• 폴리머
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• 제32권5호
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• pp.403-408
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• 2008

케라틴은 울, 머리카락, 손톱 등을 형성하는 섬유단백질의 주요성분으로 유용한 생체재료이다. 골수간염 줄기세포를 이용한 조직공학 적용을 위해 Poly(4-lactide-co-glycolide) (PLGA)에 함량별로 케라틴을 함유한 지지체를 용매 캐스팅/염 추출법을 이용하여 제조하였다. 제조된 지지체의 표면과 단면의 형태를 전자현미경(SEM)으로 관찰하고 특성분석을 위해 다공도, 표면 적심성, 물 흡수성, 그리고 열적성질을 분석하였다. 이 후 쥐에서 분리한 골수간엽줄기세포를 지지체에 파종하여 세포의 증식율을 (4,5-dimethylthiazol-2-yl)-2.5-diphenyl-tetrazolium bromide(MTT) 분석방법을 이용하여 측정하였다. 천연/합성 하이브리드 담체인 케라틴/PLGA 지지체는 PLGA 단독으로 제조된 지지체와 비교 시 골수간엽줄기세포의 생장에 유익한 환경을 제공함을 확인하였다.

• 대한핵의학회지
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• 제31권4호
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• pp.440-451
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• 1997

본 연구에서는 항암제인 taxol의 $^{111}In$ 방사성표지화합물을 합성하여 암진단제로서의 이용 가능성을 보기 위한 기초 연구를 수행하였다. Taxol의 $^{111}In$ 표지화합물을 얻기 위해 taxol구조에서 C-13의 곁가지에 있는 C-2' 부분의 hydroxyl기를 DTPA anhydride 및 succinic anhydride와 반응시켜 taxol-DTPA와 2'-hemisuccinyltaxol을 합성하였다. 반응수율은 taxol-DTPA 접합체의 경우 34%이었으며, 2'-hemisuccinyltaxol은 80%이었다. MTT법을 사용하여 HT29, B16, P388, CT26 세포주에서 taxol-DTPA와 2'-hemisuccinyltaxol의 세포독성능실험에서는 taxol 보다는 못미치나 그 세포독성이 유지됨을 확인하였다. 합성된 taxol 유도체들을 리간드 교환법과 직접법을 사용하여 In-111을 표지하였다. Taxol-DTPA 접합체의 In-111 표지반응의 경우, 리간드교환법은 반응도중 침전이 생겨 반응이 어려워 직접법으로 In-111 표지화합물을 얻을 수 있었으며 그 표지수율은 100%이었다. 2'-hemisuccinyltaxol은 두 방법을 모두 시도하였으나 반응이 진행되지 않음을 확인하였다. In-111의 taxol-DTPA 접합체 및 2'-hemisuccinyltaxol에 대한 표지반응 수율은 HPLC, paper, instant thin-layer chromatography를 실시하여 결정하였다. Li-pophilicity의 실험에서는 친수성임이 확인되었으며, 세포결합능의 실험에서는 HT29, B16, P388, CT26 세포주와의 결합이 매우 낮음을 나타내었다. 혈청단백 질과의 결합능을 보기위하여 30% trichloroacetic acid 법을 수행하였으며, 약 30%정도만이 혈청단백질과 결합하여 그 값이 크지 않았다.

• Journal of Acupuncture Research
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• 제19권4호
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• pp.74-88
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• 2002

Objectives : This study is aimed to investigate the effects of bee venom and melittin on cell death in synovial cell line. Methods : It was evaluated by using MTT assay, morphologic method, DNA fragmenation, NO generation, flow cytometry, immunocytochemistry analysis, RT-PCR, Western blot. Results : The obtained results are summarized as follows: 1. The MTT assay demonstrated that synovial cell viability was significantly inhibitted dose-dependently by treatment with bee venom and melittin in comparison with control. 2. The morphologic study demonstrated that synovial cell showed apoptosis after treatment with bee venom and melittin for 6 hours using microscope. 3. In case of NO generation bee venom group and melittin group showed significant inhibition in comparison with control. 4. The Flow cytometry demonstrated that apoptosis of synovial cell treated with bee venom and melittin was related with stop of cell cycle in stage of $G_0/G_1$. 5. DNA fragmenation demonstrated that synovial cell treated with bee venom and melittin showed DNA ladder below l Kbp. 6. Immunocytochemistry assay demonstrated that COX-II and PLA2 were strongly down-regulated by treatment with bee venom and melittin whereas iNOS was almostly not expressed by bee venom treatment and slightly expressed by melittin treatment. 7. RT-PCR analysis demonstrated that iNOS were strongly down-regulated by treatment with bee venom and melittin whereas COX-II was almostly not expressed by bee venom treatment and slightly expressed by melittin treatment. 8. Western blot demonstrated that iNOS were strongly down-regulated by treatment with $15{\mu}g/ml$ bee venom whereas COX-II was strongly down-regulated from $5{\mu}g/ml$ bee venom. Conclusions : These results suggest that bee venom and melittin have significant effect on cell death in synovial cell line and further study is needed in vivo.

