Glucocorticoids induce femoral head necrosis in rats through the HIF-1α/VEGF signaling pathway
Abstract Glucocorticoid-induced osteoblast dysfunction is the primary cause of steroid-induced osteonecrosis of the femoral head (SONFH). However, the specific underlying biological mechanisms of glucocorticoids’ effect on osteoblasts remain undetermined. Recently, the role of hypoxia-inducible fact...
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Nature Portfolio
2025-08-01
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| Online Access: | https://doi.org/10.1038/s41598-025-15018-4 |
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| author | HaoXu Tang Lingli Yuan ZhiYuan Xu GuiFen Jiang YingJie Liang Ce Li PengLin Ding MinLong Qian |
| author_facet | HaoXu Tang Lingli Yuan ZhiYuan Xu GuiFen Jiang YingJie Liang Ce Li PengLin Ding MinLong Qian |
| author_sort | HaoXu Tang |
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| description | Abstract Glucocorticoid-induced osteoblast dysfunction is the primary cause of steroid-induced osteonecrosis of the femoral head (SONFH). However, the specific underlying biological mechanisms of glucocorticoids’ effect on osteoblasts remain undetermined. Recently, the role of hypoxia-inducible factor 1-alpha (HIF-1α)/vascular endothelial growth factor (VEGF) signaling pathway in modulating bone formation has been studied. This study aimed to investigate the association and mechanism of the HIF-1α/VEGF signaling pathway in glucocorticoid-induced osteogenesis suppression in MC3T3-E1 cells. This study performed CCK8 and live/dead staining assays by stimulating MC3T3-E1 cells with varying dexamethasone (DEX) doses to elucidate its influence on cell proliferation and activity. Furthermore, Western blotting was carried out to investigate the expression of HIF-1α, runt-related transcription factor 2 (RUNX2), VEGF, osteopontin (OPN), and alkaline phosphatase (ALP) proteins to identify the optimal DEX concentration for simulating steroid-induced osteonecrosis cell models. Moreover, the osteogenic differentiation of cells was assessed by transfecting them with control or HIF-1α overexpression lentiviral vectors. Similarly, in vivo, hematoxylin and eosin staining, immunohistochemical staining, and micro-computed tomography were performed to validate in vitro results in the SONFH rat model. In vitro analyses revealed that a 10− 6 M concentration of DEX significantly suppressed cell viability and osteogenesis by decreasing HIF-1α and VEGF levels. Furthermore, HIF-1α upregulation increased osteogenic activity and VEGF expression in MC3T3-E1 cells. However, the HIF-1α antagonist 3-(5’-hydroxymethyl-2’-furyl) -1-benzylindazole (YC-1) indicated opposite effects in DEX-treated MC3T3-E1 cells. Moreover, SONFH femoral heads had reduced bone density, bone tissue content, and femoral head integrity, as well as increased bone cell lacunae, while decreased HIF-1α, OPN, VEGF, and ALP levels in bone tissue compared to normal rats. This study indicated that DEX suppresses osteoblast differentiation via the HIF-1α/VEGF pathway, thus promoting SONFH. |
| format | Article |
| id | doaj-art-431ba0e3cda94f8584b4affbb2e84e0f |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-08-01 |
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| spelling | doaj-art-431ba0e3cda94f8584b4affbb2e84e0f2025-08-20T03:42:27ZengNature PortfolioScientific Reports2045-23222025-08-0115111910.1038/s41598-025-15018-4Glucocorticoids induce femoral head necrosis in rats through the HIF-1α/VEGF signaling pathwayHaoXu Tang0Lingli Yuan1ZhiYuan Xu2GuiFen Jiang3YingJie Liang4Ce Li5PengLin Ding6MinLong Qian7Department, Bengbu Medical UniversityDepartment of the Second, Affiliated Hospital of Bengbu Medical UniversityDepartment of the Bozhou People’s HospitalDepartment of Pathology, The Sixth Affiliated Hospital, School