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Application of BMP-2/FGF-2 gene–activated scaffolds for dental pulp capping

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Clinical Oral Investigations Aims and scope Submit manuscript

Abstract

Objectives

To evaluate the effect of non-viral gene therapy on human dental pulp stem cells (DPSCs) in an in vitro and an ex vivo model.

Materials and methods

Nanoplexes comprising polyethyleneimine (PEI) and plasmid DNA (pDNA) encoding for fibroblast growth factor-2 (pFGF-2) and bone morphogenic protein-2 (pBMP-2) were cultured with DPSCs to evaluate cytotoxicity, protein expression, and mineralization activity. Collagen scaffolds loaded with these nanoplexes or mineral trioxide aggregate (MTA) were utilized in an ex vivo tooth culture model to assess pulp response, over a period of 14 days. All nanoplex formulations were characterized for size and zeta potential by measuring dynamic light scattering and electrophoretic mobility, respectively.

Results

DPSCs treated with the nanoplexes showed increased cell proliferation and enhanced expression of BMP-2 and FGF-2 proteins. Collagen scaffolds containing PEI-pBMP-2 and/or pFGF-2 nanoplexes significantly increased cell proliferation, BMP-2 and FGF-2 expression, and mineralization when compared to MTA. Ex vivo histology showed a well-preserved pulp and healthy tissue in both the MTA and scaffold groups. Connective tissue in contact with the scaffold was dense and homogeneous, with some cells present in contact and within the scaffold.

Conclusion

Transfection of DPSCs with pBMP-2/pFGF-2 nanoplexes resulted in increased expression of BMP-2 and FGF-2, enhanced proliferation, and mineralization properties compared to MTA. These findings were supported by the ex vivo observations.

Clinical relevance

This biological approach in pulp capping brings new insights into the effective management of engineered pulp tissues, mainly those generated by the transplantation of DPSCs in empty root canals.

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References

  1. Witherspoon DE (2008) Vital pulp therapy with new materials: new directions and treatment perspectives—permanent teeth. Pediatr Dent 30:220–224

    PubMed  Google Scholar 

  2. Lanza R, Langer R, Vacanti J (2014) Principles of tissue engineering. Academic Press, San Diego

    Google Scholar 

  3. Diogenes A, Hargreaves KM (2017) Microbial modulation of stem cells and future directions in regenerative endodontics. J Endod 43:S95–S101

    Article  Google Scholar 

  4. Nakashima M, Iohara K (2017) Recent progress in translation from bench to a pilot clinical study on total pulp regeneration. J Endod 43:S82–S86

    Article  Google Scholar 

  5. Cavalcanti BN and Nör JE (2019) Current and future views on pulpal tissue engineering. Book title. Springer

  6. Graham L, Cooper PR, Cassidy N, Nor JE, Sloan AJ, Smith AJ (2006) The effect of calcium hydroxide on solubilisation of bio-active dentine matrix components. Biomaterials 27:2865–2873

    Article  Google Scholar 

  7. Murray PE, Windsor LJ, Smyth TW, Hafez AA, Cox CF (2002) Analysis of pulpal reactions to restorative procedures, materials, pulp capping, and future therapies. Crit Rev Oral Biol Med 13:509–520

    Article  Google Scholar 

  8. Yildirim S, Can A, Arican M, Embree MC, Mao JJ (2011) Characterization of dental pulp defect and repair in a canine model. Am J Dent 24:331

    PubMed  Google Scholar 

  9. Li Z, Cao L, Fan M, Xu Q (2015) Direct pulp capping with calcium hydroxide or mineral trioxide aggregate: a meta-analysis. J Endod 41:1412–1417

    Article  Google Scholar 

  10. Tomson P, Lumley P, Smith A, Cooper P (2017) Growth factor release from dentine matrix by pulp-capping agents promotes pulp tissue repair-associated events. Int Endod J 50:281–292

    Article  Google Scholar 

  11. Torabinejad M, Parirokh M, Dummer P (2018) Mineral trioxide aggregate and other bioactive endodontic cements: an updated overview–part II: other clinical applications and complications. Int Endod J 51:284–317

    Article  Google Scholar 

  12. Nakashima M, Reddi AH (2003) The application of bone morphogenetic proteins to dental tissue engineering. Nat Biotechnol 21:1025–1032

    Article  Google Scholar 

  13. Bakopoulou A, Papachristou E, Bousnaki M, Hadjichristou C, Kontonasaki E, Theocharidou A, Papadopoulou L, Kantiranis N, Zachariadis G, Leyhausen G (2016) Human treated dentin matrices combined with Zn-doped, mg-based bioceramic scaffolds and human dental pulp stem cells towards targeted dentin regeneration. Dent Mater 32:e159–e175

    Article  Google Scholar 

  14. Atluri K, Seabold D, Hong L, Elangovan S, Salem AK (2015) Nanoplex-mediated codelivery of fibroblast growth factor and bone morphogenetic protein genes promotes osteogenesis in human adipocyte-derived mesenchymal stem cells. Mol Pharm 12:3032–3042

    Article  Google Scholar 

  15. Khorsand B, Nicholson N, Do A-V, Femino JE, Martin JA, Petersen E, Guetschow B, Fredericks DC, Salem AK (2017) Regeneration of bone using nanoplex delivery of FGF-2 and BMP-2 genes in diaphyseal long bone radial defects in a diabetic rabbit model. J Control Release 248:53–59

