Hans J. Kaper

Characterization of poly(ethylene oxide) brushes on glass surfaces and adhesion of Staphylococcus epidermidis

Poly(ethylene oxide) brushes have been covalently bound to glass surfaces and their presence was demonstrated by an increase in water contact angles from fully wettable on glass to advancing contact angles of 54°, with a hysteresis of 32°. In addition, electrophoretic mobilities of glass and brush-coated glass were determined using streaming potential measurements. The dependence of the electrophoretic mobilities on the ionic strength was analyzed in terms of a soft-layer model, yielding an electrophoretic softness and fixed charge density of the layer. Brush-coated glass could be distinguished from glass by a 2-3-fold decrease in fixed charge density, while both surfaces were about equally soft. Adhesion of Staphylococcus epidermidis HBH276 to glass in a parallel plate flow chamber was extremely high and after 4 h, 19.0 × 106 bacteria were adhering per cm2. In contrast, the organisms did not adhere to brush-coated glass, with numbers below the detection limit, i.e. 0.1 × 106 per cm2. These results attest to the great potential of polymer brushes in preventing bacterial adhesion to surfaces.

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

ABSTRACT 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.

Nanoscopic Vibrations of Bacteria with Different Cell-Wall Properties Adhering to Surfaces under Flow and Static Conditions

Bacteria adhering to surfaces demonstrate random, nanoscopic vibrations around their equilibrium positions. This paper compares vibrational amplitudes of bacteria adhering to glass. Spring constants of the bond are derived from vibrational amplitudes and related to the electrophoretic softness of the cell surfaces and dissipation shifts measured upon bacterial adhesion in a quartz-crystal-microbalance (QCM-D). Experiments were conducted with six bacterial strains with pairwise differences in cell surface characteristics. Vibrational amplitudes were highest in low ionic strength suspensions. Under fluid flow, vibrational amplitudes were lower in the direction of flow than perpendicular to it because stretching of cell surface polymers in the direction of flow causes stiffening of the polyelectrolyte network surrounding a bacterium. Under static conditions (0.57 mM), vibrational amplitudes of fibrillated Streptococcus salivarius HB7 (145 nm) were higher than that of a bald mutant HB-C12 (76 nm). Amplitudes of moderately extracellular polymeric substance (EPS) producing Staphylococcus epidermidis ATCC35983 (47 nm) were more than twice the amplitudes of strongly EPS producing S. epidermidis ATCC35984 (21 nm). No differences were found between Staphylococcus aureus strains differing in membrane cross-linking. High vibrational amplitudes corresponded with low dissipation shifts in QCM-D. In streptococci, the polyelectrolyte network surrounding a bacterium is formed by fibrillar surface appendages and spring constants derived from vibrational amplitudes decreased with increasing fibrillar density. In staphylococci, EPS constitutes the main network component, and larger amounts of EPS yielded higher spring constants. Spring constants increased with increasing ionic strength and strains with smaller electrophoretically derived bacterial cell surface softnesses possessed the highest spring constants.

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.

An in vitro study of cartilage-meniscus tribology to understand the changes caused by a meniscus implant

Active lifestyles increase the risk of meniscal injury. A permanent meniscus implant of polycarbonate urethane (PCU) is a promising treatment to postpone/prevent total knee arthroplasty. Study of the changes in articular cartilage tribology in the presence of PCU is essential in developing the optimum meniscus implant. Therefore, a cartilage-meniscus reciprocating, sliding model was developed in vitro, mimicking the stance and swing phases of the gait cycle. The meniscus was further replaced with PCU and surface-modified PCUs (with C18 chains, mono-functional polydimethylsiloxane groups and mono-functional polytetrafluoroethylene groups) to study the changes. The coefficient of friction (COF) was calculated, and cartilage wear was determined and quantified histologically. The cartilage-meniscus sliding resulted in low COF during both stance and swing (0.01< COF <0.12) and low wear of cartilage (scores <1). The cartilage-PCU sliding, during stance, revealed similar low COFs. But during swing, the COFs were high (average ∼1, maximum 1.6), indicating a breakdown in interstitial fluid pressurization lubrication and non-effective activation of the boundary lubrication. This may lead to wear of cartilage in long term. However, under the tested conditions the wear of cartilage against PCUs was not higher than its wear against meniscus, and the cartilage was occasionally damaged. The COF decreased with increasing the contact pressure (as-per a power equation) up to 1MPa. The changes in the surface modification of PCU did not affect PCUs tribological performance.

Notochordal cell matrix as a bioactive lubricant for the osteoarthritic joint

Notochordal cell derived matrix (NCM) can induce regenerative effects on nucleus pulposus cells and may exert such effects on chondrocytes as well. Furthermore, when dissolved at low concentrations, NCM forms a viscous fluid with potential lubricating properties. Therefore, this study tests the feasibility of the use of NCM as a regenerative lubricant for the osteoarthritic joint. Chondrocyte-seeded alginate beads were cultured in base medium (BM), BM with NCM (NCM), or BM with TGF-β1 (TGF), as well as BM and NCM treated with IL-1β. NCM increased GAG deposition and cell proliferation (stronger than TGF), and GAG/DNA ratio and hydroxyproline content (similar to TGF). These effects were maintained in the presence of IL-1β. Moreover, NCM mitigated expression of IL-1β-induced IL-6, IL-8, ADAMTS-5 and MMP-13. Reciprocating sliding friction tests of cartilage on glass were performed to test NCM’s lubricating properties relative to hyaluronic acid (HA), and showed a dose-dependent reduction in coefficient of friction with NCM, similar to HA. NCM has anabolic and anti-inflammatory effects on chondrocytes, as well as lubricating properties. Therefore, intra-articular NCM injection may have potential as a treatment to minimize pain while restoring the affected cartilage tissue in the osteoarthritic joint.

An ex vivo salivary lubrication system to mimic xerostomic conditions and to predict the lubricating properties of xerostomia relieving agents

Advances in medical research has resulted in successful treatment of many life-threatening infectious diseases as well as autoimmune and lifestyle-related diseases, increasing life-expectancy of both the developed and developing world. As a result of a growing ageing population, the focus has also turned on chronic diseases which seriously affect the quality of older patient life. Xerostomia (dry mouth) is one such condition, which leads to bad oral health and difficulty in consumption of dry foods and speech. Saliva substitutes are used to ease symptoms. However, they often don’t work properly and objective comparison of saliva substitutes to mimic natural salivary functions does not exist. The study thus aims to develop an ex vivo friction assay simulating dry mouth conditions and facilitating objective comparison of saliva substitutes. A reciprocating sliding tongue-enamel system was developed and compared to a PDMS (polydimethylsiloxane)-PDMS friction system. The tongue-enamel system, but not the PDMS-PDMS model, showed high mucin-containing saliva (unstimulated and submandibular/sublingual saliva) to give higher Relief than mucin-poor lubricants (water, parotid saliva, Dentaid Xeros) and correlated well (r = 0.97) with in vivo mouth feel. The tongue-enamel friction system mimicked dry mouth conditions and relief and seems suited to test agents meant to lubricate desiccated oral surfaces.