Mobile and immobile adhesion of staphylococcal strains to hydrophilic and hydrophobic surfaces.

Staphylococcus epidermidis adheres to hydrophilic glass and hydrophobic dimethyldichlorosilane (DDS)-coated glass in similar numbers, but in different modes. Real-time observation of staphylococcal adhesion under a shear rate of 15 s(-1) revealed different adhesion dynamics on both substrata. The number of adsorption and desorption events to achieve a similar number of adhering bacteria was twofold higher on hydrophilic than on hydrophobic DDS-coated glass. Moreover, 22% of all staphylococci on glass slid over the surface prior to adhering on a fixed site ("mobile adhesion mode"), but mobile adhesion was virtually absent (1%) on DDS-coated glass. Sliding preceded desorption on hydrophilic glass in about 20% of all desorption events, while on hydrophobic DDS-coated glass 2% of all staphylococci desorbed straight from their adhesion site. Since acid-base interactions between the staphylococci and a hydrophobic DDS-coating are attractive, it is suggested that these interactions facilitate a closer approach of the bacteria and therewith enhance immobile adhesion at local, high affinity sites. Alternatively, if the local site is low affinity, this may lead to desorption. In the absence of attractive acid-base interactions, as on hydrophilic glass, bacteria can be captured in the minimum of the DLVO-interaction energy curve, but this does not prevent them from sliding under flow at a fixed distance from a substratum surface until immobilization or desorption at or from a local high or low affinity site, respectively.

Staphylococcus aureus-fibronectin interactions with and without fibronectin-binding proteins and their role in adhesion and desorption.

Adhesion and residence-time-dependent desorption of two Staphylococcus aureus strains with and without fibronectin (Fn) binding proteins (FnBPs) on Fn-coated glass were compared under flow conditions. To obtain a better understanding of the role of Fn-FnBP binding, the adsorption enthalpies of Fn with staphylococcal cell surfaces were determined using isothermal titration calorimetry (ITC). Interaction forces between staphylococci and Fn coatings were measured using atomic force microscopy (AFM). The strain with FnBPs adhered faster and initially stronger to an Fn coating than the strain without FnBPs, and its Fn adsorption enthalpies were higher. The initial desorption was high for both strains but decreased substantially within 2 s. These time scales of staphylococcal bond ageing were confirmed by AFM adhesion force measurement. After exposure of either Fn coating or staphylococcal cell surfaces to bovine serum albumin (BSA), the adhesion of both strains to Fn coatings was reduced, suggesting that BSA suppresses not only nonspecific but also specific Fn-FnBP interactions. Adhesion forces and adsorption enthalpies were only slightly affected by BSA adsorption. This implies that under the mild contact conditions of convective diffusion in a flow chamber, adsorbed BSA prevents specific interactions but does allow forced Fn-FnBP binding during AFM or stirring in ITC. The bond strength energies calculated from retraction force-distance curves from AFM were orders of magnitude higher than those calculated from desorption data, confirming that a penetrating Fn-coated AFM tip probes multiple adhesins in the outermost cell surface that remain hidden during mild landing of an organism on an Fn-coated substratum, like that during convective diffusional flow.

Residence time dependent desorption of Staphylococcus epidermidis from hydrophobic and hydrophilic substrata.

Adhesion and desorption are simultaneous events during bacterial adhesion to surfaces, although desorption is far less studied than adhesion. Here, desorption of Staphylococcus epidermidis from substratum surfaces is demonstrated to be residence time dependent. Initial desorption rate coefficients were similar for hydrophilic and hydrophobic dimethyldichlorosilane (DDS)-coated glass, likely because initial desorption is controlled by attractive Lifshitz-Van der Waals interactions, which are comparable on both substratum surfaces. However, significantly slower decay times of the desorption rate coefficients are found for hydrophilic glass than for hydrophobic DDS-coated glass. This difference is suggested to be due to the acid-base interactions between staphylococci and these surfaces, which are repulsive on hydrophilic glass and attractive on hydrophobic DDS-coated glass. Final desorption rate coefficients are higher on hydrophilic glass than on hydrophobic DDS-coated glass, due to the so called hydrophobic effect, facilitating a closer contact on hydrophobic DDS-coated glass.

Bond-strengthening in staphylococcal adhesion to hydrophilic and hydrophobic surfaces using atomic force microscopy.

Time-dependent bacterial adhesion forces of four strains of Staphylococcus epidermidis to hydrophobic and hydrophilic surfaces were investigated. Initial adhesion forces differed significantly between the two surfaces and hovered around -0.4 nN. No unambiguous effect of substratum surface hydrophobicity on initial adhesion forces for the four different S. epidermidis strains was observed. Over time, strengthening of the adhesion forces was virtually absent on hydrophobic dimethyldichlorosilane (DDS)-coated glass, although in a few cases multiple adhesion peaks developed in the retract curves. Bond-strengthening on hydrophilic glass occurred within 5-35 s to maximum adhesion forces of -1.9 +/- 0.7 nN and was concurrent with the development of multiple adhesion peaks upon retract. Poisson analysis of the multiple adhesion peaks allowed separation of contributions of hydrogen bonding from other nonspecific interaction forces and revealed a force contribution of -0.8 nN for hydrogen bonding and +0.3 nN for other nonspecific interaction forces. Time-dependent bacterial adhesion forces were comparable for all four staphylococcal strains. It is concluded that, on DDS-coated glass, the hydrophobic effect causes instantaneous adhesion, while strengthening of the bonds on hydrophilic glass is dominated by noninstantaneous hydrogen bond formation.

