银离子产品的含量特性及细胞毒性检测  

Content characteristics and cytotoxicity detection of silver ion products

作　　者：苏雪荣 甘俊英 薛玉英[1] 李强 Su Xuerong;Gan Junying;Xue Yuying;Li Qiang(Key Laboratory of Environmental Medicine and Engineering, Ministry of Education, School of Public Health & Collaborative Innovation Center of Suzhou Nano Science and Technology, Southeast University, Jiangsu key Laboratory for Biomaterials and Devices, Nanjing 210009, China;Changshu Food and Drug Administration, Changshu 215500, China)

机构地区：[1]环境医学工程教育部重点实验室,东南大学公共卫生学院&苏州纳米科技协同创新中心,江苏省生物材料与器件重点实验室,南京210009 [2]常熟市食品药品监督管理局,常熟215500

出　　处：《中华烧伤杂志》2019年第1期12-17,共6页Chinese Journal of Burns

基　　金：国家自然科学基金(81573186);江苏省普通高校研究生科研创新计划 (SJCX17_0067、SJCX18_0078).

摘　　要：目的分析含银产品的银含量、均匀性及细胞毒性。方法(1)从市场上购买的5种含银产品A、B、C、D、E,其中前4种为液体或凝胶形式、含银产品E为敷料形式。采用火焰法测定各产品银含量及产品E的均匀性,样本数为3。(2)选用人肝癌细胞株(HepG2)作为评价模型,将A、B、C、D4种含银产品分别用DMEM高糖培养液进行1∶100、1∶200、1∶400、1∶800的倍比稀释后培养细胞,以含20μg/mL硝酸银的DMEM高糖培养液培养细胞为阳性对照、DMEM高糖培养液培养细胞为空白对照,采用噻唑蓝法测定细胞存活率,样本数均为5。(3)将含银产品A配制成0.0313、0.0625、0.1250、0.2500μg/mL4种质量浓度处理HepG2细胞,采用含294μg/mL重铬酸钾、胎牛血清的DMEM高糖培养液处理细胞为阳性对照、采用含胎牛血清的DMEM高糖培养液处理细胞为空白对照,采用Hoechst33258染色法检测细胞凋亡率,采用彗星实验检测细胞彗星尾矩、尾长及尾部DNA百分比判断DNA损伤,样本数均为3。对数据行单因素方差分析、LSD-t检验。结果(1)A、B、C、D含银产品的银含量依次为(256.5±1.5)μg/mL、(271.5±1.3)μg/mL、(652.4±2.6)μg/g、(330.0±2.1)μg/g,与标示含量吻合。含银产品E的银含量为(0.158±0.013)mg/g、单片产品E的银含量为(0.125±0.017)mg/g,均匀性良好。(2)与空白对照处理比较,产品A在稀释比1∶100、产品B在稀释比1∶100及产品C在稀释比1∶100、1∶200处理时细胞存活率均显著降低(t=35.506、8.914、37.594、30.693,P<0.01)。与阳性对照处理比较,产品A在稀释比1∶200、1∶400、1∶800及产品C在稀释比1∶400、1∶800与产品B、D在各稀释比处理时细胞存活率均显著升高(t=27.537、18.262、18.709,26.333、41.762,15.776、19.759、20.443、15.715,26.792、24.963、31.803、30.537,P<0.01)。(3)0.2500μg/mL产品A及阳性对照处理细胞的凋亡率分别为(6.1±0.4)%、(62.2±3.9)%,明显高于空白对照处理细胞的(3.3Objective To analyze the silver content, homogeneity, and cytotoxicity of silver-containing products. Methods (1) Five kinds of silver-containing products A, B, C, D, and E were purchased from the market, and products A, B, C, and D are liquid or gel form while product E was dressing form. The silver content of each product and the homogeneity of product E were determined by flame method. The sample number was 3. (2) Human hepatocellular carcinoma cell line (HepG2) was selected as the evaluation model. Four silver-containing products A, B, C, and D were diluted with high-glucose dulbecco′s modified eagle medium (DMEM) at multiple ratios of 1∶100, 1∶200, 1∶400, and 1∶800, and then they were used for cell culture. Cells cultured with high-glucose DMEM and high-glucose DMEM containing 20 μg/mL silver nitrate were used as blank control and positive control, respectively. The cell viability was determined by methyl thiazolyl tetrazolium assay, and each sample number was 5. (3) Four mass concentrations of 0.031 3, 0.062 5, 0.125 0, and 0.250 0 μg/mL were prepared from silver-containing product A, and then they were used to culture HepG2 cell. Cells cultured with high-glucose DMEM containing fetal calf serum and 294 μg/mL potassium dichromate were used as positive control, while those containing fetal calf serum were used as blank control. Hoechst 33258 staining method was used to detect apoptosis rate of cells. The tail moment, tail length, and the percentage of DNA in the tail of cells were observed by comet assay to evaluate DNA damage. The sample numbers were all 3. Data were processed with one-way analysis of variance and least significant difference-t test. Results The silver content of products A, B, C, and D was (256.5±1.5) μg/mL, (271.5±1.3) μg/mL, (652.4±2.6) μg/g , (330.0±2.1) μg/g, which was in accordance with labelled amount. The silver content of product E was (0.158±0.013) mg/g, and the silver content of each piece of product E was (0.125±0.017) mg/g, showing good uniformity of prod

关 键 词：银 药物稳定性 毒性试验 HEPG2细胞 

分 类 号：R318.08[医药卫生—生物医学工程]