Department of Periodontology, Dental Research Division, Guarulhos University, Guarulhos, Brazil.
J Periodontal Res. 2019 Apr;54(2):174-179. doi: 10.1111/jre.12617. Epub 2018 Oct 26.
Little is known about the correspondence of cytokines detected among the different biological matrices used in periodontal research and whether locally-detected cytokine mRNA can also be identified at the translated protein level. The aims of this study were to compare: (a) the detection of classic cytokines at protein level in corresponding samples of serum, gingival tissue and gingival crevicular fluid (GCF) in subjects with periodontitis and (b) the detection of cytokines at protein level in GCF and tissue with their detection at mRNA level in tissue.
Protein concentrations of IL-1β, IL-6 and TNF-α were measured in corresponding samples of GCF, gingival tissue and serum from 125 subjects with chronic periodontitis by multiplex immunoassay. The mRNA levels of these cytokines were evaluated in the same gingival tissue by qPCR.
The mean protein concentrations of IL-1ß and TNF-α were higher in tissue and GCF than in serum (P < 0.05). The mean protein concentration of IL-6 was higher in tissue, followed by serum and GCF, respectively (P < 0.05). Using serum as the gold standard, detection of IL-1ß in GCF and IL-6 in tissue had the highest sensitivity (100%), while detection of IL-6 in the GCF had the lowest sensitivity (77.9%). Using tissue as the gold standard, the detection sensitivities of IL-1ß, IL-6 and TNF-α were 93.6%, 78.4% and 94.2%, respectively, in GCF. Using mRNA as the gold standard, the detection of IL-1ß and IL-6 at the protein level in tissue (100.0%) had the highest sensitivity, while the detection of IL-6 in GCF (77.9%) had the lowest sensitivity.
Tissue and GCF exhibited overall higher protein concentrations of cytokines than serum. GCF presented good reliability for identifying the target cytokines at protein level, when compared with detecting tissue cytokines at protein level. GCF and tissue presented high sensitivities for detecting cytokines at the protein level, when compared with detecting tissue cytokines at the mRNA level. The sensitivity for the identification of cytokines in the GCF depended on the target cytokine, where IL-6 was the least likely cytokine to be detected in this matrix.
目前对于牙周研究中不同生物基质中检测到的细胞因子之间的一致性，以及局部检测到的细胞因子 mRNA 是否也能在翻译蛋白水平上被识别知之甚少。本研究的目的是比较：（a）在患有牙周炎的受试者的血清、牙龈组织和龈沟液（GCF）的相应样本中检测经典细胞因子的蛋白水平，以及（b）在 GCF 和组织中检测细胞因子的蛋白水平与其在组织中的 mRNA 水平的检测结果之间的一致性。
通过多重免疫分析检测 125 例慢性牙周炎患者的 GCF、牙龈组织和血清中相应样本的 IL-1β、IL-6 和 TNF-α 的蛋白浓度。通过 qPCR 评估相同牙龈组织中这些细胞因子的 mRNA 水平。
IL-1ß 和 TNF-α 的平均蛋白浓度在组织和 GCF 中均高于血清（P<0.05）。IL-6 的平均蛋白浓度依次为组织、血清和 GCF（P<0.05）。以血清为金标准，GCF 中检测 IL-1ß 和组织中检测 IL-6 的敏感性最高（100%），而 GCF 中检测 IL-6 的敏感性最低（77.9%）。以组织为金标准，GCF 中 IL-1ß、IL-6 和 TNF-α 的检测灵敏度分别为 93.6%、78.4%和 94.2%。以 mRNA 为金标准，组织中检测 IL-1ß 和 IL-6 的蛋白水平（100.0%）的敏感性最高，而 GCF 中检测 IL-6 的蛋白水平（77.9%）的敏感性最低。
组织和 GCF 中细胞因子的蛋白浓度总体上高于血清。与检测组织细胞因子的蛋白水平相比，GCF 在检测细胞因子的蛋白水平方面具有良好的可靠性。与检测组织细胞因子的 mRNA 水平相比，GCF 和组织在检测细胞因子的蛋白水平方面具有较高的灵敏度。GCF 中细胞因子的识别灵敏度取决于目标细胞因子，其中 IL-6 是最不可能在该基质中被检测到的细胞因子。
