miR-1246 regulates odontogenic differentiation of human dental pulp stem cells in an inflammatory microenvironment by targeting AXIN2: an in vitro study
DOI:
https://doi.org/10.2340/aos.v85.46788Keywords:
Dental pulp stem cells, inflammatory microenvironment, miR-1246, odontogenic differentiationAbstract
Objective: This study aimed to investigate the modulatory function of microRNA-1246 (miR-1246) in the odontogenic differentiation of human dental pulp stem cells (hDPSCs) under inflammatory conditions in vitro and to elucidate its underlying molecular mechanism.
Methods: An inflammatory state was replicated in vitro by treating hDPSCs with lipopolysaccharide (LPS). miR-1246 expression was modulated via transfection with its mimics or inhibitors. Cell proliferation, movement, and apoptosis were assessed. Odontogenic differentiation capacity was assessed via alkaline phosphatase (ALP) activity, Alizarin Red S (ARS) staining, and transcript levels by quantitative real-time quantitative real-time polymerase chain reaction (qRT-PCR), along with Western blotting. To validate the direct interaction between miR-1246 and AXIN2, a dual-luciferase reporter assay was employed. All experiments were performed in three independent biological replicates.
Results: miR-1246 overexpression significantly reduced inflammatory cytokine levels (IL-1β, P < 0.05; IL-6, P < 0.05; TNF-α, P < 0.001) and partially restored hDPSC proliferation (P < 0.01), migration (P < 0.001), and survival (reduced apoptosis rate, P < 0.001). It also significantly increased ALP activity (P < 0.001), mineralized nodule formation (P < 0.001), and the expression of odontogenic markers (NRP1, DSPP, DMP1). Mechanistically, miR-1246 directly targeted AXIN2, preventing AXIN2-mediated β-catenin degradation and promoting the Wnt/β-catenin pathway, elevating downstream factors such as Cyclin D1 and c-Myc. AXIN2 knockdown reproduced the differentiation-promoting effects of miR-1246, whereas AXIN2 overexpression diminished them.
Conclusion: miR-1246 promotes the odontogenic differentiation of hDPSCs and partially counteracts the detrimental effects of inflammation in vitro by directly targeting and inhibiting AXIN2 expression, thereby stimulating the Wnt/β-catenin signaling cascade.
Downloads
References
Bar JK, Lis-Nawara A, Grelewski PG. Dental pulp stem cell-derived secretome and its regenerative potential. Int J Mol Sci. 2021;22:12018. DOI: https://doi.org/10.3390/ijms222112018
Sloan AJ, Smith AJ. Stem cells and the dental pulp: potential roles in dentine regeneration and repair. Oral Dis. 2007;13:151–7. DOI: https://doi.org/10.1111/j.1601-0825.2006.01346.x
Bugueno IM, Alastra G, Balic A, Stadlinger B, Mitsiadis TA. Limited adipogenic differentiation potential of human dental pulp stem cells compared to human bone marrow stem cells. Int J Mol Sci. 2024;25:11105. DOI: https://doi.org/10.3390/ijms252011105
Chmilewsky F, Liang R, Kanazawa M, About I, Cooper LF, George A. C5L2 regulates DMP1 expression during odontoblastic differentiation. J Dent Res. 2019;98:597–604. DOI: https://doi.org/10.1177/0022034518820461
Galler KM, Weber M, Korkmaz Y, Widbiller M, Feuerer M. Inflammatory response mechanisms of the dentine-pulp complex and the periapical tissues. Int J Mol Sci. 2021;22:1480. DOI: https://doi.org/10.3390/ijms22031480
Colombo JS, Moore AN, Hartgerink JD, D’Souza RN. Scaffolds to control inflammation and facilitate dental pulp regeneration. J Endod. 2014;40(4 Suppl):S6–12. DOI: https://doi.org/10.1016/j.joen.2014.01.019
