牙周炎是一种慢性传染病,其特征是进行性炎症和牙槽骨丢失。叉头箱O1(FoxO1),一个重要的监管者,间充质干细胞(MSCs)在维持骨稳态、调节巨噬细胞能量代谢和成骨分化等方面起着至关重要的作用。在这项研究中,使用工程技术将FoxO1过表达到小的细胞外囊泡(sEV)中,研究了FoxO1过表达sEV对牙周组织再生的影响及其机制。
■使用超速离心分离人牙周膜干细胞(hPDLSCs)来源的sEV(hPDLSCs-sEV)。然后用透射电子显微镜对它们进行了表征,Nanosight,和西方印迹分析。脂多糖刺激后,用hPDLSCs-sEV体外处理hPDLSCs,并评估成骨作用。还评价了hPDLSCs-sEV对THP-1巨噬细胞的极化表型的影响。此外,我们测量了活性氧(ROS)水平,三磷酸腺苷(ATP)生产,线粒体特征,以及hPDLSCs和THP-1细胞的代谢。在小鼠体内建立实验性牙周炎。将HPDLSCs-sEV或磷酸盐缓冲盐水(PBS)注射入牙周组织4周,上颌骨被收集并通过显微计算机断层扫描进行评估,组织学染色,和小动物体内成像。
■体外,FoxO1过表达的sEV促进炎症环境中hPDLSCs的成骨分化,并将THP-1细胞从M1表型极化为M2表型。此外,FoxO1过表达的sEV调节ROS水平,ATP生产,线粒体特征,以及hPDLSCs和THP-1细胞在炎症环境中的代谢。在体内分析中,FoxO1过表达sEV可有效促进骨形成并抑制炎症。
■过表达FoxO1的sEV可以调节成骨和免疫调节。FoxO1过表达的sEV调节炎症和成骨的能力可以为建立牙周炎的治疗方法铺平道路。
UNASSIGNED: Periodontitis is a chronic infectious disease characterized by progressive inflammation and alveolar bone loss. Forkhead box O1 (FoxO1), an important regulator, plays a crucial role in maintaining bone homeostasis and regulating macrophage energy metabolism and osteogenic differentiation of mesenchymal stem cells (MSCs). In this study, FoxO1 was overexpressed into small extracellular vesicles (sEV) using engineering technology, and effects of FoxO1-overexpressed sEV on periodontal tissue regeneration as well as the underlying mechanisms were investigated.
UNASSIGNED: Human periodontal ligament stem cell (hPDLSCs)-derived sEV (hPDLSCs-sEV) were isolated using ultracentrifugation. They were then characterized using transmission electron microscopy, Nanosight, and Western blotting analyses. hPDLSCs were treated with hPDLSCs-sEV in vitro after stimulation with lipopolysaccharide, and osteogenesis was evaluated. The effect of hPDLSCs-sEV on the polarization phenotype of THP-1 macrophages was also evaluated. In addition, we measured the reactive oxygen species (ROS) levels, adenosine triphosphate (ATP) production, mitochondrial characteristics, and metabolism of hPDLSCs and THP-1 cells. Experimental periodontitis was established in vivo in mice. HPDLSCs-sEV or phosphate-buffered saline (PBS) were injected into periodontal tissues for four weeks, and the maxillae were collected and assessed by micro-computed tomography, histological staining, and small animal in vivo imaging.
UNASSIGNED: In vitro, FoxO1-overexpressed sEV promoted osteogenic differentiation of hPDLSCs in the inflammatory environment and polarized THP-1 cells from the M1 phenotype to the M2 phenotype. Furthermore, FoxO1-overexpressed sEV regulated the ROS level, ATP production, mitochondrial characteristics, and metabolism of hPDLSCs and THP-1 cells in the inflammatory environment. In the in vivo analyses, FoxO1-overexpressed sEV effectively promoted bone formation and inhibited inflammation.
UNASSIGNED: FoxO1-overexpressed sEV can regulate osteogenesis and immunomodulation. The ability of FoxO1-overexpressed sEV to regulate inflammation and osteogenesis can pave the way for the establishment of a therapeutic approach for periodontitis.