Forces involved in bacterial adhesion to hydrophilic and hydrophobic surfaces.

Using a parallel-plate flow chamber, the hydrodynamic shear forces to prevent bacterial adhesion (F(prev)) and to detach adhering bacteria (F(det)) were evaluated for hydrophilic glass, hydrophobic, dimethyldichlorosilane (DDS)-coated glass and six different bacterial strains, in order to test the following three hypotheses. 1. A strong hydrodynamic shear force to prevent adhesion relates to a strong hydrodynamic shear force to detach an adhering organism. 2. A weak hydrodynamic shear force to detach adhering bacteria implies that more bacteria will be stimulated to detach by passing an air-liquid interface (an air bubble) through the flow chamber. 3. DLVO (Derjaguin, Landau, Verwey, Overbeek) interactions determine the characteristic hydrodynamic shear forces to prevent adhesion and to detach adhering micro-organisms as well as the detachment induced by a passing air-liquid interface. F(prev) varied from 0.03 to 0.70 pN, while F(det) varied from 0.31 to over 19.64 pN, suggesting that after initial contact, strengthening of the bond occurs. Generally, it was more difficult to detach bacteria from DDS-coated glass than from hydrophilic glass, which was confirmed by air bubble detachment studies. Calculated attractive forces based on the DLVO theory (F(DLVO)) towards the secondary interaction minimum were higher on glass than on DDS-coated glass. In general, all three hypotheses had to be rejected, showing that it is important to distinguish between forces acting parallel (hydrodynamic shear) and perpendicular (DLVO, air-liquid interface passages) to the substratum surface.

Porous hybrid structures based on P(DLLA-co-TMC) and collagen for tissue engineering of small-diameter blood vessels.

Poly (D,L-lactide)-7co-(1,3-trimethylene carbonate) [P(DLLA-co-TMC)] (83 mol % DLLA) was used to produce matrices suitable for tissue engineering of small-diameter blood vessels. The copolymer was processed into tubular structures with a porosity of approximately 98% by melt spinning and fiber winding, thus obviating the need of organic solvents that may compromise subsequent cell culture. Unexpectedly, incubation in culture medium at 37 degrees C resulted in disconnection of the contact points between the polymer fibers. To improve the structural stability of these P(DLLA-co-TMC) scaffolds, a collagen microsponge was formed inside the pores of the synthetic matrix by dip coating and freeze drying. Hybrid structures with a porosity of 97% and an average pore size of 102 mum were obtained. Structural stability was preserved during incubation in culture medium at 37 degrees C. Smooth-muscle cells (SMCs) were seeded in these hybrid scaffolds and cultured under pulsatile flow conditions in a bioreactor (120 beats/min, 80-120 mmHg). After 7 days of culture in a dynamic environment viable SMCs were homogeneously distributed throughout the constructs, which were five times stronger and stiffer than noncultured scaffolds. Values for yield stress (2.8 +/- 0.6 MPa), stiffness (1.6 +/- 0.4 MPa), and yield strain (120% +/- 20%) were comparable to those of the human artery mesenterica.

Influence of shear on microbial adhesion to PEO-brushes and glass by convective-diffusion and sedimentation in a parallel plate flow chamber.

Microbial adhesion to surfaces often occurs despite high wall shear rates acting on the adhering microorganisms. In this paper, we compare the wall shear rates needed to prevent microbial adhesion to bare glass and poly(ethylene oxide) (PEO)-brush coated glass in a parallel plate flow chamber. Initial microbial deposition rates were determined for different wall shear rates between 4 and 1600 s(-1) on the top and bottom plates of the flow chamber. Deposition efficiencies alpha(SL), based on the Smoluchowski-Levich approach, for Pseudomonas aeruginosa D1, Escherichia coli O2K2 and Candida tropicalis GB 9/9 decreased with increasing wall shear rates and were lower for PEO-brush coated glass than for bare glass. Characteristic shear rates preventing adhesion to the bottom plate were around 10 and 1.0 s(-1) for the bacteria on glass and the PEO-brush and 36 and 3.4s(-1) for the yeast strain on glass and the PEO-brush, respectively. This demonstrates that the adhesive forces between microorganisms and a PEO-brush are comparatively weak, although some strains may have the ability to adhere to a PEO-brush under low shear conditions. Microbial deposition efficiencies alpha(SL) were much larger, however, than unity for bottom plate deposition, but could be reduced to realistic values by averaging the deposition rates found for the top (negative contribution of sedimentation) and bottom (positive contribution of sedimentation) plates.