Looking at the micro-topography of polished and blasted Ti-based biomaterials using atomic force microscopy and contact angle goniometry

Surface topography of polished and blasted samples of a Ti6Al4V biomaterial has been studied using an atomic force microscope. Surface RMS roughness and surface area have been measured at different scales, from 1 to 50 μm, while at distances below 10 μm the surface RMS roughness in both kinds of samples is not very different, this difference becomes significant at larger scanning sizes. This means that the surface roughness scale that could have a main role in cell adhesion varies depending on the size, shape and flexibility of participating cells. This consideration suggests that in cell–material interaction studies, surface roughness should not be considered as an absolute and independent property of the material, but should be measured at scales in the order of the cell sizes, at least if a microscopic interpretation of the influence of roughness on the adhesion is intended. The microscopic information is contrasted with that coming from a macroscopic approach obtained by contact angle measurements for polar and non-polar liquids whose surface tension is comprised in a broad range. Despite the very large differences of contact angles among liquids for each surface condition, a similar increase for the blasted surface with respect to the polished has been found. Interpretation of these results are in accordance with the microscopic analysis done through the use of a functional roughness parameter, namely the valley fluid retention index, evaluated from the AFM images, which has been shown not to correlate with the RMS roughness, one of the most commonly used roughness parameter.     

Does the nanometre scale topography of titanium influence protein adsorption and cell proliferation?

To investigate the influence of titanium films with nanometre scale topography on protein adsorption and cell growth, three different model titanium films were utilized in the present study. The chemical compositions, surface topographies and wettability were investigated by using X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM) and water contact angle measurement, respectively. The films share the same surface chemistry but exhibit different topographies on a nanometre scale. Thus, they act as model systems for biological studies regarding surface topography effects. The films were obtained by varying the deposition rate and the film thickness, respectively. These films displayed nanometre scale surface roughness (root mean square roughness, R_rms) from 2 to 21 nm over areas of 50 μm × 50 μm, with different grain sizes at their surfaces. Albumin and fibrinogen adsorption on these model titanium films were performed in this study. Bicinchoninic acid assay was employed to determine the amount of adsorbed protein on titanium film surfaces. No statistically significant differences, however, were observed for either albumin or fibrinogen adsorption between the different groups of titanium films. No statistically significant influence of surface roughness on osteoblast proliferation and cell viability was detected in the present study.     

Enhanced nucleation, smoothness and conformality of ultrananocrystalline diamond (UNCD) ultrathin films via tungsten interlayers

Extremely smooth (6 nm RMS roughness over 4 μm²), thin (100 nm), and continuous ultrananocrystalline diamond (UNCD) films were synthesized by microwave plasma chemical vapor deposition using a 10 nm tungsten (W) interlayer between the silicon substrate and the diamond film. These UNCD films possess a high content of sp³-bonded carbon. The W interlayer significantly increased the initial diamond nucleation density, thereby lowering the surface roughness, eliminating interfacial voids, and allowing thinner UNCD films to be grown. This structural optimization enhances the films’ properties and enables its integration with a wide variety of substrate materials.     

Adhesion as an interplay between particle size and surface roughness

Surface roughness plays an important role in the adhesion of small particles. In this paper we have investigated adhesion as a geometrical effect taking into account both the particle size and the size of the surface features. Adhesion is studied using blunt model particles on surfaces up to 10 nm root-mean-square (RMS) roughness. Measurements with particles both smaller and larger than surface features are presented. Results indicate different behavior in these areas. Adhesion of particles smaller than or similar in size to the asperities depend mainly on the size and shape of the asperities and only weakly on the size of the particle. For large particles also the particle size has a significant effect on the adhesion. A new model, which takes the relative size of particles and asperities into account, is also derived and compared to the experimental data. The proposed model predicts adhesion well over a wide range of particle/asperity length scales.     

