Rusu Darian, Stratul Stefan-Ioan, Calniceanu Horia, Boariu Marius, Ogodescu Alexandru, Milicescu Stefan, Didilescu Andreea, Roman Alexandra, Surlin Petra, Locovei Cosmin, Chiperi Monica, Solomon Sorina, Nica Luminita
Department of Periodontology, 'Victor Babes' University of Medicine and Pharmacy, 300041 Timisoara, Romania.
Department of Endodontics, 'Victor Babes' University of Medicine and Pharmacy, 300041 Timisoara, Romania.
Exp Ther Med. 2020 Dec;20(6):201. doi: 10.3892/etm.2020.9331. Epub 2020 Oct 14.
Over the last decades, scanning electron microscopy (SEM) proved to be invaluable for ultrastructural investigation, allowing imaging of the overall appearance and/or specific features of oral biofilms, e.g., microbial colonies and individual cells, glycocalyx, the presence of inorganic products. The aim of this study was the observation and evaluation of the morphology of the biofilm of endodontic-periodontal lesions (EPL) with a modified protocol involving a simplified histologic sample preparation and a low-vacuum SEM examination method. Twenty-one teeth with endodontic-periodontal involvement, extracted for periodontal reasons, were carefully washed with saline, underwent fixation in modified Karnovsky solution and were dehydrated in alcohol series. Samples were examined under low-vacuum SEM. Radicular surfaces were evaluated qualitatively and semiquantitatively for several characteristics, including the presence of bacterial types, the biofilm morphology and the content of root resorptions. Radicular surfaces were divided in four conventional zones Surfaces were evaluated for several characteristics: Presence of bacterial types, biofilm morphology, presence of root resorptions. High-quality images, relevant for endodontic-periodontal biofilms were collected. Continuous, established biofilm was found on all examined surfaces, its detection varying from 19% of the samples on the wall of cemental cone to 52.3% on the radicular surface of the periodontal pocket. Observed microorganisms included cocci, rods an filaments. Spirils and motile bacteria were only accidentally found. SEM investigation of surfaces involved in EPL revealed less surfaces covered by mature biofilm (in only 28.5% of the samples in the 'transition zone'), especially rods and filaments associated with cemental resorptions and calculus. Biofilm elements were better represented in periodontal pockets than in other zones of EPL (detected in up to 81% of the samples). A strong correlation between mature biofilm and the presence of cocci appears on all investigated zones P<0.01). Microbiota appeared to be morphologically similar in apical and periodontal areas, especially in old EPL.
在过去几十年中，扫描电子显微镜（SEM）已被证明在超微结构研究中具有极高价值，它能够对口腔生物膜的整体外观和/或特定特征进行成像，例如微生物菌落和单个细胞、糖萼、无机产物的存在情况。本研究的目的是通过一种改良方案观察和评估牙髓-牙周联合病变（EPL）生物膜的形态，该方案包括简化的组织学样本制备和低真空扫描电子显微镜检查方法。因牙周原因拔除的21颗患有牙髓-牙周联合病变的牙齿，先用生理盐水仔细冲洗，然后在改良的卡诺夫斯基溶液中固定，并在系列乙醇中脱水。样本在低真空扫描电子显微镜下进行检查。对牙根表面的几种特征进行定性和半定量评估，包括细菌类型的存在、生物膜形态以及牙根吸收的情况。牙根表面分为四个常规区域，对表面的几种特征进行评估：细菌类型的存在、生物膜形态、牙根吸收的存在。收集到了与牙髓-牙周生物膜相关的高质量图像。在所有检查的表面都发现了连续的、已形成的生物膜，其检出率从牙骨质锥壁样本的19%到牙周袋牙根表面的52.3%不等。观察到的微生物包括球菌、杆菌和丝状菌。螺旋菌和运动性细菌只是偶尔发现。对牙髓-牙周联合病变涉及的表面进行扫描电子显微镜研究发现，成熟生物膜覆盖的表面较少（在“过渡区”仅28.5%的样本中），尤其是与牙骨质吸收和牙石相关的杆菌和丝状菌。生物膜成分在牙周袋中比在牙髓-牙周联合病变的其他区域表现得更明显（在高达81%的样本中被检测到）。在所有研究区域，成熟生物膜与球菌的存在之间都存在很强的相关性（P<0.01）。微生物群落在根尖和牙周区域的形态似乎相似，尤其是在陈旧的牙髓-牙周联合病变中。
