Small molecule drug TD-198946 enhances osteogenic differentiation of rat bone marrow mesenchymal stem cells

doi:10.12307/2025.060

Abstract

Abstract: BACKGROUND: TD-198946 is a small molecule drug known to induce stem cells to form cartilage; however, its effect on osteogenic differentiation remains unclear.
OBJECTIVE: To investigate the effects of the small molecule drug TD-198946 on promoting osteoblastic differentiation of rat bone marrow mesenchymal stem cells and its mechanism of action.
METHODS: Bone marrow mesenchymal stem cells (BMSCs) were extracted from SD rats. CCK-8 assay was used to evaluate the effect of different concentrations of TD-198946 on the proliferation of bone marrow mesenchymal stem cells to determine the optimal concentration of TD-198946. Then, the optimal concentration of TD-198946 and osteogenic medium were added to induce osteogenic differentiation of bone marrow mesenchymal stem cells. On day 3, qRT-PCR was used to detect the expression of alkaline phosphatase, Runt-related transcription factor 2, osteopontin, osteocalcin, and type I collagen genes. On day 7 of osteogenic induction, alkaline phosphatase staining and western blot assay were performed to detect the expression of Runt-related transcription factor 2, type I collagen, AKT, p-AKT, PI3K, and p-PI3K proteins. On day 21 of osteogenic induction, Alizarin red staining was conducted.
RESULTS AND CONCLUSION: (1) CCK-8 assay results revealed that 100 nmol/L TD-198946 could promote the proliferation of bone marrow mesenchymal stem cells. (2) The results of alkaline phosphatase staining and alizarin red staining showed that TD-198946 could promote osteogenic differentiation of rat bone marrow mesenchymal stem cells. (3) qRT-PCR results showed that TD-198946 could promote the expression of osteogenic genes alkaline phosphatase, Runt-related transcription factor 2, osteopontin, osteocalcin, and type I collagen. (4) Western blot assay results showed that TD-198946 could enhance the expression of Runt-related transcription factor 2, type I collagen, p-PI3K, and p-AKT. The results indicate that the small molecule drug TD-198946 may induce osteogenic differentiation of rat bone marrow mesenchymal stem cells by activating the PI3K/AKT signaling pathway.

Key words: bone marrow mesenchymal stem cell, TD-198946, osteogenic differentiation, PI3K/AKT signaling pathway, AKT, p-AKT

CLC Number:

Yang Chao, Luo Zongping. Small molecule drug TD-198946 enhances osteogenic differentiation of rat bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(13): 2648-2654.

