Curr Microbiol (2015) 70:846–853 DOI 10.1007/s00284-015-0796-8

The Role of Plasma, Albumin, and Fibronectin in Staphylococcus epidermidis Adhesion to Polystyrene Surface Daria Eroshenko • Ilya Morozov • Vladimir Korobov

Received: 6 October 2014 / Accepted: 20 January 2015 / Published online: 7 March 2015 Ó Springer Science+Business Media New York 2015

Abstract The influence of soluble and immobilized plasma, albumin, and fibronectin (Fn) on the adhesion of three Staphylococcus epidermidis strains to polystyrene was investigated. Both soluble and immobilized plasma and albumin cause to 7-fold reduction of the amounts of adhered cells, regardless of the strain used. The soluble Fn exhibited the adhesion for one strain and did not affect the bacterial sorption for remaining strains, whereas on Fncoated polystyrene two of the three strains showed about 1.5-fold increase in the number of adsorbed bacteria. The plasma- and albumin-coated surfaces became much more hydrophilic as the contact angle changed from 78 ± 2° for control to 18 ± 2° for plasma and 21 ± 3° for albumin. The ligand–receptor specific interactions strains S. epidermidis with Fn-coated surfaces were proved by measuring the adhesion forces between cell surface and Fncoated AFM tip. The surface roughness measured using AFM after the plasma and proteins immobilization was changed within 10 nm and not correlate with changes in bacterial adhesion.

D. Eroshenko (&)  V. Korobov Institute of Ecology and Genetics of Microorganisms UB RAS, Perm, Russia e-mail: [email protected] I. Morozov Institute of Continuous Media Mechanics UB RAS, Perm, Russia I. Morozov Perm National State Research University, Perm, Russia

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Introduction Bacterial adhesion to various polymer surfaces is considered to be a critical step in the development of infections associated with indwelling medical devices. It is known that both a specific (ligand–receptor) and nonspecific interactions (electrostatic or hydrophobic bonds) play an important role in the ability of bacterial cells to attach to polymeric surfaces and resist separation from them [1]. The certain contribution of specific and nonspecific interactions in the ‘surface-bacterium’ interactions is probably determined by the physic-chemical properties of the bacterial cell surface or the attacked polymer surface. Coagulase-negative staphylococci, particularly Staphylococcus epidermidis are among microorganisms responsible for biomaterial-associated infections. Almost 50 % of infections associated with catheters, artificial joints, and heart valves are caused by S. epidermidis [20]. Largely, this is due to their non-specific binding to the polymer surface [1, 4]. But a contact of medical devices with the blood promotes a rapid adsorption of plasma proteins on the polymeric surface with the formation a ‘conditional film’ [8]. Adsorbed serum proteins can influence on the surface properties of the material, such as the surface charge, roughness, and hydrophobicity, each of them in turn determine the intensity of bacterial adhesion [5, 12]. In studies in vitro demonstrated that the presence of plasma and its major component albumin suppresses initial bacterial adhesion due to lack of specific interactions between bacteria and albumin [2, 4, 21, 27], but there are the controversial data about the effect of another serum protein– fibronectin (Fn). In several papers reported that the surface coating by Fn promotes adherence of selected staphylococcal strains [7, 17, 24, 31, 38] at the same time there are

D. Eroshenko et al.: Role of Plasma, Albumin and Fibronectin in S. epidermidis

studies in which this protein had no effect or repressed S. epidermidis adhesion [4, 9, 13, 23]. It was found that the increase in the number of bacteria associated with the Fn-coating surface is the result of a specific receptor–ligand interactions between this protein and the surface proteins of the bacterial cell wall, known as ‘microbial surface components recognizing adhesive matrix molecules, MSCRAMM’[3, 37]. In fact, protein Embp with high affinity to Fn was discovered on the surface of S. epidermidis cell [37]. Consequently, there is a need to further characterize the adhesion of various strains of S. epidermidis on the surface coating by serum proteins like albumin and Fn. In this paper, adherence of S. epidermidis cells to polystyrene surface and influence on this process of soluble and surface-bound plasma, albumin, and Fn were analyzed.

