Characterization of Extracellular Vesicles from Human Saliva: Effects of Age and Isolation Techniques

Salivary extracellular vesicles (EVs) represent an attractive source of biomarkers due to the accessibility of saliva and its non-invasive sampling methods. However, the lack of comparative studies assessing the efficacy of different EV isolation techniques hampers the use of salivary EVs in clinica...

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Bibliographic Details
Published in:Cells (Basel, Switzerland) Switzerland), 2024-01, Vol.13 (1), p.95
Main Authors: Reseco, Lucia, Molina-Crespo, Angela, Atienza, Mercedes, Gonzalez, Esperanza, Falcon-Perez, Juan Manuel, Cantero, Jose L
Format: Article
Language:English
Subjects:
Age
aging
Biomarkers
CD63 antigen
CD81 antigen
CD9 antigen
Cell organelles
Collaboration
Composition
Disease
Exocrine glands
exosomes
Experiments
Extracellular vesicles
Life cycles
Life span
magnetic bead immunocapture
Measurement
Medical research
MicroRNAs
Morphology
Older people
Physiological aspects
Physiology
Proteins
Saliva
Salivary glands
secretions
TEM
Testing
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Summary:Salivary extracellular vesicles (EVs) represent an attractive source of biomarkers due to the accessibility of saliva and its non-invasive sampling methods. However, the lack of comparative studies assessing the efficacy of different EV isolation techniques hampers the use of salivary EVs in clinical settings. Moreover, the effects of age on salivary EVs are largely unknown, hindering the identification of salivary EV-associated biomarkers across the lifespan. To address these questions, we compared salivary EV concentration, size mode, protein concentration, and purity using eight EV isolation techniques before and after magnetic bead immunocapture with antibodies against CD9, CD63, and CD81. The effects of age on salivary EVs obtained with each isolation technique were further investigated. Results showed higher expression of CD63 on isolated salivary EVs compared to the expression of CD81 and flotillin-1. Overall, magnetic bead immunocapture was more efficient in recovering salivary EVs with Norgen's Saliva Exosome Purification Kit and ExoQuick-TC ULTRA at the cost of EV yield. Regardless of age, Invitrogen Total Exosome Isolation Solution showed the highest level of protein concentration, whereas Izon qEVOriginal-70nm columns revealed the highest purity. This study provides the first comprehensive comparison of salivary EVs in younger and older adults using different EV isolation techniques, which represents a step forward for assessing salivary EVs as a source of potential biomarkers of tissue-specific diseases throughout the life cycle.
ISSN:2073-4409
2073-4409
DOI:10.3390/cells13010095