    Article  Google Scholar 

  16. McMillan A, Nguyen MK, Gonzalez-Fernandez T, Ge P, Yu X, Murphy WL, Kelly DJ, Alsberg E (2018) Dual non-viral gene delivery from microparticles within 3D high-density stem cell constructs for enhanced bone tissue engineering. Biomaterials 161:240–255

    Article  Google Scholar 

  17. Cleland JL, Daugherty A, Mrsny R (2001) Emerging protein delivery methods. Curr Opin Biotechnol 12:212–219

    Article  Google Scholar 

  18. D'Mello S, Salem AK, Hong L, Elangovan S (2016) Characterization and evaluation of the efficacy of cationic complex mediated plasmid DNA delivery in human embryonic palatal mesenchyme cells. J Tissue Eng Regen Med 10:927–937

    Article  Google Scholar 

  19. Téclès O, Laurent P, Aubut V, About I (2008) Human tooth culture: a study model for reparative dentinogenesis and direct pulp capping materials biocompatibility. J Biomed Mater Res B 85:180–187

    Article  Google Scholar 

  20. Breunig M, Lungwitz U, Liebl R, Goepferich A (2007) Breaking up the correlation between efficacy and toxicity for nonviral gene delivery. Proc Natl Acad Sci 104:14454–14459

    Article  Google Scholar 

  21. Yamano S, Dai J, Moursi AM (2010) Comparison of transfection efficiency of nonviral gene transfer reagents. Mol Biotechnol 46:287–300

    Article  Google Scholar 

  22. Jaidev L, Chatterjee K (2019) Surface functionalization of 3D printed polymer scaffolds to augment stem cell response. Mater Des 161:44–54

    Article  Google Scholar 

  23. Kumar S, Raj S, Sarkar K, Chatterjee K (2016) Engineering a multi-biofunctional composite using poly (ethylenimine) decorated graphene oxide for bone tissue regeneration. Nanoscale 8:6820–6836

    Article  Google Scholar 

  24. Jaidev L, Chellappan DR, Bhavsar DV, Ranganathan R, Sivanantham B, Subramanian A, Sharma U, Jagannathan NR, Krishnan UM, Sethuraman S (2017) Multi-functional nanoparticles as theranostic agents for the treatment & imaging of pancreatic cancer. Acta Biomater 49:422–433

    Article  Google Scholar 

  25. Oliveira R, Tavares W, Reis A, Silva V, Vieira L, Ribeiro Sobrinho A (2018) Cytokine expression in response to root repair agents. Int Endod J 51:1253–1260

    Article  Google Scholar 

  26. Chmilewsky F, Jeanneau C, Laurent P, About I (2014) Pulp fibroblasts synthesize functional complement proteins involved in initiating dentin–pulp regeneration. Am J Pathol 184:1991–2000

    Article  Google Scholar 

  27. de Souza Costa CA, Duarte PT, de Souza PP, Giro EM, Hebling J (2008) Cytotoxic effects and pulpal response caused by a mineral trioxide aggregate formulation and calcium hydroxide. Am J Dent 21:255–261

    PubMed  Google Scholar 

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Acknowledgments

The authors thank the American Association of Endodontics–Foundation for Endodontics for the financial support in the competitive research grant category. Aliasger Salem acknowledges the Lyle and Sharon Bighley Professorship for the Bighley Chair.

Funding

This study was conducted with the financial support from the American Association for Endodontics Foundation, in the competitive research grant submission.

Author information

Author notes

  1. Leela Raghava Jaidev Chakka and Jered Vislisel contributed equally for the first authorship in this manuscript

    Aliasger Salem and Bruno Neves Cavalcanti contributed equally as senior authors in this manuscript

Authors and Affiliations

  1. University of Iowa College of Pharmacy, 180 S. Grand Avenue, Iowa City, IA, 52242, USA

    Leela Raghava Jaidev Chakka & Aliasger K. Salem

  2. Department of Endodontics, University of Iowa College of Dentistry and Dental Clinics, W344 DSB, 801 Newton Rd., Iowa City, IA, 52242, USA

    Jered Vislisel & Bruno Neves Cavalcanti

  3. Department of Operative Dentistry, University of Iowa College of Dentistry and Dental Clinics, Iowa City, IA, USA

    Cristina de Mattos Pimenta Vidal

  4. Departamento de Ciências Morfológicas, Universidade Federal de Santa Catarina, Florianopolis, SC, Brazil

    Michelle Tillmann Biz

Authors
  1. Leela Raghava Jaidev Chakka
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  2. Jered Vislisel
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  3. Cristina de Mattos Pimenta Vidal
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  4. Michelle Tillmann Biz
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  5. Aliasger K. Salem
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  6. Bruno Neves Cavalcanti
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Corresponding authors

Correspondence to Aliasger K. Salem or Bruno Neves Cavalcanti.

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Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

The research protocol (no. 201711716) was submitted to the University of Iowa Institutional Review Board, which determined that it did not meet the regulatory definition of human subject research thus not requiring review.

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For this type of study, formal consent is not required.

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Chakka, L.R.J., Vislisel, J., Vidal, C.d.P. et al. Application of BMP-2/FGF-2 gene–activated scaffolds for dental pulp capping. Clin Oral Invest 24, 4427–4437 (2020). https://doi.org/10.1007/s00784-020-03308-2

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  • DOI: https://doi.org/10.1007/s00784-020-03308-2

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