Effect of Crystal Size of Calcium Oxalate Dihydrate on Cytotoxicity of African Green Monkey Kidney Epithelial Cells

doi:10.15541/jim20150332

Abstract

Abstract:

Cytotoxcity differences of submicron (700 nm) and micron (15 μm) 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 proliferation 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.

Key words: crystal adhesion, size effect, cytotoxicity, calcium oxalate

CLC Number:

TQ174

GAN Qiong-Zhi, ZHANG Chong-Yu, OUYANG Jian-Ming. Effect of Crystal Size of Calcium Oxalate Dihydrate on Cytotoxicity of African Green Monkey Kidney Epithelial Cells[J]. Journal of Inorganic Materials, 2016, 31(3): 317-323.

Figures/Tables 7

Fig. 1 XRD patterns (a, b) and SEM images (c, d) of submicron/micron COD crystalsScale bar in (c) is 1 μm, in (d) 10 μm

Fig. 2 Changes in cell viability (a) and LDH release amount (b) of Vero cells after exposure to submicron/micron COD crystals

Fig. 3 PI staining results of Vero cells after exposure to submicron/micron COD crystals for 6 h(a) Control group; (b) Micron COD crystals treated group; (c) Submicron COD crystals treated group. Crystal concentration: 200 μg/mL. Magnification×600

Fig. 4 Intracellular ROS level of Vero cells after exposure to submicron/micron COD crystals for 6 h(a) Control group; (b) Micron COD crystals treated group; (c) Submicron COD crystals treated group

Fig. 5 OPN expression in Vero cells after exposure to submicron/micron COD crystals for 6 h(a) Control group; (b) Micron COD crystals treated group; (c) Submicron COD crystals treated group Green color represents OPN

Fig. 6 SEM images of submicron/micron COD crystals adhered to Vero cells for 6 h(a) Control group; (b) Micron COD crystals treated group; (c) Submicron COD crystals treated group Crystal concentration: 200 μg/mL

Fig. 7 Schematic representation of morphology and crystal face (100) and (101) variation of COD crystals with decreased size

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