A novel in vitro model to study staphylococcal biofilm formation on intraocular lenses under hydrodynamic conditions.

PURPOSE: To develop a novel in vitro model to study the formation of Staphylococcus epidermidis biofilm on intraocular lenses (IOLs) from the primary-attachment phase to the biofilm-accumulation phase. The model was designed to replicate intraocular conditions especially by taking into account intraocular hydrodynamics. METHODS: The model consisted of Tygon tubing connected to a vial containing acrylic hydrophobic IOLs. Three septa, placed along the tubing, allowed, respectively, the artificial aqueous humor's arrival and its elimination and the bacterial suspension's inoculation. A first pump allowed the aqueous humor's movement along the circuit, whereas a second one regulated the flow at which the nutritive environment was regenerated. The whole circuit was placed in a 34 degrees C water bath. Every 2 to 4 hours, lenses were taken from this environment. Bound bacteria were removed by scraping of optical faces and counted. All data are presented as the mean, SD, and coefficient of variation (CV). Comparisons among experiments were performed by one-way analysis of variance (ANOVA). RESULTS: Calculated CVs were close to 30, showing that biofilm formation was homogeneous. Differences between experiments were nonsignificant for each removal time. The model provided the full kinetics of S. epidermidis biofilm growth on acrylic hydrophobic IOLs, with a stationary phase reached after 28 hours of incubation. CONCLUSIONS: Biofilm development is modulated by many variables, including environmental factors. The findings in the present study of bacterial colonization of IOLs under intraocular physiological conditions allow understanding and more accurate targeting of biomedical device-related infections such as endophthalmitis.

In vitro influence of vancomycin on adhesion of a Staphylococcus epidermidis strain encoding intercellular adhesion locus ica to intraocular lenses.

PURPOSE: To assess anti-adhesion and/or bactericidal properties of vancomycin in vitro and to determine when these effects are detectable to estimate its relevance to perioperative antibiotic prophylaxis and analyze the efficacy of a newly designed vancomycin insert prototype for endophthalmitis prevention. SETTING: University research laboratory, Lyon, France. METHODS: Staphylococcus epidermidis clinical strain N890074 containing the intercellular adhesion locus ica was used as the infectious agent. Vancomycin was used at 20 microg/mL. A sterile biocompatible, biodegradable vancomycin insert, releasing 230 microg of antibiotics over 100 minutes, was designed especially for this study. To obtain bacterial killing curves, experiments were first performed in a 103 colony-forming units (CFU/mL) bacterial suspension containing no intraocular lenses (IOL). Then IOLs were incubated in the suspension, and bacterial adherence was determined using bacterial counting with and without antibiotic. RESULTS: Vancomycin (solution and insert) had an anti-adhesion effect after 1 hour and a relevant bactericidal effect after 6 hours of incubation. CONCLUSIONS: Vancomycin used with irrigating solutions does not remain in the anterior chamber long enough to develop bactericidal effect. Even if it initially reduces bacterial adhesion, used at a drug level dropping below the bacterial minimal inhibitory concentration, it could result in a secondary increase of the adhesion of slime-producing bacteria. A sufficiently high concentration was obtained in vitro by the new sustained-release system, thereby overcoming the theoretical drawback of a short half-life within the anterior chamber. Anti-adhesion and bactericidal action of vancomycin inserts remains to be confirmed in clinical studies.

Biofilm formation on intraocular lenses by a clinical strain encoding the ica locus: a scanning electron microscopy study.

PURPOSE: To determine whether the Staphylococcus epidermidis strain carries the intercellular adhesion (ica) locus, which encodes production of adhesins mediating adherence to biomaterials and to study, with scanning electron microscopy, the morphologic features of this coagulase-negative Staphylococcus strain that adheres to intraocular lenses (IOLs). METHODS: Polymerase chain reaction amplification was used to investigate whether the isolate under study (S. epidermidis clinical strain N890074) carries the ica locus. Sterile intraocular lenses (IOLs) were incubated in bacterial suspension either for 5 minutes or 1 hour. IOLs were then examined by scanning electron microscopy. RESULTS: Polymerase chain reaction amplification revealed that S. epidermidis N890074 contained the ica locus. The bacteria appeared to be anchored to the surface of the lenses by several different means-particularly by leglike appendages and a slime layer-which probably came into play step by step. CONCLUSIONS: For the first time in ophthalmology, to the authors' knowledge, photographs showing leglike appendages involved in the first phase of adhesion have been obtained. They also clearly visualize the slime layer containing the embedded bacteria. This study provides information about the nature and the genesis of these attachment processes. Adherence is known to be greater when the bacterial DNA contain the ica locus. Full knowledge of the pathogenesis of bacterial adhesion is necessary to gain a better understanding of IOL infection and endophthalmitis.