Chemically surface-modified carbon nanoparticle carrier for phenolic pollutants: Extraction and electrochemical determination of benzophenone-3 and triclosan

Chemically surface-modified (tosyl-functionalized) carbon nanoparticles (Emperor 2000 from Cabot Corp.) are employed for the extraction and electrochemical determination of phenolic impurities such as benzophenone-3 (2-hydroxy-4-methoxybenzophenone) or triclosan (5-chloro-2-(2,4-dichlorophenoxy)phenol). The hydrophilic carbon nanoparticles are readily suspended and separated by centrifugation prior to deposition onto suitable electrode surfaces and voltammetric analysis. Voltammetric peaks provide concentration information over a 10–100 μM range and an estimated limit of detection of ca. 10 μM (or 2.3 ppm) for benzophenone-3 and ca. 20 μM (or 5.8 ppm) for triclosan.

Alternatively, analyte-free carbon nanoparticles immobilized at a graphite or glassy carbon electrode surface and directly immersed in analyte solution bind benzophenone-3 and triclosan (both with an estimated Langmuirian binding constants of K ≈ 6000 mol⁻¹ dm³ at pH 9.5) and they also give characteristic voltammetric responses (anodic for triclosan and cathodic for benzophenone-3) with a linear range of ca. 1–120 μM. The estimated limit of detection is improved to ca.5 μM (or 1.2 ppm) for benzophenone-3 and ca. 10 μM (or 2.3 ppm) for triclosan. Surface functionalization is discussed as the key to further improvements in extraction and detection efficiency.     

Correction of random surface roughness on colloidal probes in measuring adhesion

Atomic force microscopes (AFM) are commonly used to measure adhesion at nanoscale between two surfaces. To avoid uncertainties in the contact areas between the tip and the surface, colloidal probes have been used for adhesion measurements. We measured adhesion between glass spheres and silicon (100) surface using colloidal probes of different radii under controlled conditions (relative humidity of < 3%, temperature of 25 +/- 1 degrees C). Results showed that the adhesion forces did not correlate with the radii of the spheres as suggested by elastic contact mechanics theories. Surface roughness and random surface features were found on the surfaces of the colloidal probes. We evaluated various roughness parameters, Rumpf and Rabinovich models, and a load-bearing area correction model in an attempt to correct for the roughness effects on adhesion, but the results were unsatisfactory. We developed a new multiscale contact model taking into account elastic as well as plastic deformation in a successive contacting mode. The new model was able to correct for most of the surface roughness features except for surface ridges with sharp angular features, limited by the spherical asperity assumption made in the model.     

The deformation and adhesion of randomly rough and patterned surfaces

Using a surface forces apparatus (SFA) and an atomic force microscope (AFM) we have studied the effects of surface roughness (root-mean-square (RMS) roughness between 0.3 and 220 nm) on the "contact mechanics", which describes the deformations and loading and unloading adhesion forces, of various polymeric surfaces. For randomly rough, moderately stiff, elastomeric surfaces, the force-distance curves on approach and separation are nearly reversible and almost perfectly exponentially repulsive, with an adhesion on separation that decreases only slightly with increasing RMS. Additionally, the magnitude of the preload force is seen to play a large role in determining the measured adhesion. The exponential repulsion likely arises from the local compressions (fine-grained nano- or submicron-scale deformations) of the surface asperities. The resulting characteristic decay lengths of the repulsion scale with the RMS roughness and correlate very well with a simple finite element method (FEM) analysis based on actual AFM topographical images of the surfaces. For "patterned" surfaces, with a nonrandom terraced structure, no similar exponential repulsion is observed, suggesting that asperity height variability or random roughness is required for the exponential behavior. However, the adhesion force or energy between two "patterned" surfaces fell off dramatically and roughly exponentially as the RMS increased, likely owing to a significant decrease in the contact area which in turn determines their adhesion. For both types of rough surfaces, random and patterned, the coarse-grained (global, meso- or macroscopic) deformations of the initially curved surfaces appear to be Hertzian.     

