The decrease in zinc-finger E-box-binding homeobox-1 could accelerate steroid-induced osteonecrosis of the femoral head by repressing type-H vessel formation via Wnt/β-catenin pathway

Abstract

Background: Zinc-finger E-box-binding homeobox-1 (ZEB1) is predominantly found in type-H vessels. However, the roles of ZEB1 and type-H vessels in steroid-induced osteonecrosis of the femoral head (SONFH) are unclear.

Methods: Human femoral heads were collected to detect the expression of ZEB1 and the levels of type-H vessels. Then, the SONFH model was developed by injecting C57BL/6 mice with lipopolysaccharide and methylprednisolone. Micro-computed tomography, angiography, double calcein labeling, immunofluorescence, immunohistochemistry, quantitative real-time polymerase chain reaction, and Western blotting were performed to detect the expression of ZEB1, the Wnt/β-catenin pathway, type-H vessels, and the extent to which ZEB1 mediates angiogenesis and osteogenesis. Human umbilical vein endothelial cells were also used to explore the relationship between ZEB1 and the Wnt/β-catenin pathway.

Results: We found that ZEB1 expression and the formation of type-H vessels decreased in SONFH patients and in a mouse model. The number of vascular endothelial growth factors in the femoral heads also decreased. Moreover, the bone mineral density, trabecular number, mineral apposition rate, and expression of genes related to osteogenesis decreased. After ZEB1 knockdown, angiogenesis and osteogenesis decreased. However, the numbers of type-H vessels and the extent of angiogenesis and osteogenesis improved after activation of the Wnt/β-catenin pathway.

Conclusions: The ZEB1 expression decreased in SONFH, causing a decrease in type-H vessel, and it mediated angiogenesis and osteogenesis by regulating the Wnt/β-catenin pathway, ultimately accelerating the process of SONFH.

Keywords

steroid-induced osteonecrosis of the femoral head / type-H vessel / Wnt/β-catenin pathway / zinc-finger E-box-binding homeobox-1

References

[1] MontMA, Cherian JJ, SierraRJ, JonesLC, Lieberman JR. Nontraumatic osteonecrosis of the femoral head: where do we stand today? A ten-year update. J Bone Joint Surg Am. 2015;97:1604-1627.

[2] ZhaoD-W, YuM, HuK, et al. Prevalence of nontraumatic osteonecrosis of the femoral head and its associated risk factors in the Chinese population: results from a nationally representative survey. Chin Med J. 2015;128:2843-2850.

[3] WangA, RenM, WangJ. The pathogenesis of steroid-induced osteonecrosis of the femoral head: a systematic review of the literature. Gene. 2018;671:103-109.

[4] PengY, WuS, LiY, CraneJL. Type-H blood vessels in bone modeling and remodeling. Theranostics. 2020;10:426-436.

[5] ChenY, YinY, LuoM, et al. Occlusal force maintains alveolar bone homeostasis via type-H angiogenesis. J Dent Res. 2023;102(12):1356-1365.

[6] LiJ, WuG, XuC, et al. SLIT guidance ligand 3 (SLIT3) loaded in hydrogel microparticles enhances the tendon-bone healing through promotion of type-H vessel formation: an experimental study in mice. Int J Mol Sci. 2;13638.

[7] RuanZ, YinH, WanT-F, et al. Metformin accelerates bone fracture healing by promoting type-H vessel formation through inhibition of YAP1/TAZ expression. Bone Res. 2023;11:45.

[8] QiuM, LiC, CaiZ, et al. 3D biomimetic calcified cartilaginous callus that induces type-H vessels formation and Osteoclastogenesis. Adv Sci (Weinh). 2023;10:e2207089.

[9] HuangL, WangY, JiangY, Wu Y, HuC, OuyangH. High levels of GSK-3β signalling reduce osteogenic differentiation of stem cells in osteonecrosis of femoral head. J Biochem. 2018;163:243-251.

[10] XuH, FangL, ZengQ, et al. Glycyrrhizic acid alters the hyperoxidative stress-induced differentiation commitment of MSCs by activating the Wnt/β-catenin pathway to prevent SONFH. Food Funct. 2023;14:946-960.

[11] ZhangS, DongK, ZengX, Wang F, LuM. Astragalus polysaccharide ameliorates steroid-induced osteonecrosis of the femoral head by regulating miR-200b-3p-mediated Wnt/β-catenin signaling pathway via inhibiting SP1 expression: astragalus polysaccharide regulates SONFH via SP1. BMC Musculoskelet Disord. 2023;24:369.

[12] ShenJ, SunY, LiuX, et al. EGFL6 regulates angiogenesis and osteogenesis in distraction osteogenesis via Wnt/β-catenin signaling. Stem Cell Res Ther. 2021;12:415.

[13] WangX, MaY, ChenJ, et al. A novel decellularized matrix of Wnt signaling-activated osteocytes accelerates the repair of critical-sized parietal bone defects with osteoclastogenesis, angiogenesis, and neurogenesis. Bioact Mater. 2023;21:110-128.

