AccScience Publishing / IJB / Volume 12 / Issue 2 / DOI: 10.36922/IJB026060050
Submit to IJB
Cite this article
38
Download
422
Views
Journal Browser
Volume | Year
Issue
Search
Forthcoming Issue
Current Issue
View All
News and Announcements
View All
RESEARCH ARTICLE

Tiny bricks for oral bioprinting: Exploring gingiva and dental pulp-derived organ building blocks

Lidiia R. Grinchevskaia1 ,  Anna V. Kardosh1 ,  Vitalia R. Izhbulatova1 ,  Nastasia V. Kosheleva1 ,  Daria S. Kuznetsova1 ,  Artem M. Mozherov1 ,  Yuri M. Efremov1 ,  Alexey L. Fayzullin1 ,  Polina Y. Bikmulina1 ,  Svetlana L. Kotova1 ,  Anastasia I. Shpichka1 ,  Oleg O. Pavlov2 ,  Boris P. Yakimov1,2 ,  Peter S. Timashev1*
Show Less
1 Institute for Regenerative Medicine, I. M. Sechenov First Moscow State Medical University, Moscow, Russia
2 Faculty of Physics, M. V. Lomonosov Moscow State University, Moscow, Russia
IJB 2026, 12(2), 026060050 https://doi.org/10.36922/IJB026060050
Received: 3 September 2025 | Revised: 4 February 2026 | Accepted: 5 February 2026 | Published online: 19 February 2026
© 2026 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
Download PDF
XML
Cite
Abstract

Today, organ building blocks (OBBs) serve as important tools for in vitro tissue modeling, personalized medicine, and regenerative approaches. Despite substantial advances in dental reconstruction methods, tissues of the oral cavity remain challenging to regenerate due to their complex structure and microenvironment. However, effective regeneration of the periodontal complex, e.g., in diseases such as periodontitis, persists, as current methods do not achieve complete tissue restoration. Cells from the gingiva and dental pulp are accessible sources of mesenchymal stem cells with high regenerative potential, making them promising materials for creating OBBs. These cells can serve as fundamental units for restoring the periodontal complex using techniques such as 3D bioprinting. This study aims to characterize and compare OBBs derived from gingival cells, pulp cells, and their combinations by assessing key parameters, including morphology, extracellular matrix composition, biomechanical properties, histology, and metabolic activity. Combining the two cell types improved the structural, mechanical, and functional properties of OBBs, making them more suitable for bioprinting than those derived from a single cell type. Moreover, all types of OBBs from the two cell cultures may be suitable as components of bioinks, depending on the specific purposes. The results provide insights into the potential use of these cell sources for tissue engineering and the development of personalized periodontal bio-constructs that may significantly improve treatment approaches for oral diseases.

Graphical abstract
Keywords
Organ building blocks
Tissue engineering
Regenerative medicine
Oral tissues
Gingiva cells
Dental pulp cells
Periodontal complex regeneration
Funding
The work was carried out with financial support from the Ministry of Science and Higher Education of the Russian Federation under grant number 075-15-2024-640 (Sechenov University).
Conflict of interest
The authors declare they have no competing interests.
References
  1. De Lauretis A, Ovrebo O, Romandini M, Lyngstadaas SP, Rossi F, Haugen HJ. From Basic Science to Clinical Practice: A Review of Current Periodontal/Mucogingival Regenerative Biomaterials. Adv Sci. 2024;11(17):2308848. doi: 10.1002/advs.202308848
  2. Wolf KJ, Weiss JD, Uzel SGM, Skylar-Scott MA, Lewis JA. Biomanufacturing human tissues via organ building blocks. Cell Stem Cell. 2022;29(5):667-677. doi: 10.1016/j.stem.2022.04.012
  3. Bhise NS, Manoharan V, Massa S, et al. A liver-on-a-chip platform with bioprinted hepatic spheroids. Biofabrication. 2016;8(1):014101. doi: 10.1088/1758-5090/8/1/014101