Microspreading studies on rough surfaces by scanning electron microscopy

The use of scanning electron microscopy for direct observation of the effects of surface roughness on the spreading of liquids is described, making it possible to view moving liquid drops at distances less than 1 μm from the advancing contact line. Various surfaces were examined including several with simple forms of roughness which can assist in explaining the behavior of more complex surfaces. Spreading is shown to be highly dependent on the orientation and texture of the roughness; in particular, the presence of sharp edges of step height 0.05 μm are shown to influence spreading significantly. These observations reinforce our previously stated doubts of the significance of conventionally measured macroscopic contact angles.     

Importance of surface tension for physical paint properties

Surface tension is a parameter of decisive importance for characterizing painted and unpainted surfaces related to wetting and adhesion phenomena. This article presents measurements of the surface tension of solids by means of an automatic contact angle measurement device. The evaluation of the surface tension is based on a separation into polar and disperse components. In addition, this paper briefly touches on other, more far-reaching approaches (acid/base) and discusses a method for the determination of dynamic surface tension of liquids.     

Cell-cell contact and membrane spreading in an ultrasound trap

An ultrasonic standing wave trap [Langmuir 19 (2003) 3635] in which the morphologies of 2-D latex–microparticle aggregates, forming a pressure node plane, were characterised has been applied here to different cell suspensions with increasing order of specificity of cross-linking molecule, i.e. polylysine with chondrocytes; wheat germ agglutinin (WGA) with erythrocytes and surface receptors on neural cells. The outcome of initial cell–cell contact, i.e. whether the cells stuck at the point of contact (collision efficiency=1) or rolled around each other (collision efficiency=0), was monitored in situ by video-microscopy. The perimeter fractal dimensions (FD) of 2-D hexagonally symmetric, closely packed aggregates of control erythrocytes and chondrocytes were 1.16 and 1.18, respectively while those for the dendrititc aggregates formed initially by erythrocytes in 0.5 μg/ml WGA and chondrocytes in 20 μg/ml polylysine were 1.49 and 1.66. The FDs for control and molecularly cross-linked cells were typical of reaction-limited aggregation (RLA) and transport diffusion-limited aggregation (DLA), respectively. The FDs of the aggregates of cross-linked cells decreased with time to give more closely packed aggregates without clear hexagonal symmetry. Suspensions of neural cells formed dendritic aggregates. Spreading of inter-cellular membrane contact area occurred over 15 min for both erythrocyte and neural cell dendritic aggregates. The potential of the technique to characterise and control the progression of cell adhesion in suspension away from solid substrata is discussed.     

Surface chemistry of nanoporous carbon and the effect of pH on adsorption from aqueous phenol and 2,3,4-trichlorophenol solutions

Granular nanoporous activated carbon prepared from polyacrylonitrile (PAN) was investigated as a means of removing weak aromatic acids from aqueous solutions. This carbon is highly nanoporous, the BET surface area being 544 m²/g with V_tot=0.278 cm³/g, and V_micro=0.266 cm³/g. Aqueous treatment reduced the surface area to 364 m²/g.

Granular nanoporous carbon prepared from PAN contains O and N related surface functional groups. The surface concentration of both oxygen and nitrogen atoms was found by XPS to be 5.3%. Surface groups containing these hetero atoms are responsible for the acid–base character of this carbon in aqueous solutions. The pH_PZC of the carbon is 8.4. The microporous pore network produces a wide hysteresis loop, observed when the granular carbon was studied by continuous titration. This loop, which was found to diminish, but not to disappear completely when the particle size was reduced, is attributed to irreversible hydrolysis of surface esters and/or lactones. The surface concentration of the functional groups titrated by the equilibrium Boehm method is 449.2 μeq./g, of which 112.9 μeq./g have acidic and 336.3 μeq./g have basic character. Within the acidic species three subgroups, namely the regions pK_a<6.37, 6.37K_a<10.25 and 10.25K_a<15.74.

The acid–base behavior influences the adsorption performance of the granular carbon in aqueous solutions of weak aromatic acids, in this case phenol and 2,3,4-trichlorophenol. Both the adsorption capacity and the overall interaction parameter, K (both derived from a fit to the Langmuir equation) depend on the adsorbed species and on the pH. The former is a consequence of the different water solubilities of the solute molecules, while the latter stems from the pH sensitivity of both the surface functional groups and these weak acids. The K values show a sequence pH=3K values indicate different adsorption mechanisms. The effect of pH is more marked for 2,3,4-trichlorophenol.     

