Quantitative proteomic analysis in symptomatic and asymptomatic apical periodontitis

C Loureiro, M A R Buzalaf, F R N Moraes, T M O Ventura, V T Pelá, J P Pessan, R C Jacinto
International Endodontic Journal 2021 January 22

AIM: To quantitatively and qualitatively compare the host proteomic profile in samples of symptomatic and asymptomatic apical periodontitis (AP) using nano Liquid Chromatography Electron Spray Tandem Mass Spectrometry.

METHODOLOGY: Samples were obtained from 18 patients with radiographically evident AP, divided into symptomatic and asymptomatic groups (nine per group) according to clinical characteristics. After sample collection, protein extraction, purification and quantification of the samples were performed, which were analysed by reverse phase liquid chromatography coupled to mass spectrometry. Label-free quantitative proteomic analysis was performed by Protein Lynx Global Service software. Differences in expression of proteins between the groups were calculated using the Monte Carlo algorithm, considering p < 0.05 for down-regulated proteins and 1-p > 0.95 for up-regulated proteins. Proteins were identified with the embedded ion accounting algorithm in the software and a search of the Homo sapiens UniProt database.

RESULTS: A total of 853 individual human proteins were identified. In the quantitative analysis, common proteins to both groups accounted for 143 proteins. Differences in expression between groups resulted in 51 up-regulated proteins (1-p > 0.95) in the symptomatic group, including Alpha-1-antitrypsin, Protein S100-A8, Myeloperoxidase, Peroxiredoxin, and Lactotransferrin. This group also had 43 down-regulated proteins (p < 0.05), comprising Immunoglobulin, Neutrophil defensin, Pyruvate kinase, and Alpha-enolase. The qualitative analysis considered only the exclusive proteins of each group. For the symptomatic group, 318 complete proteins and 29 fragments were identified, such as Dedicator of cytokinesis protein, Intersectin, Prostaglandin, Phospholipase DDHD2, and Superoxide dismutase. For the asymptomatic group, 326 complete proteins and 37 fragments were identified, including Azurocidin, C-reactive protein, Collagen alpha, Cathepsin, Heat Shock, and Laminin.

CONCLUSIONS: Quantitative differences in the expression of common proteins in cases of symptomatic and asymptomatic AP were found, which were mostly related to host immune response in both groups. Exclusive proteins in the symptomatic group were mainly related to the host response to the presence of viruses in endodontic infections, oxidative stress, and proteolytic enzymes. The results provide a basis for a better understanding of cellular and molecular pathways involved in AP, establishing specific proteomic profiles for symptomatic and asymptomatic conditions.

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