drying after addition 1wt% HA solution into fabricated SIS sponge. Sponge was crosslinked with 1-ethyl-(3-3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC) solution with 100mM concentration for 24 hrs and lyophilized. SIS-HA sponge was characterized by scanning electron microscopy and fourier transform infrared spectrometer. And water absorption ability of sponge was evaluated. We seeded NIH/3T3 cells in SIS-HA sponge and cellular attachment was assayed by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltertazolium-bromide (MTT) test. We demonstrated presence of HA in SIS-HA sponge from C-O functional group observed by the FT-IR analysis. Moreover, we confirmed low cytotoxicity and high cell viability of the SIS-HA sponges. Therefore, we could expect that SIS- HA scaffolds are applicable for the tissue regeneration.

Cytotoxic Effect of Bee (A. mellifera) Venom on Cancer Cell Lines

  • Borojeni, Sima Khalilifard;Zolfagharian, Hossein;Babaie, Mahdi;Javadi, Iraj
    • Journal of Pharmacopuncture
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    • v.23 no.4
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    • pp.212-219
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    • 2020
  • Objectives: Nowadays cancer treatment is an important challenge in the medical world that needs better therapies. Many active secretions produced by insects such as honey bees used to discover new anticancer drugs. Bee venom (BV) has a potent anti inflammatory, anti cancer and tumor effects. The aim of present study is evaluation of anticancer effects induced by Apis mellifera venom (AmV) on cell Lines. Methods: AmV was selected for study on cancer cell lines. Total protein, molecular weight and LD50 of crude venom were determined. Then, cells were grown in Dulbecco's Modified Eagle medium supplemented with 10% fetal bovine serum and 1% antibiotics. The A549, HeLa and MDA-MB-231 cell Lines were exposed by different concentration of AmV. The morphology of cells was determined and cell viability was studed by MTT assay. Evaluation of cell death was determined by and DNA fragmentation. Results: The results from MTT assay showed that 3.125 ㎍/mL of A549, 12.5 for HeLa and 6.25 ㎍/mL of MDA-MB-231 killed 50% of cells (p < 0.05). Morphological analysis and the results from hoescht staining and DNA fragmentation indicated that cell death induced by AmV was significantly apoptosis. Conclusion: The data showed that using lower dosage of AmV during treatment period cause inhibition of proliferation in time and dose dependant manner. Findings indicated that some ingredients of AmV have anticancer effects and with further investigation it can be used in production of anticancer drugs.

The Antioxidant Activities and Neuroprotective Effects of Hot Water Extracts from Torreyae Semen (비자 열수 추출물의 항산화 활성 및 뇌신경세포 보호효과 연구)

  • Lee, Soong-In;Choi, Chan-Hun;Kim, Jeong-Sang;Lim, Seong-Soo;Jung, Hyun-Woo
    • The Korea Journal of Herbology
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    • v.32 no.6
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    • pp.41-48
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    • 2017
  • Objectives : This study was designed to estimate the antioxidative and neuroprotective effects of Torreyae Semen hot water extracts (TS). Methods : Torreyae Semen was extracted by hot water for 2 hours with a temperature of 105 degrees. Polyphenols and total flavonoid were measured and LC-MS/MS was used to certificate anticipated antioxidative compounds. The antioxidant activities of TS were measured as scavenging effects of 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and Nitrite Oxides (NO). Cell viability and proliferation rate was measured MTT assay. The toxicities to thymocytes and splenocytes were evaluated by the proliferation rate of primary cultured cells of 7 weeks, male Balb/c mice. The antioxidant activities of TS on C6 mouse glioma cells were measured by the analysis of total glutathione contents variation. The neuroprotective effects against oxidative stresses were measured by MTT assay. Results : Polyphenols of TS was $92.00{\pm}1.24{\mu}g/mg$, and total flavonoids was $0.36{\pm}0.14{\mu}g/mg$. TS includes gallocatechin, epigallocatechin, gallocatechin gallate and epigallocatechin gallate. TS included gallocatechin, epigallocatechin, gallocatechin gallate, epigallocatechin gallate. TS showed DPPH and NO scavenging effects as dose-dependent manner at the concentrations of $0-10mg/m{\ell}$. In MTT assay, TS shows no significant toxicity to C6 cells, primary cultured thymocytes and splenocytes of Balb/c mice. TS increased the level of total glutathiones. TS increased cell viabilities of C6 cells against oxidative stresses such as $H_2O_2$, sodium nitroprusside (SNP), Rotenone at the concentrations of $0-0.063mg/m{\ell}$. Conclusions : TS shows the antioxidant and neuroprotecitive effects in these experiments.

