ArticleFrontiers in neuroscience2026
Pre-analytical characterization of CNS-derived extracellular vesicles from human saliva: effect of room temperature and cellular origin.
Article in Frontiers in neuroscience, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.
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1 citing paper in PubMed.
- Quantification of EV-associated miRNA in liquid biopsies for biomarker signature development.Extracellular vesicles and circulating nucleic acids · 2026Review
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8 authors.
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Abstract
Introduction: Blood-derived extracellular vesicles (EVs) from neurons and astrocytes carrying Alzheimer's disease (AD) biomarkers can predict progression from mild cognitive impairment (MCI) to AD; however, their potential in saliva remains largely unexplored. Saliva-derived extracellular vesicles (sEVs) represent a promising non-invasive biomarker source for AD and other age-related dementias (ADRD), but progress has been limited by a lack of standardized protocols for saliva collection, storage, and central nervous system (CNS)-derived EV isolation. Methods: This study had two primary objectives: (1) to optimize enrichment of CNS cell-specific sEVs from the same individuals, and (2) to evaluate the impact of cellular origin and storage temperature (room temperature, 4°C, -20°C) on the stability and quantification of AD-related biomarkers and inflammatory cytokines. Saliva was collected via passive drool from participants in the Nathan Shock Healthy Aging Study (mean age 71.3 years; n = 15). EVs of neuronal, astrocytic, microglial, and oligodendrocyte origin were isolated using ExoQuick-TC precipitation followed by magnetic bead immunocapture. Executive function and attention were assessed using the NIH Toolbox Cognition Battery. Biomarkers were quantified using high-sensitivity immunoassays (MSD, SIMOA Qunaterix). Results: Astrocyte-derived EVs demonstrated significant enrichment of key AD biomarkers, including Aβ40, Aβ42, and total tau. Phosphorylated tau (p-tau217) was largely undetectable across all fractions. TDP-43 was most abundant in EV-depleted saliva, while inflammatory cytokines were broadly distributed across all fractions. Storage temperature did not consistently alter biomarker levels; however, -20°C storage yielded optimal biomarker quantification. Importantly, lower levels of inflammatory cytokines (IFN-γ, IL-10, and IL-6) in EV-depleted saliva were associated with better working memory performance. Discussion: This study provides proof-of-concept validation for the characterization and comparison of multiple CNS-derived salivary EV fractions within the same individuals. The findings support saliva as a feasible, non-invasive matrix for assessing neurodegenerative and neuroinflammatory biomarkers. Establishing a standardized methodology for salivary EV isolation and storage lays the groundwork for future longitudinal studies aimed at diagnosing and predicting AD progression using saliva-based biomarkers.
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