Evidence map›Paper›PMID 39462790›Full record

ArticleOral diseases2025

Salivary Extracellular Vesicles Separation: Analysis of Ultracentrifugation-Based Protocols.

Castillejos-García Itzel, Martínez-Martínez Eduardo, Ramírez-Amador Velia, Cisneros-Villanueva Mireya, Hidalgo-Miranda Alfredo, Ramos-Godínez María Del Pilar, Anaya-Saavedra Gabriela

Abstract read
In one paragraph

Article in Oral diseases, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

3 citing papers in PubMed.

  1. Review
  2. Review
  3. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors.

Castillejos-García ItzelDoctorado en Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana, Ciudad de México, Mexico.ORCID https://orcid.org/0000-0001-5318-3697
Martínez-Martínez EduardoLaboratorio de Vesículas Extracelulares y Comunicación Celular, Instituto Nacional de Medicina Genómica, Ciudad de México, Mexico.
Ramírez-Amador VeliaPosgrado en Patología y Medicina Oral, Universidad Autónoma Metropolitana-Xochimilco, Ciudad de México, Mexico.
Cisneros-Villanueva MireyaLaboratorio de Genómica del Cáncer, Instituto Nacional de Medicina Genómica, Ciudad de México, Mexico.ORCID https://orcid.org/0000-0002-5662-0247
Hidalgo-Miranda AlfredoLaboratorio de Genómica del Cáncer, Instituto Nacional de Medicina Genómica, Ciudad de México, Mexico.
Ramos-Godínez María Del PilarLaboratorio de Microscopía Electrónica, Instituto Nacional de Cancerología, Ciudad de México, Mexico.
Anaya-Saavedra GabrielaPosgrado en Patología y Medicina Oral, Universidad Autónoma Metropolitana-Xochimilco, Ciudad de México, Mexico.ORCID https://orcid.org/0000-0002-8881-867X

Funding

Consejo Nacional de Ciencias, Humanidades y Tecnología, MéxicoUNIVERSIDAD AUTONOMA METROPOLITANA
6 · The paper itself

Abstract

introductionThe clinical potential of extracellular vesicles (EVs) is widely acknowledged, yet the standardization and reproducibility of its separation remain challenging. This study compares three protocols: ultracentrifugation (UC), UC with purification step (UC + PS), and a combined protocol using polymer-based precipitation and UC (PBP + UC).

methodsSalivary samples were collected from healthy donors. EVs were separated (UC, UC + PS, and PBP + UC) and characterized using transmission electron microscopy, nanoparticle tracking analysis, EV purity, RNA concentration, and Western blotting. miRNA expression was evaluated by quantitative RT-PCR. Statistical analyses comparing groups were performed using ANOVA.

resultsAll methods successfully separated CD9+ and CD63+ EVs from saliva. The UC + PS and PBP + UC protocols yielded the highest concentrations of EVs, enriched in < 200 nm vesicles. EV purity and RNA recovery were comparable among all methods. Expression of miR-16, miR-27a, and miR-99a was successfully detected using all methods.

conclusionsThe UC + PS and PBP + UC protocols demonstrate comparable efficiency in separating salivary EVs. However, the combined PBP + UC protocol, with its simplified processing capability, offers a significant advantage, particularly in the initial phase of EV separation. This finding suggests its potential application in clinical settings where time-sensitive simple processing is critical. Further validation is needed to confirm its effectiveness for transcriptomic and proteomic analyses.

Indexed as

Extracellular VesiclesSalivaUltracentrifugationAdultFemaleHumansMaleMicroRNAsMicroscopy, Electron, TransmissionMicroRNAsextracellular vesiclesmiRNAsPEGsalivaultracentrifugation

Identifiers

PMID39462790
PMCPMC12021316

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.