Blanco-Pintos Triana, Regueira-Iglesias Alba, Suárez-Rodríguez Berta, Seijas-Otero Noelia, Relvas Marta, Bravo Susana B, Balsa-Castro Carlos, Tomás Inmaculada
Oral Sciences Research Group, Special Needs Unit, Department of Surgery and Medical-Surgical Specialties, School of Medicine and Dentistry, Universidade de Santiago de Compostela, Health Research Institute of Santiago (IDIS), Santiago de Compostela, Spain.
Oral Pathology and Rehabilitation Research Unit (UNIPRO), University Institute of Health Sciences (IUCS-CESPU), Gandra, Portugal.
Front Cell Infect Microbiol. 2025 May 2;15:1576906. doi: 10.3389/fcimb.2025.1576906. eCollection 2025.
Proteomic techniques are useful to analyse the periodontal proteome in gingival crevicular fluid (GCF) and saliva. However, few investigations have assessed and compared the GCF and salivary proteomes. Therefore, this research aims to analyse the proteome structure and compare protein expression in these fluids between individuals with periodontal health and those with periodontitis.
GCF and saliva were collected from 44 periodontally healthy subjects and 41 with periodontitis (stages III-IV). Samples were analysed using sequential window acquisition of all theoretical mass spectra (SWATH-MS), and proteins were identified employing the UniProt database. The periodontal proteome structure was assessed using principal component analysis (PCA). Differential protein expression was defined as an adjusted p-value <0.05 combined with a fold-change ≥2 (upregulated) or ≤0.5 (downregulated).
250 abundant proteins were quantified in GCF and 377 in saliva (238 in common). The proteome structure was different in periodontitis compared to periodontal health in both oral fluids. In GCF, 63 (25.2%) proteins were differentially expressed, with 38 upregulated and 25 downregulated in periodontitis. The most overexpressed proteins were haemoglobin subunits (Hbs) beta (fold-change of 5.06) and alpha (4.35), carbonic anhydrase 1 (4.28), and protein S100-P (4.27). Among the underexpressed proteins, 14 were keratins, with type II cytoskeletal 6B being the most downregulated (0.10), together with glyceraldehyde-3-phosphate dehydrogenase (0.12) and zymogen granule protein 16 homolog B (0.13).In saliva, 59 (15.7%) proteins were differentially expressed, with 55 upregulated and four downregulated in periodontitis. Twenty-nine proteins showed a fold-change ≥4, highlighting beta-2-microglobulin (44.14), keratin, type I cytoskeletal 13 (36.23), neutrophil defensin 1 (25.08), proteins S100-A9 (12.30), A8 (10.61), A12 (4.76), and P (4.72), annexin A1 (9.34), lysozyme C (4.98), immunoglobulin heavy constant alpha 1 (4.45), resistin (4.37), and Hbs beta (4.20) and alpha (4.06). The most downregulated protein was lipocalin-1 (0.35). Fourteen proteins were differentially expressed in GCF and saliva, where seven were keratins being underexpressed in GCF but overexpressed in saliva.
Periodontitis alters the periodontal proteome structure and the expression of numerous abundant proteins in GCF and saliva. However, proteins expressed vary qualitatively and quantitatively, indicating different expression patterns between oral fluids.
蛋白质组学技术有助于分析龈沟液（GCF）和唾液中的牙周蛋白质组。然而，很少有研究评估和比较GCF和唾液蛋白质组。因此，本研究旨在分析蛋白质组结构，并比较牙周健康个体与牙周炎个体在这些液体中的蛋白质表达情况。
从44名牙周健康受试者和41名牙周炎患者（III-IV期）中收集GCF和唾液。使用所有理论质谱的顺序窗口采集（SWATH-MS）对样品进行分析，并使用UniProt数据库鉴定蛋白质。使用主成分分析（PCA）评估牙周蛋白质组结构。差异蛋白表达定义为校正p值<0.05且倍数变化≥2（上调）或≤0.5（下调）。
在GCF中定量了250种丰富蛋白质，在唾液中定量了377种（其中238种相同）。与牙周健康相比，两种口腔液体中牙周炎的蛋白质组结构均不同。在GCF中，63种（25.2%）蛋白质差异表达，其中牙周炎中有38种上调和25种下调。表达上调最多的蛋白质是血红蛋白亚基（Hbs）β（倍数变化为5.06）和α（4.35）、碳酸酐酶1（4.28）和蛋白质S100-P（4.27）。在表达下调的蛋白质中，有14种是角蛋白，其中细胞骨架II型6B下调最多（0.10），还有甘油醛-3-磷酸脱氢酶（0.12）和酶原颗粒蛋白16同源物B（0.13）。在唾液中，59种（15.7%）蛋白质差异表达，其中牙周炎中有55种上调和4种下调。29种蛋白质的倍数变化≥4，突出的有β2-微球蛋白（44.14）、细胞骨架I型13角蛋白（36.23）、中性粒细胞防御素1（25.08）、蛋白质S100-A9（12.30）、A8（10.61）、A12（4.76）和P（).72）、膜联蛋白A1（9.34）、溶菌酶C（4.98）、免疫球蛋白重链恒定α1（4.45）、抵抗素（4.37）以及Hbsβ（4.20）和α（4.06）。下调最多的蛋白质是lipocalin-1（0.35）。14种蛋白质在GCF和唾液中差异表达，其中7种是角蛋白，在GCF中表达下调但在唾液中表达上调。
牙周炎会改变牙周蛋白质组结构以及GCF和唾液中多种丰富蛋白质的表达。然而，所表达的蛋白质在质量和数量上存在差异，表明口腔液体之间存在不同的表达模式。
