Hypoxia effects on the proliferation and differention of hydrogen peroxide-pretreated  MC3T3-E1

doi:10.3969/j.issn.2095-4344.2017.04.003

Abstract

Abstract:

Abstract
BACKGROUND: The intracellular accumulation of reactive oxygen species leads to oxidative stress. Hypoxia is widespread in physiological and pathological condition. Variation of bone proliferation and differentiation when bone tissues cultured or bone cells induced toxicity by reactive oxygen species under hypoxia have not yet been reported.
OBJECTIVE: To observe the biological characteristics of MC3T3-E1 pretreated with different concentrations of hydrogen peroxide (H2O2) in hypoxia, thus understanding the cell mechanism underlying prolonged bone healing in the elderly with osteoporosis and diabetes.
METHODS: The MC3T3-E1 cells pretreated with different concentrations of H2O2 were cultured in different oxygen concentrations. The proliferation of MC3T3-E1 was detected by cell counting kit-8. The cell differentiation was detected through alkaline phosphatase staining and alizarin red staining. Total RNAs were extracted and used for analyzing the mRNA levels of collage type 1, alkaline phosphatase and Cbfa1.
RESULTS AND CONCLUSION: When MC3T3-E1 pretreated with 200 μmol/L H2O2 for 6 hours, the cell proliferation was increased with time, but lower than that in the control group. The alkaline phosphatase activity was weakened, and the number of mineralized nodes was decreased at the early stage of differentiation. When MC3T3-E1 pretreated with 400 μmol/L H2O2 for 6 hours, the cell proliferation was decreased obviously. The alkaline phosphatase activity was still weakened, and the number of mineralized nodes was decreased further, but not affected by hypoxia. When MC3T3-E1 pretreated with 400 μmol/L H2O2 for 6 hours and then cultured in hypoxia, the mRNA expression of Cbfa1 was decreased, but the mRNA expressions of collage type 1 and alkaline phosphatase were significantly increased. These results suggest that MC3T3-E1 pretreated with low concentration of H2O2 show a significant decrease in proliferation, while MC3T3-E1 pretreated with a high concentration of H2O2 and cultured in hypoxia show a decrease in osteogenic differentiation, especially at the early stage of alkaline phosphatase formation.

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: Cell Hypoxia, Osteoblasts, Alkaline Phosphatase, Hypoxia-Inducible Factor 1, Tissue Engineering

CLC Number:

R318

Liang Jing1, 2, Wang Jun1, 2, Tang Chuan-ling3, Zhou Qi1, 2, Wei Li1, 2, Hu Fang-qiong1, 2, Wan Rong2. Hypoxia effects on the proliferation and differention of hydrogen peroxide-pretreated MC3T3-E1[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(4): 505-511.

Figures/Tables 2

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