Background:Osteoporosis arises mainly from an imbalance in bone remodeling,characterized by the impaired ability of osteoblasts(OBs)to form new bone.A sig-nificant challenge remains in identifying and validating key g...Background:Osteoporosis arises mainly from an imbalance in bone remodeling,characterized by the impaired ability of osteoblasts(OBs)to form new bone.A sig-nificant challenge remains in identifying and validating key genes involved in os-teogenic differentiation to develop effective treatments.Methods:The regulatory role of WIF1 in osteogenic differentiation stages was in-vestigated using Western blot and quantitative polymerase chain reaction assays to measure the expression levels of osteogenic-related genes in MC3T3-E1 cells.Mineralization of OBs was assessed through alizarin red S(ARS)and alkaline phos-phatase(ALP)staining assays.To explore the relationship with mitophagy,RNA se-quencing was performed to examine the effects of Wif1 overexpression on genes re-lated to osteogenic differentiation and mitophagy processes.Additionally,histological staining and micro-computed tomography were conducted on ovar-iectomized(OVX)mice treated with Wif1-overexpressing OB-derived extracellular vesicles(EVs)to evaluate their impact on bone loss and osteogenic differentiation in bone marrow stromal cells(BMSCs).Results:Wif1 was identified as a crucial marker for late-stage osteogenic differentia-tion,playing a role in regulating mitophagy to release functional EVs.In vitro ana-lyses showed that Wif1 overexpression accelerated osteogenic differentiation by ac-tivating genes related to late-stage osteogenic differentiation and mitophagy processes.In vivo experiments demonstrated that administering Wif1-overexpressing OB-derived EVs to OVX mice partially reversed bone loss and countered the sup-pression of osteogenic differentiation in BMSCs.Conclusions:Our findings shed light on the molecular mechanism of Wif1 in osteogenic differentiation and suggested the therapeutic efficacy of Wif1-overexpressing OB-derived EVs in osteoporosis.展开更多
基金supported by the Major Research Plan of the National Natural Science Foundation of China(92249303)the Key Project of the National Natural Science Foundation of China(82230071).
文摘Background:Osteoporosis arises mainly from an imbalance in bone remodeling,characterized by the impaired ability of osteoblasts(OBs)to form new bone.A sig-nificant challenge remains in identifying and validating key genes involved in os-teogenic differentiation to develop effective treatments.Methods:The regulatory role of WIF1 in osteogenic differentiation stages was in-vestigated using Western blot and quantitative polymerase chain reaction assays to measure the expression levels of osteogenic-related genes in MC3T3-E1 cells.Mineralization of OBs was assessed through alizarin red S(ARS)and alkaline phos-phatase(ALP)staining assays.To explore the relationship with mitophagy,RNA se-quencing was performed to examine the effects of Wif1 overexpression on genes re-lated to osteogenic differentiation and mitophagy processes.Additionally,histological staining and micro-computed tomography were conducted on ovar-iectomized(OVX)mice treated with Wif1-overexpressing OB-derived extracellular vesicles(EVs)to evaluate their impact on bone loss and osteogenic differentiation in bone marrow stromal cells(BMSCs).Results:Wif1 was identified as a crucial marker for late-stage osteogenic differentia-tion,playing a role in regulating mitophagy to release functional EVs.In vitro ana-lyses showed that Wif1 overexpression accelerated osteogenic differentiation by ac-tivating genes related to late-stage osteogenic differentiation and mitophagy processes.In vivo experiments demonstrated that administering Wif1-overexpressing OB-derived EVs to OVX mice partially reversed bone loss and countered the sup-pression of osteogenic differentiation in BMSCs.Conclusions:Our findings shed light on the molecular mechanism of Wif1 in osteogenic differentiation and suggested the therapeutic efficacy of Wif1-overexpressing OB-derived EVs in osteoporosis.
