• 제목/요약/키워드: in vitro cytotoxicity test

검색결과 159건 처리시간 0.024초

In Vitro에서 히스타민 측정 시 ELISA법과 HPLC-MS 분석법의 비교 (Comparison of Methods for Measuring Histamine by ELISA and HPLC-MS Assay In Vitro)

  • 이인희;김유현
    • 대한임상검사과학회지
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    • 제47권4호
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    • pp.306-312
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    • 2015
  • 알러지 반응이 일어나면 histamine이 방출되기 때문에 histamine을 정량 측정함으로써 유발된 알러지의 정도를 확인 할 수 있다. 일반적으로는 항원-항체반응으로 microplate reader를 이용하여 흡광도를 측정하여 정량 한다. 본 연구에서는 histamine 방출량을 측정함에 있어서 일반적인 항원-항체반응과 분석 화학적인 방법으로 HPLC-MS를 이용한 방법을 비교하였다. 세포주는 RBL-2H3를 사용하였고, C48/80으로 자극시켜 알러지를 유발하였다. 유발된 알러지는 ${\beta}$-hexosaminidase의 측정으로 탈 과립을 확인하였으며 실험의 정당성을 위하여 세포독성 능을 확인하였다. Histamine 정량에서 항원-항체반응에 의한 측정의 정량한계는 10.257 ppm이었고, HPLC-MS에 의한 정량한계는 0.020 ppm으로 현저한 차이를 보였다. 알러지 활성 및 항 알러지 실험에 있어서 histamine의 측정은 HPLC-MS를 이용한 분석이 더 정밀하고 정확한 실험인 것을 확인하였다.

Development of Inhibitors of $\beta$-Amyloid Plaque Formation

  • Kim, Dong-Jin
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 2006년도 Spring Conference
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    • pp.123-135
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    • 2006
  • Alzheimer's disease (AD) is the most common form of dementia in the aging population and is clinically characterized by a progressive loss of cognitive abilities. Pathologically, it is defined by the appearance of senile plaques - extracellular insoluble, congophilic protein aggregates composed of amyloid $\beta$ (A$\beta$) and neurofibrillary tangles (NFTs) - inyracellular lesions consisting of paired helical filaments from hyperphosphorylated cytoskeletal tau protein as described by Alois Alzheimer a century ago. These hallmarks still serve as the major criteria for a definite diagnosis of the disease. Consequently, one of the key strategy for drug development in this disease area focuses on reducing the concentration of cerebral A$\beta$ plaque by using substances that inhibit A$\beta$ fibril formation. We focused on developing inhibitors by synthesizing several kinds of aromatic molecules. The synthetic compounds were initially screened to evaluate the effective compound by tioflavin T fluorescence assay. The selected effective compounds were tested cytotoxicity and protective effect from A$\beta$-induced neuronal toxicity by cell based MTT assay with HT22 hippocampal neurons. The BBB permeability on effectors was also tested in in vitro co-culture model(HUVEC/C6 cell line). The behavior test wea carried out in mutant APP/PS1 transgenic mouse model of Alzheimer's disease. And inhibition of A$\beta$ fibril formation by the effective compound was monitored with transmitted electron microscopic images.

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감초 물추출물의 멜라닌 형성 억제효과 및 기전에 관한 연구 (Inhibitory Effect and Mechanism on Melanogenesis of Radix glycyrrhizae Water Extract)