Synthesis, characterization, and toxicity of multi-walled carbon nanotubes functionalized with 4-hydroxyquinazoline

  • Tahermansouri, Hasan;Mirosanloo, Atieh;Keshel, Saeed Heidari;Gardaneh, Mossa
    • Carbon letters
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    • v.17 no.1
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    • pp.45-52
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    • 2016
  • The attachment of 2-aminobenzamide to carboxylated multi-wall carbon nanotubes (MWCNTs)-COOH was achieved through the formation of amide bonds. Then, the functionalized MWCNTs, MWCNT-amide, were treated by phosphoryl chloride to produce MWCNT-quin. The products were characterized by Fourier transform infrared spectroscopy, Raman spectroscopy, scanning electron microscopy, thermogravimetric analysis, derivative thermogravimetric, steady-state fluorescence spectroscopy, and solubility testing. MWCNT-quin showed photo-electronic properties, which is due to the attachment of the 4-hydroxyquinazoline groups to them as proved by steady-state fluorescence spectroscopy. This suggests intramolecular interactions between the tubes and the attached 4-hydroxyquinazoline. The toxicity of the samples was evaluated in human embryonic kidney HEK293 and human breast cancer SKBR3 cell lines, and the viable cell numbers were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) after the cells were cultured for 24 h. Cellular investigations showed that the modified MWCNTs, particularly MWCNT-quin, have considerably significant toxic impact on SKBR3 as compared to HEK293 at the concentration of 5 µg/mL.

Selective Cytotoxicity of Novel Platinum(II) Coordination Complexes Containing DL-2-Hydroxy 3-Methylbutyric Acid (DL-2 하이드록시 3-메틸 부틸산물 배위자로 한 새로운 항암성 백금(II) 착체의 위암세포와 정상신장세포에 대한 선택적 세포독성)

  • 정지창;홍언표;최승기;장성구;육창수;노영수
    • Biomolecules & Therapeutics
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    • v.11 no.2
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    • pp.91-98
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    • 2003
  • A new series of highly water soluble platinum(II) complexes[Pt(II)(DL-2-hydroxy-3-methylbutyrate)(trans-l-1,2-dimninocyc1ohexane)] (PC-1) and [Pt(II)DL-2-hydroxy-3-methylbutyrate](cis-1,2-diaminocyclohexane)](PC-2) were synthesized and characterized by their elemental analysis and by various spectroscopic techniques [infrared(IR), $^{13}C$-nuclear magnetic resonance (NMR)]. In vitro antitumor activity of new Pt(II)complexes was tested against MKN-45, MKN/ADM and MKN/CDDP human gastric adenocarcinoma cell lines using colorimetric MTT[3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyltetrazoliumbromide] assay for cell survival and proliferation. PC-1 and PC-2 showed active against MKN-45/P, MKN/ADM and MKN/CDDP human gastric cancer cell lines, and the antitumor activity of these compounds were comparable or superior to that of cisplatin. The nephrotoxicities of PC-1 and PC-2 were found quite less then that of cisplatin using MTT and [$^3H$] thymidine uptake tests in rabbit proximal tubule cells, human kidney cortical cells human renal cortical tissues. Based on these results, these novel platinum(II) complex compounds(PC-1 & PC-2) represent a valuable lead in the development of the new anticancer chemotherapeutic agents capable of improving antitumor activity and low nephrotoxicity.

