Effect of Hydrofluoric Acid Etching Time on Titanium Topography,Chemistry, Wettability,and Cell Adhesion

Titanium implant surface etching has proven an effective method to enhance cell attachment. Despite the frequent use of hydrofluoric (HF) acid, many questions remain unresolved, including the optimal etching time and its effect on surface and biological properties. The objective of this study was to investigate the effect of HF acid etching time on Ti topography, surface chemistry, wettability, and cell adhesion. These data are useful to design improved acid treatment and obtain an improved cell response. The surface topography, chemistry, dynamic wetting, and cell adhesiveness of polished Ti surfaces were evaluated after treatment with HF acid solution for 0, 2; 3, 5, 7, or 10 min, revealing a time-dependent effect of HF acid on their topography, chemistry, and wetting. Roughness and wetting increased with longer etching time except at 10 min, when roughness increased but wetness decreased. Skewness became negative after etching and kurtosis tended to 3 with longer etching time. Highest cell adhesion was achieved after 5–7 min of etching time. Wetting and cell adhesion were reduced on the highly rough surfaces obtained after 10-min etching time.     

In situ surface electrochemical characterizations of Ti and Ti-6Al-4V alloy cultured with osteoblast-like cells

The biocompatibility of metal implants is related to their surface electrochemical characterizations. The in situ growing process of osteoblast-like U-2 OS cells on polished Ti and Ti-6Al-4V alloy during 72h incubation was monitored using the electrochemical impedance spectroscopy (EIS) measurement technique. The results showed that the presence of cells on metals led to an increase in the impedance and polarization resistance (R(p)) of metals. The impedance and R(p) increased as the cells grew (i.e., from adhesion, spreading to proliferation period). A trace amount of V element released from Ti-6Al-4V alloy led to a lower R(p) with respect to Ti metal during cell culture. In this study, a satisfactory equivalent circuit simulating the electrochemical characterizations of Ti and Ti-6Al-4V alloy cultured with cells was proposed. The EIS measurement technique was applied successfully to monitor the in situ growing process of U-2 OS cells on Ti and Ti-6Al-4V alloy.     

The relative importance of topography and RGD ligand density for endothelial cell adhesion

The morphology and function of endothelial cells depends on the physical and chemical characteristics of the extracellular environment. Here, we designed silicon surfaces on which topographical features and surface densities of the integrin binding peptide arginine-glycine-aspartic acid (RGD) could be independently controlled. We used these surfaces to investigate the relative importance of the surface chemistry of ligand presentation versus surface topography in endothelial cell adhesion. We compared cell adhesion, spreading and migration on surfaces with nano- to micro-scaled pyramids and average densities of 6×10(2)-6×10(11) RGD/mm(2). We found that fewer cells adhered onto rough than flat surfaces and that the optimal average RGD density for cell adhesion was 6×10(5) RGD/mm(2) on flat surfaces and substrata with nano-scaled roughness. Only on surfaces with micro-scaled pyramids did the topography hinder cell migration and a lower average RGD density was optimal for adhesion. In contrast, cell spreading was greatest on surfaces with 6×10(8) RGD/mm(2) irrespectively of presence of feature and their size. In summary, our data suggest that the size of pyramids predominately control the number of endothelial cells that adhere to the substratum but the average RGD density governs the degree of cell spreading and length of focal adhesion within adherent cells. The data points towards a two-step model of cell adhesion: the initial contact of cells with a substratum may be guided by the topography while the engagement of cell surface receptors is predominately controlled by the surface chemistry.     

