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Effects of Nerve Growth Factor and Basic Fibroblast Growth Factor Promote Human Dental Pulp Stem Cells to Neural Differentiation

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Abstract

Dental pulp stem cells (DPSCs) were the most widely used seed cells in the field of neural regeneration and bone tissue engineering, due to their easily isolation, lack of ethical controversy, low immunogenicity and low rates of transplantation rejection. The purpose of this study was to investigate the role of basic fibroblast growth factor (bFGF) and nerve growth factor (NGF) on neural differentiation of DPSCs in vitro. DPSCs were cultured in neural differentiation medium containing NGF and bFGF alone or combination for 7 days. Then neural genes and protein markers were analyzed using western blot and RT-PCR. Our study revealed that bFGF and NGF increased neural differentiation of DPSCs synergistically, compared with bFGF and NGF alone. The levels of Nestin, MAP-2, βIII-tubulin and GFAP were the most highest in the DPSCs + bFGF + NGF group. Our results suggested that bFGF and NGF signifiantly up-regulated the levels of Sirt1. After treatment with Sirt1 inhibitor, western blot, RT-PCR and immunofluorescence staining showed that neural genes and protein markers had markedly decreased. Additionally, the ERK and AKT signaling pathway played a key role in the neural differentiation of DPSCs stimulated with bFGF + NGF. These results suggested that manipulation of the ERK and AKT signaling pathway may be associated with the differentiation of bFGF and NGF treated DPSCs. Our date provided theoretical basis for DPSCs to treat neurological diseases and repair neuronal damage.

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Acknowledgements

This work was supported by Natural Science Foundation of China Grant (No. 81500809, 81501076); Jiangsu Natural Science Foundation (BK2011385); “Top Six Types of Talents” Financial Assistance of Jiangsu Province Grant (Nos. 2013-WSN-076, 2013-WSN-048); Nantong Health Bureau Youth Foundation of China (WQ2015016); Graduate Student Innovation of Science and Technology Projects in Jiangsu Province and in Nantong University (NO.SJLX-0588; NO.SJLX-0588); Nantong Natural Science Foundation (NO.BK2014038).

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    Authors and Affiliations

    1. Department of Spine Surgery, The Second Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People’s Republic of China

      Jinlong Zhang, Guofeng Bao, Guanhua Xu, Yuyu Sun, Lingling Wang, Jiajia Chen & Zhiming Cui

    2. Department of Stomatology, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People’s Republic of China

      Min Lian, Peipei Cao & Guijuan Feng

    3. Department of Stomatology, Wang Yi Dental Clinic of Mudu Town, Suzhou, 215000, Jiangsu, People’s Republic of China

      Yi Wang

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    1. Jinlong Zhang
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    Correspondence to Guijuan Feng or Zhiming Cui.

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    Jinlong Zhang, Min Lian and Peipei Cao contributed equally to this work.

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    Zhang, J., Lian, M., Cao, P. et al. Effects of Nerve Growth Factor and Basic Fibroblast Growth Factor Promote Human Dental Pulp Stem Cells to Neural Differentiation. Neurochem Res 42, 1015–1025 (2017). https://doi.org/10.1007/s11064-016-2134-3

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