Oral Biofilm Formation on Different Materials for Dental Implants

Thalisson S. O. Silva; Alice R. Freitas; Marília L. L. Pinheiro; Cássio do Nascimento; Evandro Watanabe; Rubens F. Albuquerque

doi:10.3791/57756

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Method Article

Oral Biofilm Formation on Different Materials for Dental Implants

11.5K views

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DOI:

10.3791/57756

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June 24th, 2018

Thalisson S. O. Silva¹ , Alice R. Freitas¹ , Marília L. L. Pinheiro¹ , Cássio do Nascimento¹ , Evandro Watanabe² , Rubens F. Albuquerque¹

¹Department of Dental Materials and Prosthodontics, School of Dentistry of Ribeirão Preto, University of São Paulo, ²Department of Restorative Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo

Summary

Here, we present a protocol to evaluate oral biofilm formation on titanium and zirconia materials for dental prosthesis abutments, including the analysis of bacterial cells viability and morphological characteristics. An in situ model associated with powerful microscopy techniques is used for the oral biofilm analysis.

Abstract

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.

Introduction

Bacterial biofilms are complex, functionally and structurally organized microbial communities, characterized by a diversity of microbial species that synthesize an extracellular, biologically active polymer matrix¹^,². The bacterial adhesion to biotic or abiotic surfaces is preceded by a formation of the acquired pellicle, mainly consisting of salivary glycoproteins¹^,³^,⁴. Weak physicochemical interactions between the microorganisms and the pellicle are initially established and followed by stronger interactions between....