Materials and Methods Bacterial Strains, Growth and Harvesting Staphylococcus epidermidis strain 33 (GISK 33), S. epidermidis strain 12228 (ATCC 12228), and S. epidermidis strain 29887 (ATCC 29887) were used in this study. Bacteria were cultured to exponential phase in Luria– Bertani broth (10 g tryptone, 5 g yeast extract, 6.4 g KCl l-1; Sigma–Aldrich, USA) at 37 °C and shaking 160 rpm. Following growth, cells were pelleted by centrifugation (13,000 rpm, 5 min), the supernatant medium discarded, and cells were washed twice with 0.85 % NaCl, pH 7.0, and were diluted to 1 9 108 CFU ml-1 in 0.85 % NaCl, pH 7.0. Pretreatment of Polystyrene Surface by Plasma or Proteins The polystyrene petri dishes (diameter 40 mm; Medpolymer, Russia) were immersed in 2 ml of 0.85 % NaCl containing either 10 % (v/v) plasma, 5 mg ml-1 bovine serum albumin (Sigma–Aldrich, USA), or 25 lg ml-1 human Fn (Sigma–Aldrich, USA) at 37 °C for one h. The control group was immersed in 2 ml 0.85 % NaCl without proteins. Human whole plasma was prepared from blood drawn by venapuncture from three healthy donors. Blood was collected in vacuum tubes with EDTA (Improvacuter, China) and centrifuged (4000 rpm, 5 min), and the supernatant was diluted in 10-fold by 0.85 % NaCl. Albumin and Fn were used in concentrations similarly to those present in 10 % human blood (5 mg ml-1 and 25 lg ml-1, respectively).

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Adhesion Assay Bacterial adhesion was studied in static conditions [10] on polystyrene petri dishes after coating by protein or plasma solutions or non-coated dishes in the same solutions. In first case, 2 ml of fresh bacterial suspensions in 0.85 % NaCl (1 9 107 CFU ml-1) were exposed on each protein-coated dish. In second case, non-protein-coated dishes were incubated concomitantly with freshly washed bacteria suspended in 2 ml of 0.85 % NaCl containing either 10 % (v/v) plasma, 5 mg ml-1 BSA, or 25 lg ml-1 Fn to 1 9 107 CFU ml-1. Bacterial suspensions (1 9 107 CFU ml-1) in 0.85 % NaCl without proteins were used as control. In all cases, S. epidermidis cells allow to adhere in static conditions at 37 °C for 30 min. After incubation, dishes were rinsed three times with PBS to remove any unbound bacteria. The adherent cells were fixed by heating at 60 °C for 30 min, stained with 0.1 % crystal violet, and directly imaged using digital optical microscope (lViso-103; Lomo, Russia). In order to determine the mean of the number of adherent bacteria, the adsorb cells were counted at least in ten field-of-view (FOV) in each dish at magnification 92500. The results reflect the average [±standard deviations (SD)] of three independent experiments. Determination of Number of Viable Cells The density of the colony-forming units (CFU ml-1) in suspension after adhesion was measuring by drop plate method on the LB-agar medium after a series of 10-fold dilutions in 0.85 % NaCl. The number of CFU was counted after incubation for 18–24 h at 37 °C. This experiment was repeated three times using 10 droplets in each repeat (n = 30). Measurement of Bacterial Cell and Surface Hydrophobicity The relative bacterial cell surface hydrophobicity was determined with the BATH method with hexadecane [29] and expressed as the percentage of bacteria adhering to hydrocarbon. Hexadecane (0.2 ml) was added to 1.2 ml of bacterial suspension in 0.85 % NaCl (*108 CFU ml-1) and in 10 min vortexed at highest speed for 120 s. Suspensions were allowed to stand for 15 min to allow phase separation prior to measurement of the absorbance of the aqueous phase at 600 nm. The percentage hexadecane adherence was calculated as the difference in the initial absorbance of the bacterial suspension (before adding hexadecane) minus the absorbance of the hexadecanetreated suspension divided by the initial absorbance of the