Roughness assessment and wetting behavior of fluorocarbon surfaces

The wetting behavior of fluorocarbon materials has been studied with the aim of assessing the influence of the surface chemical composition and surface roughness on the water advancing and receding contact angles. Diamond like carbon and two fluorocarbon materials with different fluorine content have been prepared by plasma enhanced chemical vapor deposition and characterized by X-ray photoemission, Raman and FT-IR spectroscopies. Very rough surfaces have been obtained by deposition of thin films of these materials on polymer substrates previously subjected to plasma etching to increase their roughness. A direct correlation has been found between roughness and water contact angles while a superhydrophobic behavior (i.e., water contact angles higher than 150° and relatively low adhesion energy) was found for the films with the highest fluorine content deposited on very rough substrates. A critical evaluation of the methods currently used to assess the roughness of these surfaces by atomic force microscopy (AFM) has evidenced that calculated RMS roughness values and actual surface areas are quite dependent on both the scale of observation and image resolution. A critical discussion is carried out about the application of the Wenzel model to account for the wetting behavior of this type of surfaces.     

Effect of various treatment and glazing (coating) techniques on the roughness and wettability of ceramic dental restorative surfaces

Surface treatment procedures such as grinding and polishing are needed to provide the ceramic dental restorative materials with proper fitting and occlusion. The treated surfaces are customarily glazed to improve the strength and smoothness. Though smoothness and wetting of the dental surfaces are important to minimize bacterial plaque retention, influence of the surface treatment and glazing procedures on the final surface roughness and its correlation to wettability are overlooked.

In this work, effect of various treatment (diamond fraising, stoning, sanding and aluminum oxide and rubber polishing) and glazing (auto and overglazing) techniques on the final roughness and the resulting wettability of dental ceramic surfaces were investigated using scanning electron microscopy (SEM) observations and atomic force microscopy (AFM) scans, 75 scans per sample. The surfaces were characterized and assigned an average roughness measure, R_a. The wettability of the same surfaces was evaluated using micro-contact angle measurements (25 micro-bubbles placed on a grid on each surface) to correlate the final surface roughness and wettability.

The results show that overglazing prevails over surface irregularities from different treatment procedures and provides homegeneously smooth surfaces with mean R_a < 10 nm. It also produces uniformly wetted surfaces with low contact angles around 20°. The autoglazed surfaces are less smooth (mean R_a around 50 nm) and displays sporadic topographic irregularities. They display larger and less uniform contact angles ranging between 35° and 50°. The results suggest that overglazing should be preferred after surface treatment to obtain a smooth and well-wetted dental ceramic surface.     

Surface modification of low density polyethylene (LDPE) film by low pressure O2 plasma treatment

In this work, low pressure glow discharge O₂ plasma has been used to increase wettability in a LDPE film in order to improve adhesion properties and make it useful for technical applications. Surface energy values have been estimated using contact angle measurements for different exposure times and different test liquids. In addition, plasma-treated samples have been subjected to an aging process to determine the durability of the plasma treatment. Characterization of the surface changes due to the plasma treatment has been carried out by means of Fourier transformed infrared spectroscopy (FTIR) to determine the presence of polar species such as carbonyl, carboxyl and hydroxyl groups. In addition to this, atomic force microscopy (AFM) analysis has been used to evaluate changes in surface morphology and roughness. Furthermore, and considering the semicrystalline nature of the LDPE film, a calorimetric study using differential scanning calorimetry (DSC) has been carried out to determine changes in crystallinity and degradation temperatures induced by the plasma treatment. The results show that low pressure O₂ plasma improves wettability in LDPE films and no significant changes can be observed at longer exposure times. Nevertheless, we can observe that short exposure times to low pressure O₂ plasma promote the formation of some polar species on the exposed surface and longer exposure times cause slight abrasion on LDPE films as observed by the little increase in surface roughness.     