• 한국재료학회지
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• 제20권6호
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• pp.312-318
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• 2010

Multilayer Poly methyl methacrylate (PMMA)/ Poly vinyl alcohol (PVA) bone plates were fabricated using electrospinning and in vitro investigations were carried out for pre-clinical biocompatibility studies. The initial cellular cytotoxicity of the methacrylate (PMMA)/ Poly vinyl alcohol (PVA) bone plates was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay using fibroblast-like L-929 cells. Cellular adhesion and differentiation studies were carried out using osteoblast-like MG-63 cells. As simulated body fluid (SBF) contains the same ionic concentration of body fluid and any bioactive material tends to deposit bone-like apatite on the samples surfaces into the SBF, in vitro bioactivity of the multilayer bone plates were investigated using SBF. We also studied the internal organization and tensile strength of the multilayer PMMA/PVA bone plates using micro-computed topography (${\mu}$-CT) and universal testing instrument (UTI, Korea) respectively. The cellular cytotoxicity study with MTT confirmed that the cellular viability was 78 to 90% which indicates good cyto-compatibility. Scanning electron microscopic findings revealed a good attachment and adhesion phenomenon of MG-63 cells onto the surfaces of the samples. Cellular differentiation studies also showed that osteogenic differentiation was switched on in a timely manner and affirmed along with that of the control group. Bone-like apatite formation on the surfaces was confirmed within 14 days of SBF incubation. Initial organizations of the multilayer PMMA/PVA bone plates were characterized as dense and uniform. The tensile strength of the post-pressing electronspun mat was higher than that of the pre-electronspun mat. These results suggest that a multilayer PMMA/PVA bone plate system is biocompatible, bioactive and a very good alternative bone plate system.

• 한국전기전자재료학회논문지
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• 제20권1호
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• pp.92-97
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• 2007

This paper performed the basic study for developing the Photodynamic Therapy Equipment for medical treatment. We developed the equipment palpating cell proliferation using a high brightness LED. This equipment was fabricated using a micro-controller and a high brightness LED, and designed to enable us to control light irradiation time, intensity, frequency and so on. Especially, to control the light irradiation frequency, FPGA was used, and to control the change of output value, TLC5941 was used. Control stage is divided into 30 levels by program. Consequently, the current value could be controlled by the change of level in Continue Wave(CW) and Pulse Width Modulation(PWM), and the output of a high brightness LED could be controlled stage by stage. And then, each experiment was performed to irradiation group and non-irradiation group for both Rat bone marrow and Rat tissue cells. MTT assay method was chosen to verify the cell increase of two groups and the effect of irradiation on cell proliferation was examined by measuring 590 nm transmittance of ELISA reader. As a result, the cell increase of Rat bone marrow and tissue cells was verified in irradiation group as compared to non-irradiation group. The fact that specific wavelength irradiation has an effect on cell vitality and proliferation is known through this study.

• 한국약용작물학회지
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• 제14권2호
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• pp.77-81
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• 2006

As a result of cytotoxic compounds against cancer cell lines from natural sources, senven compounds were isolated from the leaf and twig of Acer okamotoanum Nakai. The compounds (1-7) were identified as ethyl gallate (1), methyl gallate (2), gallic acid (3), trans $resveratrol-3-O-{\beta}-D-glucopyranoside$ (4), acertannin (5), nikoenoside (6), and fraxin (7) by physicochemical and spectroscopic data (including mp, UV, IR, MS, $^1H-NMR,\;^{13}C-NMR$, DEPT, and HMBC) in comparison with those of published papers. All the compounds were tested for their cytotoxic activity against L1210, HL-60, K562, and B16F10 cancer cell lines in vitro by MTT assay method. Compounds 1-3 and 5 showed cytotoxic activity against all tested cell lines with $IC_{50}$ values ranged from 12.5 to $72.2\;{\mu}M$. Of the compounds, methyl gallate (2) exhibited the most potent cytotoxic activity against L1210, HL-60, K562, and B16F10 tumor cells with $IC_{50}$ values of 12.5, 48.3, 22.8, and $22.8\;{\mu}M$, respectively. Other compounds did not show any cytotoxic activity against four cancer cell lines.

• 대한한방종양학회지
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• 제2권1호
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• pp.75-90
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• 1996

This experiment was designed to study the antitumor effects and Activity of Natural Killer Cell of semen Tiglii plus Rhizoma Rhei. The cytotoxic and antitumor effects were evaluated on human cell lines(A549, Caki-1, LL2, Sarcoma 180, NIH/3T3) after exposure to prebrewed Semen Tiglii plus Rhizoma Rhei water extract 0.1, 0.2, 0.4, 0.8, 1.6mg/ml using in MTT assay, LDH, colony forming efficency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. From the result of MTT assay, the cytotoxicity of ST(生巴豆霜), ST+RR(生巴豆霜加大黃) were concentration-dependently increased in both group of the ST and ST+RR, the cytotoxicity of ST+RR(生巴豆霜加大黃) was similar to that of ST(生巴豆霜). 2. From the result of LDH, the cytotoxicity of ST, ST +RR were concentrati -on-dependently increased in both group of the ST and ST+RR, the cytotoxicity of ST+RR was similar to that of ST. 3. The antitumor effect on A549 tumor cell from the result of colony forming efficiency showed the inhibitory effect on the growth in both group of the ST and ST+RR, the inhibitory effect on growth was low slightly in the ST+RR. 4. From the result of SRB assay, the antitumor effect on caki-1 tumor cell of ST, ST+RR showed the inhibitory effect on the growth in both group of the ST and ST+RR, the antitumor effect of ST+RR was similar to that of ST. 5. Median survival time and increased life span were increased slightly in both group of the ST and ST+RR. 6. The inhibitory effect on the growth of Sarcoma 180 and Lewis lung carcinoma tumor cell were increased slightly in both group of the ST and ST+RR. 7. The activity of NK cell was increased in the ST+RR.