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bacterial suspension. The results reflect the average (±SD) of three independent measurements. The hydrophobicity of the polystyrene surface with and without adsorbed proteins was assessed by static captive bubble method. The surfaces were inverted (active layer to the bottom) in 0.85 % NaCl at room temperature. An air bubble (50 ll, n = 3) was injected from a syringe with a stainless steel needle onto the sample surface water. The air bubble profile was photographed using digital photo camera (D90; Nikon, Japan) during 60 s after the contact. The radius (r) and height (h) of each drops in pixels were manually measured using GIMP2.0 software. The contact angle (H, ) was calculated by formula [40]  H ¼ 2  argtch h  r1 : ð1Þ The results reported the average (±SD) of three independent experiments with three droplets on one sample (nine measurement in total per surface). Atomic Force Microscopy Surface properties (roughness and adhesion force) of the polystyrene with and without adsorbed proteins were measured using Dimension Icon AFM (Veeco, USA). Images (15 9 15 lm2, 512 9 512 lines) of samples were captured in 10 mM phosphate-buffered saline (PBS; containing 0.85 % NaCl at pH 7.2), at room temperature (22–24 °C), in PeakForce QNM (quantitative nanomechanical mapping) mode using silicon nitride CSG30 (NTMDT, Russia) with a nominal spring constant of 0.7 Nm-1 and the tip radius *10 nm. The maximum applied force was set to 5 nN, and the scan rate was set to 1 Hz for the all adhesion force measurements. Three reading were made of each surface on two prepared samples, and average roughness (Ra) and mean roughness profile depth (Rz) were used to characterize the roughness of the surface. The results are expressed as mean ± SD of all measurements. To create Fn-coated AFM tip, the silicon nitride AFM cantilevers were cleaned in ethanol, rinsed with deionized water, immersed in a 100 lg ml-1 Fn PBS solution for 45 min, and then rinsed four times in PBS [39]. For certainty of measurements, each Fn-coated tip was checked for tip coating by probing a clean polystyrene dish. A small volume of washed cells, as stated above, was transferred onto a polystyrene Petri dish and allowed to sit for 5 min without drying. Nonadherent cells were washed away with PBS. The polystyrene Petri dish with attached cells was then transferred to the atomic force microscope. Force measurements between Fn-coated tip and bacterial surface were performed in PBS using Dimension Icon AFM (Veeco, USA) as stated above. For experiments, three samples of cells from different days were used; at least forty-five force curves from each sample were generated. Every time freshly Fn-coated tip was used.

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Statistical Analysis The mean ± SD of the adherent cells (n = 30), viable bacteria (n = 30), bacterial cell surface hydrophobicity (n = 3), contact angles (n = 9), and topographic parameters of surfaces (n = 6) were analyzed by Prism 6 software. Statistical significance was calculated using oneway analysis of variance (ANOVA). The value of statistical significance was set at P \ 0.05.

Results Bacteria of S. epidermidis are characterized by high sorption capacity to the various polymer surfaces [1, 2]. To get insight into what the role proteins can play in the bacterial adherence to polystyrene in vivo, adherence of three S. epidermidis strains to polystyrene following treatment by plasma or one of the serum proteins, albumin or Fn, was tested. On the second variant, bacterial adhesion to untreated polystyrene was carried out in 0.85 % NaCl containing BSA or Fn, or was tested in plasma solutions. Topography of Polystyrene Protein-Coated Surfaces As mentioned above, the treatment of the polymer surface with serum proteins or plasma causes a change in its physical–chemical characteristics [5, 12], so after immersion in protein solutions hydrophobicity, surface topography and adhesion force between the surface and silicon cantilever tip were tested. AFM images of the control and coated polystyrene surfaces are represented in Fig. 1. The topographic images show small islands distributed on the surface after treatment with plasma solution (Fig. 1b) and smooth surfaces with the BSA (Fig. 1c) or Fn (Fig. 1d) coating. The roughness measurements (the Ra and mean roughness profile depth) for these samples in nanometers are represented in Table 1. Immersion in plasma solutions led to near 2-fold increasing average roughness (Ra 4.9 ± 1.5 nm) comparing to control (Ra 2.9 ± 0.4 nm) (P \ 0.05), while the BSA or Fn coating had no significant effect on the roughness (Ra 2.6 ± 0.5 and 3.6 ± 0.3 nm, respectively). Also it was noted that the adhesion force between the cantilever and surfaces changed in the case of all protein coating (Table 1). The treatment with plasma shown the maximum effect (2-fold increase), followed by Fn (almost 1.5-fold increase) and finally BSA has about 1.3-fold reduction in adhesion force. Contact Angle Measurements The untreated polystyrene surface was hydrophobic with a contact angle of 78 ± 2° (Table 1). The plasma and

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Fig. 1 AFM topographic images of polystyrene surfaces following treatment a 0.85 % NaCl, b 10 % plasma, c 5 mg ml-1 BSA and d 25 lg ml-1 Fn. Scan size is 15 9 7.5 mm (images represent typical fields of view). 340 9 170 mm (150 9 150 DPI)

Table 1 Surface roughness, adhesion force characteristics, and hydrophobicity of polystyrene surfaces after treatment with plasma or protein solutions during 1 h (mean ± SD) Medium

