Development and Characterization of Multilayered Cu/HA/ZnS + PEEK Coating System by Hybrid Technology

Zirconium alloys belong to a group of metallic biomaterials that are commonly used for bone implants. Due to their exceptional electrochemical corrosion resistance, wear resistance and low modulus of elasticity (roughly 90 GPa), which is lower than that of stainless steels, cobalt alloys and two-phase α + β titanium alloys, they are widely studied and used as orthopedic and dental implants.^[1,2] In addition, zirconium alloys display higher biocompatibility than steels, cobalt and titanium alloys.^[3] One of the most perspective alloys is Zr–2.5Nb, which reveals favorable biocompatibility, high electrochemical corrosion resistance and acceptable mechanical properties.^[4] Although zirconium alloys are coated with a passive layer of ZrO₂, which exhibits rather moderate bioactivity, their osteoinductivity must be enhanced.^[5‐7] This can be achieved by employing composite coatings with a bioactive component.

One of the excellent composite coating matrix materials is polyetheretherketone (PEEK). It is characterized by remarkable resistance to chemicals and radicals, high stability, mechanical strength and wear resistance.^[8] In addition, PEEK exhibits an elasticity modulus close to that of human trabecular bone, much less than that of metallic biomaterials. Therefore, its use in the coating can decrease the effects of stress shielding on the surrounding bone. Due to its superior properties, it is often used as spinal, dental and orthopedic implants or medical equipment.^[9‐11] Although it is characterized by biological inertia.^[12] Due to this drawback, it is necessary to incorporate bioactive and antiseptic agents in PEEK-based coatings.

One of the most well-known biologically active factors is synthetically prepared hydroxyapatite (HA), with a chemical composition of Ca₁₀(PO₄)₆(OH)₂.^[13] This ceramic is one of the main components of natural bones. HA is widely applied in bioactive coatings and scaffolds because it can form a chemical bond with the bone.^[14] Due to its bioactivity, osteoconductivity, biocompatibility and ability to form a stable connection with osseous tissue, it is widely used for bone repair.^[15,16] Although there are numerous advantages, HA is easily agglomerated and brittle. Therefore, it is often used in composite coatings as an additional bioactive component. When incorporated into the coating, it is able to increase the concentration of local Ca²⁺ ions concentration, stimulating bone formation.^[17,18] Despite this advantage, HA induces a bone growth effect, but does not prevent the formation of bacterial microfilms. Hence, antibacterial ingredients are often applied in composite biomaterials containing HA. During osseointegration processes, it is very important to inhibit the development of biofilm, which can cause serious infection. This behavior can not only be a cause of implant repudiation, but also be a threat to patient health.^[19,20] The currently investigated and promising antibacterial ingredients include ZnS and Cu nanoparticles. ZnS is characterized by antimicrobial activity, relatively high chemical stability, and non-toxicity. Therefore, it is suitable for use in coatings.^[21,22] Furthermore, as reported in our previous study,^[23] ZnS provides a source of sulfur for the thermal sulfonation process of PEEK. Sulfonated PEEK (S-PEEK) enhances bone cell formation on the coatings,^[24,25] as well as exhibits antibacterial activity.^[26] Accordingly, the use of ZnS particles in the coating composition is doubly beneficial. Copper is one of the most important microelements in the human body. The antibacterial activity of this element is known and widely used.^[27‐29] Despite the negligible reactivity of Cu to human tissues, it is highly effective against harmful microorganisms. From the perspective of the decline in the effectiveness of the most widely used antibiotics, the use of Cu nanoparticles is a promising alternative.^[30,31]

