Understanding the Fundamental Basis for Biofilm Formation on Plastic Surfaces: Role of Conditioning Films

Graphical AbstractThe formation and composition of conditioning films.

The Influence of Surface Topography and Wettability on Escherichia coli Removal from Polymeric Materials in the Presence of a Blood Conditioning Film

The reduction of biofouling and the reduction of cross-contamination in the food industry are important aspects of safety management systems. Polymeric surfaces are used extensively throughout the food production industry and therefore ensuring that effective cleaning regimes are conducted is vital. Throughout this study, the influence of the surface characteristics of three different polymeric surfaces, polytetrafluoroethylene (PTFE), poly(methyl methacrylate) (PMMA) and polyethylene terephthalate (PET), on the removal of Escherichia coli using a wipe clean method utilising 3% sodium hypochlorite was determined. The PTFE surfaces were the roughest and demonstrated the least wettable surface (118.8°), followed by the PMMA (75.2°) and PET surfaces (53.9°). Following cleaning with a 3% sodium hypochlorite solution, bacteria were completely removed from the PTFE surfaces, whilst the PMMA and PET surfaces still had high numbers of bacteria recovered (1.2 × 107 CFU/mL and 6.3 × 107 CFU/mL, respectively). When bacterial suspensions were applied to the surfaces in the presence of a blood conditioning film, cleaning with sodium hypochlorite demonstrated that no bacteria were recovered from the PMMA surface. However, on both the PTFE and PET surfaces, bacteria were recovered at lower concentrations (2.0 × 102 CFU/mL and 1.3 × 103 CFU/mL, respectively). ATP bioluminescence results demonstrated significantly different ATP concentrations on the surfaces when soiled (PTFE: 132 relative light units (RLU), PMMA: 80 RLU and PET: 99 RLU). Following cleaning, both in the presence and absence of a blood conditioning film, all the surfaces were considered clean, producing ATP concentrations in the range of 0–2 RLU. The results generated in this study demonstrated that the presence of a blood conditioning film significantly altered the removal of bacteria from the polymeric surfaces following a standard cleaning regime. Conditioning films which represent the environment where the surface is intended to be used should be a vital part of the test regime to ensure an effective disinfection process.

Impact of locking solutions on conditioning biofilm formation in tunnelled haemodialysis catheters and inflammatory response activation

Introduction: The surface of tunnelled cuffed catheters provides an optimal environment for the development of biofilms, which have recently been described as conditioning films because of the presence of adherent biological materials. These biofilms are associated with infection and thrombosis and potentially increase patients’ inflammatory response. These complications could be reduced by the use of locking solutions. Objective: To analyse biofilm formation, using confocal and electron microscopy, in tunnelled cuffed catheters locked with three different solutions and to determine the relationship between these solutions and inflammatory response. Study design: This prospective study included 35 haemodialysis patients with tunnelled cuffed catheter removal for non–infection-related reasons. The participants were divided into three groups according to the lock solution used: (1) heparin 1: 5000 IU; (2) citrate 4%; and (3) taurolidine 1.35%, citrate 4% and heparin 500 IU (taurolock); in the latter group, 25,000 IU taurolidine–urokinase was used in the last weekly session. All tunnelled cuffed catheters were cultured, and the inner surface was evaluated with confocal and electron microscopy. The inflammatory profile of included patients was determined at tunnelled cuffed catheter removal. Results: There were no differences in clinical or demographic variables between the three subgroups. Biofilm thickness was lower in the taurolidine group than in the citrate 4% and heparin groups (28.85 ± 6.86 vs 49.99 ± 16.56 vs 56.2 ± 15.67 µm, respectively; p < 0.001), as was biofilm volume (1.01 ±1.18 vs 3.7 ± 2.15 vs 5.55 ±2.44, µm3, respectively; p < 0.001). The mean interleukin-6 value was 39%, which was 50% lower than in the citrate and heparin groups, but without significance differences. Conclusion: Our results show that biofilms were found in all tunnelled cuffed catheters, but the thickness and volume were significantly lower in tunnelled cuffed catheters locked with taurolidine solution. Therefore, the type of locking solution used in tunnelled cuffed catheters should maintain tunnelled cuffed catheter sterility and prevent catheter-related bloodstream infections. No significant difference was observed in the inflammatory profile according to the type of locking solution.