Ra (nm)

Rz (nm)

Fa (nN)

H (°)

Control

2.9 ± 0.4

2.4 ± 3.5

0.47 ± 0.02

78 ± 2

Plasma

4.9 ± 1.5*

3.4 ± 3.8

0.96 ± 0.02*

18 ± 2*

BSA

2.6 ± 0.5

2.1 ± 0.4

0.34 ± 0.002*

21 ± 3*

Fn

3.6 ± 0.3

2.9 ± 0.4

0.64 ± 0.002*

73 ± 4

Ra arithmetic mean of the absolute values of the surface height deviations measured from the mean plane, Rz absolute distance between the highest peaks and valleys relative to the mean plane, Fa adhesion force between the cantilever tip and the surface, H contact angle * Significance (P \ 0.05)

compared to control for

each characteristics

not have a significant effect on the sorption cell of S. epidermidis 33, but for strains 12228 and 29887 there was a small (1.3–1.7 fold) increase in the number of adsorbed cells compared with control (P \ 0.05). The adhesion of S. epidermidis strains in protein-containing media are shown in Fig. 2b. In the plasma or BSA containing media, the adherence of all tested strains to untreated polystyrene was up to 87 % less versus the control. The presence of Fn did not affect the intensity of the bacteria sorption for strains 33 and 29887. At the same time, the presence of this protein in the incubation medium had a stimulating effect on the adhesion of the bacteria S. epidermidis 12228. Determination of Number of Viable Cells

albumin coating caused significant alterations on the contact angle. When plasma was used, a decrease to 18 ± 2° was observed (P \ 0.05). When BSA was coated on polystyrene, the surface became much more hydrophilic with contact angle of 21 ± 3° (P \ 0.05). When the treatment was performed with Fn, contact angle is not significantly changed (73 ± 4°) and the surface hydrophobicity remained similar to control. S. epidermidis Adhesion Figure 2a shows the results of the S. epidermidis cells adherence to the protein-coated polystyrene. When the treatment was performed with plasma or BSA, the 2–3 fold decrease in the number of adsorbed cells for all tested strains was observed. Pretreatment of surface with Fn did

It was noted that incubation of the bacteria in the presence of the plasma or proteins had no effect on the cell viability level (P [ 0.05), which was shown by the determination of number of viable cell (CFU ml-1) (Table 2). Measurement of Bacterial Cell Hydrophobicity Table 3 shows that all studied S. epidermidis strains have high affinity with hexadecane within the limits of 65–78 %. All of the studied bacteria were classified as strongly hydrophobic since the surface hydrophobicity was higher than 55 % [26]. In addition, there is no significant difference (P [ 0.05) in the bacterial cell surface hydrophobicity among S. epidermidis strains.

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D. Eroshenko et al.: Role of Plasma, Albumin and Fibronectin in S. epidermidis Table 3 The bacterial cell surface hydrophobicity of S. epidermidis strains (mean ± SD) S. epidermidis strain

33

12228

29887

Hydrophobicity (%)

65.5 ± 16.0

75.8 ± 11.5

77.6 ± 9.1

[25]. For comparison, Fig. 3 also shows force curves for Fn-coated tip on the polystyrene dish. Figure 4 summarize the all adhesion force for bare polystyrene surface and each of the three S. epidermidis strains. Specific binding events of a least 0.5 nN were observed with greater frequency in the retraction curves associated with the 12228 (frequency = 0.54 ± 0.07) and 29887 (frequency = 0.48 ± 0.05), and with lower frequency in the retraction curves collected from the S. epidermidis 33 (frequency = 0.35 ± 0.07) which have not increased adhesion on Fn-coated surface (Fig. 2). Comparison of groups using Student’s t test demonstrates significant differences between the strains 12228 and 33 (P \ 0.05) and the strains 29887 and 33 (P \ 0.05), but not between the strains 12228 and 29887 (P = 0.45).