Bacterial adherence of Staphylococcus epidermidis to intraocular lenses: a bioluminescence and scanning electron microscopy study.

PURPOSE: To analyze and compare the adherence of Staphylococcus epidermidis to intraocular lenses (IOLs) made of five different biomaterials (native or heparinized polymethylmethacrylate, silicone, hydrophilic acrylic, or hydrogel) and to detail the different steps and mechanisms of bacterial adhesion to a polymer. METHODS: A clinical strain carrying the intercellular adhesion (ica) locus was used. In a previous study, the extent of bacterial binding was measured by counting. In this study, two different techniques, bioluminescence and scanning electron microscopy (SEM), were used to analyze the accuracy of each one, to obtain a comparison between the various IOLs, and to complete previous observations. The results were compared using both the Kruskal-Wallis and the Mann-Whitney nonparametric tests. RESULTS: Bacterial adhesion was statistically weakest on hydrogel and then on hydrophilic acrylic polymer. Adhesion depended on the hydrophobicity or hydrophilicity of the biomaterials. Slight differences were found between the two methods, and these differences are explained. Furthermore, SEM observations highlighted two different patterns of bacterial adhesion (isolated bacteria and clusters of bacteria), assuming that hydrophobic IOLs (silicone and PMMA) probably facilitate bacterial colonization and biofilm production. CONCLUSIONS: Attachment mechanisms may be different in each case, depending on the polymer material and the infecting organism, because there are various types of behavior among S. epidermidis strains. Hydrophilic polymer surfaces (hydrogel and probably hydrophilic acrylic) seem to be useful in avoiding the development of bacterial colonies and hence in preventing endophthalmitis. Fewer bacteria were attached, demonstrating inhibition or delay in bacterial colonization.

In vivo study of bacterial adhesion to five types of intraocular lenses.

PURPOSE: To determine in vivo behavior of the ability of the Staphylococcus epidermidis strain (American Type Culture Collection [ATCC] 14990) to attach to 120 intraocular lenses (IOLs) made of five different biomaterials: fluorine polymethylmethacrylate (PMMA), heparinized PMMA, silicone, hydrophobic acrylic, and hydrogel. The pig was chosen as an animal model of endophthalmitis, after a bibliographical analysis and a personal study of its aqueous humor composition. METHODS: Crystalline lenses from 90 domestic pigs were removed aseptically and replaced with previously infected IOLs. The animals were killed 24 hours, 72 hours, and 1 week after implantation of the IOLs. The extent of bacterial binding was then measured by counting. Results were compared with a two-factor analysis of variance (ANOVA 2), confirmed by the Kruskal-Wallis nonparametric test. RESULTS: The extent of bacterial binding (expressed as bound bacteria per area unit) was found to range in increasing order from hydrogel, to fluorine PMMA, to hydrophobic acrylic, to heparinized PMMA, to silicone polymer. Comparison of pairs of materials showed statistically significant differences, except between hydrogel and fluorine PMMA. CONCLUSIONS: To the authors' knowledge, no study has been published so far concerning the in vivo evolution of populations of bacteria adhering to different intraocular materials. Bacterial adhesion to the implant surface must therefore depend on the hydrophobicity or hydrophilicity of the biomaterial. Adhesion is also affected by the nature of the surrounding medium. Because of its complexity, the latter appears to be very difficult to model, thus making in vivo studies essential.

In-vitro study of bacterial adherence to different types of intraocular lenses.

The aim of this study was to determine the adherence of Staphylococcus epidermidis to intraocular lenses made of five different biomaterials: polymethylmethacrylate (PMMA), heparinized PMMA, silicone, hydrophilic acrylic, and hydrogel. The extent of bacterial binding was measured by counting. The results were compared using a one-factor variance analysis. Adherence was weakest on hydrogel and strongest on the silicone polymer. Bacterial adherence to the implant surface must therefore depend on the hydrophobicity or hydrophilicity of the biomaterial.