Mechanism of Chir99021 regulating Wnt/β-catenin signaling pathway inhibiting osteogenic differentiation of rat dental pulp stem cells

XIE Hui-Lan; FANG Fang; LIN Yi

doi:10.16098/j.issn.0529-1356.2024.01.010

PDF(4367 KB)

Acta Anatomica Sinica ›› 2024, Vol. 55 ›› Issue (1) : 67-72. DOI: 10.16098/j.issn.0529-1356.2024.01.010

Mechanism of Chir99021 regulating Wnt/β-catenin signaling pathway inhibiting osteogenic differentiation of rat dental pulp stem cells

XIE Hui-lan^1,2* FANG Fang^1,2 LIN Yi^1,2

Author information +

History +

Abstract

Objective To explore the effect and mechanism of Chir99021 on osteogenic differentiation of rat dental pulp stem cells. Methods Primary rat dental pulp stem cells were isolated from rat dental pulp and verified by fluorescence immunoassay. Different concentrations of Chir99021 were set, and the cell proliferation was detected by CCK-8 to select the optimal concentration. Osteogenic differentiation was detected by alizarin red staining. The expression of osteogenic differentiation related genes and proteins recombinant wingless type MMTV integration site famity member 1(Wnt1), Wnt3a and Wnt3a β-expression of catenin, axis inhibition protein 2(Axin 2), dentin sialophosphoprotein(OCN) and dentin matrix acidic phosphoprotein 1(DMP1) was detected by Real-time PCR and Western blotting. Results The positive expression of dentin sialophosphoprotein(DSPP) and vimentin indicated that rat dental pulp stem cells were successfully isolated. After osteogenic induction of rat dental pulp stem cells, calcium deposits significantly increased with the addition of glycogen synthase kinase-3β(GSK-3β) inhibitor Chir99021, calcium deposits were significanted reduced. After osteogenic differentiation of rat dental pulp stem cells, the expression of Wnt1, Wnt3a, β-catenin, Axin2, OCN and DMP1 increased, while the expression of Wnt1, Axin2, OCN and DMP1 decreased with the addition of Chir99021. Conclusion Chir99021 can inhibit the osteogenic differentiation of rat dental pulp stem cells after 7 days of induction.

Key words

Dental pulp stem cell / Osteogenic differentiation / Chir99021 / Real-time PCR / Rat

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

XIE Hui-lan FANG Fang LIN Yi. Mechanism of Chir99021 regulating Wnt/β-catenin signaling pathway inhibiting osteogenic differentiation of rat dental pulp stem cells[J]. Acta Anatomica Sinica. 2024, 55(1): 67-72 https://doi.org/10.16098/j.issn.0529-1356.2024.01.010

References

［1］Ding G, Liu Y, Wang SL. Research status of dental pulpstem cells［J］. Journal of Capital Medical University, 2008,5:660-663. (in Chinese)
丁刚,刘怡,王松灵.牙髓干细胞研究现状［J］.首都医科大学学报,2008,5:660-663.
［2］Uribe-Etxebarria V, Pineda JR, García-Gallastegi P, et al. Notch and Wnt signaling modulation to enhance DPSC stemness and therapeutic potential［J］. Int J Mol Sci, 2023, 24(8):7389.
［3］Damrongsri D, Nowwarote N, Sonpoung O, et al. Differential expression of Notch related genes in dental pulp stem cells and stem cells isolated from apical papilla［J］. J Oral Biol Craniofac Res, 2021, 11(3):379-385.
［4］Feng G, Zheng K, Song D, et al. SIRT1 was involved in TNF-α-promoted osteogenic differentiation of human DPSCs through Wnt/β-catenin signal［J］. In Vitro Cell Dev Biol Anim, 2016,52(10):1001-1011.
［5］Gronthos S, Mankani M, Brahim J, et al. Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo［J］. Proc Natl Acad Sci USA, 2000,97(25):13625-13630.
［6］Xi J, Liu Y, Liu H, et al. Specification of midbrain dopamine neurons from primate pluripotent stem cells［J］. Stem Cells,2012,30(8):1655-1663.
［7］Kolios G, Moodley Y. Introduction to stem cells and regenerative medicine［J］. Respiration, 2013, 85(1):3-10.