  4. Lee J, Van Der Valk WH, Serdy SA, et al. Generation and characterization of hair-bearing skin organoids from human pluripotent stem cells. Nat Protoc. 2022;17(5):1266-1305. doi: 10.1038/s41596-022-00681-y
  5. Birey F, Andersen J, Makinson CD, et al. Assembly of functionally integrated human forebrain spheroids. Nature. 2017;545(7652):54-59. doi: 10.1038/nature22330
  6. Hofer M, Lutolf MP. Engineering organoids. Nat Rev Mater. 2021;6(5):402-420. doi: 10.1038/s41578-021-00279-y
  7. Workman MJ, Mahe MM, Trisno S, et al. Engineered human pluripotent-stem-cell-derived intestinal tissues with a functional enteric nervous system. Nat Med. 2017;23(1):49- 59. doi: 10.1038/nm.4233
  8. Picado-Tejero D, Mendoza-Cerezo L, Rodríguez-Rego JM, Carrasco-Amador JP, Marcos-Romero AC. Recent Advances in 3D Bioprinting of Porous Scaffolds for Tissue Engineering: A Narrative and Critical Review. J Funct Biomater. 2025;16(9):328. doi: 10.3390/jfb16090328
  9. Michelutti L, Tel A, Robiony M, et al. The Properties and Applicability of Bioprinting in the Field of Maxillofacial Surgery. Bioengineering. 2025;12(3):251. doi: 10.3390/bioengineering12030251
  10. Xu H, Zhang Y, Zhang Y, et al. 3D bioprinting advanced biomaterials for craniofacial and dental tissue engineering – A review. Mater Des. 2024;241:112886. doi: 10.1016/j.matdes.2024.112886
  11. Smirani R, Médina C, Becker J, et al. In vivo vessel connection of pre-vascularised 3D-bioprinted gingival connective tissue substitutes. Biofabrication. 2025;17(2):025009. doi: 10.1088/1758-5090/adac90
  12. Almela T, Al-Sahaf S, Brook IM, et al. 3D printed tissue engineered model for bone invasion of oral cancer. Tissue Cell. 2018;52:71-77. doi: 10.1016/j.tice.2018.03.009
  13. Huang GTJ, Gronthos S, Shi S. Mesenchymal Stem Cells Derived from Dental Tissues vs. Those from Other Sources: Their Biology and Role in Regenerative Medicine. J Dent Res. 2009;88(9):792-806. doi: 10.1177/0022034509340867
  14. Stefańska K, Volponi AA, Kulus M, et al. Dental pulp stem cells – A basic research and future application in regenerative medicine. Biomed Pharmacother. 2024;178:116990. doi: 10.1016/j.biopha.2024.116990
  15. Miura M, Gronthos S, Zhao M, et al. SHED: Stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci USA. 2003;100(10):5807-5812. doi: 10.1073/pnas.0937635100
  16. Zhu W, Liang M. Periodontal Ligament Stem Cells: Current Status, Concerns, and Future Prospects. Stem Cells Int. 2015;2015:972313. doi: 10.1155/2015/972313
  17. Kang J, Fan W, Deng Q, He H, Huang F. Stem Cells from the Apical Papilla: A Promising Source for Stem Cell-Based Therapy. Biomed Res Int. 2019;2019:6104738. doi: 10.1155/2019/6104738
  18. Bi R, Lyu P, Song Y, et al. Function of Dental Follicle Progenitor/Stem Cells and Their Potential in Regenerative Medicine: From Mechanisms to Applications. Biomolecules. 2021;11(7):997. doi: 10.3390/biom11070997
  19. Cabaña-Muñoz ME, Pelaz Fernández MJ, Parmigiani- Cabaña JM, Parmigiani-Izquierdo JM, Merino JJ. Adult Mesenchymal Stem Cells from Oral Cavity and Surrounding Areas: Types and Biomedical Applications. Pharmaceutics. 2023;15(8):2109. doi: 10.3390/pharmaceutics15082109
  20. Li P, Ou Q, Shi S, Shao C. Immunomodulatory properties of mesenchymal stem cells/dental stem cells and their therapeutic applications. Cell Mol Immunol. 2023;20(6):558- 569. doi: 10.1038/s41423-023-00998-y
  21. Awais S, Balouch SS, Riaz N, Choudhery MS. Human Dental Pulp Stem Cells Exhibit Osteogenic Differentiation Potential. Open Life Sci. 2020;15(1):229-236. doi: 10.1515/biol-2020-0023
  22. Sabbagh J, Ghassibe-Sabbagh M, Fayyad-Kazan M, et al. Differences in osteogenic and odontogenic differentiation potential of DPSCs and SHED. J Dent. 2020;101:103413. doi: 10.1016/j.jdent.2020.103413
  23. Li Z, Zhao Z, Gu B, et al. Alteration of immunomodulatory properties of locally applied gingival-derived mesenchymal stem cells by the oral inflammatory environment via Caspase-3/8 in periodontitis. Int Immunopharmacol. 2025;161:114978. doi: 10.1016/j.intimp.2025.114978