Different titanium surfaces modulate the bone phenotype of SaOS-2 osteoblast-like cells

Commercially pure titanium implants presenting a relatively smooth, machined surface or a roughened endosseous surface show a large percentage of clinical success. Surface properties of dental implants seem to affect bone cells response. Implant topography appears to modulate cell growth and differentiation of osteoblasts affecting the bone healing around the titanium implant. The aim of the present study was to examine the effects of 1cm diameter and 1mm thick titanium disks on cellular morphology, adhesion and bone phenotypic expression of human osteoblast-like cells, SaOS-2. SaOS-2 cells were cultured on commercially 1 cm pure titanium disks with three different surface roughness: smooth (S), sandblasted (SB) and titanium plasma sprayed (TPS). Differences in the cellular morphology were found when they were grown on the three different surfaces. An uniform monolayer of cells recovered the S surface, while clusters of multilayered irregularly shaped cells were distributed on the rough SB and TPS surfaces. The adhesion of SaOS-2 cells, as measured after 3h of culture, was not affected by surface roughness. ECM components such as Collagen I (CoI), Fibronectin (FN), Vitronectin (VN) and Tenascin (TN) were secreted and organized only on the SB and TPS surfaces while they remained into the cytoplasm on the S surfaces. Osteopontin and BSP-II were largely detected on the SB and TPS surfaces, while only minimal production was observed on the S ones. These data show that titanium surface roughness affects bone differentiation of osteoblast like-cells, SaOS-2, indicating that surface properties may be able to modulate the osteoblast phenotype. These observations also suggest that the bone healing response around dental implants can be affected by surface topography.     

Osteoblast-like cell behavior on plasma deposited micro/nanopatterned coatings

The behavior of cells in terms of cell-substrate and cell-cell interaction is dramatically affected by topographical characteristics as shape, height, and distance, encountered in their physiological environment. The combination of chemistry and topography of a biomaterial surface influences in turns, important biological responses as inflammatory events at tissue-implant interface, angiogenesis, and differentiation of cells. By disentangling the effect of material chemistry from the topographical one, the possibility of controlling the cell behavior can be provided. In this paper, surfaces with different roughness and morphology were produced by radiofrequency (RF, 13.56 MHz) glow discharges, fed with hexafluoropropylene oxide (C(3)F(6)O), in a single process. Coatings with different micro/nanopatterns and the same uppermost chemical composition were produced by combining two plasma deposition processes, with C(3)F(6)O and tetrafluoroethylene (C(2)F(4)), respectively. The behavior of osteoblast-like cells toward these substrates clearly shows a strict dependence of cell adhesion and proliferation on surface roughness and morphology.     

Aggregation kinetics of saccharomyces cerevisiae on solid surfaces

In the colonization experiments of yeast cells on stainless steel surfaces the analyzed minimum of the colonization rate was found in a roughness interval between a highly polished sample and a very rough surface. This phenomenon is a problem of the shear forces and of adhesion. On the basis of reaction kinetics it was possible to analyze the change between an unstable and a stable cell-cluster. The analysis of the energy needed for forming a stable cluster tends toward values known from the stochastic simulation of cluster formation in an undisturbed region. But the potential function is not clear which describes the interactions between the cells and the forming cluster in the reaction field. This is a problem of special materials-testing which is not established. For producing plants it is of high interest to note that it are not the most highly polished surfaces that are the best.     

Cytotoxicity study of high gold content Degutan surfaces of various degrees of roughness with fibroblasts (BALB 3T3) and osteoblasts (hFOB 1.19)]

The cytotoxicity of Degutan surfaces with different degrees of roughness, and the effect of surface structures on osteoblast proliferation and differentiation, was investigated with standardised cell culture systems. Fibroblast cell lines (BALB/3T3) and osteoblast cell lines (hFOB 1.19) were used. The number and variability of the cells were determined for assessment of proliferation and alkaline phosphatase activity, collagen I and osteocalcin production were used as parameters for differentiation. In the early phase, the largest numbers of cells and greatest proliferation were measured on polished Degutan surfaces. In the late phase, however, larger numbers of cells and a greater degree of proliferation were to be seen on sandblasted and sandblasted/heat-treated Degutan surfaces. No differences were found for collagen I, osteocalcin production or alkaline phosphatase activity. Neither the osteoblasts nor the fibroblasts revealed a toxic effect of Degutan. The results for osteoblast differentiation correlate with recent studies on identical structured titanium surfaces. In view of the immeasurable amount of ion release, Degutan may be considered an ideal model for an inert material surface.     

