Overexpression of Sema3A promotes osteogenic differentiation of dental pulp stem cells and MC3T3-E1

doi:10.12307/2024.122

Abstract

Abstract: BACKGROUND: Sema3A is a power secretory osteoprotective factor. However, studies about Sema3A-modified dental pulp stem cells (Sema3A-DPSCs) are rare.
OBJECTIVE: To explore the osteogenic differentiation ability of Sema3A-DPSCs and their regulatory effect on the osteogenic differentiation of the pre-osteoblast cell line MC3T3-E1.
METHODS: First, Sema3A-DPSCs were constructed using a lentivirus infection system carrying the Sema3A gene. Control lentivirus-treated DPSCs (Vector-DPSCs) were used as controls. Sema3A-DPSCs or Vector-DPSCs were co-cultured with proosteoblast line MC3T3-E1 at the ratio of 1∶1 and 1∶3 for 24 hours. Finally, the Sema3A-DPSCs, Vector-DPSCs and their co-cultured cells with MC3T3-E1 were cultured for osteogenic induction and differentiation. Osteogenic gene expression was detected by alkaline phosphatase staining, alizarin red staining and real-time quantitative RT-PCR to evaluate osteogenic differentiation ability.
RESULTS AND CONCLUSION: (1) Sema3A mRNA and protein expression levels in Sema3A-DPSCs were significantly up-regulated. The level of secreted Sema3A in cell supernatant was up-regulated. (2) Compared with the Vector-DPSCs, mRNA expressions of osteogenic genes alkaline phosphatase, Runt-related transcription factor 2, osteocalcin and Sp7 transcription factors in Sema3A-DPSCs were up-regulated; the activity of alkaline phosphatase was enhanced, and the formation of mineralized nodules increased. (3) There were no obvious differences in proliferation between Sema3A-DPSCs and Vector-DPSCs. (4) Compared with MC3T3-E1/Vector-DPSCs co-culture system, the expression of MC3T3-E1 osteogenic genes was up-regulated, and the total alkaline phosphatase activity was enhanced and more mineralized nodules were formed in the MC3T3-E1/Sema3A-DPSCs co-culture system. (5) The results suggest that overexpression of Sema3A can enhance the osteogenic differentiation of DPSCs. Overexpression of Sema3A in DPSCs can promote osteogenic differentiation of MC3T3-E1 in the DPSCs/MC3T3-E1 co-culture system.

Key words: stem cell-based therapy, gene therapy, Sema3A, dental pulp stem cell, osteoblast, osteogenic differentiation

CLC Number:

Wang Wen, Zheng Pengpeng, Meng Haohao, Liu Hao, Yuan Changyong. Overexpression of Sema3A promotes osteogenic differentiation of dental pulp stem cells and MC3T3-E1[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(7): 993-999.

Figures/Tables 4

References

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