Department of Periodontology, University of Bern, School of Dental Medicine, Bern, Switzerland.
Department of Microbiology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, Krakow, Poland.
Mol Oral Microbiol. 2018 Jun;33(3):240-248. doi: 10.1111/omi.12220. Epub 2018 Apr 17.
Porphyromonas gingivalis and Tannerella forsythia secrete proteases, gingipains and KLIKK-proteases. In addition, T. forsythia produces a serpin (miropin) with broad inhibitory spectrum. The aim of this pilot study was to determine the level of expression of miropin and individual proteases in vivo in periodontal and peri-implant health and disease conditions. Biofilm and gingival crevicular fluid (GCF)/ peri-implant sulcular fluid (PISF) samples were taken from healthy tooth and implant sites (n = 10), gingivitis and mucositis sites (n = 12), and periodontitis and peri-implantitis sites (n = 10). Concentration of interleukin-8 (IL-8), IL-1β and IL-10 in GCF was determined by enzyme-linked immunosorbent assay. Loads of P. gingivalis and T. forsythia and the presence of proteases and miropin genes were assessed in biofilm by quantitative PCR, whereas gene expression was estimated by quantitative RT-PCR. The presence of P. gingivalis and T. forsythia, as well as the level of IL-8 and IL-1β, were associated with disease severity in the periodontal and peri-implant tissues. In biofilm samples harboring T. forsythia, genes encoding proteases were found to be present at 72.4% for karilysin and 100% for other KLIKK-protease genes and miropin. At the same time, detectable mRNA expression of individual genes ranged from 20.7% to 58.6% of samples (for forsylisin and miropsin-1, respectively). In comparison with the T. forsythia proteases, miropin and the gingipains were highly expressed. The level of expression of gingipains was associated with those of miropin and certain T. forsythia proteases around teeth but not implants. Cumulatively, KLIKK-proteases and especially miropin, might play a role in pathogenesis of both periodontal and peri-implant diseases.
牙龈卟啉单胞菌和福赛斯坦纳菌分泌蛋白酶、gingipains 和 KLIKK-蛋白酶。此外，T. forsythia 产生一种具有广泛抑制谱的丝氨酸蛋白酶抑制剂（miropin）。本初步研究的目的是确定在牙周和种植体健康及疾病状态下体内 miropin 和个体蛋白酶的表达水平。从健康牙齿和种植体部位（n=10）、牙龈炎和黏膜炎部位（n=12）以及牙周炎和种植体周围炎部位（n=10）采集生物膜和龈沟液（GCF）/种植体周围沟液（PISF）样本。通过酶联免疫吸附试验测定 GCF 中白细胞介素-8（IL-8）、IL-1β 和 IL-10 的浓度。通过定量 PCR 评估生物膜中 P. gingivalis 和 T. forsythia 的负荷以及蛋白酶和 miropin 基因的存在，通过定量 RT-PCR 估计基因表达。P. gingivalis 和 T. forsythia 的存在以及 IL-8 和 IL-1β 的水平与牙周和种植体组织中的疾病严重程度相关。在含有 T. forsythia 的生物膜样本中，发现编码蛋白酶的基因存在于 72.4%的卡瑞利辛和 100%的其他 KLIKK-蛋白酶基因和 miropin。同时，单个基因的可检测 mRNA 表达范围为样本的 20.7%至 58.6%（分别为 forsylisin 和 miropsin-1）。与 T. forsythia 蛋白酶相比，miropin 和 gingipains 的表达水平较高。gingipains 的表达水平与牙齿周围的 miropin 和某些 T. forsythia 蛋白酶的表达水平相关，但与种植体无关。总体而言，KLIKK-蛋白酶，尤其是 miropin，可能在牙周和种植体疾病的发病机制中发挥作用。
