王雨飞, 夏栋林, 陈超, 李晓东, 顾海鹰. 纳米金的制备及其对体外培养细胞生长的影响研究[J]. 环境卫生学杂志, 2015, 5(3): 197-201. DOI: 10.13421/j.cnki.hjwsxzz.2015.03.003
    引用本文: 王雨飞, 夏栋林, 陈超, 李晓东, 顾海鹰. 纳米金的制备及其对体外培养细胞生长的影响研究[J]. 环境卫生学杂志, 2015, 5(3): 197-201. DOI: 10.13421/j.cnki.hjwsxzz.2015.03.003
    WANG Yufei, XIA Donglin, CHEN Chao, LI Xiaodong, GU Haiying. Preparation of Gold Nanoparticles and its Cytotoxicity in Vitro[J]. Journal of Environmental Hygiene, 2015, 5(3): 197-201. DOI: 10.13421/j.cnki.hjwsxzz.2015.03.003
    Citation: WANG Yufei, XIA Donglin, CHEN Chao, LI Xiaodong, GU Haiying. Preparation of Gold Nanoparticles and its Cytotoxicity in Vitro[J]. Journal of Environmental Hygiene, 2015, 5(3): 197-201. DOI: 10.13421/j.cnki.hjwsxzz.2015.03.003

    纳米金的制备及其对体外培养细胞生长的影响研究

    Preparation of Gold Nanoparticles and its Cytotoxicity in Vitro

    • 摘要:
      目的 合成粒径均一、高生物亲和力的纳米金颗粒(gold nanoparticles; GNPs), 评价其对体外细胞生长的影响。
      方法 以4-羟乙基哌嗪乙磺酸(HEPES)为还原剂合成GNPs, 通过紫外光谱分析(UV)及透射电镜(TEM)进行表征。研究不同浓度(0、5、10、15、20及25 μg/mL)的GNPs对鼠成纤维细胞(L-929) 的生长影响。倒置显微镜观察各组细胞形态变化, MTT比色法测定溶液吸光度值来评估细胞毒性, 细胞内活性氧(ROS)检测试剂盒测定GNPs对L929细胞内活性氧含量的影响。
      结果 浓度2.5×10-5 mmol/L氯金酸与浓度1.0×10-2 mmol/L HEPES体积比100:1, 加热沸腾下合成的纳米金溶液呈透亮的酒红色, UV结果显示紫外特征峰在525 nm, TEM表征表明该方法制备的GNPs为球型, 平均直径约为24 nm, 粒径较均一, 有良好的稳定性与分散性。细胞实验显示各组细胞ROS水平随着培养基中GNPs含量的升高而降低, 当GNPs浓度20 μg/mL时, 参考《体外细胞毒性试验标准》(ISO-10993-5), 其细胞毒级为0~1级, 具有良好的生物相容性。
      结论 成功合成了粒径均一且具有高生物亲和力的GNPs。

       

      Abstract:
      Objective To prepare gold nanoparticles (GNPs) with good dispersibility and homogeneous size, and to evaluate their biocompatibility.
      Methods Gold nanoparticles were prepared by using HEPES to reduce HAuCl4, and ultraviolet spectroscopy (UV) and transmission electron microscopy (TEM) were employed to characterize the prepared Au samples. The toxic effect and dose-effect of gold nanoparticles on L-929 mouse fibroblasts were investigated by MTT method and intracellular reactive oxygen species (ROS) detection kits at 24, 48 and 72 hours of the test.
      Results The volume ratio of HAuCl4 (2.5×10-5 mmol/L) to HEPES (1.0×10-2 mmol/L) was 100:1. The gold nanoparticle solution prepared by boiling was red and translucent with good stability and dispersibility. The characteristic absorption band for UV spectrophotometry stands at 525 nm, the average diameter of spherical gold nanoparticles was 24 nm and the size distribution was narrow under TEM observation. The absorbance value between the experimental group and the negative control group was not significantly different (P > 0.05). The cell toxicity of GNPs at low concentration (≤20 μg/mL) and negative control group were graded as 0~1 degree. The level of ROS were negatively proportional to the concentration of GNPs in DMEM in a certain range of concentration.
      Conclusion Gold nanoparticles (GNPs) prepared by this method was good at safety and biocompatibility with no evident cytotoxicity.

       

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