2023, Vol. 27 ›› Issue (19): 2960-2967

Effects of iRoot BP Plus combined with Yunnan Baiyao on mineralization of dental pulp stem cells under inflammatory conditions

Zhao Ying1, Liang Guangzhi2, Zhou Yuqi2, Wang Tianqi2, Liu Yunxia2, Liu Xiaoying3, Zhang Juanjuan2, Sun Yan2   

Abstract: BACKGROUND: As a permanent preservation of living pulp, pulpotomy has been gradually applied in clinical treatment. The selection and application of pulpotomy materials can greatly enhance its clinical efficacy and expand its application range.  
OBJECTIVE: To study the effects of Yunnan Baiyao, iRoot BP Plus and their combination on the mineralization of pulp stem cells under the condition of inflammation.
METHODS: Human primary dental pulp stem cells were cultured by tissue block method, and the effects of Yunnan Baiyao and iRoot BP Plus on the proliferation of dental pulp stem cells were determined by MTT assay to determine the optimal concentration of intervention quality. Passages 3-6 dental pulp stem cells were divided into five groups: normal control group, inflammatory control group, Yunnan Baiyao group, iRoot BP Plus group, and Yunnan Baiyao + iRoot BP Plus group. Cells in the inflammatory control group, Yunnan Baiyao group, iRoot BP Plus group, and Yunnan Baiyao + iRoot BP Plus group were treated with 1 μg/mL lipopolysaccharide for 2 hours to establish models and then stimulated with 75 μg/mL Yunnan Baiyao, 100 μg/mL iRoot BP Plus alone and their combination. Effects of drugs on cellular mineralization were determined by alkaline phosphatase assay, alizarin red S staining, real-time-PCR and western blot assay.  
RESULTS AND CONCLUSION: (1) Compared with the inflammatory control group, Yunnan Baiyao, iRoot BP Plus and their combination inhibited the mRNA expression of tumor necrosis factor-α (P < 0.001). The mRNA expression of tumor necrosis factor-α in Yunnan Baiyao + iRoot BP Plus group was significantly lower than that in Yunnan Baiyao group and iRoot BP Plus group (P < 0.001). (2) Compared with the inflammatory control group, the number of mineralized nodules and alkaline phosphatase activity were significantly increased after Yunnan Baiyao, iRoot BP Plus, and their combined application. The number of mineralized nodules and alkaline phosphatase activity in Yunnan Baiyao + iRoot BP Plus group were higher than those in Yunnan Baiyao and iRoot BP Plus groups (P < 0.001). (3) Compared with the inflammatory control group, the mRNA expression levels of DSPP, IBSP and MEPE increased after Yunnan Baiyao, iRoot BP Plus, and their combined application (P < 0.001). The mRNA expression levels of DSPP, IBSP, and MEPE in Yunnan Baiyao + iRoot BP Plus group were higher than those in Yunnan Baiyao group and iRoot BP Plus group (P < 0.001). (4) Compared with the inflammatory control group, phosphorylation of Akt and mTOR was increased in Yunnan Baiyao group and Yunnan Baiyao + iRoot BP Plus group (P < 0.001). (5) The results show that both Yunnan Baiyao and iRoot BP Plus can promote the mineralization of dental pulp stem cells, and the combined application of Yunnan Baiyao and iRoot BP Plus has a more significant mineralization effect. The effect of Yunnan Baiyao on promoting the mineralization of dental pulp stem cells is associated with the Akt/mTOR pathway.

Key words: pulpitis, Yunnan Baiyao, iRoot BP Plus, pulpotomy, inflammation, mineralization