  • 김진;권일호;임홍진;임규상;황충연
    • 한방안이비인후피부과학회지
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    • 제16권2호
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    • pp.96-118
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    • 2003
  • The effect of Glycyrrhizae Radix water extract, known as depigmenting agent, on melanin biosynthesis was investigated in cellular level by using B16 mouse melanoma cells. The inhibitory effect of Glycyrrhizae Radix water extract on melanogenesis was determined by mushroom tyrosinase assay traditionally using in vitro screening test. To determine whether Glycyrrhizae Radix water extract suppress melanin synthesis in cellular level, B16 mouse melanoma cells were cultured in the presence of different concentrations of Glycyrrhizae Radix water extract. Effects on cell proliferation, melanin biosynthesis, tyrosinase activity, DOPAchrome tautomerase activity, and expression level of mRNA for tyrosinase were examined. The maximum concentration of Glycyrrhizae Radix water extract that was not inhibitory to growth of the cells was 2 mgml. At that concentration, melanin synthesis was significantly inhibited without cytotoxicity after 5 days, compared with untreated cells. The treatment with Glycyrrhizae Radix water extract reduced tyrosinase and DOPAchrome tautomerase activity in a dose-dependent manner. However, the treatment with Glycyrrhizae Radix water extract did not affect significantly mRNA levels for tyrosinase. These results suggest that the inhibitory effect of Glycyrrhizae Radix water extract on melanogenesis is correlated with the suppression of tyrosinase and DOPAchrome tautomerase activity more than altering mRNA levels of tyrosinase.

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Combined Effects of Curcumin and Triptolide on an Ovarian Cancer Cell Line

  • Cai, Ying-Ying;Lin, Wei-Ping;Li, Ai-Ping;Xu, Jian-Yang
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권7호
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    • pp.4267-4271
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    • 2013
  • Background: As natural medicines in Asia, curcumin and triptolide extracted from different drug plants have proven to possess anticancer potential and widely used for anti-cancer research. The present study attempted to clarify that curcumin and triptolide synergistically suppress ovarian cancer cell growth in vitro. Methods: To test synergic effects, cell viability and apoptosis were analyzed after curcumin and triptolide combination treatment on ovarian cancer cell lines. Synergistic effects on apoptosis induction were determined by lactate dehydrogenase (LDH) leakage assay, intracellular reactive oxygen species (ROS) assay, mitochondrial membrane potential (MMP) loss assay and flow cytometry analysis. Critical regulators of cell proliferation and apoptosis related were analyzed by qRT-PCR and Western blotting. Results: We showed that the combination of curcumin and triptolide could synergistically inhibit ovarian cancer cell growth, and induce apoptosis, which is accompanied by HSP27 and HSP70, indicating that HSP27 and HSP70 play the important role in the synergic effect. Conclusions: From the result present here, curcumin and triptolide combination with lower concentration have a synergistic anti-tumor effect on ovarian cancer and which will have a good potential in clinical applications.

Synthesis and characterization of silk fibroin-bioactive glass hybrid xerogels

  • Wu, Xiaohong;Yan, Fuhua;Liu, Wei;Zhan, Hongbing;Yang, Wenrong
    • Biomaterials and Biomechanics in Bioengineering
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    • 제1권2호
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    • pp.63-71
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    • 2014
  • This study aimed to develop a novel bioactive hybrid xerogel consisting of silk fibroin /$SiO_2-CaO-P_2O_5$ by sol-gel process at room temperature. Scanning electron microscopy (SEM), FT-IR Spectroscopy, pore measurement, mechanical property testing, in vitro bioactivity test and cytotoxicity assay were performed to characterize the xerogel for bone tissue engineering application. We have found that the xerogel possessed excellent pore structures and mechanical property. Once immersed in a simulated fluid (SBF), the xerogel exhibited profound bioactivity by inducing hydroxyapatite layers on its surfaces. The cell toxicity study also demonstrated that there was little toxic to MC3T3-E1 cells. These results indicate that silk fibroin /$SiO_2-CaO-P_2O_5$ hybrid xerogel potentially could be used as a bone tissue engineering material.