• Asian Pacific Journal of Cancer Prevention
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• 제15권12호
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• pp.5055-5061
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• 2014

In this study, an anti-oxidant and anti-tumor protein Latcripin-3 of Lentinula edodes C91-3 was expressed in Escherichia coli. for the first time. According to the cDNA library, the full-length gene of Latcripin-3 was cloned by the methods of 3'-full rapid amplification of cDNA Ends (RACE) and 5'-full RACE. The structural domain gene of Latcripin-3 was inserted into the pET32 a(+). The functional protein of Latcripin-3 was expressed in Rosetta-gami (DE3) E. coli, evaluated by Western blotting and mass spectrometry. DPPH testing showed that the protein Latcripin-3 can scavenge free radicals remarkably well. The activity of functional protein Latcripin-3 on A549 cells was studied with flow cytometry and the MTT method. The MTT assay results showed that there was a decreases in cell viability in a dose-dependent and time-dependent manner in protein Latcripin-3 treated groups. Flow cytometry demonstrated that Latcripin-3 can induce apoptosis and block S phase dramatically in human A549 lung cancer cells as compared to the control group. At the same time, the cell ultrastructure observed by transmission electron microscopy supported the results of flow cytometry. This research offers new insights and advantages for identifying anti-oxidant and anti-tumor proteins.

• 치위생과학회지
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• 제3권1호
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• pp.11-14
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• 2003

각종 토양시료로부터 M배지와 AMP배지에서 생육하는 500여종의 방선균을 분리하였으며, 그 중 전배양배지(GMY 배지)에서 가장 뛰어난 항암활성능을 소유한 균주를 불리하고 배양하여 세포외 항암성물질을 분리하여 이 물질을 완전정제하고 MTT 정량분석을 실시하여 암세포에 대한 세포독성검사를 실시하였다. 1. 정제는 배양한 균체를 완전히 제거하고 동량의 ethylacetate를 처리하여 배양액중의 항암성분을 추출하고 무수 magnesium sulfate로 건조 후 농축, ethanol로 용해, 10배량의 water을 첨가하여 $4^{\circ}C$에서 overnight 시킨 후 추출액을 원심분리(12,000 rpm, 30분)하여 methylene chloride로 용해시켜 silica gel 60 column(${\phi}35{\times}600mm$, Merck Co.), methylene chloride-ethanol(96:4) 용매로 용출하고 sephadex LH-20 column(${\phi}15{\times}300mm$, Pharmacia LKB)에서 100% methanol로 용출하여 HPLC(Waters, ODS)를 이용하여 최종적으로 완전 정제하였다. 2. Compound는 Gram(+) 세균(6균주), Gram(-) 세균(11균주), 효모(2균주), 곰팡이(1균주)에 대하여 항균 효과를 나타내었다. 3. 완전 정제된 물질에 대한 항암효과를 측정한 결과, 모든 실험 암세포에 대해서 뛰어난 항암효과를 나타내었으며 protein성 물질은 아닌 것으로 추정되었다.

• 한국한의학연구원논문집
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• 제18권1호
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• pp.85-92
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• 2012

Objective : The purpose of this study was to investigate the anti-cancer effects of Sophorae Radix and the effects of 5-Fluorouracil (5-FU) in human gastric adenocarcinoma cells (AGS). Method : We used human gastric adenocarcinoma cell line, AGS cells. We examined cell death by MTT assay and caspase 3 assay with Sophorae Radix. To examine the inhibitory effects of Sophorae Radix, cell cycle (sub G1) analysis was done the AGS cells after three days with Sophorae Radix. The reversibility of Sophorae Radix was examined on one day to five days treatment with 100 ${\mu}g/ml$ Sophorae Radix. Result : Sophorae Radix inhibited the growth of AGS cells in a dose-dependent fashion. Also we showed that Sophorae Radix induced apoptosis in AGS cells by MTT assay, caspase 3 assay and sub-G1 analysis. Sophorae Radix combined with 5-FU markedly inhibited the growth of AGS cells compared to Sophorae Radix or 5-FU alone. After 3 days treatment of AGS cells with Sophorae Radix, the fraction of cells in sub-G1 phase was much higher than that of the control group. Conclusion : Our findings provide insight into unraveling the effects of Sophorae Radix in human gastric adenocarcinoma cells and developing therapeutic agents against gastric cancer.