中国组织工程研究

中国组织工程研究

• 组织工程口腔材料 tissue-engineered oral materials • 上一篇    下一篇

载银磷酸锆可影响293T细胞的生长与增殖

刘  晶1，孙迎春2，曾妃菲2   

  1. 1天津市红桥区口腔医院，天津市  300091 2天津医科大学口腔医院修复科，天津市  300070
  • 收稿日期:2012-08-06 修回日期:2012-09-10 出版日期:2013-05-21 发布日期:2013-05-21
  • 通讯作者: 孙迎春，副教授，科主任，天津医科大学口腔医院修复科，天津市 300070 sycem@hotmail.com 曾妃菲，天津医科大学口腔医院修复科，天津市 300070 675368995@qq.com
  • 作者简介:刘晶，男，1966年生，天津市人，汉族，1989年天津医学院毕业，院长，主要从事口腔医学临床及基础研究。 liujing8489@163.com
  • 基金资助:

    天津市卫生局科技基金项目(09kz100)“银系纳米抗菌剂对人口腔粘膜上皮细胞生长活力影响的研究”

Silver-zirconium phosphate impacts the vitality of 293T cells in vitro

Liu Jing1, Sun Ying-chun2, Zeng Fei-fei2   

  1. 1 Stomatology Hospital of Hongqiao District, Tianjin  300091, China
    2 Department of Prosthodontics, Stomatology Hospital of Tianjin Medical University, Tianjin  300070, China
  • Received:2012-08-06 Revised:2012-09-10 Online:2013-05-21 Published:2013-05-21
  • Contact: Sun Ying-chun, Associate professor, Department of Prosthodontics, Stomatology Hospital of Tianjin Medical University, Tianjin 300070, China sycem@hotmail.com Zeng Fei-fei, Department of Prosthodontics, Stomatology Hospital of Tianjin Medical University, Tianjin 300070, China 675368995@qq.com
  • About author:Liu Jing, Stomatology Hospital of Hongqiao District, Tianjin 300091, China liujing8489@163.com
  • Supported by:

    the Science and Technology Foundation of Tianjin Health Bureau, No. 09kz100

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摘要：

背景：研究表明6种国内外知名品牌口腔抗菌纳米载银无机抗菌剂中载银磷酸锆的杀菌性和安全性最佳。
目的：进一步观察载银磷酸锆对体外培养人胚肾上皮293T细胞生长、超微结构及胰岛素样生长因子1表达与分泌的影响。
方法：采用CCK-8方法检测5，2.5，1.25，0.625 g/L载银磷酸锆稀释液对293T细胞增殖的影响。根据结果选择对细胞增殖影响最大的质量浓度处理293T细胞，光学倒置显微镜观察细胞形态变化，于透射电镜下观察细胞超微结构及代谢变化，RT-PCR检测细胞胰岛素样生长因子1 mRNA的表达，ELISA法检测细胞培养上清液中胰岛素样生长因子1蛋白的水平。
结果与结论：载银磷酸锆作用于293T细胞48 h，1.25 g/L组细胞毒性为Ⅱ级，轻度细胞毒性，其余质量浓度组均为Ⅰ级，无细胞毒性。1.25 g/L载银磷酸锆处理293T细胞24，36，48 h后，细胞形态发生异常，细胞膜破损，出现轻度中毒现象，尤以48 h表现明显。1.25 g/L载银磷酸锆处理48 h后，293T细胞中胰岛素样生长因子1 mRNA的表达及上清液中胰岛素样生长因子1蛋白水平与正常培养细胞差异无显著性意义(P > 0.05)。表明载银磷酸锆影响293T细胞的生长及增殖，具有一定的细胞毒性，其在牙科材料中的应用还有待于进一步研究。

关键词: 生物材料, 组织工程口腔材料, 无机抗菌剂, 磷酸锆, 纳米, 口腔生物材料, 细胞增殖, 细胞毒性, 生物相容性；293T细胞, 省级基金

Abstract:

BACKGROUND: Studies have shown that silver-zirconium phosphate in the six kinds of well-known brands of oral nanosilver inorganic antibacterial agents has the best sterilization and safety.  
OBJECTIVE: To explore the influence of silver-zirconium phosphate antimicrobial agent FUMAT T200-4 on the cell growth, morphology, ultrastructure, insulin-like growth factor 1 expression and secretion of 293T cells in vitro.
METHODS: FUMAT T200-4 was prepared into diluents of 5, 2.5, 1.25, 0.625 g/L mass concentration. Cell Counting Kit-8 was employed to detect the impact on 293T cells proliferation with diluents of variousc oncentrations. After 293T cells were treated with1.25 g/L FUMAT T200-4, the cell morphology was observed using inverted phase contrast microscope. And further transmission electron microscope was used to observe the ultrastructure of 293T cells. In addition, reverse transcription-PCR was used to investigate cellular insulin-like growth factor mRNA expression and enzyme linked immunosorbent assay was used to detect the content of insulin-like growth factor protein in the supernatant solution.  
RESULTS AND CONCLUSION: After 293T cells were treated with FUMAT T200-4 for 48 hours, the cytotoxicity of 293T cells was graded as II for of 1.25 g/L showing mild cytotoxicity and I for the other concentrations showing no toxicity. The images of inverted phase contrast microscope showed that the cell shape after treated with    1.25 g/L for 24, 36, and 48 hours became abnormal which showed mild cell toxicity, especially after 48 hours. The ultrastructure images of transmission electron microscope showed nanoparticles entered 293T cells and damaged cell membrane. There was no significant deviation in cellular insulin-like growth factor 1 mRNA and protein expression after treated with 1.25 g/L for 48 hours in contrast to the control group (P > 0.05). Silver-zirconium phosphate can influence 293T cells proliferation and growth, exhibiting certain cytotoxicity to 293T cells, and its application in dental materials still needs further study.

Key words: biomaterials, tissue-engineered oral materials, inorganic antibacterial agents, zirconium phosphate, nano, oral biomaterials, cell proliferation, cytotoxicity, biocompatibility, 293T cells, provincial grants-supported paper

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