The Effects of Artemisia capillaries Herbal Acupuncture on Ethanol-induced Apoptosis in Neuroblastoma Cell Line (인진 약침액이 신경아세포주에서 에탄올에 의해 유발된 아폽토시스에 미치는 영향)

  • Ee-Hwa, Kim;Youn-Hee, Kim;Youn-Jung, Kim;Mi-Hyun, Jang;Joo-Ho, Chung;Chang-Ju, Kim
    • The Journal of Korean Medicine
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    • v.22 no.1
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    • pp.90-95
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    • 2001
  • 목적:인진 약침액이 SK-N-MC 신경아세포주에서 에탄올에 의해 유발된 아폽토시스에 미치는 영향을 조사하였다. 방법 : SK-N-MC cell line에서의 아폽토시스 변화를 관찰하기 위해서 MTT assay, DAPI staining 및 flow cytometric analysis 방법을 이용하였다. 결과: MTT assay를 이용하여 분석한 결과 농도에 따른 세포 독성의 효과가 에탄올 투여로부터 관찰되었다. 또한 인진 약침액으로 전처치하고 에탄올을 처치하였을 때 세포 독성이 크게 감소되었다. DAPI staining에서 인진 약침액 투여군은 에탄올 투여군에 비해서 fragmentation이 억제되었다. Flow cytometry를 통하여 인진 약침액 투여군은 에탄올 투여군에 비하여 세포주기 중 sub $G_1$ 분획의 증가가 억제되었다. 결론 : SK-N-MC 신경아세포주에서 에탄올에 의해서 유발된 아폽토시스는 전형적인 세포사별 형태를 나타내었다. 또한 인진 약침액은 에탄올에 의해서 유발된 아폽토시스에서 세포보호 효과가 있음이 확인되었다.

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Growth Inhibitory Effects of Omega-3 Unsaturated Fatty Acid against Cancer Cell Lines (Omega 3계열 불포화 지방산의 암세포주에 대한 성장 억제효과)

  • Han, Du-Seok;Choi, Hyoung-Gyu;Kang, Jeong-Il;Choi, Hwa-Jung;Baek, Seung-Hwa
    • YAKHAK HOEJI
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    • v.52 no.4
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    • pp.264-273
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    • 2008
  • The inhibitory effect of omega-3 such as linolenic acid (LNA), docosahexaenoic acid (DNA) and eicosapentaenoic acid(EPA) on the growth of normal cell lines and cancer cell lines was evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyItetrazolium bromide (MTT) and 2,3-bis-2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-caboxanilide (XTT) methods. LNA was found to decrease the cell viability of human oral epithelioid carcinoma cells (KB) in the MTT assay, whereas EPA appeared to inhibit the cell adhesion activity of human skin melanoma cells (SK-MEL-3) in the XTT assay analysis. DPPH radical scavenging activity was examined on LNA, DHA and EPA at the concentration of 100 ${\mu}M$, where they showed about 53% scavenging activity. These results suggest that omega-3 unsaturated fatty acid has a potential anticancer activity.

Synergistic Anticancer Activity of 5-Aminolevulinic Acid Photodynamic Therapy in Combination with Low-dose Cisplatin on Hela Cells

  • Wei, Xiao-Qiang;Ma, Hui-Qing;Liu, Ai-Hong;Zhang, You-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.3023-3028
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    • 2013
  • Objective: Photodynamic therapy (PDT ) is a promising modality for the treatment of various tumors. In order to assist in optimizing treatment, we applied 5-ALA/PDT in combination with low-dose cisplatin to evaluate cytotoxicity in Hela cells. Methods: Antiproliferative effects of 5-ALA/PDT and cisplatin, alone and in combination, were assessed using MTT assay. To examine levels of apoptosis, Hela cells treated with 5-ALA/PDT, and combination treatment were assessed with Annexin-V/PI by flow cytometry. To investigate the molecular mechanisms underlying alterations in cell proliferation and apoptosis, Western blot analysis was conducted to determine the expression of p53, p21, Bax and Bcl-2 proteins. Results: MTT assays indicated that combination treatment obviously decreased the viability of Hela cells compared to individual drug treatment. In addition, it was confirmed that exposure of Hela cells to 5-ALA/PDT in combination with low-dose cisplatin resulted in more apoptosis in vitro. Synergistic anticancer activity was related to upregulation p53 expression and alteration in expression of p21, Bcl-2 and Bax. Conclusion: Our findings suggest that administration of 5-ALA/PDT in combination with the low-dose cisplatin may be an effective and feasible therapy for cervical cancer.