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20 μg/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0.031 3、0.062 5、0.125 0、0.250 0μg/mL 4种质量浓度处理HepG2细胞,采用含294 μg/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)0.250 0 μg/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(3.3±0.7)％(t=13.327、30.475,P＜0.05);0.031 3、0.062 5、0.125 0μg/mL产品A处理细胞的凋亡率分别为(2.9±0.4)％、(3.1±0.4)％、(4.2±0.9)％,与空白对照处理时相近(t=1.181、0.133、1.097,P＞0.05).(4)0.125 0、0.250 0μg/mL产品A处理细胞时,尾矩、尾长、尾部DNA百分比明显高于空白对照处理(t=29.026、51.194,21.851、36.138,24.721、50.455,P ＜0.05或P＜0.01).0.031 3、0.062 5μg/mL产品A处理细胞时,尾矩、尾长、尾部DNA百分比与空白对照处理比较,差异无统计学意义(t=5.878、3.429,2.779、1.960,1.328、7.763,P＞0.05).结论 5种含银产品含量均达到各标识要求;产品C浓度较其他产品更高,导致HepG2细胞存活率下降的可能性更大,建议银离子产品在浓度设置方面以产品A、B为参考;产品浓度越高对细胞DNA可能存在更高的损伤风险,不建议相关产品为了达到更好抗菌效果而一味调高含银量.

abstractsObjective 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 mnethod 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 product E.(2) Compared with the rate of blank control,the cell survival rates of product A at the dilution ratio of 1∶100,product B at the dilution ratio of 1∶100,and product C at the dilution ratio of 1∶100 and 1∶200 were significantly reduced (t =35.506,8.914,37.594,30.693,P ＜ 0.01).Compared with the rate of positive control,the cell survival rates of product A at the dilution ratio of 1∶200,1∶400,and 1∶800,product C at the dilution ratio of 1∶400 and 1∶800,products B and D at each dilution ratio were increased significantly (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) The apoptosis rates of cells treated by 0.250 0 μg/mL product A and positive control were (6.1 ± 0.4)％ and (62.2 ± 3.9)％ respectively,which were significantly higher than the apoptosis rate of blank control [(3.3 ± 0.7) ％,t =13.327,30.475,P ＜ 0.05].The apoptosis rates of cells treated by 0.031 3,0.062 5,0.125 0 μg/mL product A were (2.9 ±0.4)％,(3.1 ±0.4)％,and (4.2 ± 0.9)％ respectively,which were close to the apoptosis rate of blank control (t =1.181,0.133,1.097,P ＞ 0.05).(4) The tail moment,tail length,and tail DNA percentage of cells cultured with 0.125 0 and 0.250 0 μg/mL product A were significantly higher than those cultured with blank control (t =29.026,51.194,21.851,36.138,24.721,50.455,P ＜0.05 orP ＜0.01).However,the tail moment,tail length,and tail DNA percentage of cells cultured with 0.031 3 and 0.062 5 μg/mL product A were close to those cultured with blank control (t =5.878,3.429,2.779,1.960,1.328,7.763,P ＞0.05).Conclusions The silver content of silver-containing products meets the requirements of the labeling.The concentration of product C is higher than that of other products,leading to a greater possibility of decreasing the survival rate of HepG2 cells.It is suggested that the products A and B should be taken as reference in the concentration setting of silver ion products.The product solution with higher concentration may have higher risk of damage to cell DNA.Therefore,it is not recommended to upregulate silver content of relevant products blindly in order to achieve better antibacterial effect.