Abstract
This pilot study aims to investigate whether salivary small extracellular vesicle (sEV)-associated microRNAs could act as potential biomarkers for periodontal disease status. Twenty-nine participants (10 who were healthy, nine with gingivitis, 10 with stage III/IV periodontitis) were recruited and unstimulated whole saliva samples were collected. Salivary sEVs were isolated using the size-exclusion chromatography (SEC) method and characterised by morphology, EV-protein and size distribution using transmission electron microscopy (TEM), Western Blot and Nanoparticle Tracking Analysis (NTA), respectively. Ten mature microRNAs (miRNAs) in salivary sEVs and saliva were evaluated using RT-qPCR. The discriminatory power of miRNAs as biomarkers in gingivitis and periodontitis versus healthy controls was evaluated by Receiver Operating Characteristics (ROC) curves. Salivary sEVs were comparable to sEVs morphology, mode, size distribution and particle concentration in healthy, gingivitis and periodontitis patients. Compared to miRNAs in whole saliva, three significantly increased miRNAs (hsa-miR-140-5p, hsa-miR-146a-5p and hsa-miR-628-5p) were only detected in sEVs in periodontitis when compared to that of healthy controls, with a good discriminatory power (area under the curve (AUC) = 0.96) for periodontitis diagnosis. Our study demonstrated that salivary sEVs are a non-invasive source of miRNAs for periodontitis diagnosis. Three miRNAs that are selectively enriched in sEVs, but not whole saliva, could be potential biomarkers for periodontal disease status.
Highlights
Periodontitis is a complex inflammatory disease, associated with a dysbiotic plaque biofilm and characterised by the destruction of periodontal tissues
One-specificity, while the area under the curve (AUC) indicates the diIsnct.rJi.mMionl.aStcoi.r2y02p0o, w21e, xr FoOf RthPeEEbRioRmEVaIrEkWers. This pilot study is the first to investigate the diagnostic potential of salivary small extracellular vesicle (sEV)-associated miRNAs in periodontal disease
The overall objective was to determine the diagnostic value of miRNAs obtained from sEVs compared to those sourced from whole saliva in terms of discriminating
Summary
Periodontitis is a complex inflammatory disease, associated with a dysbiotic plaque biofilm and characterised by the destruction of periodontal tissues. Very recent research has demonstrated that salivary CD9- and CD81-positive (two tetraspanin proteins enriched in EVs) sEVs are decreased in periodontitis compared to healthy controls [17]. According to published reviews [21,22,23,24,25,26], ten periodontitis-associated miRNAs (hsa-miR-15a-5p, hsa-miR-29b-3p, hsa-miR-124-3p, hsa-miR-140-5p, hsa-miR-146a-5p, hsa-miR-148a-3p, hsa-miR-155-5p, hsa-miR-223-3p, hsa-miR-301b, hsa-miR-628-5p) have been explored as potential periodontitis biomarkers from one or more sample sources, such as saliva, gingival tissues, or periodontium-derived cells. Whether salivary sEV-associated miRNAs have the same profile as whole saliva-associated miRNAs in periodontitis remains unknown This pilot study aimed to evaluate the miRNA expression profile of whole saliva and salivary sEVs obtained from gingivitis, periodontitis and periodontally healthy patients, and to determine the diagnostic potential of miRNAs associated with sEVs as biomarkers of periodontal status
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