The use of multiple pseudo-physiological solutions to simulate the degradation behavior of pure iron as a metallic resorbable implant: a surface-characterization study.

Understanding the interactions of a pure iron surface with biological elements, such as ions and proteins in an aqueous medium, is essential for an accurate in vitro assessment of corrosion patterns. In fact, the synergy of chlorides, carbonates, phosphates and complex organic molecules present in the body environment is a key factor affecting both in vivo and in vitro degradation of materials, especially iron and its alloys. The aim of this work was the assessment of degradation patterns of pure iron in 5 commercial pseudo-physiological solutions by a thorough study of degraded surface chemistry and morphology. It also provides a methodological basis to understand the short-term degradation mechanism of degradable iron depending on the surrounding physiological media. The standard static immersion corrosion test was modified to adapt the procedure to pseudo-physiological solutions. After a 14-day static immersion test, the surfaces of samples were investigated by scanning electron microscopy, stylus profilometry and atomic force microscopy techniques. The chemistry and phase composition of the degraded layers were evaluated, respectively, by X-ray photoelectron spectrometry and X-ray diffractometry. The morphology and composition of the degradation layers were found to be different for the test-solutions: for phosphate-rich solutions, the formation of an adherent passive layer was found; degradation mechanisms related to general corrosion were predominant for all the other solutions. In conclusion, the chemical composition of the used medium plays a fundamental role in the degradation pattern of pure iron, so that direct comparisons of solutions with different ion concentrations, as reported in the literature, need to be carefully assessed.

Characterization of Amorphous Oxide Nano-Thick Layers on 316L Stainless Steel by Electron Channeling Contrast Imaging and Electron Backscatter Diffraction.

Characterization of the topmost surface of biomaterials is crucial to understanding their properties and interactions with the local environment. In this study, the oxide layer microstructure of plasma-modified 316L stainless steel (SS316L) samples was analyzed by a combination of electron backscatter diffraction and electron channeling contrast imaging using low-energy incident electrons. Both techniques allowed clear identification of a nano-thick amorphous oxide layer, on top of the polycrystalline substrate, for the plasma-modified samples. A methodology was developed using Monte Carlo simulations combined with the experimental results to estimate thickness of the amorphous layer for different surface conditions. X-ray photoelectron spectroscopy depth profiles were used to validate these estimations.

Mesoporous Silica Nanoparticles under Sintering Conditions: A Quantitative Study.

Thin films made of mesoporous silica nanoparticles (MSNs) are finding new applications in catalysis, optics, as well as in biomedicine. The fabrication of MSNs thin films requires a precise control over the deposition and sintering of MSNs on flat substrates. In this study, MSNs of narrow size distribution (150 nm) are synthesized, and then assembled onto flat silicon substrates, by means of a dip-coating process. Using concentrated MSN colloidal solutions (19.5 mg mL(-1) SiO2), withdrawal speed of 0.01 mm s(-1), and well-controlled atmospheric conditions (ambient temperature, ∼ 70% of relative humidity), monolayers are assembled under well-structured compact patterns. The thin films are sintered up to 900 °C, and the evolution of the MSNs size distributions are compared to those of their pore volumes and densities. Particle size distributions of the sintered thin films were precisely fitted using a model specifically developed for asymmetric particle size distributions. With increasing temperature, there is first evidence of intraparticle reorganization/relaxation followed by intraparticle sintering followed by interparticle sintering. This study is the first to quantify the impact of sintering on MSNs assembled as thin films.

Rapid Nucleation of Iron Oxide Nanoclusters in Aqueous Solution by Plasma Electrochemistry.

