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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、50μmol/L),孵育10min,波长670nm半导体激光照射5min,输出功率40mW,功率密度10mW/cm2。光照结束后继续培养24h,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≤25μmol/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>25μmol/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。
Objective To study the toxic effects of methylene blue (MB) -mediated photodynamic therapy (PDT) on human periodontal ligament fibroblasts (HPDLFs) cultured in vitro. Methods HPDLFs cultured in vitro were added to two different MB formulations (5 and 10 concentrations were set at 0.1, 1.0, 10, 25 and 50 μmol / L, respectively) for 10 min and 670 nm wavelength laser irradiation 5min, output power 40mW, power density 10mW / cm2. The cells were cultured for 24 h after the light was illuminated. Cytotoxicity was detected by MTT colorimetric assay. Morphological changes were observed under light microscope. Results There was no significant difference in the toxic effects of PDT mediated by two different formulations of MB on cultured HPDLFs (P> 0.05). When the concentration of MB was less than or equal to 25μmol / L, the cytotoxicity was 1 ~ 2, no obvious morphological changes were observed under the light microscope. When the concentration was> 25μmol / L, the cytotoxicity was grade 3, Round. Conclusion The toxic effects of PDT on HPDLFs are affected by the concentration of photosensitizer. PDT induced by PDT with two different dosage forms has a slight effect on HPDLFs cultured in vitro. There is no difference between the two formulations of MB-mediated PDT in vitro cytotoxicity.