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Hydroxyapatite–collagen augments osteogenic differentiation of dental pulp stem cells

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Abstract

The objectives of this study were to isolate and culture dental pulp stem cells (DPSCs) and to investigate their proliferation and osteogenic differentiation on hydroxyapatite–collagen (HA–Col) scaffold. DPSCs were characterized by fluorescence-activated cell sorting (FACS). Cultured cells were CD73+, CD90+, CD105+ and CD31−, CD45−. A commercially available HA–Col scaffold was used for culture of DPSCs. Cell attachment and viability of DPSCs cultured on scaffold was studied by sulforhodamine assay. Osteoblast differentiation capacity was studied by alkaline phosphatase assay and the effects of growth factors such as PDGF, IGF1 and FGF2 were further studied. Scanning electron microscopy (SEM) of cell seeded scaffolds was also performed. We found that DPSCs cultured exhibited the characteristic mesenchymal stem cells (MSCs) morphology and differentiation properties. Scaffold was found to be non-cytotoxic and had good biocompatibility in vitro. Osteoblast differentiation ability was found to increase at higher concentration of scaffold and additive effects were observed with the use of growth factors. In SEM, cells appeared to cover the entire surface of the scaffold forming continuous cell layer and extending filopodial extensions. HA–Col scaffold is apt for MSCs attachment and proliferation in vitro. Their unique self-renewal and multilineage differential potential make them ideal for use in regenerative medicine. The limitations of currently available bone graft materials have led to the emergence of tissue engineering using mesenchymal stem cells (MSCs). Since, HA–Col scaffold potentiated the proliferation and osteogenic differentiation of DPSCs, this biomimetic material may be an ideal one for maxillofacial and alveolar bone regeneration.

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Acknowledgements

We would like to thank Dr. Prabhaker Mishra, Department of Biostatistics and Health Informatics, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India, for providing assistance in statistical analysis. This project was partially funded by the Department of Health Research, New Delhi, India (DHR-MRU 014).

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Authors and Affiliations

  1. DHR MRU Lab, Dept of Oral & Maxillofacial Surgery, Faculty of Dental Sciences, King George’s Medical University, Lucknow, 226003, India

    Shilpa Trivedi, Sumit Kumar & Divya Mehrotra

  2. Department of Center for Advanced Research (Stem Cell/Cell Culture Lab), King George’s Medical University, Lucknow, 226003, India

    Kamini Srivastava, Tajindra Singh Saluja, Hari Shyam, Shailendra K Saxena & Satyendra Kumar Singh

  3. Department of Biochemistry, All India Institute of Medical Sciences, Rishikesh, Rishikesh, Uttarakhand, India

    Anjana Singh

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  1. Shilpa Trivedi
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  2. Kamini Srivastava
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  4. Hari Shyam
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  5. Sumit Kumar
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  7. Shailendra K Saxena
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  9. Satyendra Kumar Singh
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Correspondence to Divya Mehrotra or Satyendra Kumar Singh.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Supplementary material 1 (DOCX 20 kb)

10266_2019_464_MOESM2_ESM.pdf

Supplementary material 2 Supplementary Fig. 1. After 10 days of osteogenic differentiation concentration of RANKL was determined by ELISA in cell lysate of DPMSCs grown with or without HA–collagen scaffold. Data are presented as mean ± SEM (n = 2). RANKL, Receptor Activator of Nuclear Factor Kappa. (PDF 5 kb)

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Trivedi, S., Srivastava, K., Saluja, T.S. et al. Hydroxyapatite–collagen augments osteogenic differentiation of dental pulp stem cells. Odontology 108, 251–259 (2020). https://doi.org/10.1007/s10266-019-00464-0

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  • DOI: https://doi.org/10.1007/s10266-019-00464-0

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