Progresses in cold atmospheric plasma technologies have made possible the synthesis of nanoparticles in aqueous solutions using plasma electrochemistry principles. In this contribution, a reactor based on microhollow cathodes and operating at atmospheric pressure was developed to synthesize iron-based nanoclusters (nanoparticles). Argon plasma discharges are generated at the tip of the microhollow cathodes, which are placed near the surface of an aqueous solution containing iron salts (FeCl2 and FeCl3) and surfactants (biocompatible dextran). Upon reaction at the plasma-liquid interface, reduction processes occur and lead to the nucleation of ultrasmall iron-based nanoclusters (IONCs). The purified IONCs were investigated by XPS and FTIR, which confirmed that the nucleated clusters contain a highly hydrated form of iron oxide, close to the stoichiometric constituents of α-FeOOH (goethite) or Fe5O3(OH)9 (ferrihydrite). Relaxivity values of r1 = 0.40 mM(-1) s(-1) and r2/r1 = 1.35 were measured (at 1.41 T); these are intermediate values between the relaxometric properties of superparamagnetic iron oxide nanoparticles used in medicine (USPIO) and those of ferritin, an endogenous contrast agent. Plasma-synthesized IONCs were injected into the mouse model and provided positive vascular signal enhancement in T1-w. MRI for a period of 10-20 min. Indications of rapid and strong elimination through the urinary and gastrointestinal tracts were also found. This study is the first to report on the development of a compact reactor suitable for the synthesis of MRI iron-based contrast media solutions, on site and upon demand.

Correlation between the plasma characteristics and the surface chemistry of plasma-treated polymers through partial least-squares analysis.

We investigated the effect of various plasma parameters (relative density of atomic N and H, plasma temperature, and vibrational temperature) and process conditions (pressure and H2/(N2 + H2) ratio) on the chemical composition of modified poly(tetrafluoroethylene) (PTFE). The plasma parameters were measured by means of near-infrared (NIR) and UV-visible emission spectroscopy with and without actinometry. The process conditions of the N2-H2 microwave discharges were set at various pressures ranging from 100 to 2000 mTorr and H2/(N2+H2) gas mixture ratios between 0 and 0.4. The surface chemical composition of the modified polymers was determined by X-ray photoelectron spectroscopy (XPS). A mathematical model was constructed using the partial least-squares regression algorithm to correlate the plasma information (process condition and plasma parameters as determined by emission spectroscopy) with the modified surface characteristics. To construct the model, a set of data input variables containing process conditions and plasma parameters were generated, as well as a response matrix containing the surface composition of the polymer. This model was used to predict the composition of PTFE surfaces subjected to N2-H2 plasma treatment. Contrary to what is generally accepted in the literature, the present data demonstrate that hydrogen is not directly involved in the defluorination of the surface but rather produces atomic nitrogen and/or NH radicals that are shown to be at the origin of fluorine atom removal from the polymer surface. The results show that process conditions alone do not suffice in predicting the surface chemical composition and that the plasma characteristics, which cannot be easily correlated with these conditions, should be considered. Process optimization and control would benefit from plasma diagnostics, particularly infrared emission spectroscopy.

Surface modification of gadolinium oxide thin films and nanoparticles using poly(ethylene glycol)-phosphate.

The performance of nanomaterials for biomedical applications is highly dependent on the nature and the quality of surface coatings. In particular, the development of functionalized nanoparticles for magnetic resonance imaging (MRI) requires the grafting of hydrophilic, nonimmunogenic, and biocompatible polymers such as poly(ethylene glycol) (PEG). Attached at the surface of nanoparticles, this polymer enhances the steric repulsion and therefore the stability of the colloids. In this study, phosphate molecules were used as an alternative to silanes or carboxylic acids, to graft PEG at the surface of ultrasmall gadolinium oxide nanoparticles (US-Gd(2)O(3), 2-3 nm diameter). This emerging, high-sensitivity "positive" contrast agent is used for signal enhancement in T(1)-weighted molecular and cellular MRI. Comparative grafting assays were performed on Gd(2)O(3) thin films, which demonstrated the strong reaction of phosphate with Gd(2)O(3) compared to silane and carboxyl groups. Therefore, PEG-phosphate was preferentially used to coat US-Gd(2)O(3) nanoparticles. The grafting of this polymer on the particles was confirmed by XPS and FTIR. These analyses also demonstrated the strong attachment of PEG-phosphate at the surface of Gd(2)O(3), forming a protective layer on the nanoparticles. The stability in aqueous solution, the relaxometric properties, and the MRI signal of PEG-phosphate-covered Gd(2)O(3) particles were also better than those from non-PEGylated nanoparticles. As a result, reacting PEG-phosphate with Gd(2)O(3) particles is a promising, rapid, one-step procedure to PEGylate US-Gd(2)O(3) nanoparticles, an emerging "positive" contrast agent for preclinical molecular and cellular applications.