Electrophoretic deposition (EPD) is widely applied and studied for the deposition of polymer-based coatings with the addition of various bioactive ceramics, including HA.^[32‐34] EPD is accomplished via the movement of particles suspended in a liquid medium under an electric field. The conductive substrate is used as the working electrode and is coated by particles of opposite charge. This method is advantageous due to the short time of deposition and the limited restriction of the substrate shape. Furthermore, for the densification of PEEK coatings, thermal treatment is necessary.^{[23,32,33,35‐38]} Multilayered coatings often require the use of multiple suspensions during EPD and repeating individual deposition operations. However, they allow for limited control of the location of individual components. PEEK-based coatings with both antimicrobial and bioactive agents were previously obtained only in a few studies.^{[23,33,39‐42]} Seuss et al.^[39] developed PEEK coatings containing HA and Ag particles. Abdulkareem et al.^[40] achieved PEEK coatings with the addition of HA and chitosan. In our previous studies, multicomponent HA/MoS₂/PEEK and HA/ZnS/PEEK coatings were successfully deposited.^[23,33] However, efficiently all of the above coatings were obtained as single-layered. Although potentially beneficial, the multilayered EPD technique was applied for the development of PEEK-based composites on a limited basis. Virk et al.^[41] developed multilayered PEEK-based coatings with curcumin, hexagonal boron nitride and bioactive glass. Moreover, Ur Rehman^[42] obtained a bioactive glass (BG)/chitosan layer loaded with lawsone on the base layer of PEEK/BG. Nevertheless, no studies have been reported on the hybrid method involving EPD and selective pulsed laser deposition (PLD) for obtaining multilayered coatings with fully controlled distribution of antibacterial and bioactive agents. Physical vapour phase deposition using masking is a very promising technique for selective deposition of Cu “island structures” on any substrate.^[43] This technique provides numerous advantages, including high precision, speed and efficiency of deposition, tight control of the deposited structure and high flexibility.^[43,44] A significant benefit of using PLD is the preservation of the phase composition and stoichiometry of the deposited materials. Shadow masking is often used as a factor to reduce the occurrence of coating defects, such as droplets, which are micrometer-sized particles that can have a negative impact on coating properties, which is related to mask displacement along the target-substrate path and verification of the deposition rate.^[44,45] In the present work, the selective Cu deposition process was carried out with the mask attached directly to the samples, which is a new approach. PLD with a shadow mask enables selective deposition of Cu "islands" of desired size, thickness and precise distribution within the bioactive coating matrix and thus can regulate the biological response of both cells and bacteria. The present work focuses on the development of multilayered Cu "islands"/HA/ZnS + PEEK coatings by a hybrid method consisting of EPD, heat treatment (HT) as well as shadow masked PLD on Zr–2.5Nb alloy substrates. The parameters of the processes, including EPD kinetics, were optimized and studied. The resulting coatings were studied in terms of morphology and microstructure, surface topography, scratch resistance, and adhesion strength.

Based on our previous studies^[23,33] CHp was used for electrosteric stabilization of the suspension and co-deposition of ZnS and PEEK particles on the cathode. Previous studies of the zeta potential (ZP) of PEEK, HA, and ZnS particles in pure ethanolic alcohol and the alcohol with the addition of 5 vol pct CHp, showed that the CHp provides positive values of ZP for a pH range of 3.0 to 12.0.^[23] It is most crucial for PEEK particles, for which the isoelectric point (slightly above 5.5 pH) in pure EtOH is very adjacent to the isoelectric point of the suspension of ZnS + PEEK with CHp (5.4 pH), which could significantly impede cathodic deposition. The EPD of the HA layer was carried out from a pure EtOH suspension due to the wide range of positive ZP values from 3.0 to 8.5 pH, with an isoelectric point of 8.75 pH. The pH value of the suspension was 8.14 at RT.

A short deposition time of 15 seconds was enough to obtain a sufficiently thick homogeneous ZnS + PEEK layer as the base for the second HA layer. The influence of voltage on the homogeneity of the layer was visible with the unaided eye, as well as with the use of SEM. The SEM investigation of the as-deposited first layer of ZnS + PEEK confirmed that the deposition voltage of 30 V during 15 seconds is not sufficient to obtain a homogenous coating that completely covers the underlying substrate (Figure 2(a)). Deposition in the voltage range of 30 to 70 V at the same time resulted in the achievement of a non-uniform thin layer. Observed with the unaided eye, the ZnS + PEEK layer deposited at a voltage between 70 and 110 V was satisfactory thick, with the presence of open pores. EPD at higher voltages (90, 150 V) led to the acquisition of continuous coatings that covered the substrate material (Figures 2(b) and (c)). The microscopic surface morphology of the coatings deposited with the voltage of 90 and 150 V was similar and contained numerous open pores with the ECD up to 20 μm, which were relatively homogeneously distributed on the surface. Furthermore, microcracks up to 100 µm long were visible on both coatings, although they were slightly more apparent on the surface of the coating developed at 150 V. The coatings deposited at voltages higher than 110 V were macroscopically inhomogeneous with numerous pores, and the number of inconsistencies multiplied with increasing voltage. During the EPD of the second layer (HA), no deposition was observed on the surface of the ZnS + PEEK below 50 V, as observed with the naked eye. The SEM–EDS investigation of the HA layer obtained with the use of 30 V revealed the deposition to a very limited extent (Figure 3(a)). Almost no change in surface morphology was observed compared to that of the base ZnS + PEEK layer. Inspecting macroscopically, in the voltage range of 50 to 70 V, single HA agglomerates were obtained. On the contrary, macroscopically, deposition above 110 V resulted in massive and uneven HA agglomeration. The SEM and SEM–EDS investigation revealed that the selective HA coating developed at 90 and 150 V on the surface of ZnS + PEEK was clearly visible (Figures 3(b) and (c)). The presence of pores on the surface of these coatings was limited. Nevertheless, microcracks with lengths up to 220 µm were visible. The surface morphology of the coating with HA deposited at 150 V was more diversely developed, with increased HA agglomeration. Therefore, EPD in the voltage range from 70 to 110 V was applied to achieve the HA layer.

