Quantitative study on cytotoxicity of methylene blue to human nucleus pulposus cells by cell counting-kit 8 assay

doi:10.3969/j.issn.2095-4344.0240

Abstract

Abstract:

BACKGROUND: Our preliminary study has shown that methylene blue exerts toxic effect on human nucleus pulposus cells in a concentration-dependent manner, but the toxic range remains unclear.
OBJECTIVE: To determine the critical range of the cytotoxicity of methylene blue to nucleus pulposus cells by cell counting-kit 8 (CCK-8) assay.
METHODS: The nucleus pulposus was from a patient with intervertebral disc herniation. The nucleus pulposus cells were extracted and cultured. Passage 1 cells mere used to make cell suspensions. The cells were divided into nine groups for culture: blank control (only the medium, CCK-8 solution), control (only medium, cells and CCK-8 solution), and methylene blue groups (1%, 0.5%, 0.1%, 0.05%, 0.01% and 0.005% of methylene blue). The absorbance values were measured by CCK-8 assay at 24, 48, 72 and 96 hours after incubation. The cell viability was calculated, and the color was observed.
RESULTS AND CONCLUSION: The color in the control group was the darkest, and the color in the methylene blue groups became lighter with the concentration of methylene blue increasing, and the 0.05%, 0.01%, 0.005% and 0.001% methylene blue groups showed darker color similar to the control group. The absorbance values in the 0.1% methylene blue group were significantly less than those in the control group (P < 0.05). There were no significant differences in the absorbance values and cell viability between 1% and 0.5% methylene blue groups (P > 0.05). The absorbance values and cell viability in the 0.1% methylene blue group were significantly higher than those in the 0.5% methylene blue group, but were significantly less than those in the 0.05% methylene blue group (P < 0.05). Thus, methylene blue exerts cytotoxicity to human nucleus pulposus cells, and the critical value of toxicity is between 0.1% and 0.05% and close to 0.05%. However, the exact value needs a further investigation.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Methylene Blue, Cytotoxicity, Immunologic, Tissue Engineering

CLC Number:

R318

Feng Hua-long1, He Sheng-hua2, Huang Fei-qiang1, Lai Ju-yi1, Sun Zhi-tao2, Xu Fu-guang1, Lan Zhi-ming1. Quantitative study on cytotoxicity of methylene blue to human nucleus pulposus cells by cell counting-kit 8 assay[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(16): 2532-2536.

Figures/Tables 2

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