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3D Porous Chitosan Scaffolds Suit Survival and Neural Differentiation of Dental Pulp Stem Cells

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Abstract

A key aspect of cell replacement therapy in brain injury treatment is construction of a suitable biomaterial scaffold that can effectively carry and transport the therapeutic cells to the target area. In the present study, we created small 3D porous chitosan scaffolds through freeze-drying, and showed that these can support and enhance the differentiation of dental pulp stem cells (DPSCs) to nerve cells in vitro. The DPSCs were collected from the dental pulp of adult human third molars. At a swelling rate of ~84.33 ± 10.92 %, the scaffold displayed high porosity and interconnectivity of pores, as revealed by SEM. Cell counting kit-8 assay established the biocompatibility of the chitosan scaffold, supporting the growth and survival of DPSCs. The successful neural differentiation of DPSCs was assayed by RT-PCR, western blotting, and immunofluorescence. We found that the scaffold-attached DPSCs showed high expression of Nestin that decreased sharply following induction of differentiation. Exposure to the differentiation media also increased the expression of neural molecular markers Microtubule-associated protein 2, glial fibrillary acidic protein, and 2′,3′-cyclic nucleotide phosphodiesterase. This study demonstrates that the granular 3D chitosan scaffolds are non-cytotoxic, biocompatible, and provide a conducive and favorable micro-environment for attachment, survival, and neural differentiation of DPSCs. These scaffolds have enormous potential to facilitate future advances in treatment of brain injury.

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Acknowledgments

This work was supported by Graduate Student Innovation of Science and Technology Projects in Jiangsu Province and in Nantong University (Grants No.YKC13084); the “Top Six Types of Talents” Financial Assistance of Jiangsu Province Grant (No. 7 and No. 10); National Natural Science Foundation of China (Grant No. 31171038), Jiangsu Natural Science Foundation (BK2011385); the Grant of Nantong University for Innovation Talent and a project funded by the Priority Academic Program Development (PAPD) of Jiangsu Higher Education Institutions. We thank Department of Human Anatomy of Medical College in Nantong University for technical assistance and equipment support.

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Authors and Affiliations

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

    Xingmei Feng, Xiaohui Lu, Dan Huang, Jing Xing & Guijuan Feng

  2. Department of Human Anatomy, Medical College, Nantong University, Nantong, 226001, Jiangsu, People’s Republic of China

    Guohua Jin, Xin Yi, Xiang Chen, Lei Zhang & Xinhua Zhang

  3. Department of Rheumatology, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People’s Republic of China

    Zhifeng Gu

  4. Department of Gastroenterology and Hepatology, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People’s Republic of China

    Liren Li

  5. Department of Cardiology Medical, Tongzhou First People’s Hospital, Nantong, 226300, Jiangsu, People’s Republic of China

    Yuanzhou Lu

  6. Department of Neurosurgery, Yancheng First People’s Hospital, Yancheng, 224005, Jiangsu, People’s Republic of China

    Dekang Nie

Authors
  1. Xingmei Feng
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  2. Xiaohui Lu
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  3. Dan Huang
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  4. Jing Xing
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  5. Guijuan Feng
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  6. Guohua Jin
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  7. Xin Yi
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  8. Liren Li
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  9. Yuanzhou Lu
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  10. Dekang Nie
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  11. Xiang Chen
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  12. Lei Zhang
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  13. Zhifeng Gu
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  14. Xinhua Zhang
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Corresponding authors

Correspondence to Zhifeng Gu or Xinhua Zhang.

Additional information

Xingmei Feng and Xiaohui Lu contributed equally to this work.

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Feng, X., Lu, X., Huang, D. et al. 3D Porous Chitosan Scaffolds Suit Survival and Neural Differentiation of Dental Pulp Stem Cells. Cell Mol Neurobiol 34, 859–870 (2014). https://doi.org/10.1007/s10571-014-0063-8

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  • DOI: https://doi.org/10.1007/s10571-014-0063-8

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