Examination of the change in current density of the ZnS + PEEK layer deposition exhibited its fluctuations for a voltage of 90 and 150 V for the first 5 seconds of the process (Figure 4(a)). After that, the process was stable. For 30 V, the change in current density was without fluctuations. For the deposition voltage of 90 V, the current density was stable at 0.54 mA/cm². The instability of the current density was reflected in the macroscopic unevenness and noticeable thickness of the coatings deposited with the voltage of 150 V. They were characterized by numerous pores and irregularities. Unlike coatings deposited at 30 V in which no inconsistencies were observed, although they were thin. Coatings deposited at 90 V had limited pores and uniformly covered the substrate. As for the HA layer, the change in current density for voltage values of 30, 90 and 150 V was relatively similar, ranging from 0.005 to 0.02 mA/cm² for the entire deposition time of 30 seconds (Figure 4(b)). The current density for the three voltages was meagre due to the presence of an insulating ZnS + PEEK layer. Interestingly, in contrast to the ZnS + PEEK layer, the change in current density was the highest for a deposition voltage of 30 V and oscillated at around 0.018 mA/cm². The current density change for deposition at 150 V peaked at 0.0085 mA/cm², but the average value most of the time was about 0.008 mA/cm². For the EPD at 90 V, the current change was in the range of 0.007 to 0.017 mA/cm².

Moreover, the EPD kinetics, deposition yield and rate of both layers were investigated. The deposition yield of the ZnS + PEEK layer was growing fast and almost linearly and peaked close to 0.06 mg/mm² at the end of the layer deposition (15 seconds) (Figure 5(a)). The deposition yield of the HA layer also increased with the linear progress of the deposition, but was slower, reaching 0.03 mg/mm² at 30 seconds (Figure 5(a)). Regarding the deposition rate of the ZnS + PEEK layer it increased promptly during the first 5 seconds of the deposition reaching 0.004 mm/mg²·s, then decreased to 0.0035 mm/mg²·s in the 10th second, reaching the final value of 0.004 mm/mg²·s in the 15th second. The deposition rate of the HA layer on the underlying ZnS + PEEK expanded significantly until the 10th second of deposition up to 0.001 mm/mg²·s and then stabilized at this value until the end of the process. The lower deposition rate and yield of the HA layer were a result of the occurrence of a non-conductive continuous ZnS + PEEK layer on the conductive substrate. It was observed that the HA nanoparticles were deposited preferably in areas of the lower thickness of the ZnS + PEEK layer, such as open pores, because of the lesser insulation effect. Similarly, Virk et al.^[41] and Ur Rehmann et al.^[42] reported that the second layers of curcumin/chitosan and bioactive glass/chitosan, respectively, filled the open pores of the PEEK-based layer.

