SIRT1 Regulates Osteogenic Differentiation of BMSCs and H-type Vessels Formation to Promote Healing of Osteoporotic Bone Defects

doi:10.13701/j.cnki.kqyxyj.2024.09.006

Abstract

Abstract: Objective: To investigate the role of silent information regulator 1 (SIRT1) on H-type vessels regeneration and bone defect healing in senile osteoporotic state by regulating osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and to explore its specific molecular mechanism. Methods: Immunohistochemical staining was used to detect age-related changes in SIRT1 expression in mouse bone tissue. BMSCs were treated with SIRT1 activator SRT1720 and inhibitor EX527, and the changes in the expression of osteogenic markers and H-type angiogenic factors were detected by RT-qPCR and Western Blot, and the changes in the Wnt/β-catenin signaling pathway in the above process were detected by immunofluorescence staining and Western Blot. A model of osteoporotic bone defects was constructed in the elderly, and SRT1720 and EX527 were administered with XAV939, an inhibitor of Wnt signaling pathway, and CHIR-99021, an activator of Wnt signaling pathway, respectively. The differences in the formation of new bone and H-vessel content of the defects were detected by Micro-CT, HE/Masson staining, and immunofluorescent staining in the respective groups. Results: The expression of SIRT1 showed an age-increasing down-regulation trend in mouse bone tissues. SIRT1 was able to promote the expression of alkaline phosphatase (ALP), Runt-related transcription factor 2 (RUNX2), and the pro-H-type angiogenic factors slit homolog 3 (SLIT3) and vascular endothelial growth factor (VEGF), and able to promote the H-type angiogenesis and bone regeneration in the region of osteoporotic bone defects in old age, which was mediated by the Wnt/β-catenin signaling pathway. Conclusion: SIRT1 can promote the osteogenic differentiation and H-type angiogenesis of BMSCs by regulating the Wnt/β-catenin signaling pathway, which in turn promotes the healing of bone defects in the osteoporotic state of aged mice.

Key words: SIRT1, Wnt/β-catenin signaling pathway, H-type vessels, senile osteoporosis, bone defect

YANG Qiheng, LIU Shibo, LIU Hanghang, LIU Yao, SHAO Jingjing, LUO En. SIRT1 Regulates Osteogenic Differentiation of BMSCs and H-type Vessels Formation to Promote Healing of Osteoporotic Bone Defects[J]. Journal of Oral Science Research, 2024, 40(9): 785-792.

References

[1] 贺丽英,孙蕴,要文娟,等. 2010-2016年中国老年人骨质疏松症患病率Meta分析[J].中国骨质疏松杂志,2016,22(12): 1590-1596.
[2] Filipowska J, Tomaszewski KA, Niedzwiedzki L, et al. The role of vasculature in bone development, regeneration and proper systemic functioning [J]. Angiogenesis, 2017, 20(3): 291-302.
[3] Fan D, Lu J, Yu N, et al. Curcumin prevents diabetic osteoporosis through promoting osteogenesis and angiogenesis coupling via NF-kappaB signaling [J]. Evid Based Complement Alternat Med, 2022, 2022: 4974343.
[4] Jing H, Liao L, Su X, et al. Declining histone acetyltransferase GCN5 represses BMSC-mediated angiogenesis during osteoporosis [J]. FASEB J, 2017, 31(10): 4422-4433.
[5] Kusumbe AP, Ramasamy SK, Adams RH. Coupling of angiogenesis and osteogenesis by a specific vessel subtype in bone [J]. Nature, 2014, 507(7492): 323-328.
[6] Avilkina V, Chauveau C, Ghali MO. Sirtuin function and metabolism: Role in pancreas, liver, and adipose tissue and their crosstalk impacting bone homeostasis [J]. Bone, 2022, 154: 116232.
[7] Chen W, Chen X, Chen AC, et al. Melatonin restores the osteoporosis-impaired osteogenic potential of bone marrow mesenchymal stem cells by preserving SIRT1-mediated intracellular antioxidant properties [J]. Free Radic Biol Med, 2020, 146: 92-106.
[8] Lv N, Zhou Z, Hou M, et al. Research progress of vascularization strategies of tissue-engineered bone [J]. Front Bioeng Biotechnol, 2023, 11: 1291969.
[9] Stahl A, Yang YP. Regenerative approaches for the treatment of large bone defects [J]. Tissue Eng Part B Rev, 2021, 27(6): 539-547.
[10] Chen J, Li M, Liu AQ, et al. Gli1⁺ cells couple with type H vessels and are required for type H vessel formation [J]. Stem Cell Reports, 2020, 15(1): 110-124.
[11] Xie H, Cui Z, Wang L, et al. PDGF-BB secreted by preosteoclasts induces angiogenesis during coupling with osteogenesis [J]. Nat Med, 2014, 20(11): 1270-1278.
[12] Mercken EM, Mitchell SJ, Martin-Montalvo A, et al. SRT2104 extends survival of male mice on a standard diet and preserves bone and muscle mass [J]. Aging Cell, 2014, 13(5): 787-796.
[13] Song J, Li J, Yang F, et al. Nicotinamide mononucleotide promotes osteogenesis and reduces adipogenesis by regulating mesenchymal stromal cells via the SIRT1 pathway in aged bone marrow [J]. Cell Death Dis, 2019, 10(5): 336.
[14] Lemieux ME, Yang X, Jardine K, et al. The Sirt1 deacetylase modulates the insulin-like growth factor signaling pathway in mammals [J]. Mech Ageing Dev, 2005, 126(10): 1097-1105.
[15] Maeda K, Kobayashi Y, Koide M, et al. The regulation of bone metabolism and disorders by Wnt signaling [J]. Int J Mol Sci, 2019, 20(22):5525.
[16] Nusse R, Clevers H. Wnt/beta-catenin signaling, disease, and emerging therapeutic modalities [J]. Cell, 2017, 169(6): 985-999.
[17] Sun Q, Liu S, Feng J, et al. Current status of microRNAs that target the Wnt signaling pathway in regulation of osteogenesis and bone metabolism: A review [J]. Med Sci Monit, 2021, 27: e929510.
[18] Wong SK, Mohamad NV, Jayusman PA, et al. A review on the crosstalk between insulin and Wnt/beta-catenin signalling for bone health [J]. Int J Mol Sci, 2023, 24(15):12441.