Long-Term Biobanked Dental Pulp Stem Cells Retain Angiogenic Potential for Vascularised Tissue Engineering-Laboratory Investigation

  • Int Endod J. 2025 Sep 23. doi: 10.1111/iej.70036.
Shuntaro Yamada  1  2 ,  Katerina Holomkova  1  3  4 ,  Åshild Johansen  1 ,  Masoumeh Jahani Kadousaraei  1 ,  Niyaz Al-Sharabi  1 ,  Francesco Torelli  1 ,  Pierfrancesco Pagella  5 ,  Ana Angelova Volponi  2 ,  Hiroshi Egusa  6  7 ,  Inge Fristad  1 ,  Kamal Mustafa  1
Affiliations
  • 1. Center of Translational Oral Research - Tissue Engineering, Department of Clinical Dentistry, University of Bergen, Bergen, Norway.
  • 2. Centre for Craniofacial & Regenerative Biology, Faculty of Dentistry, Oral & Craniofacial Sciences, King's College London, London, UK.
  • 3. Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Brno, Czechia.
  • 4. Institute of Animal Physiology and Genetics, Czech Academy of Sciences, Brno, Czechia.
  • 5. Laboratory of Molecular Materials, Division of Biophysics and Bioengineering, Department of Physics, Chemistry and Biology (IFM), Linköping University, Linköping, Sweden.
  • 6. Center for Advanced Stem Cell and Regenerative Research, Tohoku University Graduate School of Dentistry, Miyagi, Japan.
  • 7. Division of Molecular & Regenerative Prosthodontics, Tohoku University Graduate School of Dentistry, Miyagi, Japan.
PMID: 40988150 DOI: 10.1111/iej.70036
Abstract

Aim: This study aimed to evaluate whether human dental pulp stem cells (DPSCs), after long-term biobanking (7-8 years), retain their pro-angiogenic properties and can be used to engineer vascularised tissues, addressing their potential for clinical translation in regenerative dentistry.

Methodology: Cryopreserved DPSCs from adolescent donors were recovered from biobanking and characterised for chromosomal integrity, MSC immunophenotype and multipotency. After conditioning in pro-angiogenic conditions in vitro, gene and protein expression were analysed by RT-qPCR array, flow cytometry and high-throughput immunophenotyping. Functional angiogenic capacity was assessed via in vitro tube formation, ex ovo CAM implantation assay, organ-on-chip perfusion model and long-term culture (45 days) in clinical-grade GelMA hydrogels, with and without HUVECs.

Results: Biobanked DPSCs retained MSC identity and multi-lineage differentiation potential. Pro-angiogenic/endothelial conditioning enhanced the expression of angiogenic/endothelial genes (PECAM1, VEGFR2, NRP1, ACE), yet most cells maintained a pericyte-like phenotype. Both naive and endothelial-conditioned DPSCs (i.e., naiveDPSCs and endoDPSCs, respectively) significantly enhanced vascular ingrowth in the CAM model. In the organ-on-chip system, naiveDPSCs formed perfusable vasculature with HUVECs and differentiated into perivascular cell types. Most notably, endoDPSCs alone successfully generated vascularised tissue with both CD31(+) and αSMA(+) cells present in GelMA hydrogels after prolonged stimulation.

Conclusion: Long-term biobanked DPSCs preserve their angiogenic potential and, following extended endothelial induction, can independently generate vascularised tissue in 3D in vitro culture models. This is the first report demonstrating the comprehensive pro-angiogenic characterisation and the feasibility of using biobanked DPSCs for vascularised tissue engineering, highlighting their strong clinical applicability for future regenerative therapies.

Keywords
angiogenesis; dental pulp; dental research; mesenchymal stem cells; regenerative medicine; tissue engineering.
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