Arora S, Cooper PR, Friedlander LT, Rizwan S, Seo B, Rich AM, et al. Potential application of immunotherapy for modulation of pulp inflammation: opportunities for vital pulp treatment. Int Endod J. 2021;54:1263–74. DOI: https://doi.org/10.1111/iej.13524
Gronthos S, Mankani M, Brahim J, Robey PG, Shi S. Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci U S A. 2000;97:13625–30. DOI: https://doi.org/10.1073/pnas.240309797
Yang X, Li L, Xiao L, Zhang D. Recycle the dental fairy’s package: overview of dental pulp stem cells. Stem Cell Res Ther. 2018;9:347. DOI: https://doi.org/10.1186/s13287-018-1094-8
Sonmez Kaplan S, Sazak Ovecoglu H, Genc D, Akkoc T. TNF-α, IL-1B and IL-6 affect the differentiation ability of dental pulp stem cells. BMC Oral Health. 2023;23:555. DOI: https://doi.org/10.1186/s12903-023-03288-1
Pezelj-Ribaric S, Anic I, Brekalo I, Miletic I, Hasan M, Simunovic-Soskic M. Detection of tumor necrosis factor alpha in normal and inflamed human dental pulps. Arch Med Res. 2002;33:482–4. DOI: https://doi.org/10.1016/S0188-4409(02)00396-X
Bucchi C, Bucchi A, Martínez-Rodríguez P. Biological properties of dental pulp stem cells isolated from inflamed and healthy pulp and cultured in an inflammatory microenvironment. J Endod. 2023;49:395–401.e6. DOI: https://doi.org/10.1016/j.joen.2023.02.002
Inostroza C, Vega-Letter AM, Brizuela C, Castrillón L, Saint Jean N, Duran CM, et al. Mesenchymal stem cells derived from human inflamed dental pulp exhibit impaired immunomodulatory capacity in vitro. J Endod. 2020;46:1091–8.e2. DOI: https://doi.org/10.1016/j.joen.2020.05.003
Morsczeck C. Cellular senescence in dental pulp stem cells. Arch Oral Biol. 2019;99:150–5. DOI: https://doi.org/10.1016/j.archoralbio.2019.01.012
Chang C, Weiping L, Jibing C. Exosomal MiRNA therapy for central nervous system injury diseases. Cell Mol Neurobiol. 2024;45:3. DOI: https://doi.org/10.1007/s10571-024-01522-0
Ramprosand S, Govinden-Soulange J, Ranghoo-Sanmukhiya VM, Sanan-Mishra N. miRNA, phytometabolites and disease: connecting the dots. Phytother Res. 2024;38:4570–91. DOI: https://doi.org/10.1002/ptr.8287
Ye YY, Yue L, Zou XY, Wang XY. [Characteristics and microRNA expression profile of exosomes derived from odontogenic dental pulp stem cells]. Beijing Da Xue Xue Bao Yi Xue Ban. 2023;55:689–96.
Liu L, Silke J. AXIN2 is a non-redundant regulator of AXIN1 stability and β-catenin in colorectal cancer cells. FEBS J. 2025;292:990–4. DOI: https://doi.org/10.1111/febs.17336
Choi H, Jeong J-K, Adasooriya D, Cho S-W, Cho E-S. Notum as a crucial regulator of matrix integrity in dentinogenesis. J Cell Physiol. 2025;240:e70070. DOI: https://doi.org/10.1002/jcp.70070
Lan C, Chen S, Jiang S, Lei H, Cai Z, Huang X. Different expression patterns of inflammatory cytokines induced by lipopolysaccharides from Escherichia coli or Porphyromonas gingivalis in human dental pulp stem cells. BMC Oral Health. 2022;22:121. DOI: https://doi.org/10.1186/s12903-022-02161-x
Liang C, Liao L, Tian W. Stem cell-based dental pulp regeneration: insights from signaling pathways. Stem Cell Rev Rep. 2021;17(4):1251–1263. DOI: https://doi.org/10.1007/s12015-020-10117-3
Amir M, Jeevithan L, Barkat M, Fatima SH, Khan M, Israr S, et al. Advances in regenerative dentistry: a systematic review of harnessing Wnt/β-Catenin in dentin-pulp regeneration. Cells. 2024;13:1153. DOI: https://doi.org/10.3390/cells13131153
Ma Y, Liu X, Dai R, Li Q, Cao CY. LL-37 regulates odontogenic differentiation of dental pulp stem cells in an inflammatory microenvironment. Stem Cell Res Ther. 2024;15:469. DOI: https://doi.org/10.1186/s13287-024-04075-7