Based on the heat treatment parameters of single-layered multicomponent HA/ZnS/PEEK and HA/MoS₂/PEEK coatings used in our previous works,^[23,33,48] the coated substrates were heat treated at 380 °C and 450 °C for 30 minutes and then cooled with the furnace. The temperature of 380 °C was found to be not sufficient for the PEEK sulfonation and its partial crystallization was found (Figure 6(a)). In contrast, the coatings obtained at 450 °C exhibited an amorphous PEEK structure indicating that the sulfonation process took place (Figure 6(b)). Moreover, both patterns validated the presence of ZnS (hp) and ZnS (rp). Although the thermal decomposition temperature of ZnS is about 650 °C,^[49] the lower temperature of the heat treatment (450 °C) of the coatings in the air environment was selected due to the degradation of PEEK, which takes place above 575 °C.^[50] The results of the XRD investigation performed in the present work and the FTIR investigation described in our previous work^[23] indicate that PEEK sulfonation occurred under the conditions mentioned above. We suppose that this process is due to a partial decomposition of sulfides, which is probably sufficient to substitute the PEEK chemical chain with sulfur. Due to the lower temperature of applied heat treatment compared to the thermal decomposition temperature of ZnS, this phase is still present in the PEEK matrix, similar to our previous research on multicomponent n-HA/ZnS/PEEK coatings.^[23] Mid-infrared spectroscopy (MIR) analysis of n-HA/ZnS/PEEK^[23] revealed the characteristic bands of vibrations of sulfur-oxygen bonding at approx. 1240 cm⁻¹ (asymmetric O=S=O stretching) and 1035 cm⁻¹ (symmetric O=S=O stretching) in the spectrum, which did not appear in pure, non-sulfonated PEEK spectrum. Furthermore, the significant increase in the intensity of the bands at 684, 1035, 1112 and 1243 cm⁻¹ associated with vibrations of the S–O bonds was additionally observed. Similarly, Moskalewicz et al.^[48] showed sulfonation of MoS₂/PEEK coatings at temperature of 390 °C, which is much lower than the thermal decomposition temperature of MoS₂ of about 650 °C.^[51] MIR analysis revealed the appearance the band at 1224 cm⁻¹, characteristic of the stretching vibrations of S=O, as well as the splitting of the band at around 1500 cm⁻¹ characteristic for aromatic C–C bonds, which is related to the appearance of the sulfur substitutions. In both cases of PEEK coatings containing ZnS and MoS₂, XRD patterns revealed an amorphous PEEK structure, which is not observed in pure PEEK coatings or composite PEEK coatings^{[35,38,52,53]} without sulfides heat-treated under the same or similar conditions. This result also indicates sulfonation. Thus, the two-layered n-HA/PEEK + ZnS substrates were subjected to the same treatment.

The scratch resistance of the coating and the adhesion strength were investigated. First, the adhesion of the ZnS + PEEK coating to the Zr–2.5Nb alloy substrates was investigated. Unfortunately, the ZnS + PEEK coating achieved the worst class of adhesion (0B) to the as-received alloy substrate graded with 600-grit sandpaper. The ZnS + PEEK layers, despite their homogeneity, were in some cases completely detached, starting from the upper edge, without being damaged. The upper area of the layer on the sample was the main site of detachment due to the smaller thickness caused by sedimentation of the suspension during EPD. Thus, to enhance the adhesion strength of the layer to the surface of the substrate, it has undergone different thermal and chemical treatments. Thermal treatment was carried out to create an oxide layer on the alloy substrates. After thermal treatment of the alloy, the adhesion of ZnS + PEEK was not improved. Therefore, a chemical treatment of the substrate was applied. The ZnS + PEEK layer deposited on the chemically treated Zr–2.5Nb substrates exhibited a high class of adhesion (4B) (Figures 7(a) and (b)). Slight delamination of the coating occurred only near the pores in the path of the cut (Figure 7(a)). Finally, multicomponent Cu/HA/ZnS + PEEK coatings did not detach from the chemically treated substrate and exhibited the same high adhesion class (4B).

The micro-scratch test was carried out to investigate the mechanism of damage by scratching and to extend the coating adhesion analysis. The scratch results provide quantitative information on the coating adhesion to the alloy substrate. The critical load that causes characteristic coating failure is determined in the scratch test. At a critical load of L_c1, the first cohesive cracks emerge in the scratch track, L_c2 causes the first adhesion deterioration with minor disclosure of the substrate, and L_c3 is the load that causes massive delamination, with the thoroughly exposed area. The increase in the load transmitted by the stylus generates an increase in interfacial shear stress between the mobile and irregular surfaces of the coating. In addition, tensile stresses and pile-ups of the coating material occur at the edges of the scratch track.