[14] FuR, HanC-F, NiT, et al. A ZEB1/p53 signaling axis in stromal fibroblasts promotes mammary epithelial tumours. Nat Commun. 2019;10:3210.

[15] FuR, LvW-C, XuY, et al. Endothelial ZEB1 promotes angiogenesis-dependent bone formation and reverses osteoporosis. Nat Commun. 2020;11:460.

[16] ZengK, XieW, WangC, et al. USP22 upregulates ZEB1-mediated VEGFA transcription in hepatocellular carcinoma. Cell Death Dis. 2023;14:194.

[17] ZhouJ, LiY, HeJ, et al. ROS scavenging graphene-based hydrogel enhances type-H vessel formation and vascularized bone regeneration via ZEB1/Notch1 mediation. Macromol Biosci. 2023;23:e2200502.

[18] Sánchez-TillóE, de BarriosO, VallsE, DarlingDS, Castells A, PostigoA. ZEB1 and TCF4 reciprocally modulate their transcriptional activities to regulate Wnt target gene expression. Oncogene. 2015;34:5760-5770.

[19] LiL-Y, YangJ-F, RongF, et al. ZEB1 serves an oncogenic role in the tumourigenesis of HCC by promoting cell proliferation, migration, and inhibiting apoptosis via Wnt/β-catenin signaling pathway. Acta Pharmacol Sin. 2021;42:1676-1689.

[20] WangJ, XuP, ZhouL. Comparison of current treatment strategy for osteonecrosis of the femoral head from the perspective of cell therapy. Front Cell Dev Biol. 2023;11:995816.

[21] YuQC, GengA, PreuschCB, et al. Activation of Wnt/β-catenin signaling by Zeb1 in endothelial progenitors induces vascular quiescence entry. Cell Rep. 2022;41:111694.

[22] ZhaoQ, RenY, XieH, et al. ELK3 mediated by ZEB1 facilitates the growth and metastasis of pancreatic carcinoma by activating the Wnt/β-catenin pathway. Front Cell Dev Biol. 2021;9:700192.

[23] YangJ, TaoQ, ZhouY, et al. MicroRNA-708 represses hepatic stellate cells activation and proliferation by targeting ZEB1 through Wnt/β-catenin pathway. Eur J Pharmacol. 2020;871:172927.

[24] ZhuX, YanF, LiuL, HuangQ. ZEB1 regulates bone metabolism in osteoporotic rats through inducing POLDIP2 transcription. J Orthop Surg Res. 2022;17:423.

[25] TaoS-C, LiX-R, WeiW-J, et al. Polymeric coating on β-TCP scaffolds provides immobilization of small extracellular vesicles with surface-functionalization and ZEB1-loading for bone defect repair in diabetes mellitus. Biomaterials. 2022;283:121465.

[26] RuhM, Stemmler MP, FrischI, et al. The EMT transcription factor ZEB1 blocks osteoblastic differentiation in bone development and osteosarcoma. J Pathol. 2021;254:199-211.

[27] XuC, ShiH, JiangX, et al. ZEB1 mediates bone marrow mesenchymal stem cell osteogenic differentiation partly via Wnt/β-catenin signaling. Front Mol Biosci. 2021;8:682728.

[28] XuZ, Kusumbe AP, CaiH, WanQ, ChenJ. Type-H blood vessels in coupling angiogenesis-osteogenesis and its application in bone tissue engineering. J Biomed Mater Res B Appl Biomater. 2023;111:1434-1446.

[29] LiJ, WeiG, LiuG, et al. Regulating type-H vessel formation and bone metabolism via bone-targeting Oral micro/Nano-hydrogel microspheres to prevent bone loss. Adv Sci (Weinh). 2023;10:e2207381.

[30] CaoH, ShiK, LongJ, et al. PDGF-BB prevents destructive repair and promotes reparative osteogenesis of steroid-associated osteonecrosis of the femoral head in rabbits. Bone. 2023;167:116645.

[31] ZhaoY, LiD, DuanD-P, Song Q-C. The effect of endostatin on angiogenesis and osteogenesis of steroid-induced osteonecrosis of the femoral head in a rabbit model. Acta Orthop Traumatol Turc. 2022;56:178-186.

[32] LiB, LeiY, HuQ, LiD, ZhaoH, Kang P. Porous copper-and lithium-doped nano-hydroxyapatite composite scaffold promotes angiogenesis and bone regeneration in the repair of glucocorticoids-induced osteonecrosis of the femoral head. Biomed Mater. 2021;16:065012.

[33] ZhouC-H, MengJ-H, ZhaoC-C, et al. PTH[1-34] improves the effects of core decompression in early-stage steroid-associated osteonecrosis model by enhancing bone repair and revascularization. PLoS One. 2017;12:e0178781.

[34] SunH, ZhangW, YangN, et al. Activation of cannabinoid receptor 2 alleviates glucocorticoid-induced osteonecrosis of femoral head with osteogenesis and maintenance of blood supply. Cell Death Dis. 2021;12:1035.

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2024 The Author(s). Animal Models and Experimental Medicine published by John Wiley & Sons Australia, Ltd on behalf of The Chinese Association for Laboratory Animal Sciences.