effect of different concentrations of carbamide peroxide on the expression of autophagy related genes in human dental pulp stem cells

Abstract

Considering that autophagy plays an important role in the self-renewal and differentiation of stem cells, therefore, knowing the effective substances in autophagy has therapeutic benefits for a wide range of diseases. Therefore, the results of this project will show whether carbamide peroxide (different concentrations of this substance) can increase the survival potential and odontogenic differentiation of dental pulp stem cells by regulating autophagy. One of the important advantages of this method is its safety, in which the stem cell is not tampered with genetically, and the fact that this method can be used without risk in clinical medicine. Materials and methods: Dental pulp cells with cell line DPS_7 and cell bank code IBRCC10371 were purchased from the Genetic Reserve Center. Stem cells were treated with different concentrations of carbamide peroxide and their survival percentage was checked using MTT test. In order to measure the expression level of genes related to autophagy, the cells were treated for 24 hours with doses lower than the IC50 of carbamide peroxide, and total RNA of the cells was extracted using the RNA extraction kit based on the protocol of the manufacturer of the kit. Then RNA was converted into complementary DNA (cDNA) using cDNA synthesis kit. Finally, the expression level of autophagy genes (LC3, Beclin-1, p62) was investigated using Real time PCR technique. All tests were repeated three times and the group that did not receive carbamide peroxide was considered as the control group. All the results obtained from the treated groups and the control group were analyzed by Graphpad Prism software. Results: A concentration of 15 μg/mL was the IC50 concentration for carbamide peroxide and a concentration of 25 μg/mL was identified as a toxic concentration. At the concentration of 15 μg/mL, the expression level of Becline-1 and LC3 gene increased and the expression level of P62 gene decreased significantly. Dental pulp stem cells were exposed to carbamide peroxide as an oxidative agent and this process triggered the autophagy process. Conclusion: Carbamide peroxide, as an oxidative agent, stimulated the autophagy process in pulp stem cells.