삼종실유의 항돌연변이 효과 (Mutagenic and Antimutagenic Effects of Hemp Seed Oil Evaluated by Ames Salmonella Testing)

  • 전정애;조희준;전희진;이지혜;가요요;조경상;김은수;이성준
    • 한국식품과학회지
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    • 제43권3호
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    • pp.396-400
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    • 2011
  • We examined the in vitro mutagenic and antimutagenic effects of hexane-extracted hemp (Cannabis sativa L. subsp. sativa var. sativa) seed oil (HO) with and without S9-mediated metabolic activation, using the TA98 and TA100 Salmonella Typhimurium strains. The MTT assay revealed no cytotoxicity in HepG2 cells for HO quantities $\leq400g$/mL. In the mutagenicity test, revertant colonies did not exceed spontaneous colonies in number. Colony numbers did not increase in either strain after HO treatment, with or without metabolic activation. HO showed no mutagenic effects and did not induce a mutation in either strain. In the antimutagenicity test, HO reduced the number of mutated colonies induced by 4NQO in both strains. The inhibition rates of HO (TA98, 21-91%; TA100, 21-85%) indicated a potent reduction in mutagenicity induced by 4NQO. HO showed no significant mutagenicity and may have antimutagenic effects, as assessed by Ames testing.

실란처리된 하이드록시프로필 메틸셀룰로오스/상전이된 칼슘포스페이트 복합체 골시멘트의 제조 및 특성평가 (Preparation and characterization of silanized-hydroxypropyl methylcellulose/phase transformed calcium phosphate composite bone cements)

  • 정나현;김동현;조훈상;윤석영
    • 한국결정성장학회지
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    • 제26권6호
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    • pp.243-251
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    • 2016
  • 실란처리된 하이드록시프로필 메틸셀룰로오스/상전이된 칼슘포스페이트 복합체 시멘트는 향상된 생체적합성을 가지는 주입이 가능한 골시멘트로서 제조 되었다. 복합체 시멘트의 고체상과 액체상으로서 상전이된 칼슘포스페이트와 실란처리된 하이드록시프로필 메틸셀룰로오스의 결정구조 및 화학적 상태는 XRD 및 FT-IR에 의해 측정 되었고, 시멘트의 조작성 및 기계적 물성은 주입성 및 3점 굽힘 시험에 의해 측정하였다. 체외에서 진행된 골시멘트의 기계적 물성, XRD 및 SEM 결과는 골시멘트의 고유 기능으로서의 향상된 경화거동을 보였다. 또한 골시멘트의 세포독성 결과 향상된 생체적합성을 보였다. 따라서 이러한 주입이 가능한 시멘트는 생체의학 응용에 대한 칼슘포스페이트 시멘트로서 사용이 가능하리라 판단되어진다.

Development and Evaluation of Natural Hydroxyapatite Ceramics Produced by the Heat Treatment of Pig Bones

  • Lim, Ki-Taek;Kim, Jin-Woo;Kim, Jangho;Chung, Jong Hoon
    • Journal of Biosystems Engineering
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    • 제39권3호
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    • pp.227-234
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    • 2014
  • Purpose: The aim of this research was to develop and evaluate natural hydroxyapatite (HA) ceramics produced from the heat treatment of pig bones. Methods: The properties of natural HA ceramics produced from pig bones were assessed in two parts. Firstly, the raw materials were characterized. A temperature of $1,200^{\circ}C$ was chosen as the calcination temperature. Fine bone powders (BPs) were produced via calcinations and a milling process. Sintered BPs were then characterized using field emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), X-ray fluorescence spectroscopy (XRF), energy dispersive X-ray spectroscopy (EDX), Fourier transform infrared (FTIR) spectroscopy, and a 2-year in vitro degradability test. Secondly, an indirect cytotoxicity test was conducted on human osteoblast-like cells, MG63, treated with the BPs. Results: The average particle size of the BPs was $20{\pm}5{\mu}m$. FE-SEM showed a non-uniform distribution of the particle size. The phase obtained from XRD analysis confirmed the structure of HA. Elemental analysis using XRF detected phosphorus (P) and calcium (Ca) with the Ca/P ratio of 1.6. Functional groups examined by FTIR detected phosphate ($PO{_4}^{3-}$), hydroxyl ($OH^-$), and carbonate ($CO{_3}^{2-}$). The EDX, XRF, and FTIR analysis of BPs indicated the absence of organic compounds, which were completely removed after annealing at $1,200^{\circ}C$. The BPs were mostly stable in a simulated body fluid (SBF) solution for 2 years. An indirect cytotoxicity test on natural HA ceramics showed no threat to the cells. Conclusions: In conclusion, the sintering temperature of $1,200^{\circ}C$ affected the microstructure, phase, and biological characteristics of natural HA ceramics consisting of calcium phosphate. The Ca-P-based natural ceramics are bioactive materials with good biocompatibility; our results indicate that the prepared HA ceramics have great potential for agricultural and biological applications.