  24. Shetty SS, Sowmya S, Pradeep A, Jayakumar R. Gingival Mesenchymal Stem Cells: A Periodontal Regenerative Substitute. Tissue Eng Regen Med. 2025;22(1):1-21. doi: 10.1007/s13770-024-00676-8
  25. Cacciamali A, Villa R, Dotti S. 3D Cell Cultures: Evolution of an Ancient Tool for New Applications. Front Physiol. 2022;13:836480. doi: 10.3389/fphys.2022.836480
  26. Ji Q, Tang R, Liu X, et al. From microtissues to organs: the future of reconstructive surgery with organ building block-based bioprinting. Biofabrication. 2025;17(3):032010. doi: 10.1088/1758-5090/aded37
  27. Bikmulina P, Kosheleva N, Efremov Y, et al. 3D or not 3D: a guide to assess cell viability in 3D cell systems. Soft Matter. 2022;18(11):2222-2233. doi: 10.1039/D2SM00018K
  28. Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8(4):315-317. doi: 10.1080/14653240600855905
  29. Fonseca LN, Bolívar-Moná S, Agudelo T, et al. Cell surface markers for mesenchymal stem cells related to the skeletal system: A scoping review. Heliyon. 2023;9(2):e13464. doi: 10.1016/j.heliyon.2023.e13464
  30. Lertkiatmongkol P, Liao D, Mei H, Hu Y, Newman PJ. Endothelial functions of platelet/endothelial cell adhesion molecule-1 (CD31). Curr Opin Hematol. 2016;23(3):253- 259. doi: 10.1097/MOH.0000000000000239
  31. Huang MS, Christakopoulos F, Roth JG, Heilshorn SC. Organoid bioprinting: from cells to functional tissues. Nat Rev Bioeng. 2025;3(2):126-142. doi: 10.1038/s44222-024-00268-0
  32. Kuntjoro M, Hendrijantini N, Agustono B, Rahmania PN, Al Bana FR, Gabriela N. Osteogenic differentiation enhancement of human dental pulp mesenchymal stem cells: A review. World J Adv Res Rev. 2023;19(2):146-151. doi: 10.30574/wjarr.2023.19.2.1525
  33. Nowwarote N, Petit S, Ferre FC, et al. Extracellular Matrix Derived From Dental Pulp Stem Cells Promotes Mineralization. Front Bioeng Biotechnol. 2022;9:740712. doi: 10.3389/fbioe.2021.740712
  34. Kornsuthisopon C, Nowwarote N, Chansaenroj A, et al. Human dental pulp stem cells derived extracellular matrix promotes mineralization via Hippo and Wnt pathways. Sci Rep. 2024;14(1):6777. doi: 10.1038/s41598-024-56845-1
  35. Martinez EF, Araújo VC. In vitro immunoexpression of extracellular matrix proteins in dental pulpal and gingival human fibroblasts. Int Endod J. 2004;37(11):749-755. doi: 10.1111/j.1365-2591.2004.00864.x
  36. Angelopoulos I, Brizuela C, Khoury M. Gingival Mesenchymal Stem Cells Outperform Haploidentical Dental Pulp-derived Mesenchymal Stem Cells in Proliferation Rate, Migration Ability, and Angiogenic Potential. Cell Transplant. 2018;27(6):967-978. doi: 10.1177/0963689718759649
  37. Roato I, Chinigò G, Genova T, Munaron L, Mussano F. Oral Cavity as a Source of Mesenchymal Stem Cells Useful for Regenerative Medicine in Dentistry. Biomedicines. 2021;9(9):1085. doi: 10.3390/biomedicines9091085
  38. Lee H, Hwa S, Cho S, et al. Impact of Polydeoxyribonucleotides on the Morphology, Viability, and Osteogenic Differentiation of Gingiva-Derived Stem Cell Spheroids. Medicina. 2024;60(10):1610. doi: 10.3390/medicina60101610
  39. Toyoda M, Fukuda T, Fujimoto R, et al. Scaffold-free bone-like 3D structure established through osteogenic differentiation from human gingiva-derived stem cells. Biochem Biophys Rep. 2024;38:101656. doi: 10.1016/j.bbrep.2024.101656
  40. Alfonso García SL, Mira Uribe LM, Castaño López S, Parada-Sanchez MT, Arboleda-Toro D. Ultrastructural Characterization of Human Gingival Fibroblasts in 3D Culture. Cells. 2022;11(22):3647. doi: 10.3390/cells11223647
  41. Guo L, Zou Z, Freytag M, et al. Human Dental Pulp Cells form Spheroids in the Presence of Serum When Seeded on a Low-Attachment Cultural Surface. Processes. 2022;10(5):1021. doi: 10.3390/pr10051021
  42. Zheng Y, Jiang L, Yan M, et al. Optimizing Conditions for Spheroid Formation of Dental Pulp Cells in Cell Culture. In Vivo. 2021;35(4):1965-1972. doi: 10.21873/invivo.12464