Enhancement in Xerostomia Patient Salivary Lubrication Using a Mucoadhesive

Oral lubrication mediated by mucin and protein containing salivary conditioning films (SCFs) with strong water retainability can get impaired due to disease such as xerostomia, that is, a subjective dry mouth feel associated with the changed salivary composition and low salivary flow rate. Aberrant SCFs in xerostomia patient cause difficulties in speech, mastication, and dental erosion while the prescribed artificial saliva is inadequate to solve the complications on a lasting basis. With the growing aging population, it is urgently needed to propose a new strategy to restore oral lubrication. Existing saliva substitutes often overwhelm the aberrant SCFs, generating inadequate relief. Here we demonstrated that the function of aberrant SCFs in a patient with Sjögren syndrome can be boosted through mucin recruitment by a simple mucoadhesive, chitosan-catechol (Chi-C). Chi-C with different conjugation degrees (Chi-C7.6%, Chi-C14.5%, Chi-C22.4%) was obtained by carbodiimide chemistry, which induced a layered structure composed of a rigid bottom and a soft secondary SCF (S-SCF) after reflow of saliva. The higher conjugation degree of Chi-C generates a higher glycosylated S-SCF by mucin recruitment and a lower friction in vitro. The layered S-SCF extends the “relief period” for Sjögren patient saliva over 7-fold, measured on an ex vivo tongue-enamel friction system. Besides lubrication, Chi-C-treated S-SCF reduces dental erosion depths from 125 to 70 μm. Chi-C shows antimicrobial activity against Streptococcus mutans. This research provides a new key insight in restoring the functionality of conditioning film at articulating tissues in living systems.

Attachment of Listeria innocua to Polystyrene: Effects of Ionic Strength and Conditioning Films from Culture Media and Milk Proteins

It is recognized that bacterial adhesion usually occurs on conditioning films made of organic macromolecules absorbed to abiotic surfaces. The objectives of this study were to determine the extent to which milk protein–coated polystyrene (PS) pegs interfere with biofilm formation and the synergistic effect of this conditioning and hypertonic growth media on the bacterial adhesion and biofilm formation of Listeria innocua, used as a nonpathogenic surrogate for Listeria monocytogenes. PS pegs were uncoated (bare PS) or individually coated with whey proteins isolate (WPI), β-lactoglobulin, bovine serum albumin, or tryptic soy broth (TSB) and were incubated in bacterial suspensions in modified Welshimer's broth. After 4 h, the number of adherent cells was dependent on the coating, as follows: TSB (107 CFU/ml) &gt; bare PS &gt; β-lactoglobulin &gt; bovine serum albumin ≈ WPI (104 CFU/ml). The sessile cell counts increased up to 24 h, reaching &gt;107 CFU per peg for all surfaces (P &gt; 0.1), except for WPI-coated PS; this indicates that the inhibitory effects of milk protein conditioning films are transient, slowing down the adhesion process. The 4-h bacterial adhesion on milk protein–coated PS in modified Welshimer's broth supplemented with salt (0 to 10% [wt/vol]) did not vary (P &gt; 0.1), indicating that conditioning with milk proteins was the major determinant for inhibition of bacterial adhesion and that the synergetic effect of salt and milk proteins on adhesion was minimal. Moreover, the presence of 5 to 10% salt significantly inhibited 24-h biofilm formation on the TSB-coated and bare PS, with a decrease of &gt;3 log at 10% (wt/vol) NaCl and almost completely depleted viable sessile bacteria on the milk protein–coated PS.