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Transcriptomic profiling of human dental pulp cells treated with tricalcium silicate–based cements by RNA sequencing

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Abstract

Objectives

Tricalcium silicate (TCS)–based biomaterials induce differentiation of human dental pulp cells (hDPCs) into odontoblasts/osteoblasts, which is regulated by the interplay between various intracellular pathways and their resultant secretome. The aim of this study was to compare the transcriptome-wide effects by next-generation RNA sequencing of custom-prepared hDPCs stimulated with TCS-based biomaterials: ProRoot white MTA (WMTA) (Dentsply, Tulsa; Tulsa, OK) and Biodentine (Septodont, Saint Maur des Fosses, France).

Methods

Self-isolated hDPCs were seeded in a 6-well plate at a density of 5 × 105 cells per well. ProRoot white MTA and Biodentine were then placed in transwell inserts with a pore size of 0.4 μm and inserted in the well plate. RNA sequencing was performed after 3 and 7 days treatment. For post-validation, RT-PCR analyses were done on some of the RNA samples used for RNA sequencing.

Results

Our RNA sequencing results for the first time identified 7533 differentially expressed genes (DEGs) between different treatments and the number of DEGs in Biodentine was higher than ProRoot WMTA at both 3 and 7 days. Despite their differential gene expression, both the TCS-based biomaterial treatments showed gene expressions mainly involved in odontoblast differentiation, angiogenesis, neurogenesis, dentinogenesis, and tooth mineralization.

Conclusions

The results of the present study illustrate that several important signalling pathways are induced by hDPCs stimulated with TCS-based biomaterials.

Clinical relevance

The differential expression of the genes associated with odontogenesis, angiogenesis, neurogenesis, dentinogenesis, and mineralization may affect the prognosis of teeth treated with Biodentine or ProRoot white MTA.

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Acknowledgments

The authors would like to thank the VIB Nucleomics Core (www.nucleomics.be) that performed the library preparation, sequencing, and statistical data analysis.

Funding

This research was funded by Septodont, Saint Maur des Faussés, France.

Author information

Authors and Affiliations

  1. Department of Paediatric Dentistry & Special Care, PAECOMEDIS Research Cluster, Ghent University, Ghent University Hospital, 9000, Ghent, Belgium

    Elanagai Rathinam, Sivaprakash Rajasekharan & Luc Martens

  2. Department of Internal Medicine and Paediatrics, Ghent University, Ghent University Hospital, 9000, Ghent, Belgium

    Srinath Govindarajan & Dirk Elewaut

  3. Unit for Molecular Immunology and Inflammation, VIB-Center for Inflammation Research, Technologiepark 71, Zwijnaarde, 9052, Ghent, Belgium

    Srinath Govindarajan & Dirk Elewaut

  4. Tissue Engineering and Biomaterials Group, Department of Human Structure and Repair, Ghent University, Ghent University Hospital, 9000, Ghent, Belgium

    Heidi Declercq

  5. Tissue Engineering Lab, Department of Development and Regeneration, KU Leuven, 8500, Kortrijk, Belgium

    Heidi Declercq

  6. Department of Reconstructive Dentistry and Oral Biology, Dental School, Ghent University, Ghent University Hospital, 9000, Ghent, Belgium

    Peter De Coster

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  1. Elanagai Rathinam
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Contributions

Elanagai Rathinam: conceptualization, isolation of stem cells, sample preparation, conducting all experiments, result interpretation, analysis, and writing of original draft; Srinath Govindarajan: devising the methodology, sample preparation, RT-PCR experiments, result analysis, and editing the final draft; Sivaprakash Rajasekharan: result analysis and editing the final draft; Heidi Declercq: RT-PCR experiments and editing the final draft; Dirk Elewaut: RT-PCR experiments, interpretation, and editing the final draft; Peter De Coster: result interpretation, supervision, and editing the final draft; Luc Martens: result interpretation, supervision, and editing the final draft. All authors gave their final approval and agree to be accountable for all aspects of the work.

Corresponding author

Correspondence to Elanagai Rathinam.

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Conflict of interest

Dr. Govindarajan holds a post-doctoral fellowship from the Fund for Scientific Research – Flanders (FWO). The research was supported by grants from the Group-ID Multidisciplinary Platform (MRP) of Ghent University (Dr. Elewaut) and Fund for Scientific Research – Flanders (Dr. Elewaut). Dr. Rajasekahran holds a post-doctoral fellowship from the Fund for Scientific Research – Flanders (FWO). The authors would like to report research grants and non-financial support (i.e., cement materials free of charge) from Septodont, France. Outside of the submitted work, Dr. Martens and Dr. Rajasekharan report personal fees (honoraria/lecture fees/educational courses). In addition, Dr. Martens would like to report research grants from Septodont, France, negotiated with the Ghent University. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Ethical approval

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. Ethical approval was obtained from the Ethical Committee of University Hospital, Ghent, Belgium, according to laws of ICH Good Clinical Practice (GE11-LM-go-2006/57).

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Informed consent was obtained from all individual participants included in the study.

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Rathinam, E., Govindarajan, S., Rajasekharan, S. et al. Transcriptomic profiling of human dental pulp cells treated with tricalcium silicate–based cements by RNA sequencing. Clin Oral Invest 25, 3181–3195 (2021). https://doi.org/10.1007/s00784-020-03647-0

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