Extension and dentin differentiation potential of dental pulp
stem cells from human deciduous teeth on the stiff matrix surface

doi:10.3969/j.issn.2095-4344.2058

Abstract

Abstract:

BACKGROUND: Dental pulp stem cells can differentiate into dentin under appropriate induction conditions, which are important seed cells in dental tissue engineering. However, the commonly used inducers are chemical agents, which are not available for in vivo application. Mesenchymal stem cells can differentiate with the material hardness, and the physical property-induced cell differentiation is little reported.

OBJECTIVE: To observe the extension characteristics and dentin differentiation potential of dental pulp stem cells from human deciduous teeth on the stiff matrix surface.

METHODS: Dental pulp stem cells from naturally shed deciduous teeth were isolated, cultured and identified. Four solid gel matrixes with elasticity modulus of (9.12±0.94), (27.18±3.55), (59.37±4.05) and (86.45±5.33) kPa were made using low melting point agarose. The extension ability of passage 4 dental pulp stem cells on the surface of the above solid matrixes was detected by two-dimensional clone formation and cell scratch tests. The protein expression levels of dentin matrix protein-1, dentin phosphoprotein and dentin sialoprotein were detected by western blot assay.

RESULTS AND CONCLUSION: Dental pulp stem cells from human deciduous teeth seeded on the gel matrix with extremely low and low hardness almost existed as cell clones with neat edges, and cell spreading and extension were rare. When seeded on the gel matrix with moderate and high hardness, the cloned edge of deciduous dental pulp stem cells spread and extended obviously. The cell body became large and the cell edge extended significantly. The cell scratch test revealed the similar phenomenon. When seeded on the gel matrix with moderate and high hardness, dental pulp stem cells from human deciduous teeth exhibited high expression levels of of dentin matrix protein-1, dentin phosphoprotein and dentin sialoprotein. In summary, with the increase of matrix hardness, the abilities of extension and differentiation into dentin of dental pulp stem cells from human deciduous teeth are increased gradually, which provides a method for dental tissue engineering.

Key words: dental pulp stem cells, dentin, cell metastasis, cell differentiation, dentin matrix protein-1, dentin phosphoprotein, dentin sialoprotein

CLC Number:

Li Zhangyi, Liu Fengting, Huang Jianyong, Su Xiaoying, Wang Zhixing, Zheng Quan, Li Yanxia, Liu Xiaozhi. Extension and dentin differentiation potential of dental pulp stem cells from human deciduous teeth on the stiff matrix surface [J]. Chinese Journal of Tissue Engineering Research, 2020, 24(13): 2005-2010.