  43. Liu F, Wu Q, Liu Q, et al. Dental pulp stem cells-derived cannabidiol-treated organoid-like microspheroids show robust osteogenic potential via upregulation of WNT6. Commun Biol. 2024;7(1):972. doi: 10.1038/s42003-024-06655-y
  44. Van Der Geest AT, Jakobs CE, Ljubikj T, et al. Molecular pathology, developmental changes and synaptic dysfunction in (pre-) symptomatic human C9ORF72-ALS/FTD cerebral organoids. Acta Neuropathol Commun. 2024;12(1):152. doi: 10.1186/s40478-024-01857-1
  45. Yin J, VanDongen AM. Enhanced Neuronal Activity and Asynchronous Calcium Transients Revealed in a 3D Organoid Model of Alzheimer’s Disease. ACS Biomater Sci Eng. 2021;7(1):254-264. doi: 10.1021/acsbiomaterials.0c01583
  46. Kronemberger GS, Spagnuolo FD, Karam AS, Chattahy K, Storey KJ, Kelly DJ. Rapidly Degrading Hydrogels to Support Biofabrication and 3D Bioprinting Using Cartilage Microtissues. ACS Biomater Sci Eng. 2024;10(10):6441-6450. doi: 10.1021/acsbiomaterials.4c00819
  47. Tartagni O, Borók A, Mensà E, et al. Microstructured soft devices for the growth and analysis of populations of homogenous multicellular tumor spheroids. Cell Mol Life Sci. 2023;80(4):93. doi: 10.1007/s00018-023-04748-1
  48. Kosheleva NV, Efremov YM, Koteneva PI, et al. Building a tissue: Mesenchymal and epithelial cell spheroids mechanical properties at micro- and nanoscale. Acta Biomater. 2023;165:140-152. doi: 10.1016/j.actbio.2022.09.051
  49. Revokatova DP, Koteneva PI, Kosheleva NV, Shpichka AI, Timashev PS. Spheroids from Epithelial and Mesenchymal Cell Phenotypes as Building Blocks in Bioprinting (Review). Sovrem Tekhnol Med. 2025;17(1):121-132. doi: 10.17691/stm2025.17.1.11
  50. Rovere M, Reverberi D, Arnaldi P, Palamà MEF, Gentili C. Spheroid size influences cellular senescence and angiogenic potential of mesenchymal stromal cell-derived soluble factors and extracellular vesicles. Front Bioeng Biotechnol. 2023;11:1297644. doi: 10.3389/fbioe.2023.1297644
  51. Nilsson Hall G, Rutten I, Lammertyn J, et al. Cartilaginous spheroid-assembly design considerations for endochondral ossification: towards robotic-driven biomanufacturing. Biofabrication. 2021;13(4):045025. doi: 10.1088/1758-5090/ac2208
  52. Efremov YM, Zurina IM, Presniakova VS, et al. Mechanical properties of cell sheets and spheroids: the link between single cells and complex tissues. Biophys Rev. 2021;13(4):541- 561. doi: 10.1007/s12551-021-00821-w
  53. Banerjee D, Singh YP, Datta P, et al. Strategies for 3D bioprinting of spheroids: A comprehensive review. Biomaterials. 2022;291:121881. doi: 10.1016/j.biomaterials.2022.121881
  54. De Moor L, Fernandez S, Vercruysse C, et al. Hybrid Bioprinting of Chondrogenically Induced Human Mesenchymal Stem Cell Spheroids. Front Bioeng Biotechnol. 2020;8:484. doi: 10.3389/fbioe.2020.00484
  55. Bikmulina P, Kosheleva N, Efremov Y, et al. Building a tissue: gingiva- and adipose-derived mesenchymal cell spheroids’ survivability and functionality after 3D extrusion bioprinting. Bioprinting. 2023;32:e00279. doi: 10.1016/j.bprint.2023.e00279
  56. Skala MC, Riching KM, Bird DK, et al. In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia. J Biomed Opt. 2007;12(2):024014. doi: 10.1117/1.2717503
  57. Stringari C, Edwards RA, Pate KT, Waterman ML, Donovan PJ, Gratton E. Metabolic trajectory of cellular differentiation in small intestine by Phasor Fluorescence Lifetime Microscopy of NADH. Sci Rep. 2012;2(1):568. doi: 10.1038/srep00568

58. Heikal AA. Intracellular Coenzymes as Natural Biomarkers for Metabolic Activities and Mitochondrial Anomalies. Biomark Med. 2010;4(2):241-263. doi: 10.2217/bmm.10.1

Share
Back to top
International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing
We use cookies on our website to ensure you get the best experience.
Read more about our cookies here
Accept