Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (23): 3630-3635.doi: 10.12307/2021.031

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Transforming growth factor beta combined with bone morphogenetic protein-2 induces the proliferation and differentiation of mouse MC3T3-E1 cells

Zhou Wu1, Wang Binping2, Wang Yawen2, Cheng Yanan2, Huang Xieshan1   

  1. 1Stomatology Center, 2Department of oral implantology, Affiliated Haikou Hospital of Xiangya Medical School, Central South University, Haikou 570208, Hainan Province, China
  • Received:2020-06-10 Revised:2020-06-16 Accepted:2020-08-04 Online:2021-08-18 Published:2021-01-26
  • Contact: Wang Binping, Chief physician, Department of oral implantology, Affiliated Haikou Hospital of Xiangya Medical School, Central South University, Haikou 570208, Hainan Province, China
  • About author:hou Wu, Master, Stomatology Center, Affiliated Haikou Hospital of Xiangya Medical School, Central South University, Haikou 570208, Hainan Province, China
  • Supported by:
    the Natural Science Foundation of Hainan Province, No. 20168313 (to WBP)

Abstract: BACKGROUND: Growth factors are one of the key elements for bone formation, remodeling and repair. The combination of multiple growth factors to induce osteogenic differentiation of bone stem cells has certain advantages compared with single ones. However, what combination of concentrations can play the best inducing effect has not been fully documented.
OBJECTIVE: To explore the effects of transforming growth factor-β combined with bone morphogenetic protein-2 on the proliferation and differentiation of mouse MC3T3-E1 cells.
METHODS: Different concentrations of transforming growth factor-β and bone morphogenetic protein-2 were combined to cultivate MC3T3-E1 cells. The cells were then divided into six groups, including one control group and five experimental groups, and were tested at different time points. The proliferation of the cells was determined by MTT method. Real-time quantitative RT-PCR was used to detect Runx2 and osteocalcin gene expression. Alkaline phosphatase staining was used to observe the viability and function of osteoblasts. Alizarin red staining was used to observe the amount of calcium nodules.
RESULTS AND CONCLUSION: Whether transforming growth factor-β and bone morphogenetic protein-2 were used alone or in combination could significantly promote the proliferation of MC3T3-E1cells, promote the expression of Runx2 and osteocalcin, and increase the alkaline phosphatase activity and mineralization ability (P < 0.05). The osteogenic effect of two growth factors in combination was positively correlated with the concentration. Even the combination at a low concentration can achieve or surpass the effect of single use. The optimal combined concentration was 1 μg/L for transforming growth factor-β and 100 μg/L for bone morphogenetic protein-2. Therefore, transforming growth factor-β and bone morphogenetic protein-2 used alone or in combination can significantly promote the proliferation and osteogenesis of osteoblasts, which focus on different aspects and interact with each other at different stages of proliferation and differentiation. Within the concentration range of this study, 1 μg/L transforming growth factor-β plus 100 μg/L bone morphogenetic protein-2 show the best promotion effect.

Key words: MC3T3-E1 cells, transforming growth factor-β, bone morphogenetic protein-2, osteoblast, cell proliferation, cell differentiation

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