치근 천공 치료 재료의 생체친화성의 비교 (COMPARISON OF BIOCOMPATIBILITY OF FOUR ROOT PERFORATION REPAIR MATERIALS)

  • 강민경;배인호;고정태;황윤찬;황인남;오원만
    • Restorative Dentistry and Endodontics
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    • 제34권3호
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    • pp.192-198
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    • 2009
  • 이번 연구는 치근 천공의 치료 재료인 white mineral trioxide aggregate (MTA)를 흔히 사용되는 calcium hydroxide liner ($Dycal^{(R)}$), glass ionomer cement (GIC), 그리고 MTA와 유사한 성분을 가진 Portland cement와 세포독성 실험으로 생체 친화성을 평가하는 것이다. 세포독성의 정도는 MG-63 세포를 이용해 주사전자 현미경적 관찰과 수용성 tetrazolium salt를 이용한 흡광도를 측정 (XTT assay)하여 평가하였다. SEM 관찰에서, 1일과 3일째 모두에서 GIC와 MTA, Portland cement 표면에서는 잘 부착된 세포를 보여주었다. 반면에, Dycal 표면의 세포들은 둥글고 부착되지 않은 양상을 보여 주었다. XTT assay에서는 Dycal을 제외한 모든 재료에서 유사하게 높은 세포 활성도를 보여주었으며, 이는 SEM 관찰 소견과 일치하였다. 이번 연구는 MTA가 생체친화적인 재료라는 견해를 뒷받침한다. 또한 Portland cement와 GIC에서도 MTA와 유사한 세포반응을 보여주었다.

Fabrication and Characterization of Novel Electrospun PVPA/PVA Nanofiber Matrix for Bone Tissue Engineering

  • Franco, Rose-Ann;Nguyen, Thi Hiep;Lee, Byong-Taek
    • 한국재료학회:학술대회논문집
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    • 한국재료학회 2011년도 춘계학술발표대회
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    • pp.51.2-51.2
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    • 2011
  • A novel electrospun nanofiber membrane was fabricated using combined poly (vinylphosphonic acid) (PVPA) and polyvinyl alcohol (PVA) intended for bone tissue engineering applications. PVPA is a proton-conducting polymer used as primer for bone implants and dental cements to prevent corrosion and brush abrasion. The phosphonate groups of PVPA have the ability to crosslink and attach itself to the hydroxyapatite surface facilitating faster integration of the biomaterial to the bone matrix. PVA was combined with PVPA to provide hydrophilicity, biocompatibility and improve its spinnability. To improve its mechanical strength, PVPA/PVA and neat PVA mixtures were combined to produce a multilayer scaffold. The physical and chemical properties of the of the fabricated matrix was investigated by SEM and TEM morphological analyses, tensile strength test, XRD, FT-IR spectra, swelling behavior and biodegradation rates, porosity and contact angle measurements. Biocompatibility was also examined in vitro by cytotoxicity and cell proliferation studies with MTT assay and cell adhesion behavior by SEM and confocal microscopy.

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