Discussion

Fig. 2 Adhesion of three S. epidermidis strains (33, 12228, and 29887) to the polystyrene a after protein adsorption on polymer surface or b in the protein-containing media. Control-0.85 % NaCl; Plasma-10 % plasma; BSA-5 mg ml-1 albumin and Fn-25 lg ml-1 fibronectin. The results presented as mean of the number of adhered cells in field-of-view ± SD of three independent experiments. *Significance compared to control for each strain (P \ 0.05). 163 9 214 mm (300 9 300 DPI)

Force Measurements with S. epidermidis Strains Approximately 540 force curves were collected on samples prepared from the three different S. epidermidis strains. Figure 3 shows some representative force curves for each of the three S. epidermidis strains. Such nonlinear, sawtooth-shaped force-distance profile of force curve has been attributed to specific binding events mediated by proteins

Table 2 The number of viable cell of S. epidermidis 33 in incubation medium after 30 min adhesion (mean ± SD)

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Medium

The direct interaction between bacteria and bare polymer surfaces can play minimal role in vivo as after contact with blood the polymer surface almost immediately become covered with plasma and tissue proteins [8, 15], reaching a maximum saturation level after 15–60 min [8, 18, 35]. This ‘conditional film’ cause the change in physical–chemical characteristics of surface [5, 12]. There are evidences of a strong relationship between the early stages of bacterial adhesion and the surface roughness [30, 34]. But the results of this study indicated that the topographies of BSA (mean Ra 2.6 nm)- and Fn (mean Ra 3.6 nm)-coated surfaces are similar to the untreated surface (mean Ra 2.9 nm) but they differ in the ability to adsorb of S. epidermidis cells (Fig. 2). For instance, the adhesion of S. epidermidis 33 to the surface after treatment with BSA was up to 87 % less than the control while the number of adhered cells on Fn-coated surfaces does not differ from the control (23 ± 3 and 23 ± 5 adhered cells FOV-1,

The number of viable cell, 9107 CFU ml-1 After protein adsorption on polystyrene surface

In the protein-containing media

Control

1.2 ± 0.2

1.3 ± 0.4

Plasma

1.0 ± 0.2

0.8 ± 0.3

BSA

1.2 ± 0.3

1.0 ± 0.4

Fn

1.3 ± 0.2

1.2 ± 0.3

D. Eroshenko et al.: Role of Plasma, Albumin and Fibronectin in S. epidermidis

Fig. 3 Retraction force profiles collected as an Fn-coated probe was pulled from contact with the cell wall of each of three S. epidermidis strains (33, 12228, and 29887) and the polystyrene in PSB. Shown are randomly selected retraction curves from a total of 135 force profiles for each of three S. epidermidis strains. 89 9 86 mm (300 9 300 DPI)

Fig. 4 Frequence of binding and binding force (in nN) for a Fncoated probe on bare polystyrene and the three S. epidermidis strains (33, 12228 and 29887) and the in PSB. This figure includes analyses of 135 force curves for each of three S. epidermidis strains 103 9 94 mm (300 9 300 DPI)

respectively). Furthermore, a smooth untreated polystyrene surface (mean Ra 2.9 nm) is characterized by a greater bacterial adhesion versus plasma-coated one (mean Ra 4.9 nm) (Fig. 2). Thus, it is possible to agree with Shida et al. [32] that the surface roughness less than 10 nm Ra has a limited effect on the adhesion of the bacteria S.

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epidermidis because this size is characterized by absence of microcracks, which could serve as niches for microbial cells [33]. The results of this research (Fig. 2) concerning plasma and its main protein, albumin, are agreed with the wellknown fact that the plasma and albumin significantly reduce bacterial adhesion of the important pathogens such as Pseudomonas and Staphylococcus [4, 11, 16, 21]. As well as Ardehali et al. [2] observed a marked reduction in bacterial adhesion of S. epidermidis ATCC 12228 during 2 h to polyurethane surfaces in the presence of bovine/ human plasma at 0.5 % or higher concentration. Linnes et al. [23] demonstrated that S. epidermidis ATCC 12228 cells less able to adhere to a variety of biomaterials, including polyethylene, poly(ethylene terephthalate), fluorinated ethylene propylene, poly(ether urethane), silicone, and borosilicate glass after the albumin immobilizing. The analysis of the experimental data has shown that the reduction in the number of adsorbed cells during 30 min for all tested strains S. epidermidis (Fig. 2a) was not accompanied by an decrease in the number of the viable cells in suspension (Table 2) but correlated with the decrease in the water contact angle (Table 1) for albumin- and plasmacoated surfaces. It allows concluding the adhesion of all tested S. epidermidis strains depend on the hydrophobicity of the polymer surface. Indeed, experiments with 125I-labeled albumin showed that the cells do not adsorb this protein [4]. Due to this fact, most researchers have come to believe that the main mechanism of the albumin and plasma action on the bacterial cells adhesion is manifested through the sorption of proteins to polymer surfaces, which causing the reduction of surface hydrophobicity that has been shown through measuring the water contact angle of polystyrene [12] and fluorinated ethylene propylene [16]. It is important to note that this effect of the plasma and albumin was shown on various types of surfaces including polyethylene [4], Teflon, polyethylene, polycarbonate [5], nylon, polyvinylchloride [6], polyurethanes, and fluorocarbon [27]. Fn is very important glycoprotein of the extracellular matrix and plasma and it also participates in the process of bacterial adhesion to polymeric materials and host tissues. The Fn immobilization on the polystyrene surface does not change its hydrophobicity (Table 1). In this regard, all tested S. epidermidis strains with the similar hydrophobicity (Table 3) are supposed to show similar adhesion because the hydrophobicity of bacterial cells well correlated with their adhesion [13]. However, the heterogeneity in binding with Fn-coated surface among tested staphylococci strains was observed. Thus, pretreatment of polystyrene surfaces with Fn cause a stimulating effect on the sorption of bacterial cells (S. epidermidis 12228 and 29887) as well as did not show any visible effect

