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The regenerative potential of Tideglusib and CHIR99021 small molecules as potent odontogenic differentiation enhancers of human dental pulp stem cells

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Abstract

Objectives

To assess the effect of Tideglusib and CHIR99021 small molecules on the odontogenic differentiation potential of human dental pulp stem cells (hDPSCs) via Wnt/β-catenin pathway activation.

Methodology

hDPSCs were isolated from impacted third molars indicated for extraction and were characterized by flow cytometry. hDPSCs were then induced to differentiate into odontogenic lineage in the presence of Tideglusib and CHIR99021. Odontogenic differentiation was evaluated using Alizarin Red stain and RT-PCR for expression of odontogenic specific differentiation markers: DSPP, DMP1, ALP, OPN, and RUNX2 in relation to undifferentiated cells. RT-PCR was also conducted to assess the expression of Wnt/β-catenin pathway activation marker (AXIN2). One-way ANOVA Kruskal-Wallis test was used for statistical analysis.

Results

Wnt/β-catenin pathway was successfully activated by Tideglusib and CHIR99021 in hDPSCs where AXIN2 was significantly upregulated. Successful odontogenic differentiation was confirmed by Alizarin Red staining of calcified nodules. RT-PCR for odontogenic differentiation markers DSPP, DMP1, and RUNX expression by hDPSCs induced by CHIR99021 was higher than that expressed by hDPSCs induced by Tideglusib, whereas expression of OPN and ALP was higher in Tideglusib-induced cells than in CHIR99021-induced cells.

Conclusions

Both small molecules successfully induced odontogenic differentiation of hDPSCs through Wnt/β-catenin pathway activation.

Clinical relevance

These findings suggest that Tideglusib and CHIR99021 can be applied clinically in pulp regeneration to improve strategies for vital pulp regeneration and to promote dentine repair.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

  1. Murray PE, Garcia-Godoy F, Hargreaves KM, Article R (2007) Regenerative endodontics: a review of current status and a call for action. J Endod 33:377–390. https://doi.org/10.1016/j.joen.2006.09.013

    Article  PubMed  Google Scholar 

  2. Hargreaves KM, Law AS (2010) Regenerative endodontics. In: Cohen’s pathways of the pulp, Tenth Edition, Elsevier, pp 602–619

  3. Zaugg LK, Banu A, Walther AR et al (2020) Translation approach for dentine regeneration using GSK-3 antagonists. J Dent Res 99:544–551. https://doi.org/10.1177/0022034520908593

    Article  PubMed  CAS  Google Scholar 

  4. Birjandi AA, Sharpe P (2021) Wnt signalling in regenerative dentistry. Front Dent Med 2:725468

    Article  Google Scholar 

  5. Birjandi AA, Suzano FR, Sharpe PT (2020) Drug repurposing in dentistry; towards application of small molecules in dentin repair. Int J Mol Sci 21. https://doi.org/10.3390/ijms21176394

  6. Siddiqa A (2020) GSK 3 inhibitors releasing nanoparticles for dentine repair—IJSER Journal Publication. Int J Sci Eng Res 11:1714–1731

    Google Scholar 

  7. Wu D, Pan W (2010) GSK3: a multifaceted kinase in Wnt signaling. Trends Biochem Sci 35:161–168

    Article  PubMed  CAS  Google Scholar 

  8. Neves VCM, Babb R, Chandrasekaran D, Sharpe PT (2017) Promotion of natural tooth repair by small molecule GSK3 antagonists. Sci Rep 7. https://doi.org/10.1038/srep39654

  9. Tatullo M, Makeeva I, Rengo S et al (2019) Small molecule GSK-3 antagonists play a pivotal role in reducing the local inflammatory response, in promoting resident stem cell activation and in improving tissue repairing in regenerative dentistry. Histol Histopathol 34:1195–1203

    PubMed  CAS  Google Scholar 

  10. Sato N, Meijer L, Skaltsounis L et al (2004) Maintenance of pluripotency in human and mouse embryonic stem cells through activation of Wnt signaling by a pharmacological GSK-3-specific inhibitor. Nat Med 10:55–63. https://doi.org/10.1038/nm979

    Article  PubMed  CAS  Google Scholar 

  11. Hanna S, Aly R, Eldeen GN et al (2023) Small molecule GSK-3 inhibitors safely promote the proliferation and viability of human dental pulp stem cells—in vitro. Biomedicines 11. https://doi.org/10.3390/biomedicines11020542

  12. Tolosa E, Litvan I, Höglinger GU et al (2014) A phase 2 trial of the GSK-3 inhibitor tideglusib in progressive supranuclear palsy. Mov Disord 29:470–478. https://doi.org/10.1002/mds.25824