Different target surfaces for the analysis of peptides, peptide mixtures and peptide mass fingerprints by AP-MALDI ion trap-mass spectrometry

The desorption/ionization behavior of individual peptides, an equimolare peptide mixture and a tryptic digest was investigated by AP-MALDI-IT-MS using four different target materials (gold-covered stainless steel (SS), titanium nitride-covered SS, hand-polished SS, and microdiamond-covered hardmetal) under identical conditions. Gold-covered as well as polished SS targets yielded comparable mass spectra for peptides and peptide mixture in the low pMol-range. The first target exhibited superior data down to the 10fMol-range. In contrast, titanium nitride-covered SS and microdiamond-covered hardmetal AP-MALDI-targets yielded poor sensitivity. These observations could be correlated with the surface roughness of the targets determined by 3D-confocal-white-light-microscopy. The roughest surfaces were found for titanium nitride-covered SS and microdiamond-covered hardmetal material showing both poor MS sensitivity. A less rough surface could be determined for the hand-polished SS target and the smoothest surface was found for the gold-covered target yielding the best sensitivity of all surfaces. These differences in the roughness having a strong impact on the ultimate sensitivity obtainable for peptide samples could be corroborated by electron microscopy. A peptide mixture covering a wide range of molecular weights and a tryptic protein digest (from 2-DE) exhibit the same behavior. This clearly indicates that the smooth gold-covered SS target is the surface of choice in AP-MALDI MS proteomics.     

Surface topography of kenaf (Hibiscus cannabinus) sized papers

Bleached kenaf handsheets sized with different polymers such as chitosan, polyvinyl alcohol and cationic starch, were used for the determination of surface topography. A non-contact profilometer, the AltiSur 500, was used to characterize the topography of structural details in the paper surface. Numerical and visual characterization of surface roughness indicated that the surface of chitosan-sized paper was less rough than other surfaces, and all the sized papers were commonly smoother than the unsized paper. One property of bio-polymer of chitosan is its ability to form films that improve the surface properties of paper when it is applied to the surface of the sheet.     

Surface roughness parameters as predictors of anchorage strength in bone: a critical analysis

The surface roughness of a bone implant was defined parametrically. The values of the parameters defining the surface were varied. Some traditionally used surface roughness parameters were calculated. By means of a theoretical model the bone-implant interfacial shear strength was estimated. No simple correlation between the values of the surface roughness parameters and the estimated interfacial shear strength was found. It was concluded that the value of the traditional surface roughness parameters as predictors of interfacial shear strength is limited. If however a change of the surface topography of an implant is restricted to scale a positive correlation was found between the theoretical interfacial shear strength and some surface roughness parameters. It is suggested that the bone-implant interfacial shear strength in the general case be estimated by means of strength analyses based upon a study of the size, shape and density of the individual elements constituting the rough surface.     

The influence of bone damage on press-fit mechanics

Press-fitting is used to anchor uncemented implants in bone. It relies in part on friction resistance to relative motion at the implant–bone interface to allow bone ingrowth and long-term stability. Frictional shear capacity is related to the interference fit of the implant and the roughness of its surface. It was hypothesised here that a rough implant could generate trabecular bone damage during implantation, which would reduce its stability. A device was constructed to simulate implantation by displacement of angled platens with varying surface finishes (polished, beaded and flaked) onto the surface of an embedded trabecular bone cube, to different nominal interferences. Push-in (implantation) and Pull-out forces were measured and micro-CT scans were made before and after testing to assess permanent bone deformation. Depth of permanent trabecular bone deformation (‘damage’), Pull-out force and Radial force all increased with implantation displacement and with implantation force, for all surface roughnesses. The proposed hypothesis was rejected, since primary stability did not decrease with trabecular bone damage. In fact, Pull-out force linearly increased with push-in force, independently of trabecular bone damage or implant surface. This similar behaviour for the different surfaces might be explained by the compaction of bone into the surfaces during push-in so that Pull-out resistance is governed by bone-on-bone, rather than implant surface-on-bone friction. The data suggest that maximum stability is achieved for the maximum implantation force possible (regardless of trabecular bone damage or surface roughness), but this must be limited to prevent periprosthetic cortical bone fracture, patient damage and component malpositioning.     

