Yamaguchi Arisa, Tsuruya Yuto, Igarashi Kazuma, Jin Zhenyu, Yamazaki-Takai Mizuho, Takai Hideki, Nakayama Yohei, Ogata Yorimasa
Department of Periodontology, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba, Japan.
Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba, Japan.
J Periodontal Implant Sci. 2023 Oct;53(5):347-361. doi: 10.5051/jpis.2203700185. Epub 2023 Jan 26.
Exosomes are membrane vesicles that are present in body fluids and contain proteins, lipids, and microRNA (miRNA). Periodontal tissue examinations assess the degree of periodontal tissue destruction according to the probing depth (PD), clinical attachment loss (CAL), bleeding on probing, and X-ray examinations. However, the accurate evaluation of the prognosis of periodontitis is limited. In this study, we collected saliva from patients before and after initial periodontal therapy (IPT) and compared changes in the clinical parameters of periodontitis with changes in the components of salivary exosomes.
Saliva was collected from patients with stage III and IV periodontitis at the first visit and post-IPT. Exosomes were purified from the saliva, and total protein and RNA were extracted. Changes in expression levels of C6, CD81, TSG101, HSP70, and 6 kinds of miRNA were analyzed by western blots and real-time polymerase chain reaction.
Patients with increased C6 expression after IPT had significantly higher levels of periodontal inflamed surface area (PISA), miR-142, and miR-144 before and after IPT than patients with decreased C6 expression after IPT. Patients with decreased and unchanged CD81 expression after IPT showed significantly higher PD, CAL, and PISA before IPT than after IPT. Patients with decreased and unchanged TSG101 expression after IPT had significantly higher PD before IPT than after IPT. Patients with increased HSP70 expression after IPT had significantly higher PD and PISA before and after IPT than patients with unchanged HSP70 after IPT. The expression levels of miR-142, miR-144, miR-200b, and miR-223 changed with changes in the levels of C6, CD81, TSG101, and HSP70 in the salivary exosomes of periodontitis patients before and after IPT.
The expression levels of proteins and miRNAs in salivary exosomes significantly changed after IPT in periodontitis patients, suggesting that the components of exosomes could serve as biomarkers for periodontitis.
外泌体是存在于体液中的膜性囊泡，含有蛋白质、脂质和微小RNA（miRNA）。牙周组织检查根据探诊深度（PD）、临床附着丧失（CAL）、探诊出血和X线检查来评估牙周组织破坏程度。然而，牙周炎预后的准确评估存在局限性。在本研究中，我们收集了患者在初始牙周治疗（IPT）前后的唾液，并比较了牙周炎临床参数的变化与唾液外泌体成分的变化。
收集III期和IV期牙周炎患者初诊时及IPT后的唾液。从唾液中纯化外泌体，提取总蛋白和RNA。通过蛋白质免疫印迹法和实时聚合酶链反应分析C6、CD81、TSG101、HSP70及6种miRNA表达水平的变化。
IPT后C6表达增加的患者，其牙周炎症表面积（PISA）、miR-142和miR-144在IPT前后的水平显著高于IPT后C6表达降低的患者。IPT后CD81表达降低和未改变的患者，IPT前的PD、CAL和PISA显著高于IPT后。IPT后TSG101表达降低和未改变的患者，IPT前的PD显著高于IPT后。IPT后HSP70表达增加的患者，其IPT前后的PD和PISA显著高于IPT后HSP70未改变的患者。牙周炎患者IPT前后唾液外泌体中，miR-142、miR-144、miR-200b和miR-223的表达水平随C6、CD81、TSG101和HSP70水平的变化而改变。
牙周炎患者IPT后唾液外泌体中蛋白质和miRNA的表达水平显著变化，提示外泌体成分可作为牙周炎的生物标志物。
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