Figures/Tables 6

References

[1] WILLIAMS AR, HARE JM. Mesenchymal stem cells: biology, pathophysiology, translational findings, and therapeutic implications for cardiac disease. Circ Res. 2011;109(8): 923-940.
[2] UCCELLI A, MORETTA L, PISTOIA V. Mesenchymal stem cells in health and disease. Nat Rev Immunol. 2008;8(9): 726-736.
[3] KUO CK, TUAN RS. Tissue engineering with mesenchymal stem cells. IEEE Eng Med Biol Mag. 2003;22(5):51-56.
[4] ZHANG YD, CHEN Z, SONG YQ, et al. Making a tooth: growth factors, transcription factors, and stem cells. Cell Res. 2005;15(5):301-316.
[5] VICTOR AK, REITER LT. Dental pulp stem cells for the study of neurogenetic disorders. Hum Mol Genet. 2017;26(R2): R166-R171.
[6] WANG X, WU TT, JIANG L, et al. Deferoxamine-Induced Migration and Odontoblast Differentiation via ROS-Dependent Autophagy in Dental Pulp Stem Cells. Cell Physiol Biochem. 2017;43(6):2535-2547.
[7] HASTURK O, ERMIS M, DEMIRCI U, et al. Square prism micropillars on poly(methyl methacrylate) surfaces modulate the morphology and differentiation of human dental pulp mesenchymal stem cells. Colloids Surf B Biointerfaces. 2019; 178:44-55.
[8] OSATHANON T, SAWANGMAKE C, NOWWAROTE N, et al. Neurogenic differentiation of human dental pulp stem cells using different induction protocols. Oral Dis. 2014;20(4): 352-358.
[9] KRIFKA S, SPAGNUOLO G, SCHMALZ G, et al. A review of adaptive mechanisms in cell responses towards oxidative stress caused by dental resin monomers. Biomaterials. 2013; 34(19):4555-4563.
[10] NUTI N, CORALLO C, CHAN BM, et al. Multipotent Differentiation of Human Dental Pulp Stem Cells: a Literature Review. Stem Cell Rev Rep. 2016;12(5):511-523.
[11] MAO AS, SHIN JW, MOONEY DJ. Effects of substrate stiffness and cell-cell contact on mesenchymal stem cell differentiation. Biomaterials. 2016;98:184-191.
[12] LIN X, SHI Y, CAO Y, et al. Recent progress in stem cell differentiation directed by material and mechanical cues. Biomed Mater. 2016;11(1):014109.
[13] JIANG L, SUN Z, CHEN X, et al. Cells Sensing Mechanical Cues: Stiffness Influences the Lifetime of Cell-Extracellular Matrix Interactions by Affecting the Loading Rate. ACS Nano. 2016;10(1):207-217.
[14] TSE JR, ENGLER AJ. Stiffness gradients mimicking in vivo tissue variation regulate mesenchymal stem cell fate. PLoS One. 2011;6(1):e15978.
[15] 刘晓智,刘振林,姜忠敏,等.不同物理密度介质诱导干细胞表达多种组织细胞标记蛋白[J].中华实验外科杂志,2012,29(2):341.
[16] 刘晓智.蛋白质SUMO化修饰在低温与高热应激条件下的细胞保护功能研究[D].天津:天津医科大学,2017.
[17] GRAZIANO A, D'AQUINO R, LAINO G, et al. Dental pulp stem cells: a promising tool for bone regeneration. Stem Cell Rev. 2008;4(1):21-26.
[18] GU L, SHAN T, MA YX, et al. Novel Biomedical Applications of Crosslinked Collagen. Trends Biotechnol. 2019;37(5): 464-491.
[19] SULIMAN S, MUSTAFA K, KRUEGER A, et al. Nanodiamond modified copolymer scaffolds affects tumour progression of early neoplastic oral keratinocytes. Biomaterials. 2016;95: 11-21.
[20] LI G, ZHOU T, LIN S, et al. Nanomaterials for Craniofacial and Dental Tissue Engineering. J Dent Res. 2017;96(7):725-732.
[21] TAN J, XU X, LIN J, et al. Dental Stem Cell in Tooth Development and Advances of Adult Dental Stem Cell in Regenerative Therapies. Curr Stem Cell Res Ther. 2015; 10(5): 375-383.
[22] BIANCO P."Mesenchymal" stem cells. Annu Rev Cell Dev Biol. 2014;30:677-704.
[23] ANKRUM JA, ONG JF, KARP JM. Mesenchymal stem cells: immune evasive, not immune privileged. Nat Biotechnol. 2014;32(3):252-260.
[24] UCCELLI A, MORETTA L, PISTOIA V. Mesenchymal stem cells in health and disease. Nat Rev Immunol. 2008;8(9): 726-736.
[25] FIORE EJ, DOMÍNGUEZ LM, BAYO J, et al. Taking advantage of the potential of mesenchymal stromal cells in liver regeneration: Cells and extracellular vesicles as therapeutic strategies. World J Gastroenterol. 2018;24(23): 2427-2440.
[26] MURAKAMI M, HAYASHI Y, IOHARA K, et al. Trophic Effects and Regenerative Potential of Mobilized Mesenchymal Stem Cells From Bone Marrow and Adipose Tissue as Alternative Cell Sources for Pulp/Dentin Regeneration. Cell Transplant. 2015;24(9):1753-1765.
[27] ZHANG W, YANG G, WANG X, et al. Magnetically Controlled Growth-Factor-Immobilized Multilayer Cell Sheets for Complex Tissue Regeneration. Adv Mater. 2017;29(43): 1703795.
[28] VINING KH, SCHERBA JC, BEVER AM, et al. Synthetic Light-Curable Polymeric Materials Provide a Supportive Niche for Dental Pulp Stem Cells. Adv Mater. 2018;30(4).
[29] BOTELHO J, CAVACAS MA, MACHADO V, et al. Dental stem cells: recent progresses in tissue engineering and regenerative medicine. Ann Med. 2017;49(8):644-651.
[30] XUAN K, LI B, GUO H, et al. Deciduous autologous tooth stem cells regenerate dental pulp after implantation into injured teeth. Sci Transl Med. 2018;10(455):eaaf3227.