He W, Wang Z, Luo Z, Yu Q, Jiang Y, Zhang Y, et al. LPS promote the odontoblastic differentiation of human dental pulp stem cells via MAPK signaling pathway. J Cell Physiol. 2015;230:554–61. DOI: https://doi.org/10.1002/jcp.24732
Xia Y, Cheng T, Zhang C, Zhou M, Hu Z, Kang F, et al. Human bone marrow mesenchymal stem cell-derived extracellular vesicles restore Th17/Treg homeostasis in periodontitis via miR-1246. FASEB J. 2023;37:e23226. DOI: https://doi.org/10.1096/fj.202300674RR
Kim J-H, Irfan M, Hossain MA, Shin S, George A, Chung S. LPS-induced inflammation potentiates dental pulp stem cell odontogenic differentiation through C5aR and p38. Connect Tissue Res. 2023;64:505–15. DOI: https://doi.org/10.1080/03008207.2023.2218944
Yu S, Liu X-M, Liu Y, Tang L, Lei S, Geng C, et al. Inflammatory microenvironment of moderate pulpitis enhances the osteo-/odontogenic potential of dental pulp stem cells by autophagy. Int Endod J. 2024;57:1465–77. DOI: https://doi.org/10.1111/iej.14108
Kornsuthisopon C, Photichailert S, Nowwarote N, Tompkins KA, Osathanon T. Wnt signaling in dental pulp homeostasis and dentin regeneration. Arch Oral Biol. 2022;134:105322. DOI: https://doi.org/10.1016/j.archoralbio.2021.105322
Xie K, Liu L, Chen J, Liu F. Exosomes derived from human umbilical cord blood mesenchymal stem cells improve hepatic ischemia reperfusion injury via delivering miR-1246. Cell Cycle. 2019;18:3491–501. DOI: https://doi.org/10.1080/15384101.2019.1689480
Wu S, Xu X, Gao S, Huo S, Wan M, Zhou X, et al. MicroRNA-93-5p regulates odontogenic differentiation and dentin formation via KDM6B. J Transl Med. 2024;22:54. DOI: https://doi.org/10.1186/s12967-024-04862-z
Liu P, Zhang Q, Mi J, Wang S, Xu Q, Zhuang D, et al. Exosomes derived from stem cells of human deciduous exfoliated teeth inhibit angiogenesis in vivo and in vitro via the transfer of miR-100-5p and miR-1246. Stem Cell Res Ther. 2022;13:89. DOI: https://doi.org/10.1186/s13287-022-02764-9
Liu S, Sun J, Yuan S, Yang Y, Gong Y, Wang Y, et al. Treated dentin matrix induces odontogenic differentiation of dental pulp stem cells via regulation of Wnt/β-catenin signaling. Bioact Mater. 2022;7:85–97. DOI: https://doi.org/10.1016/j.bioactmat.2021.05.026
Babb R, Chandrasekaran D, Vitor CMN, Sharpe PT. Axin2-expressing cells differentiate into reparative odontoblasts via autocrine Wnt/β-catenin signaling in response to tooth damage. Sci Rep. 2017;7:3102. DOI: https://doi.org/10.1038/s41598-017-03145-6
Godoi-Lopes J, Ribeiro-Silva VHA, Candido Do Prado LG, Petean IBF, Magri LV, Lopes-Olhê FC, et al. Efficacy of dental stem cell–derived exosomes for pulp regeneration: a systematic review of clinical, animal, and in vitro studies. Mol Biol Rep. 2026;53:426. DOI: https://doi.org/10.1007/s11033-026-11547-x
Nasiri K, Jahri M, Kolahdouz S, Soleimani M, Makiya A, Saini RS, et al. MicroRNAs function in dental stem cells as a promising biomarker and therapeutic target for dental diseases. Mol Diagn Ther. 2023;27:703–22. DOI: https://doi.org/10.1007/s40291-023-00675-w
Downloads
Published
Issue
Section
License
Copyright (c) 2026 Shunying Wu, Yueyan Wang, Jiayi Yang
This work is licensed under a Creative Commons Attribution 4.0 International License.
Acta Odontologica Scandinavica publishes high quality original research papers as well as critical reviews relevant on diseases of the oral cavity, their treatment, epidemiology and associated biomaterials and their development.
