■自体干细胞移植已成为一种有希望的骨修复策略。然而,来自糖尿病患者的间充质干细胞的成骨潜能受损,可能是由于高血糖引起的衰老。这项研究的目的是评估源自H2O2刺激的脂肪干细胞(ADSCs)和非修饰的ADSCs的细胞外囊泡对糖尿病骨髓间充质干细胞(BMSCs)成骨潜能的预处理作用。
■Sprague-Dawley(SD)大鼠通过高脂饮食,然后注射链脲佐菌素,从这些大鼠的骨髓中收集糖尿病BMSCs。细胞外囊泡(EV)从ADSC的条件培养基中分离,有或没有过氧化氢(H2O2)预处理,使用密度梯度离心。H2O2预处理对形貌的影响,标记表达,并对电动汽车的粒径进行了分析。此外,EV预处理对生存能力的影响,生存能力,迁移能力,成骨,细胞衰老,并检测糖尿病BMSCs的氧化应激。此外,还评估了Nrf2/HO-1途径的表达以探索潜在的机制。此外,我们将EV预处理的BMSCs移植到糖尿病大鼠的颅骨缺损中,以评估其体内骨形成和抗衰老能力.
■我们的研究表明,用来自ADSCs的EV预处理显着提高了生存力,衰老,糖尿病BMSCs的成骨分化潜能。此外,体外实验表明,用H2O2激活的EV处理的糖尿病BMSCs表现出增加的活力,减少衰老,与未改良的电动汽车相比,成骨分化增强。此外,当移植到大鼠骨缺损中时,与未修饰的EV相比,用H2O2激活的EV治疗的糖尿病BMSCs显示出改善的骨再生潜能和增强的抗衰老功能。H2O2激活的EV和未修饰的EV均上调糖尿病BMSCs中Nrf2/HO-1通路的表达,然而,H2O2活化的电动汽车的促进作用比未改性的电动汽车更明显。
■来自H2O2预处理的ADSC的细胞外囊泡通过激活Nrf2/HO-1途径减轻糖尿病BMSCs的衰老并增强其骨再生功能。
UNASSIGNED: Autologous stem cell transplantation has emerged as a promising strategy for bone repair. However, the osteogenic potential of mesenchymal stem cells derived from diabetic patients is compromised, possibly due to hyperglycemia-induced senescence. The objective of this study was to assess the preconditioning effects of extracellular vesicles derived from H2O2-stimulated adipose-derived stem cells (
ADSCs) and non-modified
ADSCs on the osteogenic potential of diabetic bone marrow mesenchymal stem cells (BMSCs).
UNASSIGNED: Sprague-Dawley (SD) rats were experimentally induced into a diabetic state through a high-fat diet followed by an injection of streptozotocin, and diabetic BMSCs were collected from the bone marrow of these rats. Extracellular vesicles (EVs) were isolated from the conditioned media of
ADSCs, with or without hydrogen peroxide (H2O2) preconditioning, using density gradient centrifugation. The effects of H2O2 preconditioning on the morphology, marker expression, and particle size of the EVs were analyzed. Furthermore, the impact of EV-pretreatment on the viability, survivability, migration ability, osteogenesis, cellular senescence, and oxidative stress of diabetic BMSCs was examined. Moreover, the expression of the Nrf2/HO-1 pathway was also assessed to explore the underlying mechanism. Additionally, we transplanted EV-pretreated BMSCs into calvarial defects in diabetic rats to assess their in vivo bone formation and anti-senescence capabilities.
UNASSIGNED: Our study demonstrated that pretreatment with EVs from
ADSCs significantly improved the viability, senescence, and osteogenic differentiation potential of diabetic BMSCs. Moreover, in-vitro experiments revealed that diabetic BMSCs treated with H2O2-activated EVs exhibited increased viability, reduced senescence, and enhanced osteogenic differentiation compared to those treated with non-modified EVs. Furthermore, when transplanted into rat bone defects, diabetic BMSCs treated with H2O2-activated EVs showed improved bone regeneration potential and enhanced anti-senescence function t compared to those treated with non-modified EVs. Both H2O2-activated EVs and non-modified EVs upregulated the expression of the Nrf2/HO-1 pathway in diabetic BMSCs, however, the promoting effect of H2O2-activated EVs was more pronounced than that of non-modified EVs.
UNASSIGNED: Extracellular vesicles derived from H2O2-preconditioned
ADSCs mitigated senescence in diabetic BMSCs and enhanced their bone regenerative functions via the activation of the Nrf2/HO-1 pathway.