    Article  PubMed  CAS  Google Scholar 

  13. Comeau-Gauthier M, Tarchala M, Luna JLRG et al (2020) Unleashing β-catenin with a new anti-Alzheimer drug for bone tissue regeneration. Injury 51:2449–2459. https://doi.org/10.1016/j.injury.2020.07.035

    Article  PubMed  Google Scholar 

  14. Ring DB, Johnson KW, Henriksen EJ et al (2003) Selective glycogen synthase kinase 3 inhibitors potentiate insulin activation of glucose transport and utilization in vitro and in vivo. Diabetes 52:588–595. https://doi.org/10.2337/diabetes.52.3.588

    Article  PubMed  CAS  Google Scholar 

  15. Heng BC, Jiang S, Yi B et al (2019) Small molecules enhance neurogenic differentiation of dental-derived adult stem cells. Arch Oral Biol 102:26–38. https://doi.org/10.1016/j.archoralbio.2019.03.024

    Article  PubMed  CAS  Google Scholar 

  16. Govarthanan K, Vidyasekar P, Gupta PK et al (2020) Glycogen synthase kinase 3β inhibitor-CHIR 99021 augments the differentiation potential of mesenchymal stem cells. Cytotherapy 22:91–105. https://doi.org/10.1016/j.jcyt.2019.12.007

    Article  PubMed  CAS  Google Scholar 

  17. Cao N, Liang H, Huang J et al (2013) Highly efficient induction and long-term maintenance of multipotent cardiovascular progenitors from human pluripotent stem cells under defined conditions. Cell Res 23:1119–1132. https://doi.org/10.1038/cr.2013.102

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  18. Dang Le Q, Rodprasert W, Kuncorojakti S et al (2022) In vitro generation of transplantable insulin-producing cells from canine adipose-derived mesenchymal stem cells. Sci Reports 12(11):1–18. https://doi.org/10.1038/s41598-022-13114-3

    Article  CAS  Google Scholar 

  19. Hoglinger GU, Huppertz H-J, Wagenpfeil S et al (2014) Tideglusib reduces progression of brain atrophy in progressive supranuclear palsy in a randomized trial. Mov Disord 29:479–487. https://doi.org/10.1002/mds.25815

    Article  PubMed  CAS  Google Scholar 

  20. Del Ser T, Steinwachs KC, Gertz HJ et al (2013) Treatment of Alzheimer’s disease with the GSK-3 inhibitor tideglusib: a pilot study. J Alzheimer’s Dis 33:205–215. https://doi.org/10.3233/JAD-2012-120805

    Article  CAS  Google Scholar 

  21. Lektemur Alpan A, Çalişir M, Kizildağ A et al (2020) Effects of a glycogen synthase kinase 3 inhibitor tideglusib on bone regeneration with calvarial defects. J Craniofac Surg 31:1477–1482. https://doi.org/10.1097/SCS.0000000000006326

    Article  PubMed  Google Scholar 

  22. Gronthos S, Mankani M, Brahim J et al (2000) Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci USA 97:13625–13630. https://doi.org/10.1073/pnas.240309797

  23. Ahmed NE-MB, Aly RM, El-Massieh PMA, El Azeem AFA (2018) Comparing the stemness properties of dental pulp cells (Dpscs), collected, processed, and cryopreserved under different conditions. J Stem Cells 13:95–106

    CAS  Google Scholar 

  24. Huang GTJ, Gronthos S, Shi S (2009) Critical reviews in oral biology & medicine: mesenchymal stem cells derived from dental tissues vs. those from other sources: their biology and role in regenerative medicine. J Dent Res 88:792–806

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  25. Shi X, Mao J, Liu Y (2020) Pulp stem cells derived from human permanent and deciduous teeth: biological characteristics and therapeutic applications. Stem Cells Transl Med 9:445–464. https://doi.org/10.1002/SCTM.19-0398

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  26. Aly RM, Aglan HA, Eldeen GN, Ahmed HH (2022) Optimization of differentiation protocols of dental tissues stem cells to pancreatic β-cells. BMC Mol Cell Biol 23. https://doi.org/10.1186/s12860-022-00441-6

  27. Ledesma-Martínez E, Mendoza-Núñez VM, Santiago-Osorio E (2015) Mesenchymal stem cells derived from dental pulp: a review. Stem Cells Int 2016:4709572–4709572

  28. Nakashima M, Iohara K (2014) Mobilized dental pulp stem cells for pulp regeneration: initiation of clinical trial. J Endod 40:26–32. https://doi.org/10.1016/j.joen.2014.01.020