Plasma-immersion ion implantation surface oxidation on a cobalt-chromium alloy for biomedical applications.

Co-Cr alloys such as L605 are widely applied for the manufacture of medical devices, including tiny cardiovascular stents. The presence of potentially toxic and allergenic release of Ni, Co, and Cr ions from these devices remains an unsolved concern. Surface modification by oxygen plasma immersion implantation (PIII) could be an excellent technique to create a dense and thin passive oxide layer on a relatively complex shape of a tiny device, such as a stent, thus reducing the potential release of metallic ions. The effect of oxygen PIII was investigated on L605 alloy specimens, from 5 to 50 mTorr gas pressures, and under pulsed bias voltages from -0.1 to -10 kV. The surface chemistry was investigated by x-ray photoelectron spectroscopy, while its morphology and surface energy were evaluated, respectively, by atomic force microscopy and scanning electron microscopy and by a sessile drop static contact angle. Electrochemical characterization was performed by potentiodynamic tests in the saline solution. Mechanical properties of the modified surface layer, specifically film adhesion and hardness (H), were assessed by scratch and nanoindentation tests. Results shown that the oxidized layers were composed of a mixture of Co and Cr oxides and hydroxides and were rich in Co. The corrosion rate was considerably reduced after O PIII, even for treatments using low bias voltage (-0.1 kV) and with consequent low oxygen implantation depth. Moreover, O PIII also improved surface hardness. The oxidized layers were found to have good adhesion and to be scratch resistant.

Specific shortcomings of endograft design.

Further to the rapid enlargement of an aneurysm to 5.6 cm in diameter after 3 years of surveillance, a 79-year-old patient was fitted with a Vanguard modular stent graft and monitored on a regular basis for 6 years. Two years later, the aneurysmal sac ruptured. The patient died 1 month after an open surgery. The device was devoid of any encapsulation and the ipsilateral limb was detached from the body. The Nitinol skeleton was mostly maintained, however, some polypropylene sutures were broken. The resulting motion of the sharp-angled Nitinol wires caused abrasion and resulted in a few localized holes that were sufficient to permit blood to percolate through the textile wall. Some polyester yarns in the warp direction were ruptured. The Nitinol wire used in this device was shown to be corrosion resistant but the selection of the polypropylene suture was inappropriate. Because this technology is maturing rapidly, these weaknesses can be avoided in the future generations of endovascular devices. It is recommended that these Nitinol wires be sutured to the fabric and that polyester yarns stronger than 68 decitex in tubes 8 mm in diameter are selected.

Persistent type II endoleak unrelated to an Anaconda aortic stent graft fulfilling the 3Bs requirements of biofunctionality, biodurability, and biocompatibility.

A patient was fitted with an Anaconda stent graft for which there was a persistent type II endoleak. Two subsequent attempts at embolization were unable to resolve the endoleak. The diameter of the aneurysm varied initially from 5.5 cm in diameter down to 4.8 cm but then later re-dilated to 6.1 cm, with evidence of persistent flow into the aneurysmal sac from the inferior mesenteric artery. Results from serial computed tomography scans demonstrated clear evidence of a type II endoleak that originated from the inferior mesenteric artery with outflow to a distal lumbar artery. The harvested stent graft did not show evidence of a device-related failure. The stent graft and its modular segments were found to have been properly deployed. Only a thin external capsule was evident at explantation. The internal wall of the device showed irregular and thin encapsulation with scattered mural thrombi, which were more prominent at the bifurcation of the main body of the device. Blood deposits and tissue development were sufficient to prevent blood oozing through the wall. The explanted Anaconda stent graft was devoid of any construction flaws or damage (fatigue of the textile or corrosion of the Nitinol wires) after implantation.

Characterization of an endovascular prosthesis using the 3Bs rule (biocompatibility, biofunctionality and biodurability): a recommended protocol to investigate a device harvested at necropsy.