Influence of Chemical Heterogeneity and Nanoscale Roughness on the DLVO Energy Interaction by Spherical Coordinates

In this work, the influence of chemical heterogeneity on the stability of nanocolloidal systems is surveyed with a new method. Zone of influence as a very important parameter for chemical patch surveying is modeled for sphere and flat surface. Surface chemical heterogeneity with specified properties, size, and position are created by spherical coordinate integration method. Rippled sphere model is used to create roughness and the flat surface is created by changing two sphere radius ratio. Using the spherical coordinate system for modeling of surface roughness and chemical heterogeneity is very accurate and fast. Results show that the patches could destabilize the colloidal system at very small sizes. Surface roughness reduces the effect of chemical patches for destabilizing the colloidal system, and with increasing the size of roughness the total DLVO energy interaction increases.     

Investigation of the hydrothermal stability of cross-linked liquid silicone rubber (LSR)

We investigate the hydrothermal stability of cross-linked liquid silicone rubber (LSR) in water at 100 °C up to period of two years. Optical microscopy of cross-sections of the exposed samples reveal that only the outer 100 μm of the surface layer is affected after two years. However, the surface chemistry of the material after prolonged exposure becomes significantly modified, as monitored by X-ray photoelectron spectroscopy (XPS) and attenuated total reflectance Fourier transform infrared (ATR-FTIR), which probes depths of 10 nm and 1 μm, respectively. In addition, changes to the bulk physical properties of the rubber samples, prior to and after the exposure, were investigated by thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC). Micro-hardness analysis showed that surface roughness of the two year exposed sample increased from 60 (IRHD) to 75 (IRHD). Furthermore, the volume change (%) measurement showed a significant decrease in the course of exposure at prolonged time. The results provide the experimental basis for development of LSR materials suitable for numerous technical applications.     

Surface free energy effect on bacterial retention

Bacterial infection is one of the most frequent and severe complications in the long-term effectiveness of medical implants and devices, greatly increasing treatment cost and inconvenience to the patient. Surface physical and chemical properties are known to influence the extent and form of bacterial infection, although the exact correlation with specific properties is difficult due to the complexity of the system. One approach in the attempt to reduce the bacterial colonisation is to modify the surface energy and chemistry, so as to influence the interactions between the surface and the bacteria that come into contact with it. Five types of coatings were investigated in this study, together with silicone, and polished and non-polished stainless steel 316L. Surfaces were tested for retention of Pseudomonas aeruginosa AK1 after 1 h. A good correlation (>90%) was found between P. aeruginosa AK1 retention and total surface free energy, as well as its polar and dispersive components. The minimum level of P. aeruginosa AK1 retention was found for a range of total surface free energy in the range 20–27 mN/m.     

Effect of laser modified surface microtopochemistry on endothelial cell growth

The introduction of microelectronics technology in the area of biological sciences has brought forth previously unforeseeable applications such as DNA or protein biochips, miniaturized, multiparametric biosensors for high performance multianalyte assays, DNA sequencing, biocomputers, and substrates for controlled cell growth (i.e. tissue engineering). We developed and investigated a new method using “cold” excimer laser beam technology combined with microlithographical techniques to create surfaces with well defined 3D microdomains in order to delineate critical microscopic surface features governing cell–material interactions. Microfabricated surfaces with microgrooves 30–3 μm deep, 10–1 μm wide spaced 30 μm apart were obtained with micron resolution, by “microsculpturing” polymer model surfaces using a computer controlled laser KrF excimer beam coupled with a microlithographic projection technique. The laser beam after exiting a mask was focused onto the polymer target surface via an optical setup allowing for a 10-fold reduction of the mask pattern. Various 3D micropatterned features were obtained at the micron level. Reproducible submicron features could also be obtained using this method. Subsequently, model human umbilical endothelial cells (HUVEC) were cultured on the laser microfabricated surfaces in order to study the effects of specific microscopic surface features on cell deposition and orientation. Cell deposition patterns were found to be microstructure dependant, and showed cell orientation dependency for features in the cell range dimension, a behaviour significantly different from that of a previously studied cell model (osteoprogenitor cell). This model may be a promising in so far as it is very rapid (a time frame less than a second per square centimeter of micropatterned surface) and provides further insights into the effects of surface microtopography on cell response with possible applications in the field of biosensors, biomedical and/or pharmaceutical engineering sciences.     

