中国实用口腔科杂志

中国实用口腔科杂志 ›› 2012, Vol. 5 ›› Issue (04): 214-218.

• 论著 • 上一篇    下一篇

不同剂型亚甲基蓝光动力疗法对人牙周膜成纤维细胞的体外毒性研究

  

  1. 1. 福建医科大学口腔医学院，福州  350002；2. 福建省高校口腔医学重点实验室， 福州  350002
  • 收稿日期:2012-01-09 修回日期:2012-03-02 出版日期:2012-04-15 发布日期:2012-05-16

  • Received:2012-01-09 Revised:2012-03-02 Online:2012-04-15 Published:2012-05-16

摘要：

目的    研究不同剂型亚甲基蓝（methylene blue，MB）介导的光动力疗法（photodynamic therapy，PDT）对体外培养的人牙周膜成纤维细胞（human periodontal ligament fibroblasts，HPDLFs）的毒性作用。方法    将体外培养的HPDLFs分别加入两种不同剂型MB中（水剂型和混合剂型，均设置5组浓度，分别为0.1、1.0、10、25、50 μmol/L），孵育10 min，波长670 nm半导体激光照射5 min，输出功率40 mW，功率密度10 mW/cm2。光照结束后继续培养24 h，用噻唑蓝（MTT）比色法检测细胞毒性，光镜下观察细胞形态学变化。 结果    两种不同剂型MB介导的PDT对体外培养的HPDLFs的毒性作用未见明显差异（P > 0.05）。当MB浓度≤ 25 μmol/L时，细胞毒性为1 ~ 2级，光镜下观察细胞未发现明显形态学改变；浓度> 25 μmol/L时，细胞毒性为3级，并可观察到细胞皱缩、变圆。结论    PDT对HPDLFs的毒性作用受光敏剂浓度影响，适当浓度的两种剂型MB介导的PDT对体外培养的HPDLFs影响轻微，且两种剂型MB介导的PDT对体外细胞毒性无差异。

关键词: 亚甲基蓝, 光动力疗法, 人牙周膜成纤维细胞, 细胞毒性

Abstract:

Objective    To investigate the photodynamic effect of two different formulations of methylene blue（MB）on the killing ability to human periodontal ligament fibroblasts（HPDLFs）in vitro. Methods    Cultured HPDLFs were incubated in a medium containing two different formulations of MB with 5 different concentration（0.1，1.0，10，25，50 μmol/L）for 10 minutes，and irradiated 5 minutes using a semiconductor laser（wavelength 670 nm，output power 40 mW，energy density 10 mW/cm2）. After treatment，the cells continued to be cultivated for 24 hours，and then the cytotoxicity was investigated by light microscope and MTT assay. Results    There was no significant difference in the cytotoxicity of the two different formulations of MB（P > 0.05）. When the concentration of the photosensitizers was no more than 25 μmol/L，the cells were normal in appearance and cytotoxicity score was at 1-2 grade；when the concentration was more than 25 μmol/L，the cells shrank and became round，and cytotoxicity score was grade 3. Conclusion    There is no significant difference in the cytotoxicity of the two different formulations of MB. The cytotoxicity is affected by the concentration of the photosensitizers. When using appropriate drug concentration，the cell has a lower toxicity.

Key words: methylene blue；photodynamic therapy；human periodontal ligament fibroblasts；cytotoxicity