Quantification of initial adhesion of Enterococcus faecalis to medical grade polymers using a DNA-based fluorescence assay

Abstract

This paper reports on the use of a DNA-based fluorescence assay to study and quantify the initial interactions of the uropathogen Enterococcus faecalis with different polymers commonly used for the fabrication of medical devices and implants, including polyurethane (PU), silicone (SI), high-density polyethylene (HDPE), polyamide (PA), poly(methyl methacrylate) (PMMA) and polytetrafluoroethylene (PTFE). To follow the kinetics of E. faecalis adhesion, polymer samples were incubated in bacterial solution for various times and the relative concentration of adhered bacteria was obtained using two methods: commonly used CFU plate counting and a DNA quantification assay. Results obtained from DNA-based fluorescence assays showed that E. faecalis adhesion on PU is 3-times higher than that on PTFE following exposure to bacteria for 180 min. Neither surface wettability nor surface roughness of the studied polymers was found to correlate with E. faecalis adhesion, suggesting the involvement of much more complex adhesion mechanisms of bacteria onto surfaces. SEM micrographs of adhered bacteria illustrated that adhesion was different depending on the type of polymeric substrate: adhesion on PU samples was characterized by the aggregation of bacterial cells in dense clusters, as well as by the presence of fimbriae between cells and the substrate, which could explain the high adhesion to PU compared to the other polymers. This work demonstrated that the bacterial adhesion to polymers occurs at an early stage of the contact and suggests that the initial adhesion stage should be controlled, in order to prevent subsequent biofilm formation and, thus, reduce the risk of implant-associated infections.

Keywords:

• INITIAL BACTERIAL ADHESION
• MEDICAL GRADE POLYMERS
• DNA-BASED FLUORESCENCE ASSAY
• SURFACE PROPERTIES
• SEM
• IMPLANT-ASSOCIATED INFECTION