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Volume 14
Issue 20
10.3390/cells14201603
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Dental Pulp Stem Cell-Derived Organoids: Advancing the Development of 3D Structures

by
Loreto Lancia
1,†,
Fanny Pulcini
1,†,
Emanuela Mari
2,
Luca Piccoli
3,
Leda Assunta Biordi
1,
Luciano Mutti
1,5,
Claudio Festuccia
1,
Giovanni Luca Gravina
1,6,
Vincenzo Mattei
2,
Annunziata Mauro
4,
Valentina Notarstefano
4 and
Simona Delle Monache
1,*
1
Department of Biotechnological and Applied Clinical Sciences, University of L’Aquila, 67100 L’Aquila, Italy
2
Department of Life Science, Health, and Health Professions, Link Campus University, 00165 Rome, Italy
3
Department of Science Dentistry and Maxillofacial, Sapienza University of Rome, 00185 Rome, Italy
4
Department of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo, 64100 Teramo, Italy
5
Center for Biotechnology, Sbarro Institute for Cancer Research and Molecular Medicine, College of Science and Technology, Temple University, Philadelphia, PA 19122, USA
6
Division of Radiation Oncology, University of L’Aquila, 67100 L’Aquila, Italy
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Cells 2025, 14(20), 1603; https://doi.org/10.3390/cells14201603
Submission received: 11 September 2025 / Revised: 9 October 2025 / Accepted: 10 October 2025 / Published: 15 October 2025
(This article belongs to the Special Issue 3D Cultures and Organ-on-a-Chip in Cell and Tissue Cultures)
Versions Notes

Abstract

Two-dimensional cell cultures are crucial research tools, and they have been widely used, although they are not completely representative of biological processes in vivo due to the lack of tissue architecture and complexity. Recent advances in organoid technology have addressed these limitations and are revolutionizing the tools available for in vitro culture. Although there are no unified protocols for generating organoids, they can be obtained with various techniques, leading to cell aggregation by promoting cell adhesion. This work aims to generate and characterise organoid models of dental pulp from dental pulp stem cells (DPSCs), a type of mesenchymal stem/stromal cells known for their high regenerative potential and ease of accessibility, to establish a model for translational studies. The organoids were subjected to osteogenic differentiation conditions. Cell viability was evaluated using a CCK-8 assay, while osteogenic morphology and mineralization were confirmed by Alizarin red analysis, Raman microspectroscopy, and by immunofluorescence for the lineage markers expression. The Alizarin red analysis indicated a higher presence of calcium phosphate deposits in the differentiated organoids than in the control group (CTR). These results were confirmed by spectral profiles obtained using Raman microspectroscopy, which were attributable to a hydroxyapatite-based biomaterial. Immunofluorescence analysis also revealed increased expression of odonto/osteogenic markers (RUNX and OSX), alongside reduced expression of stemness markers. In conclusion, the organoids appeared to have successfully differentiated into an osteogenic lineage, forming a mineralized matrix containing hydroxyapatite and showing increased expression of relevant lineage markers.
Keywords: organoids; mesenchymal stem/stromal cells; dental pulp stem cells; 3D models; cells differentiation organoids; mesenchymal stem/stromal cells; dental pulp stem cells; 3D models; cells differentiation

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MDPI and ACS Style

Lancia, L.; Pulcini, F.; Mari, E.; Piccoli, L.; Biordi, L.A.; Mutti, L.; Festuccia, C.; Gravina, G.L.; Mattei, V.; Mauro, A.; et al. Dental Pulp Stem Cell-Derived Organoids: Advancing the Development of 3D Structures. Cells 2025, 14, 1603. https://doi.org/10.3390/cells14201603

AMA Style

Lancia L, Pulcini F, Mari E, Piccoli L, Biordi LA, Mutti L, Festuccia C, Gravina GL, Mattei V, Mauro A, et al. Dental Pulp Stem Cell-Derived Organoids: Advancing the Development of 3D Structures. Cells. 2025; 14(20):1603. https://doi.org/10.3390/cells14201603

Chicago/Turabian Style

Lancia, Loreto, Fanny Pulcini, Emanuela Mari, Luca Piccoli, Leda Assunta Biordi, Luciano Mutti, Claudio Festuccia, Giovanni Luca Gravina, Vincenzo Mattei, Annunziata Mauro, and et al. 2025. "Dental Pulp Stem Cell-Derived Organoids: Advancing the Development of 3D Structures" Cells 14, no. 20: 1603. https://doi.org/10.3390/cells14201603

APA Style

Lancia, L., Pulcini, F., Mari, E., Piccoli, L., Biordi, L. A., Mutti, L., Festuccia, C., Gravina, G. L., Mattei, V., Mauro, A., Notarstefano, V., & Monache, S. D. (2025). Dental Pulp Stem Cell-Derived Organoids: Advancing the Development of 3D Structures. Cells, 14(20), 1603. https://doi.org/10.3390/cells14201603

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