噻唑蓝试验(MTT)法评价纳米TiO2的细胞毒性
EVALUATION ON THE CYTOTOXICITY OF TiO2 NANOPARTICLES BY MTT ASSAY IN MACROPHAGES AND LYMPHOCYTES
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摘要: 探讨纳米TiO2颗粒对原代培养的巨噬细胞、淋巴细胞的毒性效应及作用机制.选用了平均粒径为15nm左右的纳米TiO2制备颗粒悬液,设立6个剂量组(0、2、20、60、100、200μg·ml-1)分别对小鼠巨噬细胞、淋巴细胞进行24、48h染毒培养,利用细胞形态学观察和噻唑蓝试验(MTT比色法)检测纳米TiO2颗粒对巨噬细胞和淋巴细胞活性的影响.结果表明,纳米TiO2颗粒均能明显影响巨噬细胞和淋巴细胞的生长形态,染毒24h后,巨噬细胞和淋巴细胞都呈现不同程度的回缩变形,细胞间隙增大,巨噬细胞内颗粒物随纳米TiO2颗粒浓度的增大而增多,细胞折光性下降;纳米TiO2颗粒在24h内对巨噬细胞具有明显的抑制作用,而对淋巴细胞却有一定的增殖作用;在48h内都呈现一定的抑制作用,且存在剂量-效应关系,随着纳米TiO2浓度的升高,其对巨噬细胞的毒性也显著增大.Abstract: This study was initiated to explore the toxic effects and mechanism of titanium dioxide nanoparticles to macrophages and lymphocytes in vitro, cells were exposed to the average size of the titanium dioxide nanoparticles was 15 nm at six different concentrations(0、2、20、60、100、200 μg·ml-1). Cellular morphology and MTT assay were carried out to evaluate the viability of particle-treated cells after 24h and 48h exposure. Results showed that the titanium dioxide nanoparticles impacted the morphology of macrophages and lymphocytes evidently. Macrophages and lymphcytes were presented vary degree retraction distortion and increased cell spaces and sparse arrangement after 24h exposure, as well as increased intracellular particles and reduced cellular transparency. The particles in macrophages increased with the increasing concentrations of titanium dioxide nanoparticles. The titanium dioxide nanoparticles inhibited the growth of macrophages, while promoted the lymphocytes in 24h. However, it inhibited the macrophages and lymphocytes growth markedly in 48h. The effect of titanium dioxide nanoparticles on viability inhibition was significantly enhanced in a dose-dependent manner and showed the toxicity was strengthed with the concentrations of titanium dioxide nanoparticles increased.
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Key words:
- titanium dioxide nanoparticles /
- macrophage /
- lymphocyte /
- MTT
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