中国组织工程研究

中国组织工程研究 ›› 2013, Vol. 17 ›› Issue (3): 392-399.doi: 10.3969/j.issn.2095-4344.2013.03.003

• 纳米生物材料 nanobiomaterials • 上一篇    下一篇

纳米Ag-SiO2聚氨酯的体外细胞毒性评价

杨六成1，徐 帅1，李师思2，吴 凯1，王键俊1   

  1. 南方医科大学珠江医院普通外科，广东省广州市  510282
  • 收稿日期:2012-11-13 修回日期:2012-12-04 出版日期:2013-01-15 发布日期:2013-01-15
  • 通讯作者: 杨六成，教授，主任医师，南方医科大学珠江医院普通外科，广东省广州市 510282 sdylc@yahoo.com.cn
  • 作者简介:杨六成☆，男，1964年生，汉族，南方医科大学毕业，博士，硕士生导师，主任医师，教授，主要从事普通外科基础与临床研究。

In vitro cytotoxicity of nanoAg-SiO2 polyurethane materials

Yang Liu-cheng1, Xu Shuai1, Li Shi-si2, Wu Kai1, Wang Jian-jun1   

  1. 1 Department of General Surgery, Zhujiang Hospital of Southern Medical University, Guangzhou 510282, Guangdong Province, China
    2 Department of Nuclear Medicine, Zhujiang Hospital of Southern Medical University, Guangzhou 510282, Guangdong Province, China
  • Received:2012-11-13 Revised:2012-12-04 Online:2013-01-15 Published:2013-01-15
  • Contact: Yang Liu-cheng, Department of General Surgery, Zhujiang Hospital of Southern Medical University, Guangzhou 510282, Guangdong Province, China sdylc@yahoo.com.cn
  • About author:Yang Liu-cheng☆, Doctor, Master’s supervisor, Chief physician, Professor, Department of General Surgery, Zhujiang Hospital of Southern Medical University, Guangzhou 510282, Guangdong Province, China

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摘要：

背景：为改善聚氨酯材料的抗菌性能，前期研究中合成了纳米Ag-SiO2聚氨酯材料。
目的：比较7种含不同质量分数纳米Ag-SiO2聚氨酯材料的体外细胞毒性。
方法：将纳米Ag-SiO2与聚氨酯以熔融共混方式制备成含纳米Ag-SiO2，质量分数分别为0，0.5%，1.0%，1.5%，2.0%，2.5%，5.0%的聚氨酯材料。以上述7种聚氨酯材料浸提液、高密度聚乙烯浸提液(阴性对照)、0.1%苯酚液浸提液(阳性对照)分别培养L929细胞24，48，72 h后，并设置试剂对照(含体积分数10%胎牛血清的RPMI 1640液体培养基)和空白对照(不加细胞，只加含体积分数10%胎牛血清的RPMI 1640液体培养基)。采用MTT比色法定量检测各组细胞相对增殖率，并进行毒性反应分级；同时在显微镜下观察细胞形态。
结果与结论：7种含不同质量分数纳米Ag-SiO2聚氨酯材料组、试剂对照组、阴性对照组细胞贴壁良好，形态正常，胞体丰满，胞质、核质分布均匀，细胞相对增殖率均大于80%，毒性反应分级均为1级，且随着纳米Ag-SiO2质量分数的降低，体外细胞毒性渐小、生物相容性更好。随培养时间延长，阳性对照组细胞相对增殖率逐渐降低，72 h后细胞相对增殖率降至8.7%，与其他组比较差异有显著性意义(P < 0.05)，细胞萎缩、变圆、漂浮、片状脱落。表明7种含不同质量分数纳米Ag-SiO2聚氨酯材料均具有良好的体外细胞相容性，毒性反应分级均为1级，符合医用材料体外实验要求。

关键词: 生物材料, 纳米生物材料, 纳米Ag-SiO2, 聚氨酯, 体外, L929细胞, 细胞相对增殖率, 细胞毒性, 抗菌性能, MTT比色法, 省级基金, 生物材料图片文章

Abstract:

BACKGROUND: We synthesized nanoAg-SiO2 polyurethane during preliminary research for improvement of antibacterial property of polyurethane materials.
OBJECTIVE: To compare the in vitro cytotoxicity of seven kinds of polyurethane materials with different content of nanoAg-SiO2.
METHODS: We melted nanoAg-SiO2 into polyurethane to prepare polyurethane materials containing 0%, 0.5%, 1.0%, 1.5%, 2.0%, 2.5% and 5.0% nanoAg-SiO2, respectively. Then we prepared the leaching liquor of high density polyethylene (negative control), 0.1% phenol fluid (positive control) and these nanoAg-SiO2 polyurethane materials to culture L929 cells for 24, 48 and 72 hours. Cells cultured in RPMI 1640 medium containing 10% fetal bovine serum served as reagent control group, and blank control group was also set. Then MTT colorimetric method was used to quantitatively detect cell relative growth rate and conduct toxicity reaction grading. Cell morphology was observed under microscope.
RESULTS AND CONCLUSION: Cell relative growth rate was greater than 80% and the toxicity reaction ranked level 1 in all polyurethane groups, reagent control group and negative control group. In the above-mentioned groups, cells adhered to the cavity wall with normal appearance, plump soma and cytoplasm under microscope; in vitro cytotoxicity of these polyurethane materials reduced and their biocompatibility improved when the nanoAg-SiO2 content decreased. Cell relative growth rate became lower (P < 0.05) as culture-time prolonged and reduced to 8.7% after 72 hours in the positive control group. Cells cultured in positive control group drifted in the leaching liquor with their trophy and round shape. Seven kinds of polyurethane materials with different contents of nanoAg-SiO2 have good in vitro cellular compatibility and comply with the requirements of in vitro experiments for medical materials with their toxicity reaction ranking level 1.

Key words: biomaterials, nano-biological materials, nanoAg-SiO2, polyurethane, in vitro, L929 cells, relative growth rate, cytotoxicity, antibacterial properties, MTT colorimetric method, provincial grants-supported paper, biomaterial photographs-containing paper

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