Characterization of the properties of differentially modified titanium surfaces for dental implantology. 2: In vitro biocompatibility studies]

The aim of the present study was to determine whether specific surface modifications are capable of improving the biocompatibility of a titanium surface, and whether there is a correlation between the physico-chemical properties of the implant material and its biocompatibility. To this end, the properties of titanium surfaces were modified using various methods or the latter were coated with various materials. Plasma treatments under different atmospheric conditions (N2-plasma, SO2-plasma, acetylene plasma) as well as plasma polymerization were used to affect the biological response. Characterization of the physico-chemical surface properties by means of X-ray photoelectron spectroscopy (XPS), contact angle measurements and the calculation of surface tensions or surface energy provided important information on the interactions at the interface between the implant material and the aqueous environment. The influence of the respective surface modification on cell proliferation, cell viability and the activity of mitochondrial dehydrogenases was evaluated in specific in vitro tests with human gingiva fibroblasts. It was show that different modifications of the titanium samples induce different biological responses of the gingiva fibroblasts. The results confirm the existence of correlations between thermodynamic surface properties and cellular reactions under in vitro conditions.     

Stem surface roughness alters creep induced subsidence and 'taper-lock' in a cemented femoral hip prosthesis

The clinical success of polished tapered stems has been widely reported in numerous long term studies. The mechanical environment that exists for polished tapered stems, however, is not fully understood. In this investigation, a collarless, tapered femoral total hip stem with an unsupported distal tip was evaluated using a 'physiological' three-dimensional (3D) finite element analysis. It was hypothesized that stem-cement interface friction, which alters the magnitude and orientation of the cement mantle stress, would subsequently influence stem 'taper-lock' and viscoelastic relaxation of bone cement stresses. The hypothesis that creep-induced subsidence would result in increases to stem-cement normal (radial) interface stresses was also examined. Utilizing a viscoelastic material model for the bone cement in the analysis, three different stem-cement interface conditions were considered: debonded stem with zero friction coefficient (mu=0) (frictionless), debonded stem with stem-cement interface friction (mu=0.22) ('smooth' or polished) and a completely bonded stem ('rough'). Stem roughness had a profound influence on cement mantle stress, stem subsidence and cement mantle stress relaxation over the 24-h test period. The frictionless and smooth tapered stems generated compressive normal stress at the stem-cement interface creating a mechanical environment indicative of 'taper-lock'. The normal stress increased with decreasing stem-cement interface friction but decreased proximally with time and stem subsidence. Stem subsidence also increased with decreasing stem-cement interface friction. We conclude that polished stems have a greater potential to develop 'taper-lock' fixation than do rough stems. However, subsidence is not an important determinant of the maintenance of 'taper-lock'. Rather subsidence is a function of stem-cement interface friction and bone cement creep.     

Wetting control through topography and surface hydrophilic/hydrophobic property changes by coarse grained simulation

Abstract

The changes of wetting state of water droplet on the solid surface featuring pillared structures are quantitatively studied by Coarse Grained simulation. Our results demonstrate that wetting state changes with the different topography (surface roughness), and it depends on the intrinsic hydrophilic/hydrophobic property of surface as well. Only if the contact angle of water droplet on the smooth surface is larger than 93.13°, the wetting state translates from the Wenzel state to the Cassie state on the rough surface with certain pillar height and width, and the contact angle climb up to the highest point and then remain almost unchanged with the increasing of pillar height and the same pillar distance. However, the wetting state does not change if the contact angle on the smooth surface is 85.1° or less, no matter what pillar structure the surface has. Additionally, the contact angles will remain almost unchanged if the pillar height is higher than a certain value. Our simulation results provide a quantitative understanding about the wetting state of water droplet on solid rough surfaces, and the results show the wetting state can be controlled by combining rough structure design and hydrophilic/hydrophobic property change of surfaces.     