Langmuir monolayer properties of the fluorinated-hydrogenated hybrid amphiphiles with dipalmitoylphosphatidylcholine (DPPC)

Surface pressure–area (π–A), surface potential–area (ΔV–A), and dipole moment–area (μ–A) isotherms were obtained for the Langmuir monolayer of two fluorinated-hydrogenated hybrid amphiphiles (sodium phenyl 1-[(4-perfluorohexyl)-phenyl]-1-hexylphosphate (F6PH5PPhNa) and (sodium phenyl 1-[(4-perfluorooctyl)-phenyl]-1-hexylphosphate (F8PH5PPhNa)), DPPC and their two-component systems at the air/water interface. Monolayers spread on 0.02 M Tris buffer solution (pH 7.4) with 0.13 M NaCl at 298.2 K were investigated by the Wilhelmy method, ionizing electrode method and fluorescence microscopy. Moreover, the miscibility of two components was examined by plotting the variation of the molecular area and the surface potential as a function of the molar fraction for the fluorinated-hydrogenated hybrid amphiphiles on the basis of the additivity rule. The miscibility of the monlayers was also examined by construction of two-dimensional phase diagrams. Furthermore, assuming the regular surface mixture, the Joos equation for analysis of the collapse pressure of two-component monolayers allowed calculation of the interaction parameter (ξ) and the interaction energy (−Δ) between the fluorinated-hydrogenated hybrid amphiphiles and DPPC. The observations by a fluorescence microscopy also supported our interpretation as for the miscibility in the monolayer state. Comparing the monolayer behavior between the two binary systems, no remarkable difference was found among various aspects. Among the two combinations, the mole fraction dependence in monlayer properties was commonly classified into two ranges: 0 ≤ X ≤ 0.3 and 0.3 < X ≤ 1. Dependence of the chain length of fluorinated part was reflected for the molecular packing and surface potential.     

The role of few-asperity contacts in adhesion

The surface roughness of a few asperities and their influence on the work of adhesion is of scientific interest. Macroscale and nanoscale adhesion data have seemingly given inconsistent results. Despite the importance of bridging the gap between the two regimes, little experimental work has been done, presumably due to the difficulty of the experiment needed to determine how small amounts of surface roughness might influence adhesion data lying in between the two scales. To investigate the role of few-asperity contacts in adhesion, the pull-off force was measured between different sized atomic-force microscope (AFM) tips (with different roughnesses) and sample surfaces that had well-controlled material properties. There were seventeen tips of four different types, with radii from 200 nm to 60 microm. The samples were unpatterned single crystal silicon with a chemical silicon dioxide surface resulting from a standard silicon wafer clean. Some of the samples were treated with a few angstroms of vapor deposited diphenylsiloxane. We observed that the uncorrected (for surface roughness) pull-off force was independent of the radius of the AFM tip, which was contrary to all continuum-mechanics model predictions. To explain this behavior, we assumed that the interactions between the AFM tip and sample were additive, material properties were constant, and that the AFM tip, asperities, and sample surfaces were of uniform density. Based on these assumptions, we calculated a simple correction due to the measured root mean square (RMS) surface roughness of the AFM tips. The simple correction for the RMS surface roughness resulted in the expected dependence of the pull-off force on radius, but the magnitudes were higher than expected. Commercial and heat-treated AFM tips have minimal surface roughness and result in magnitudes that are more reliable. The relative uncertainty for the pull-off force was estimated to be 10%. In this paper, we derive how the cantilever and tip parameters contribute to the measured pull-off force and show how the corrected results compare with theory. Although much work is still needed, the work presented here should advance the understanding of adhesion between the macroscale and nanoscale regimes.