［8］Zhan XW, Yang L, Zheng MR, et al. Research progress in the regulation of lipogenic balance of bone marrow mesenchymal stem cells ［J］. Acta Antica Sinica,2019,50(3):400-404. (in Chinese)

占秀文,杨磊,郑美蓉,等.骨髓间充质干细胞成脂成骨平衡调控研究进展［J］.解剖学报,2019,50(3):400-404.
［9］Mortada I, Mortada R, Al Bazzal M. Dental pulp stem cells and neurogenesis［J］. Adv Exp Med Biol,2018,1083: 63-75.
［10］Anitua E, Troya M, Zalduendo M. Progress in the use of dental pulp stem cells in regenerative medicine［J］. Cytotherapy,2018,20(4):479-498.
［11］Li WL, Zhang GY, Sun PX. Chemical biology in stem cell research: a new interdisciplinary frontier ［J］. Journal of Second Military Medical University, 2017, 38(2): 133-141. (in Chinese)
李文林,张冠宇,孙平新.干细胞研究中的化学生物学:学科交叉的新前沿［J］.第二军医大学学报,2017,38(2):133-141.
［12］von Wangenheim KH, Peterson HP. The role of cell differentiation in controlling cell multiplication and cancer［J］. J Cancer Res Clin Oncol,2008,134(7):725-741.
［13］Hu H, Zhao P, Liu J, et al. Lanthanum phosphate/chitosan scaffolds enhance cytocompatibility and osteogenic efficiency via the Wnt/β-catenin pathway［J］. J Nanobiotechnology,2018,16(1):98.
［14］Xu YX, Wu CL, Wu Y, et al. Epimedium-derived flavonoids modulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats via Wnt/β-catenin signal pathway activation［J］. Chin J Integr Med, 2012,18(12):909-917.
［15］Tóth F, T?zsér J, Hegedüs C. Effect of inducible BMP-7 expression on the osteogenic differentiation of human dental pulp stem cells［J］. Int J Mol Sci, 2021, 22(12):6182.
［16］He QJ, Zhang NN, Yin YCh, et al. Wnt/β-catenin signaling pathway promotes differentiation of mouse embryonic stem cells into insulin-secreting cells ［J］. Journal of Anhui Medical University, 2019, 54(12): 1841-1848. (in Chinese)
何巧娟,张娜娜,殷应传,等.Wnt/β-catenin信号通路促进小鼠胚胎干细胞向胰岛素分泌细胞分化［J］.安徽医科大学学报,2019,54(12):1841-1848.
［17］Cai M, Li J, Yue R, et al. Glycosylation of DMP1 maintains cranial sutures in mice［J］. J Oral Rehabil, 2020, 47（Suppl 1）:19-28.
［18］Padovano JD, Ramachandran A, Bahmanyar S, et al. Bone-specific overexpression of DMP1 influences osteogenic gene expression during endochondral and intramembranous ossification［J］. Connect Tissue Res,2014,55(Suppl 1):121-124.
［19］Zhao Q, Wang ZX, He JT. Effects of β-ecdysterone on proliferation, differentiation and apoptosis of rat osteoblasts induced by high glucose ［J］. Chinese Journal of Clinical Pharmacology, 2021, 37(4): 443-446. (in Chinese)
赵强,王字兴,何江涛.β-蜕皮甾酮对高糖诱导的大鼠成骨细胞增殖、分化和凋亡的影响［J］.中国临床药理学杂志,2021,37(4):443-446.

［20］Wei J, Karsenty G. An overview of the metabolic functions of osteocalcin［J］. Rev Endocr Metab Disord, 2015,16(2):93-98.

［21］Sun T, Annunziato S, Bergling S, et al. ZNRF3 and RNF43 cooperate to safeguard metabolic liver zonation and hepatocyte proliferation［J］. Cell Stem Cell, 2021, 28(10):1822-1837.