Engineered antifouling microtopographies - effect of feature size, geometry, and roughness on settlement of zoospores of the green alga Ulva

The effect of feature size, geometry, and roughness on the settlement of zoospores of the ship fouling alga Ulva was evaluated using engineered microtopographies in polydimethylsiloxane elastomer. The topographies studied were designed at a feature spacing of 2 microm and all significantly reduced spore settlement compared to a smooth surface. An indirect correlation between spore settlement and a newly described engineered roughness index (ERI) was identified. ERI is a dimensionless ratio based on Wenzel's roughness factor, depressed surface fraction, and the degree of freedom of spore movement. Uniform surfaces of either 2 mum diameter circular pillars (ERI=5.0) or 2 microm wide ridges (ERI=6.1) reduced settlement by 36% and 31%, respectively. A novel multi-feature topography consisting of 2 mum diameter circular pillars and 10 microm equilateral triangles (ERI=8.7) reduced spore settlement by 58%. The largest reduction in spore settlement, 77%, was obtained with the Sharklet AF topography (ERI=9.5).     

Investigation of interfacial capacitance of Pt,Ti and TiN coated electrodes by electrochemical impedance spectroscopy

Electrochemical processes at the electrode-electrolyte (body fluid) interface are of ultimate importance for stimulating/sensing electrode function. A high electrode surface area is desirable for safe stimulation through double-layer charging and discharging. Pt and Pt-Ir alloys have been the most common electrode materials. The use of TiN coating as the surface layer on the electrode has found increasing interest because of its metal-like conductivity, excellent mechanical and chemical properties, and the fact that it can be deposited with a high surface area. In this work, electrochemical impedance spectroscopy (EIS), which is a sensitive and non-destructive technique and widely used for characterization of electrical properties of electrode-electrolyte interfaces, was applied to investigate pure Pt and Ti, and TiN coated electrodes exposed to a phosphate-buffered-saline (PBS) solution. Platinized Pt and Ti were also studied for comparison. The capacitance value of the electrodes in PBS was obtained through quantitative analysis of the EIS spectra. The results reveal that the capacitance of the TiN coated electrodes with a rough surface is several hundreds times higher than that of a smooth Pt surface. Platinization of Ti can also increase the capacitance to the same extent as platina. EIS has been shown to be a powerful technique for characterization of stimulating/sensing electrodes.     

Collagen Self-Assembly on Orthopedic Magnesium Biomaterials Surface and Subsequent Bone Cell Attachment

Magnesium (Mg) biomaterials are a new generation of biodegradable materials and have promising potential for orthopedic applications. After implantation in bone tissues, these materials will directly interact with extracellular matrix (ECM) biomolecules and bone cells. Type I collagen, the major component of bone ECM, forms the architecture scaffold that provides physical support for bone cell attachment. However, it is still unknown how Mg substrate affects collagen assembly on top of it as well as subsequent cell attachment and growth. Here, we studied the effects of collagen monomer concentration, pH, assembly time, and surface roughness of two Mg materials (pure Mg and AZ31) on collagen fibril formation. Results showed that formation of fibrils would not initiate until the monomer concentration reached a certain level depending on the type of Mg material. The thickness of collagen fibril increased with the increase of assembly time. The structures of collagen fibrils formed on semi-rough surfaces of Mg materials have a high similarity to that of native bone collagen. Next, cell attachment and growth after collagen assembly were examined. Materials with rough surface showed higher collagen adsorption but compromised bone cell attachment. Interestingly, surface roughness and collagen structure did not affect cell growth on AZ31 for up to a week. Findings from this work provide some insightful information on Mg-tissue interaction at the interface and guidance for future surface modifications of Mg biomaterials.     