Numerous endovascular stent grafts to treat intrarenal aortic aneurysms are now commercially available, and many new concepts are currently in development worldwide. In order to objectively quantify their outcomes, we propose a detailed protocol to examine a reference device that was harvested from a patient who died a few hours after endovascular stent-graft deployment for an abdominal aortic aneurysm according to the 3Bs rule (biocompatibility, biofunctionality, and biodurability). Relevant patient history of this 63-year-old man included radiotherapy treatment for lung cancer. Following the patient's death, the device was harvested en bloc together with the aneurysmal sac. The analysis of the device was conducted using nondestructive testing (X-rays, CT scan, magnetic resonance imaging [MRI], and endoscopy) and destructive testing (dissection, histology, and fabric and wire component analyses). Results from the gross examination demonstrated that the outer layer of the aneurysm sac was white, stiff, and continuous without any disruption. The Xray analysis, CT scan, and MRI confirmed that the device together with its modular segments was properly deployed at implantation. Endoscopy showed that the device was deployed securely immediately distal to the renal arteries. As anticipated, thin scattered mural thrombi at the blood/foreign material interface were observed on the blood tight flow surface. There were no tears in the fabric, and the dimensions and textile structure were well preserved. The metallic wires were intact. This fatality had no association with the stent graft as the patient's death was caused by the rupture of the pulmonary artery following intensive radiotherapy. In conclusion, autopsy, nondestructive testing, and destructive testing are therefore the necessary steps to validate any explanted endovascular stent graft in terms of biocompatibility, biofunctionality, and biodurability. In this specific case, the endovascular device fulfills the 3Bs rule. The authors recommend this protocol to investigate explanted endovascular devices.

Effect of grain sizes on mechanical properties and biodegradation behavior of pure iron for cardiovascular stent application.

Pure iron has been demonstrated as a potential candidate for biodegradable metal stents due to its appropriate biocompatibility, suitable mechanical properties and uniform biodegradation behavior. The competing parameters that control the safety and the performance of BMS include proper strength-ductility combination, biocompatibility along with matching rate of corrosion with healing rate of arteries. Being a micrometre-scale biomedical device, the mentioned variables have been found to be governed by the average grain size of the bulk material. Thermo-mechanical processing techniques of the cold rolling and annealing were used to grain-refine the pure iron. Pure Fe samples were unidirectionally cold rolled and then isochronally annealed at different temperatures with the intention of inducing different ranges of grain size. The effect of thermo-mechanical treatment on mechanical properties and corrosion rates of the samples were investigated, correspondingly. Mechanical properties of pure Fe samples improved significantly with decrease in grain size while the corrosion rate decreased marginally with decrease in the average grain sizes. These findings could lead to the optimization of the properties to attain an adequate biodegradation-strength-ductility balance.

Influence of cross-rolling on the micro-texture and biodegradation of pure iron as biodegradable material for medical implants.

Iron-based biodegradable metals have been shown to present high potential in cardiac, vascular, orthopaedic and dental in adults, as well as paediatric, applications. These require suitable mechanical properties, adequate biocompatibility while guaranteeing a low toxicity of degradation products. For example, in cardiac applications, stents need to be made by homogeneous and isotropic materials in order to prevent sudden failures which would impair the deployment site. Besides, the presence of precipitates and pores, chemical inhomogeneity or other anisotropic microstructural defects may trigger stress concentration phenomena responsible for the early collapse of the device. Metal manufacturing processes play a fundamental role towards the final microstructure and mechanical properties of the materials. The present work assesses the effect of mode of rolling on the micro-texture evolution, mechanical properties and biodegradation behaviour of polycrystalline pure iron. Results indicated that cross-rolled samples recrystallized with lower rates than the straight-rolled ones due to a reduction in dislocation density content and an increase in intensity of {100} crystallographic plane which stores less energy of deformation responsible for primary recrystallization. The degradation resulted to be more uniform for cross-rolled samples, while the corrosion rates of cross-rolled and straight-rolled samples did not show relevant differences in simulated body solution. Finally, this work shows that an adequate compromise between biodegradation rate, strength and ductility could be achieved by modulating the deformation mode during cold rolling.

On the long term antibacterial features of silver-doped diamondlike carbon coatings deposited via a hybrid plasma process.

