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.

中文翻译：

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牙齿和植入物的牙周病原体在体内表达蛋白酶和蛋白酶抑制剂丝氨酸蛋白酶抑制剂。

牙龈卟啉单胞菌和连翘藻（Tannerella forsythia）分泌蛋白酶，牙龈蛋白酶和KLIKK蛋白酶。此外，连翘产生的丝氨酸蛋白酶抑制剂（米洛平）具有广泛的抑制谱。这项初步研究的目的是确定在牙周和种植体周围的健康状况和疾病状况下，米洛平和个别蛋白酶在体内的表达水平。生物膜和龈沟液（GCF）/种植体周围龈沟液（PISF）样本取自健康的牙齿和种植体部位（n = 10），牙龈炎和粘膜炎部位（n = 12），牙周炎和种植体周围炎部位（ n = 10）。通过酶联免疫吸附测定法测定了GCF中白细胞介素8（IL-8），IL-1β和IL-10的浓度。负荷牙龈和通过定量PCR在生物膜中评估了连翘的T.连翘以及蛋白酶和促肌动蛋白基因的存在，而通过定量RT-PCR估计了基因表达。牙龈卟啉单胞菌和连翘T.连翘的存在以及IL-8和IL-1β的水平与牙周组织和种植体周围组织的疾病严重程度有关。在含有连翘的生物膜样品中，发现编码蛋白酶的基因的溶菌素含量为72.4 ％，其他KLIKK-蛋白酶基因和miropin含量为100％。同时，单个基因的可检测到的mRNA表达范围为样品的20.7％至58.6％（对于连翘蛋白和miropsin-1， 分别）。与连翘蛋白酶相比，米洛平和牙龈蛋白酶高表达。牙龈蛋白酶的表达水平与米洛宾和牙齿周围的某些连翘蛋白酶的表达水平相关，但与植入物无关。累计而言，KLIKK蛋白酶，尤其是米洛平可能在牙周病和种植体周围疾病的发病机理中起作用。