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(S. epidermidis 33) (Fig. 2). Similar data were obtained by Brokke et al. [4] when Fn was immobilized on the surfaces of the polyethylene catheters. Most likely, this effect is related to the specificity of the interaction between bacterial cell of analyzed strains and Fn. Figure 3 shows attractive, nonlinear force-distance profiles (or waveforms) when a bond forms between a Fn-coated probe and S. epidermidis strains 12228 and 29887 that have showed the highest adhesion on Fn-coated polystyrene. Sawtooth-shaped force spectra such as these have been attributed to the mechanical unraveling of protein molecules that have formed a bond between two surfaces [24]. Our force measurements indicate that the binding force between the Fn and cell wall of S. epidermidis strains 12228 and 29887 is remarkably strong, near 2 nN (Fig. 3), which is similar to the strength of a covalent bond [14]. The peak of adhesion force curve between Fn-coated tip and these two strains was observed near 170 nm of separation. According to Xu et al. [38] the interactions of Fn and bacterial cell with a rupture length above 93.6 nm were considered as specific interactions. Furthermore, these force-distance profiles differ from the one observed when AFM is used on S. epidermidis 33 that adhere to Fn-coated surface as well as to untreated polystyrene (Fig. 2a) or when a Fn probe is retracted from an inert surface (Fig. 3). Moreover, Western blots confirmed low level synthesis and production of Embp, the protein with high affinity for binding to Fn [37], in cell wall extracts of various S. epidermidis strains, including ATCC 12228 [22]. Therefore, the increase in bacterial adhesion of S. epidermidis 12228 and 29887 on the polystyrene Fn-coated surfaces and the sawtooth-shaped force signature allow making an assumption about a presence of protein like Embp that forms bond between Fn on a substrate and the cell wall of bacterial cells of strains 12228 and 29887, and the absence of such structures on S. epidermidis 33 cell surface. The number of adhered cells for all tested S. epidermidis strains is almost identical both on the plasma and BSAcoated surface and in their presence in solution (Fig. 2). This can be explained by the fact that the albumin is first adsorbed on the surface after the contact of the polymer with blood [36]. At the same time, the different binding characteristics of S. epidermidis 29887 for soluble versus immobilized Fn (Fig. 2) presumably reflect conformational differences, since soluble Fn appears to be a globular protein, whereas during immobilization on polymer surfaces, the N-terminus domain of Fn which is preferred for the binding by S. epidermidis cell [19] becomes available [28].

Conclusion The plasma and serum proteins play an important role in the S. epidermidis biofilm formation starting from the early

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stages (30 min) of the formation of these specific bacterial communities. When polystyrene surface was pretreated with plasma or BSA, main role belongs a change in the surface hydrophobicity, whereas in the case of Fn-coated polystyrene, the S. epidermidis adhesion is controlled by the specific types of interaction (ligand–receptor). The alterations in the surface roughness within limits of 10 nm after immobilizing of both proteins or plasma have no significant importance on the adhesion process. Acknowledgments This work was supported by the RFBR under Grants 12-04-01431-a and 14-04-00687; and UB RAS under Grants 12-P-4-1002, 12-I-4-1003 and 12-M-14-2035.

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The role of plasma, albumin, and fibronectin in Staphylococcus epidermidis adhesion to polystyrene surface.

The influence of soluble and immobilized plasma, albumin, and fibronectin (Fn) on the adhesion of three Staphylococcus epidermidis strains to polystyr...
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