Environmental surfaces are increasingly recognized as important sources of transmission of hospital-acquired infections. The use of antibacterial surface coatings may constitute an effective solution to reduce the spread of contamination in healthcare settings, provided that they exhibit sufficient stability and a long-term antibacterial effect. In this study, silver-incorporated diamondlike carbon films (Ag-DLC) were prepared in a continuous, single-step plasma process using a hybrid, inductively coupled plasma reactor combined with a very-low-frequency sputtering setup. The average Ag concentration in the films, ranging from 0 to 2.4 at. %, was controlled by varying the sputtering bias on the silver target. The authors found that the activity of Escherichia coli was reduced by 2.5 orders of magnitude, compared with the control surface, after a 4-h contact with a 2.4 at. % Ag-DLC coating. The coatings displayed slow release kinetics, with a total silver ion release in the sub-ppb range after 4 h in solution, as measured by graphite furnace-atomic absorption spectroscopy. This was confirmed by Kirby-Bauer diffusion tests, which showed limited diffusion of biocidal silver with a localized antibacterial effect. As a slow and continuous release is mandatory to ensure a lasting antibacterial effect, the newly developed Ag-DLC coatings appears as promising materials for environmental hospital surfaces.

Characterization of multilayer anti-fog coatings.

Fog formation on transparent substrates constitutes a major challenge in several optical applications requiring excellent light transmission characteristics. Anti-fog coatings are hydrophilic, enabling water to spread uniformly on the surface rather than form dispersed droplets. Despite the development of several anti-fog coating strategies, the long-term stability, adherence to the underlying substrate, and resistance to cleaning procedures are not yet optimal. We report on a polymer-based anti-fog coating covalently grafted onto glass surfaces by means of a multistep process. Glass substrates were first activated by plasma functionalization to provide amino groups on the surface, resulting in the subsequent covalent bonding of the polymeric layers. The anti-fog coating was then created by the successive spin coating of (poly(ethylene-maleic anhydride) (PEMA) and poly(vinyl alcohol) (PVA) layers. PEMA acted as an interface by covalently reacting with both the glass surface amino functionalities and the PVA hydroxyl groups, while PVA added the necessary surface hydrophilicity to provide anti-fog properties. Each step of the procedure was monitored by XPS, which confirmed the successful grafting of the coating. Coating thickness was evaluated by profilometry, nanoindentation, and UV visible light transmission. The hydrophilic nature of the anti-fog coating was assessed by water contact angle (CA), and its anti-fog efficiency was determined visually and tested quantitatively for the first time using an ASTM standard protocol. Results show that the PEMA/PVA coating not only delayed the initial period required for fog formation but also decreased the rate of light transmission decay. Finally, following a 24 hour immersion in water, these PEMA/PVA coatings remained stable and preserved their anti-fog properties.

Influence of the 316 L stainless steel interface on the stability and barrier properties of plasma fluorocarbon films.

Coatings are known to be one of the more suited strategies to tailor the interface between medical devices and the surrounding cells and tissues once implanted. The development of coatings and the optimization of their adhesion and stability are of major importance. In this work, the influence of plasma etching of the substrate on a plasma fluorocarbon ultrathin coating has been investigated with the aim of improving the stability and the corrosion properties of coated medical devices. The 316 L stainless steel interface was subjected to two different etching sequences prior to the plasma deposition. These plasma etchings, with H(2) and C(2)F(6) as gas precursors, modified the chemical composition and the thickness of the oxide layer and influenced the subsequent polymerization. The coating properties were evaluated using flat substrates submitted to deformation, aging into aqueous medium and corrosion tests. X-ray photoelectron spectroscopy (XPS), time of flight-secondary ion mass spectrometry (ToF-SIMS), ellipsometry, and atomic force microscopy (AFM) were performed to determine the effects of the deformation and the aging on the chemistry and morphology of the coated samples. Analyses showed that plasma etchings were essential to promote reproducible polymerization and film growth. However, the oxide layer thinning due to the etching lowered the corrosion resistance of the substrate and affected the stability of the interface. Still, the deformed samples did not exhibited adhesion and cohesion failure before and after the aging.

Effects of chemical composition and the addition of H2 in a N2 atmospheric pressure dielectric barrier discharge on polymer surface functionalization.