of Medicine, South China University of TechnologyDepartment of the Second, Affiliated Hospital of Bengbu Medical UniversityDepartment of the Second, Affiliated Hospital of Bengbu Medical UniversityDepartment of the Second, Affiliated Hospital of Bengbu Medical UniversityDepartment of the Second, Affiliated Hospital of Bengbu Medical UniversityAbstract Glucocorticoid-induced osteoblast dysfunction is the primary cause of steroid-induced osteonecrosis of the femoral head (SONFH). However, the specific underlying biological mechanisms of glucocorticoids’ effect on osteoblasts remain undetermined. Recently, the role of hypoxia-inducible factor 1-alpha (HIF-1α)/vascular endothelial growth factor (VEGF) signaling pathway in modulating bone formation has been studied. This study aimed to investigate the association and mechanism of the HIF-1α/VEGF signaling pathway in glucocorticoid-induced osteogenesis suppression in MC3T3-E1 cells. This study performed CCK8 and live/dead staining assays by stimulating MC3T3-E1 cells with varying dexamethasone (DEX) doses to elucidate its influence on cell proliferation and activity. Furthermore, Western blotting was carried out to investigate the expression of HIF-1α, runt-related transcription factor 2 (RUNX2), VEGF, osteopontin (OPN), and alkaline phosphatase (ALP) proteins to identify the optimal DEX concentration for simulating steroid-induced osteonecrosis cell models. Moreover, the osteogenic differentiation of cells was assessed by transfecting them with control or HIF-1α overexpression lentiviral vectors. Similarly, in vivo, hematoxylin and eosin staining, immunohistochemical staining, and micro-computed tomography were performed to validate in vitro results in the SONFH rat model. In vitro analyses revealed that a 10− 6 M concentration of DEX significantly suppressed cell viability and osteogenesis by decreasing HIF-1α and VEGF levels. Furthermore, HIF-1α upregulation increased osteogenic activity and VEGF expression in MC3T3-E1 cells. However, the HIF-1α antagonist 3-(5’-hydroxymethyl-2’-furyl) -1-benzylindazole (YC-1) indicated opposite effects in DEX-treated MC3T3-E1 cells. Moreover, SONFH femoral heads had reduced bone density, bone tissue content, and femoral head integrity, as well as increased bone cell lacunae, while decreased HIF-1α, OPN, VEGF, and ALP levels in bone tissue compared to normal rats. This study indicated that DEX suppresses osteoblast differentiation via the HIF-1α/VEGF pathway, thus promoting SONFH.https://doi.org/10.1038/s41598-025-15018-4Steroid-induced osteonecrosis of the femoral headHIF-1αVEGFOsteoblastsOsteogenic differentiation |
| spellingShingle | HaoXu Tang Lingli Yuan ZhiYuan Xu GuiFen Jiang YingJie Liang Ce Li PengLin Ding MinLong Qian Glucocorticoids induce femoral head necrosis in rats through the HIF-1α/VEGF signaling pathway Scientific Reports Steroid-induced osteonecrosis of the femoral head HIF-1α VEGF Osteoblasts Osteogenic differentiation |
| title | Glucocorticoids induce femoral head necrosis in rats through the HIF-1α/VEGF signaling pathway |
| title_full | Glucocorticoids induce femoral head necrosis in rats through the HIF-1α/VEGF signaling pathway |
| title_fullStr | Glucocorticoids induce femoral head necrosis in rats through the HIF-1α/VEGF signaling pathway |
| title_full_unstemmed | Glucocorticoids induce femoral head necrosis in rats through the HIF-1α/VEGF signaling pathway |
| title_short | Glucocorticoids induce femoral head necrosis in rats through the HIF-1α/VEGF signaling pathway |
| title_sort | glucocorticoids induce femoral head necrosis in rats through the hif 1α vegf signaling pathway |
| topic | Steroid-induced osteonecrosis of the femoral head HIF-1α VEGF Osteoblasts Osteogenic differentiation |
| url | https://doi.org/10.1038/s41598-025-15018-4 |
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