Figures/Tables 6

References

[1] ZHAO H, DING Y, YANG J, et al. Efficacy and safety of bisphosphonates on childhood osteoporosis secondary to chronic illness or its treatment: a meta-analysis. Ther Adv Chronic Dis. 2022;13:20406223221129163.
[2] CHANG X, XU S, ZHANG H. Regulation of bone health through physical exercise: Mechanisms and types. Front Endocrinol (Lausanne). 2022; 13:1029475.
[3] PRADA D, CRANDALL CJ, KUPSCO A, et al. Air pollution and decreased bone mineral density among Women’s Health Initiative participants. EClinicalMedicine. 2023;57:101864.
[4] DE SIRE A, LIPPI L, APRILE V, et al. Pharmacological, Nutritional, and Rehabilitative Interventions to Improve the Complex Management of Osteoporosis in Patients with Chronic Obstructive Pulmonary Disease: A Narrative Review. J Pers Med. 2022;12(10):1626.
[5] WANG W, LIU H, LIU T, et al. Insights into the Role of Macrophage Polarization in the Pathogenesis of Osteoporosis. Oxid Med Cell Longev. 2022;2022:2485959.
[6] ZHOU J, CHENG J, LIU L, et al. Lactobacillus acidophilus (LA) Fermenting Astragalus Polysaccharides (APS) Improves Calcium Absorption and Osteoporosis by Altering Gut Microbiota. Foods. 2023;12(2):275.
[7] TONK CH, SHOUSHRAH SH, BABCZYK P, et al. Therapeutic Treatments for Osteoporosis-Which Combination of Pills Is the Best among the Bad? Int J Mol Sci. 2022;23(3):1393.
[8] JAFARI A, QANIE D, ANDERSEN TL, et al. Legumain Regulates Differentiation Fate of Human Bone Marrow Stromal Cells and Is Altered in Postmenopausal Osteoporosis. Stem Cell Reports. 2017; 8(2):373-386.
[9] MASTROLIA I, GIORGINI A, MURGIA A, et al. Autologous Marrow Mesenchymal Stem Cell Driving Bone Regeneration in a Rabbit Model of Femoral Head Osteonecrosis. Pharmaceutics. 2022;14(10):2127.
[10] CAMPBELL TM, DILWORTH FJ, ALLAN DS, et al. The Hunt Is On! In Pursuit of the Ideal Stem Cell Population for Cartilage Regeneration. Front Bioeng Biotechnol. 2022;10:866148.
[11] YANO F, HOJO H, OHBA S, et al. A novel disease-modifying osteoarthritis drug candidate targeting Runx1. Ann Rheum Dis. 2013; 72(5):748-753.
[12] YANO F, HOJO H, OHBA S, et al. Cell-sheet technology combined with a thienoindazole derivative small compound TD-198946 for cartilage regeneration. Biomaterials. 2013;34(22):5581-5587.
[13] KOBAYASHI M, CHIJIMATSU R, HART DA, et al. Evidence that TD-198946 enhances the chondrogenic potential of human synovium-derived stem cells through the NOTCH3 signaling pathway. J Tissue Eng Regen Med. 2021;15(2):103-115.
[14] KUSHIOKA J, KAITO T, CHIJIMATSU R, et al. The small compound, TD-198946, protects against intervertebral degeneration by enhancing glycosaminoglycan synthesis in nucleus pulposus cells. Sci Rep. 2020; 10(1):14190.
[15] WANG J, WU H, PENG Y, et al. Hypoxia adipose stem cell-derived exosomes promote high-quality healing of diabetic wound involves activation of PI3K/Akt pathways. J Nanobiotechnology. 2021;19(1): 202.
[16] LIU X, WANG J, FAN Y, et al. Particulate Matter Exposure History Affects Antioxidant Defense Response of Mouse Lung to Haze Episodes. Environ Sci Technol. 2019;53(16):9789-9799.
[17] KENT LN, RUMI MA, KUBOTA K, et al. FOSL1 is integral to establishing the maternal-fetal interface. Mol Cell Biol. 2011;31(23):4801-4813.
[18] ZHANG W, LI X, ZHANG W, et al. The LncRNA CASC11 Promotes Colorectal Cancer Cell Proliferation and Migration by Adsorbing miR-646 and miR-381-3p to Upregulate Their Target RAB11FIP2. Front Oncol. 2021;11:657650.
[19] ZHAO B, PENG Q, POON EHL, et al. Leonurine Promotes the Osteoblast Differentiation of Rat BMSCs by Activation of Autophagy via the PI3K/Akt/mTOR Pathway. Front Bioeng Biotechnol. 2021;9:615191.
[20] SUN K, LUO J, GUO J, et al. The PI3K/AKT/mTOR signaling pathway in osteoarthritis: a narrative review. Osteoarthritis Cartilage. 2020; 28(4):400-409.
[21] CHEN X, CHEN W, AUNG ZM, et al. LY3023414 inhibits both osteogenesis and osteoclastogenesis through the PI3K/Akt/GSK3 signalling pathway. Bone Joint Res. 2021;10(4):237-249.
[22] CHO SW, SUN HJ, YANG JY, et al. Transplantation of mesenchymal stem cells overexpressing RANK-Fc or CXCR4 prevents bone loss in ovariectomized mice. Mol Ther. 2009;17(11):1979-1987.
[23] HUANG J, PARK J, JUNG N, et al. Hydrothermally treated coral scaffold promotes proliferation of mesenchymal stem cells and enhances segmental bone defect healing. Front Bioeng Biotechnol. 2023;11:1332138.
[24] WAESE EY, KANDEL RA, STANFORD WL. Application of stem cells in bone repair. Skeletal Radiol. 2008;37(7):601-608.
[25] PIPINO C, PANDOLFI A. Osteogenic differentiation of amniotic fluid mesenchymal stromal cells and their bone regeneration potential. World J Stem Cells. 2015;7(4):681-690.
[26] 马明明,徐宏光,张艺凡,等.小分子药物TD-198946对终板软骨退变具有保护作用[J].中国临床药理学与治疗学,2015,20(8): 871-875.
[27] YE C, ZHANG W, HANG K, et al. Extracellular IL-37 promotes osteogenic differentiation of human bone marrow mesenchymal stem cells via activation of the PI3K/AKT signaling pathway. Cell Death Dis. 2019; 10(10):753.
[28] LIU X, BRUXVOORT KJ, ZYLSTRA CR, et al. Lifelong accumulation of bone in mice lacking Pten in osteoblasts. Proc Natl Acad Sci U S A. 2007;104(7):2259-2264.
[29] LI H, LI T, FAN J, et al. miR-216a rescues dexamethasone suppression of osteogenesis, promotes osteoblast differentiation and enhances bone formation, by regulating c-Cbl-mediated PI3K/AKT pathway. Cell Death Differ. 2015;22(12):1935-1945.
[30] ZHOU X, XI K, BIAN J, et al. Injectable engineered micro/nano-complexes trigger the reprogramming of bone immune epigenetics. Chem Eng J. 2023;462:142158.
[31] SHANG J, YU Z, XIONG C, et al. Resistin targets TAZ to promote osteogenic differentiation through PI3K/AKT/mTOR pathway. iScience. 2023;26(7):107025.