    Article  Google Scholar 

  29. Nakashima M, Iohara K, Murakami M et al (2017) Pulp regeneration by transplantation of dental pulp stem cells in pulpitis: a pilot clinical study. Stem Cell Res Ther 43:S82–S86. https://doi.org/10.1186/s13287-017-0506-5

    Article  CAS  Google Scholar 

  30. Szubarga A, Kamińska M, Kotlarz W et al (2021) Human stem cells-sources, sourcing and in vitro methods. Med J Cell Biol 9:73–85. https://doi.org/10.2478/acb-2021-0011

    Article  Google Scholar 

  31. Rezai Rad M, Hosseinpour S, Ye Q, Yao S (2021) Dental tissues originated stem cells for tissue regeneration. In: Regenerative approaches in dentistry. Springer International Publishing, pp 9–33

    Chapter  Google Scholar 

  32. Atari M, Gil-Recio C, Fabregat M et al (2012) Dental pulp of the third molar: a new source of pluripotent-like stem cells. J Cell Sci 125:3343–3356. https://doi.org/10.1242/JCS.096537

    Article  PubMed  CAS  Google Scholar 

  33. El-Moataz N, Ahmed B, Aly RM et al (2018) Comparing the stemness properties of dental pulp cells (DPSCs), collected, processed, and cryopreserved under different conditions. J Stem Cells 13:95–107

    Google Scholar 

  34. Bernkopf DB, Brückner M, Hadjihannas MV, Behrens J (2019) An aggregon in conductin/axin2 regulates Wnt/β-catenin signaling and holds potential for cancer therapy. Nat Commun 10(1):1–14. https://doi.org/10.1038/s41467-019-12203-8

    Article  CAS  Google Scholar 

  35. Tziafas D, Kodonas K (2010) Differentiation potential of dental papilla, dental pulp, and apical papilla progenitor cells. J Endod 36:781–789. https://doi.org/10.1016/J.JOEN.2010.02.006

  36. Sabbagh J, Ghassibe-Sabbagh M, Fayyad-Kazan M et al (2020) Differences in osteogenic and odontogenic differentiation potential of DPSCs and SHED. J Dent 101:103413. https://doi.org/10.1016/J.JDENT.2020.103413

    Article  PubMed  CAS  Google Scholar 

  37. Deng P, Huang J, Zhang Q et al (2023) The role of EMILIN-1 in the osteo/odontogenic differentiation of dental pulp stem cells. BMC Oral Health 23. https://doi.org/10.1186/s12903-023-02905-3

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Acknowledgements

We acknowledge the help of Dr Adham Abdel Azim in revising the manuscript and providing valuable advice.

Funding

Study was self-funded.

Author information

Authors and Affiliations

  1. Endodontics Department, Universidad Europea De Madrid (UEM), Madrid, Spain

    Samer Hanna & Ruth Perez Alfayate

  2. Molecular Genetics & Enzymology Department, Human Genetic & Genome Research Institute, National Research Centre, Dokki, Giza, Egypt

    Ghada Nour Eldeen

  3. Basic Dental Science Department, Oral Medicine & Dentistry Research Institute, National Research Centre, Dokki, Giza, Egypt

    Riham Aly

  4. Stem Cells Lab, Center of Excellence for Advanced Sciences, National Research Centre, 33 El Bohouth St., Dokki, Giza, 12622, Egypt

    Riham Aly

Authors
  1. Samer Hanna
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  2. Ghada Nour Eldeen
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  3. Ruth Perez Alfayate
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  4. Riham Aly
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Contributions

SH: contributed to the conception of the design, data acquisition, analysis, drafted the manuscript, critically revised the manuscript and giving final approval. RA: contributed to the conception of the design, data acquisition, analysis, or interpretation; critically revised the manuscript; gave final approval. GN: responsible for the molecular studies, critically revised the manuscript, and gave final approval. RP: Contributed to conception or design, analysis, or interpretation; critically revised the manuscript; gave final approval.

Corresponding author

Correspondence to Riham Aly.

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Ethics approval and consent to participate

The authors are accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. All methods were performed in accordance with the guidelines and regulations of the Declaration of Helsinki. Informed consent has been obtained from all participants. The experimental protocol was reviewed by the Ethical Committee of the Medical Research of the National Research Centre, Egypt and granted approval under the number 5435062021.

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The authors declare no competing interests.

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Hanna, S., Eldeen, G.N., Alfayate, R.P. et al. The regenerative potential of Tideglusib and CHIR99021 small molecules as potent odontogenic differentiation enhancers of human dental pulp stem cells. Clin Oral Invest 28, 48 (2024). https://doi.org/10.1007/s00784-023-05452-x

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