采用扫描电子显微镜、激光扫描共聚焦显微镜、电感耦合等离子体发射光谱仪、流式细胞仪、酶标仪、细胞活性分析试剂盒和乳酸脱氢酶试剂盒(LDH)等方法比较研究了尺寸分别约为700 nm和15μm的二水草酸钙(COD)晶体对非洲绿猴肾上皮细胞(Vero)损伤的差异.结果显示这两种不同尺寸的COD晶体都能引起Vero活力下降、LDH释放量升高以及碘化丙啶染色增强,并表达带负电荷的骨桥蛋白,说明它们对Vero都具有损伤作用,且700 nm COD对Vero的细胞毒性及在细胞表面的粘附量均大于15μm COD晶体.本研究从尺寸减小后COD的晶面变化、表面电荷变化、表面粘附位点、细胞与COD晶体间氢键作用等方面解释了亚微米COD毒性显著增强的原因.
Cytotoxcity differences of submicron (700 nm) and micron (15μm) calcium oxalate dihydrate (COD) on African green monkey kidney epithelial (Vero) cells were investigated by scanning electron microscope, laser scanning confocal microscope, inductively coupled plasma emission spectrometer, flow cytometry, microplate reader, cell pro-liferation assay kit and lactate dehydrogenase (LDH) release assay kit. The results revealed that both submicron and micron COD could decrease cell viability, increase LDH release amount, enhance propidium iodide staining, and up-regulate negatively-charged osteopontin expression, which indicated that both submicron and micron COD crystals could damage Vero cells. Submicron COD exhibited higher cytotoxicity and more adhesion amount than micron COD. The higher cytoxicity of submicron COD was discussed from change of crystal face, surface charge, surface adhesion sites, and hydrogen bonding between cells and crystals.
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