Alzheimer’s disease (AD) has emerged as a global health threat that demands early detection to seize the optimal intervention opportunity. Central nervous system (CNS)-derived extracellular vesicles (EVs), lipid-bilayer nanoparticles released by CNS cells, carry key biomolecules involved in AD pathology, positioning them as a promising source of biomarkers for early detection. Current breakthroughs in EV-based isolation and detection technologies have opened up the possibility of early, accurate AD diagnosis. This review summarizes their multifaceted roles in AD pathogenesis, including amyloid-β (Aβ) aggregation, tau propagation, neuroinflammation, and synaptic dysfunction, and highlights neuron- and glia-derived EV biomarkers with translational potential. We further outline recent advances in EV isolation techniques—including density-, size-, charge/dielectric-, immunoaffinity-, and acoustics-based approaches—and emerging detection platforms such as fluorescence, surface plasmon resonance (SPR), surface-enhanced Raman spectroscopy (SERS), electrochemical, and nanomechanical sensors for sensitive, multiplex AD diagnostics. Finally, we discuss key challenges, including standardization, sensitivity, and high-throughput adaptation, and explore future directions such as automated microfluidics and single-vesicle analysis. CNS-derived EVs hold significant promise as minimally invasive, next-generation tools for early AD detection and precision medicine.

Objectives: Diabetes aggravates the risk and severity of periodontitis, but the specific mechanism remains confused. Complement 3 (C3) is closely related to complications of type 2 diabetes (T2DM). In the present study, we concentrated on whether C3 mediates the development of periodontitis in T2DM.Materials and Methods: Levels of C3 in blood and gingival crevicular fluid (GCF) of patients were measured first. A C3‐knockout diabetic mouse model was established, real‐time PCR, Western blotting and histological investigation were performed to evaluate the progress of periodontitis. Microcomputed tomography (micro‐CT) and TRAP staining were performed to detect alveolar bone resorption. Immunofluorescence was performed to detect polarization of macrophages.Results: Our data showed that C3 levels were elevated in the blood and GCF of T2DM patients compared with non‐diabetic individuals. Increased C3 was closely related to the upregulation of inflammatory cytokines including interleukin (IL)‐1, IL‐6 and tumour necrosis factor‐alpha (TNF‐α), as well as the decline of the bone volume density (BMD) and bone volume over total volume (BV/TV) of the alveolar bones in diabetic mice. The deletion of C3 inhibited inflammatory cytokines and rescued the decreased BMD and BV/TV of the alveolar bones. C3‐mediated polarization of macrophages was responsible for the damage.Conclusion: T2DM‐related upregulation of C3 contributes to the development of periodontitis by promoting macrophages M1 polarization and inhibiting M2 polarization, triggering a pro‐inflammatory effect on periodontal tissues.

Tooth eruption as a crucial part in tooth development and regeneration is accompanied by ongoing osteogenesis and osteoclast activity. The dental follicle (DF) surrounding the developing tooth harbors dental follicle stem cells (DFSCs) which play a crucial role in maintaining bone remodeling. However, the mechanisms through which they regulate the balance between osteogenesis and osteoclast activity during tooth eruption remain poorly understood. Notably, extracellular vesicles (EVs) in bone homeostasis are considered essential. Our study revealed that the DFSCs could modulate tooth eruption by inhibiting osteoclast differentiation via EVs. Further investigation showed that EVs from DFSCs could inhibit osteoclast differentiation through the ANXA1-PPARγ-CEBPα pathway. Animal experiments indicated that EVs from DFSCs and the cargo ANXA1 affected tooth eruption. In summary, this study suggests the critical role of the dental follicle in tooth eruption through EVs, which may provide therapeutic targets for abnormal tooth eruption and effective approaches for the eruption of regenerated teeth.

Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) are emerging as promising treatments for immunomodulation and tissue regeneration. However, the scalable production of functionally enhanced EVs remains a critical challenge. This study introduces microgravity culture using a three-dimensional (3D) rotating cell culture system as a novel strategy to optimize MSC-EVs yield and bioactivity. We first investigated the effects of microgravity on the proliferation and stemness of human umbilical cord-derived MSCs (UCMSCs). The yield of microgravity-derived EVs (µg-EVs) was quantified by nanoparticle tracking analysis. The function of µg-EVs was analyzed by proteomic profiling and further assessed by macrophage polarization and osteogenic differentiation of periodontal ligament stem cells (PDLSCs). Proteomic analysis of UCMSCs was performed to further explore the underlying mechanisms of EVs biogenesis and functional activity under microgravity condition. Microgravity culture significantly enhanced UCMSCs proliferation and stemness. Compared with conventional static culture, EVs production increased by 7.7-fold under microgravity. Functionally, µg-EVs more effectively promoted macrophage polarization toward the anti-inflammatory M2 phenotype and significantly enhanced the osteogenic differentiation capacity of PDLSCs. Mechanistically, Rab27B upregulation in microgravity-cultured UCMSCs was associated with increased EVs secretion and enhanced therapeutic efficacy. This study identifies microgravity as an effective platform for the large-scale production of high-quality UCMSC-EVs, addressing key manufacturing barriers and accelerating the clinical translation of EVs-based therapies.

Extracellular vesicles (EVs) derived from mesenchymal stem cells (MSC-EVs) are promising therapeutic agents for various diseases. However, current methods to improve MSC-EV production are insufficient to meet the clinical demands. Although various strategies have been investigated to enhance MSC-EV production, they are often hampered by limited scalability, loss of stemness, or suboptimal therapeutic outcomes. Our study identified three key stimulators that significantly boosted MSC-EV production: epidermal growth factor (EGF), tumor necrosis factor-α (TNF-α), and hypoxia. Employing an orthogonal design, we developed an optimized cell culture condition, subsequently referred to as ETH (EGF 10 ng/mL, TNF-α 50 ng/mL, and a hypoxic environment of 1% O₂) preconditioning. This approach led to a remarkable 4- to 5-fold increase in MSC-EV yield while preserving the stemness of MSCs. Through proteomic analysis, we elucidated the underlying mechanisms of ETH preconditioning, providing insight into the complex processes driving enhanced MSC-EV production. Notably, MSC-EVs generated through ETH preconditioning demonstrated enhanced therapeutic potential including superior angiogenesis, collagen deposition, and regulation of inflammation. These findings present a scalable and effective strategy for elevating MSC-EV production, paving the way for its broader clinical application in regenerative medicine.