We examined the effect of hydrogen content in various polymers in a N2/H2 discharge for surface amine functionalization. Three polymers (polyethylene (PE), polyvinylidene fluoride (PVDF), and poly(tetrafluoroethylene) (PTFE)) containing various amounts of hydrogen and fluorine were treated with an atmospheric pressure dielectric barrier discharge (DBD). While surface modification was observed on the PE and the PVDF in a pure N2 discharge, adding H2 in a N2 discharge was necessary to observe the fluorine etching on the surface of the PVDF and PTFE polymers. The presence of a slight amount of hydrogen in the gas mixture was also a prerequisite to the formation of amino groups on the surface of all three polymers (max NH2/C approximately 5%). Aging revealed that the modified polymer surfaces treated in a N2-H2 discharge were less prone to hydrophobic recovery than were surfaces treated in pure N2, due primarily to the presence of a higher density of polar groups on the surfaces. We demonstrated that H atoms in the discharge are necessary for the surface amine functionalization of polymers in a N2 atmospheric pressure DBD, regardless of polymer chemical composition. It is therefore possible to control the plasma functionalization process and to optimize the concentration and specificity of NH2 grafted onto polymer surfaces by varying the H2 concentration in a N2 atmospheric pressure DBD.

X-ray photoelectron emission microscopy and time-of-flight secondary ion mass spectrometry analysis of ultrathin fluoropolymer coatings for stent applications.

Fluoropolymer plasma coatings have been investigated for application as stent coatings due to their chemical stability, conformability, and hydrophobic properties. The challenge resides in the capacity for these coatings to remain adherent, stable, and cohesive after the in vivo stent expansion, which can generate local plastic deformation of up to 25%. Plasma-coated samples have been prepared by a multistep process on 316L stainless steel substrates, and some coated samples were plastically deformed to mimic a stent expansion. Analyses were then performed by X-ray photoelectron spectroscopy (XPS), X-ray photoelectron emission microscopy (X-PEEM), and time-of-flight secondary ion mass spectrometry (TOF-SIMS) to determine the chemical and physical effects of such a deformation on both the coating and the interfacial region. While XPS analyses always showed a continuous coating with no significant effect of the deformation, TOF-SIMS and near-edge X-ray absorption fine structure (derived from X-PEEM) data indicated the presence of a certain density of porosity and pinholes in all coatings as well as sparse fissures and molecular fragmentation in the deformed ones. The smallness of the area fraction affected by the defects and the subtlety of the chemical changes could only be evidenced through the higher chemical sensitivity of these latter techniques.

AFM imaging of immobilized fibronectin: does the surface conjugation scheme affect the protein orientation/conformation?

Covalent grafting of biomolecules could potentially improve the biocompatibility of materials. However, these molecules have to be grafted in an active conformation to play their biological roles. The present work aims at verifying if the surface conjugation scheme of fibronectin (FN) affects the protein orientation/conformation and activity. FN was grafted onto plasma-treated fused silica using two different crosslinkers, glutaric anhydride (GA) or sulfosuccinimidyl 4-(p-maleimidophenyl)butyrate (SMPB). Fused silica was chosen as a model surface material because it presents a roughness well below the dimensions of FN, therefore allowing AFM analyses with appropriate depth resolution. Cell adhesion assays were performed to evaluate the bioactivity of grafted FN. Cell adhesion was found to be higher on GA-FN than on SMPB-FN. Since FN-radiolabeling assays allowed us to rule out a surface concentration effect (approximately 80 ng/cm2 of FN on both crosslinkers), it was hypothesized that FN adopted a more active conformation when grafted via GA. In this context, the FN conformation on both crosslinkers was investigated through AFM and contact angle analyses. Before FN grafting, GA- and SMPB-modified surfaces had a similar water contact angle, topography, and roughness. However, water contact angles of GA-FN and SMPB-FN surfaces clearly show differences in surface hydrophilicity, therefore indicating a dependence of protein organization toward the conjugation strategy. Furthermore, AFM results demonstrated that surface topography and roughness of both FN-conjugated surfaces were significantly different. Distribution analysis of FN height and diameter confirmed this observation as the protein dimensions were significantly larger on GA than SMPB. This study confirmed that the covalent immobilization scheme of biomolecules influences their conformation and, hence, their activity. Consequently, selecting the appropriate conjugation strategy is of paramount importance in retaining molecule bioactivity.