Silva Thalisson S O, Freitas Alice R, Pinheiro Marília L L, do Nascimento Cássio, Watanabe Evandro, Albuquerque Rubens F
Department of Dental Materials and Prosthodontics, School of Dentistry of Ribeirão Preto, University of São Paulo.
Department of Restorative Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo.
J Vis Exp. 2018 Jun 24(136):57756. doi: 10.3791/57756.
Dental implants and their prosthetic components are prone to bacterial colonization and biofilm formation. The use of materials that provides low microbial adhesion may reduce the prevalence and progression of peri-implant diseases. In view of the oral environment complexity and oral biofilm heterogeneity, microscopy techniques are needed that can enable a biofilm analysis of the surfaces of teeth and dental materials. This article describes a series of protocols implemented for comparing oral biofilm formation on titanium and ceramic materials for prosthetic abutments, as well as the methods involved in oral biofilms analyses at the morphological and cellular levels. The in situ model to evaluate oral biofilm formation on titanium and zirconia materials for dental prosthesis abutments as described in this study provides a satisfactory preservation of the 48 h biofilm, thereby demonstrating methodological adequacy. Multiphoton microscopy allows the analysis of an area representative of the biofilm formed on the test materials. In addition, the use of fluorophores and the processing of the images using multiphoton microscopy allows the analysis of the bacterial viability in a very heterogeneous population of microorganisms. The preparation of biological specimens for electron microscopy promotes the structural preservation of biofilm, images with good resolution, and no artifacts.
牙种植体及其修复组件容易受到细菌定植和生物膜形成的影响。使用具有低微生物粘附性的材料可能会降低种植体周围疾病的发生率和进展。鉴于口腔环境的复杂性和口腔生物膜的异质性,需要能够对牙齿和牙科材料表面进行生物膜分析的显微镜技术。本文描述了一系列用于比较口腔修复基台钛材料和陶瓷材料上口腔生物膜形成的方案,以及在形态学和细胞水平上进行口腔生物膜分析所涉及的方法。本研究中描述的用于评估牙种植体基台钛材料和氧化锆材料上口腔生物膜形成的原位模型能够令人满意地保存48小时的生物膜,从而证明了方法的充分性。多光子显微镜允许对测试材料上形成的生物膜的代表性区域进行分析。此外,使用荧光团并通过多光子显微镜对图像进行处理,可以分析非常异质的微生物群体中的细菌活力。用于电子显微镜检查的生物标本制备可促进生物膜的结构保存、获得具有良好分辨率且无伪影的图像。