Conditioning film formation and its influence on the initial adhesion and biofilm formation by a cyanobacterium on photobioreactor materials

Abstract

Although cyanobacteria are a common group of microorganisms well-suited to utilization in photobioreactors (PBRs), studies of cyanobacteria fouling and its prevention are scarce. Using a cyanobacterium, Anabaena sp. PCC 7120, which had been genetically modified to enhance linalool production, the formation of conditioning films and the effects of these on the physico-chemical surface properties of various PBR materials during initial adhesion and biofilm formation were investigated. The adhesion assay revealed that the overall attachment of Anabaena was substratum dependent and no correlation between the hydrophobicity/roughness of clean material and cell attachment was found. Surface hydrophilicity/hydrophobicity of all the materials changed within 12?h due to formation of conditioning films. ATR-FTIR spectroscopy revealed that the fractional change in protein deposition between 12 to 96?h was consistent with Anabaena cell attachment but polysaccharide deposition was material specific and did not correlate with cell attachment on the PBR materials. Also, the delay in conditioning film proteins on PVC and PTFE indicated that components other than proteins may be responsible for the decrease in contact angles on these surfaces within 12?h. This indicates the important role of the chemical nature of adsorbed conditioning films in determining the initial attachment of Anabaena to PBR materials. The lower rate of attachment of Anabaena on the hydrophilic surfaces (glass and PMMA) between 72?h to 96?h (regime 3) showed that these surfaces could potentially have low fouling characteristics at extended time scales and should be considered for further research.     

The role of nano-roughness in antifouling

Nano-engineered superhydrophobic surfaces have been investigated for potential fouling resistance properties. Integrating hydrophobic materials with nanoscale roughness generates surfaces with superhydrophobicity that have water contact angles (θ) >150° and concomitant low hysteresis (<10°). Three superhydrophobic coatings (SHCs) differing in their chemical composition and architecture were tested against major fouling species (Amphora sp., Ulva rigida, Polysiphonia sphaerocarpa, Bugula neritina, Amphibalanus amphitrite) in settlement assays. The SHC which had nanoscale roughness alone (SHC 3) deterred the settlement of all the tested fouling organisms, compared to selective settlement on the SHCs with nano- and micro-scale architectures. The presence of air incursions or nanobubbles at the interface of the SHCs when immersed was characterized using small angle X-ray scattering, a technique sensitive to local changes in electron density contrast resulting from partial or complete wetting of a rough interface. The coating with broad spectrum antifouling properties (SHC 3) had a noticeably larger amount of unwetted interface when immersed, likely due to the comparatively high work of adhesion (60.77 mJ m^?2 for SHC 3 compared to 5.78 mJ m^?2 for the other two SHCs) required for creating solid/liquid interface from the solid/vapour interface. This is the first example of a non-toxic, fouling resistant surface against a broad spectrum of fouling organisms ranging from plant cells and non-motile spores, to complex invertebrate larvae with highly selective sensory mechanisms. The only physical property differentiating the immersed surfaces is the nano-architectured roughness which supports longer standing air incursions providing a novel non-toxic broad spectrum mechanism for the prevention of biofouling.     

Influence of physicochemical properties on the hygienic status of stainless steel with various finishes

The relative hygienic status of 16 stainless steel surfaces, characterised by topography and surface free energy was investigated. B. thuringiensis spores suspended in Bechamel sauce was chosen as the test fouling suspension. Surface topography was assessed using 10 standardised roughness parameters, along with scanning electron microscope observations. The number of residual adhering spores after a fouling and cleaning in place procedure was found to be influenced by the topography of the stainless steel surface, but not by the surface free energy. Among the various roughness parameters, R_A, R_RR R_PK and R_VK were shown to be related to the hygienic status. Microscopic observations demonstrated the influence of the shape and size of surface irregularities on the level of residual soil after cleaning. This confirms that the use of only one roughness parameter, usually R_A, is not sufficient in defining the hygienic status of stainless steel surfaces.