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The Effect of Bone and Dentin Matrix Derivatives on the Differentiation of Human Dental Pulp Stem Cells for Osteogenesis and Dentinogenesis in a Scaffold-Free Culture

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Abstract

Purpose

The most important challenges in tissue engineering are to find an effective growth/differentiating factor for stem cell induction while retaining high cell vitality and convert to correct end cell linage. In this study, for the first time, self-administered dentin matrix derivatives were used as a growth/differentiation factor for dentin regeneration.

Methods

In the fourth passage, colonies of human dental pulp stem cells (hDPSCs) were extracted; then, the purified cells were evaluated based on the percentage of vital cells by the MTT method. Afterward, hDPSCs in the first group were cultured with self-administered dentin derivative signal (DDS), while those in the second group with demineralized freeze-dried bone allograft (DFDBA) as a signal for 3, 7, and 10 days, and then compared with group 3 (negative control) that contained only hDPSC colonies. Three groups were evaluated based on cell differentiation and expression of osteoblast and odontoblast cell markers by immunocytochemistry (ICC) staining.

Results

In the first group, hDPSCs were differentiated into odontoblast and in the second group into osteoblast. The mean expression percentage of the dentin sialophosphoprotein (DSPP) marker in differentiated cells in the first group was significant at days 3, 7, and 10 (p ≤ 0.0001). Also, the mean expression of the bone sialoprotein (BSP) marker was significant in the second group on days 3, 7, and 10 (p ≤ 0.0001).

Conclusion

hDPSCs in scaffold-free culture enable to differentiate to odontoblasts and osteoblasts through DDS and DFDBA. Both the DSPP and BSP markers were increasingly expressed with time and effective in cell induction. Future in vivo studies can recommend to investigate the role of such natural growth factors in regenerative endodontic and dentistry in clinics.

Lay Summary

This study evaluated the differentiation capability of dental pulp stem cells by two bioactive materials, self-administered dentin derivative signal (DDS) and demineralized freeze-dried bone allograft (DFDBA), as induction signals. The challenge in this regard is to find a simple, suitable, and efficient method of DDS preparation, retaining proteins undamaged and ensuring enough disinfected particles for cell exposure. For this purpose, we investigated freeze-dried methods that are commonly applied in material preparation, and tested DDS with the scaffold-free stem cell’s culture comparing with DFDBA that exhibits great induction potential to be used for tissue engineering in dentistry.

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Data Availability

Data generated or analyzed data during this study are included in this manuscript and are available from the corresponding author upon reasonable request.

Abbreviations

hDPSCs:

Human dental pulp stem cells

DFDBA:

Demineralized freeze-dried bone allograft

ICC:

Immunocytochemistry

DSPP:

Dentin sialophosphoprotein

BSP:

Bon sialoprotein

PDL:

Periodontal ligament

SD:

Standard deviation

SDF-1:

Stromal cell-derived factor-1

EMD:

Enamel matrix derivative

COL1A1:

Collagen type 1

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Funding

In this research, we used the financial support of Semnan University of Medical Sciences Research and Technology Vice-Chancellor with the code 1359, as well as the executive support of the Nervous System Stem Cell Research Center.

Author information

Authors and Affiliations

  1. Oral and Maxillofacial Medicine Department, Dental School, Semnan University of Medical Sciences, Semnan, Iran

    Maryam Jalili Sadrabad

  2. Cancer Research Center, Semnan University of Medical Sciences, Semnan, Iran

    Maryam Jalili Sadrabad

  3. Department of Tissue Engineering and Applied cell Sciences, School of Medicine ,Semnan University of Medical Science, Semnan, Iran

    Maryam Jalili Sadrabad

  4. Nervous System Stem Cells Research Center and Dept. of Anatomical Sciences, School of Medicine, Semnan University of Medical Sciences, Semnan, Iran

    Hamid-Reza Sameni & Sam Zarbakhsh

  5. Social Determinants of Health Research Center, Semnan University of Medical Science, Semnan, Iran

    Raheb Ghorbani

  6. Dental Faculty, Semnan University of Medical Sciences, Semnan, Iran

    Amin Naghipoor & Alireza Jarahi

Authors
  1. Maryam Jalili Sadrabad
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  2. Hamid-Reza Sameni
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  3. Sam Zarbakhsh
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  4. Raheb Ghorbani
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  5. Amin Naghipoor
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Contributions

M.J. designed the study, performed and wrote the manuscript draft, and edited and submitted it. H.R. S. assisted in research design, performed and supervised the laboratory process, and edited the final manuscript. S.Z. supervised the laboratory process and edited the final manuscript. R.G.H. analyzed the data and kindly adjusted the manuscript. A.N. and A.J. searched the literature, gathered data, and cooperated in writing the manuscript. Moreover, all authors have read and approved the manuscript.

Corresponding author

Correspondence to Hamid-Reza Sameni.

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Ethics Approval

This research project (ref no: IR.SEMUMS.REC.1396.246) was approved by the ethics committee of Semnan University of Medical Science, Semnan, Iran.

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Not applicable.

Competing Interests

The authors declare no competing interests.

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Jalili Sadrabad, M., Sameni, HR., Zarbakhsh, S. et al. The Effect of Bone and Dentin Matrix Derivatives on the Differentiation of Human Dental Pulp Stem Cells for Osteogenesis and Dentinogenesis in a Scaffold-Free Culture. Regen. Eng. Transl. Med. 9, 416–423 (2023). https://doi.org/10.1007/s40883-022-00291-w

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  • DOI: https://doi.org/10.1007/s40883-022-00291-w

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