It should be mentioned that the ZnS + PEEK base layers were much more resistant to scratch than previously reported multicomponent n-HA/ZnS/S-PEEK coatings.^[23] In the ZnS + PEEK layers, only cohesive cracks were found under the load of L_c1 = 23 N and up to a load of 30 N no adhesive failures were observed (Figure 8). In contrast, in the multicomponent coatings, cohesive cracks appeared under the load L_c1 of 16 N and the disclosure of the substrate under L_c2 of 27 N. The difference in scratch resistance may be caused by differences in the microstructure of the coatings and the geometrical surface structure.

Electrophoretically deposited multilayered coatings contained numerous visible HA agglomerates and separate HA nanoparticles selectively embedded in the S-PEEK matrix, which formed a uniform solid matrix of the 1st layer (Figure 9). Thus, heat treatment caused a partial immersion of the HA in the PEEK matrix, which contributed to the high adhesion of the 2nd layer to the 1st one.

The as-deposited ZnS + PEEK layer thickness was in the range of 75 to 100 μm. After deposition of HA, the thickness of the as-deposited double layered coatings increased to 85 to 190 μm, reaching a thickness of about 120 μm in the most parts of the coating. After heat treatment, the final thickness of the two-layered coating was mostly in the range of 80 to 110 μm, due to the filling of pores and cracks in the coating, and the melting of the polymer. Despite this, there were fragments of much greater thickness in the coating, reaching up to about 180 μm, especially at the lower edge of the coating (Figure 10). SEM analysis of the cross-section (Figure 10) of the hybrid Cu/HA/ZnS + PEEK coating indicates that HA during the heat treatment was submerged at different depths in the coating. All components formed a dense and homogeneous coating. HA agglomerates were found on the surface and in-depth of the coating.

As a result of PLD with evenly distributed shadow masking, Cu “islands” were obtained on the surface of heat-treated multilayers (Figure 11). The HA was distributed non-uniformly on the surface of the coating. The ZnS distribution also varied on the coating surface. Such a distribution of components on the coating surface potentially allows for simultaneous antibacterial (Cu, ZnS) and bioactive (HA + S-PEEK) effects.

Moreover, the coating microstructure was examined by TEM on cross-section (Figures 12 and 13). The selective Cu layer at the top of the coating was from 200 to 500 nm thick. It was continuous, mostly uniform, with limited protrusions. No delamination was observed in the investigated area. The HA agglomerates were deeply inserted in the amorphous sulfonated PEEK matrix, reaching from the coating surface up to the alloy substrate. The agglomerates were shaped in various ways as submerged elongated “waves” and irregular particles with ECD from 50 nm to 8 μm. The differential location of the HA agglomerates resulted from the embedment of heavy ceramics in the soft polymer matrix during thermal treatment. Sporadically, ZnS particles were present in the PEEK matrix or in the HA agglomerates.

No closed porosity was observed on the cross-section of the coating. The element distribution images confirmed the presence of Cu in the outer “islands” layer, Ca and P in the HA, as well as Zn in the ZnS particles (Figures 11 and 13).

The roughness of the ZnS + PEEK, HA/ZnS + PEEK and Cu/HA/ZnS + PEEK layers was measured for particular layers. The ZnS + PEEK layer had the lowest roughness of the average arithmetical mean deviation (R_a) of 0.41 ± 0.01 µm, root mean square roughness (R_q) of 0.53 ± 0.01 µm and the maximum peak to valley height (R_t) of 6.9 ± 0.3 μm. The HA/ZnS + PEEK multilayer had higher roughness, values of R_a = 4.74 ± 0.07 μm, R_q = 5.94 ± 0.10 μm and R_t = 49.2 ± 7.6 μm, due to the presence of HA agglomerates on the layer surface (Figures 10, 11, 13 and 14).

The thin Cu layer did not have a significant impact on the surface roughness of the multilayered coating. Representative images of the surface of the 1st ZnS + PEEK layer, the 2nd HA layer on the top of ZnS + PEEK layer and the final multilayered coating are shown in Figures 14(a) through (c). This relatively high surface roughness of the multilayered coating may be important due to the potentially better adhesion of bone cells. It is known,^[54,55] that high roughness and surface area stimulate the formation of the extracellular matrix.

The surface properties, wetting angle (WA) and interfacial free energy (IFE) of the developed materials were investigated and their values are present in Table I. During our previous study^[23] the WA and IFE for the Zr–2.5Nb substrate and the PEEK coating without additional components were characterized. The WA for the water of Zr–2.5Nb (53 ± 8 deg) was lower in comparison to the PEEK coated alloy (71.1 ± 9.0 deg). The IFE of the alloy was marginally higher (53.6 ± 6.0 mN/m) than that of the PEEK (51.3 ± 4.1 mN/m) coating, these values were higher than for each layer of the three-layer coating. The WA and IFE of the 1st ZnS + PEEK layer of 70.7 ± 4.5 deg and 40.8 ± 5.0 mN/m, respectively, were comparable to those of the pure PEEK coating. Interestingly, both multilayer coatings, with and without Cu, exhibited mild hydrophobic WA at a level above 90 deg, with the highest WA of 109.7 ± 6.6 deg for Cu/HA/ZnS + S-PEEK coatings with IFE of 34.8 ± 1.1 mN/m. Although selective deposition of Cu did not have a considerable impact on surface roughness, it influenced the WA of the multilayered coating, increasing its hydrophobicity. It is well known that copper oxides form at a rapid rate when exposed to ambient air and that the oxide layer is not self-protective for further oxidation.^[56,57] Generally, copper oxides are also considered hydrophilic,^[58,59] but after exposure to ambient air, they become hydrophobic over time. This phenomenon was explained by Shirazy et al.^[60] They proved that volatile organic compounds in the air were responsible for the decrease in the hydrophilic properties of copper surfaces of various types. Similarly, Yin et al.^[61] studied the same dependence with the loss of hydrophilicity of TiO₂ in air.

In our study, the deposition of the Cu layer on HA/ZnS + PEEK coatings by PLD was performed in a vacuum, but the multilayered samples produced were not stored in a protective environment. Therefore, the potential contamination of volatile organic compounds may be responsible for the loss of hydrophilicity. Although the addition of HA could potentially lower WA due to an increase in the hydrophilic hydroxyl groups characteristic of HA on the surface of the composite,^[62,63] it was not observed in the investigated coatings. High WA indicates hydrophobicity of the coating, similar to HA/PEEK coatings electrophoretically deposited by Bastan et al.^[64] Although moderate hydrophilicity is preferred in cell adhesion studies,^[65‐67] the results of the research vary. Hydrophilic cells are commonly approved to adhere to hydrophilic surfaces, whereas hydrophobic cells adhere to hydrophobic surfaces.^[68] Mild hydrophobicity may not adversely affect cell adsorption, because of the thermodynamic preference for protein adsorption from aqueous solutions.^[66] The surface of the coating has a significant influence on its wettability. In general, high hydrophobicity combined with a high surface area and high roughness of the coatings results in primal cellular adhesion.^[69]

Hardness (H_IT) and modulus of elasticity (E_IT) of the base 1st layer of ZnS + PEEK mainly responsible for the mechanical properties of the multilayered coating were investigated using of instrumented indentation under different loads (P_max) equal to 100, 200, 400 and 1000 mN. Only data acquired for 100 mN were analyzed because the standardized penetration depth of 10 pct of the coating thickness was not achieved at this load. Exceeding this depth causes disturbance of the results related to the interaction between the coating and alloy properties. The hardness value obtained was 0.21 ± 0.02 GPa and the modulus of elasticity was 3.7 ± 0.2 GPa. The hardness of the ZnS + PEEK coatings was marginally higher than that of the pure amorphous PEEK coating (0.19 ± 0.01 GPa). Although, the Young modulus was lower than that of the amorphous PEEK coating (4.50 ± 0.20 GPa) reported by Kruk et al.^[70]

The hybrid technology developed in this work may be used to obtain coatings with controlled bioactive and antibacterial properties. By developing Cu “islands”, the coating can obtain an initial potential antibacterial response against the formation of bacterial microfilm, especially in the early stage of implantation. Moreover, the antimicrobial activity of Cu can be enhanced by ZnS particles. However, due to selective PLD, the contact of bioactive HA with bone is not limited. Further optimization of the antibacterial and bioactive phases on the top of the coatings, as well as advanced antibacterial and bioactive studies, are necessary.