Protein Corona Formation on Magnetite Nanoparticles: Effects of Culture Medium Composition, and Its Consequences on Superparamagnetic Nanoparticle Cytotoxicity.

The physicochemical properties and potential cytotoxicity of nanoparticles (NPs) are significantly influenced by their inter- action with proteins, which results in corona formation. Here, we have determined whether corona formation, resulting from interactions between superparamagnetic iron oxide nanoparticles (SPIONs) and different cell culture media, may have consequences for driving NP toxic effects. To address this issue, complementary methods were used. The deter- mination of the hydrodynamic size distribution by ζ (zeta) potential measurement indicated that SPIONs were negatively charged under all conditions but that the actual charge was differed with the cell culture medium used. In vitro protein adsorption studies were carried out using the Bradford protein assay and Fourier transform infrared spectroscopy (FTIR). The Bradford assay revealed that the concentration of unadsorbed proteins and other biomolecules decreased when the SPION concentration increased. FTIR showed that the proteins were, indeed, adsorbed onto the NP surface. This was followed by matrix-assisted laser desorption/ionization time-of-flight secondary ion mass spectrometry (MALDI TOF-SIMS), to identify the adsorbed proteins. Ultimately, three different cell viability assays led to the conclusion that the SPIONs were not toxic for all the concentrations used here. In summary, we found that corona formation on the SPIONs depends on the composition of the culture media but has no consequence for nanotoxicity. We have shown that the application of complementary methods has provided novel insights into SPION/protein interactions.

Core-shell nanoparticles as prodrugs: possible cytotoxicological and biomedical impacts of batch-to-batch inconsistencies.

Numerous samples of magnetite@silica and magnetite@silica@silane core-shell nanoparticles, previously used as prodrugs, were prepared by an experienced chemist, using the same identical equipment and the same lots of reagents. Their surface analyses showed batch-to-batch chemical variations: no two batches were found to have the same surface chemistries, showing unexpected Si-O bond scission and amine oxidation. Because the preparations used reactions recognized to be mild, and bond scission and oxidation were never previously reported for similar reactions on larger surfaces, the Fe(3)O(4) nanoparticles that form the nanoparticle core appear to have acted as catalysts for these side reactions. The intended use of these nanoparticles, as drug carriers, is discussed in terms of cytotoxicological and biomedical consequences.

Nitric oxide delivery by core/shell superparamagnetic nanoparticle vehicles with enhanced biocompatibility.

We report the synthesis of Fe(3)O(4)/silica core/shell nanoparticles and their functionalization with S-nitrosothiols. These nanoparticles are of immense interest because of their nitric oxide (NO) release capabilities in human alveolar epithelial cells. Moreover, they act as large storage reservoirs of NO that can be targeted magnetically to the specific site with a sustainable release of NO for up to 50 h. Such nanoparticles provide an enhancement of the biocompatibility with released NO while allowing intracellular accumulation ascribed to their small size.

Reduced stress shielding with limited micromotions using a carbon fibre composite biomimetic hip stem: a finite element model.

Total hip arthroplasty (THA) enjoys excellent rates of success in older patients, but younger patients are still at risk of aseptic loosening and bone resorption from stress shielding. One solution to the stress shielding problem is to use a hip stem with mechanical properties matching those of cortical bone. The objective of the present study was to investigate numerically the biomechanical performance of such a biomimetic hip stem based on a hydroxyapatite (HA)-coated carbon fibre composite. A finite element model (FEM) of the biomimetic stem was constructed. Contact elements were studied to model the bone-implant interface in a non-osseointegrated and osseointegrated state in the best way. Three static load cases representing slow walking, stair climbing, and gait in a healthy individual were considered. Stress shielding and bone-implant interface micromotions were evaluated and compared with the results of a similar FEM based on titanium alloy (Ti-6Al-4V). The composite stems allowed for reduced stress shielding when compared with a traditional Ti-6Al-4V stem. Micromotions were slightly higher with the composite stem, but remained below 40 microm on most of the HA-coated surface. It is concluded that a biomimetic composite stem might offer a better compromise between stress shielding and micromotions than the Ti-6Al-4V stem with the same external geometry.

Biocompatibility and physicochemical characteristics of alginate-polycation microcapsules.

There is a need for better understanding of the biocompatibility of alginate-polycation microcapsules based on their physicochemical characteristics. Microcapsules composed of alginate with 44% (IntG) or 71% (HiG) guluronate, gelled with calcium (Ca) or barium (Ba) and coated with poly-L-lysine (PLL) or poly-l-ornithine (PLO), followed by IntG alginate were compared. For microcapsules with an IntG(Ca) gel core, using PLO instead of PLL resulted in less immune cell adhesion after 2 days in C57BL/6J mice. The PLO microcapsules were also characterized by greater hydrophilicity and superior resistance to swelling and damage under osmotic stress. For microcapsules with a PLL membrane, replacing the IntG(Ca) gel core with IntG(Ba) or HiG(Ca) gel resulted in stronger immune responses (p<0.05). This was explained by poor penetration of PLL into the gel, as demonstrated by Fourier transform infrared spectroscopy analyses and membrane rupturing during osmotic swelling. X-ray photoelectron spectroscopy analyses show that all microcapsules had the same amount of polycation at their surface. Moreover, alginate coatings had non-significant effects on the biocompatibility and physicochemical properties of the microcapsules. Thus, alginate-polycation interactions for membrane formation are more important for biocompatibility than either the quantity of polycation at the surface or the alginate coating.

The effect of varying Al2O3 percentage in hydroxyapatite/Al2O3 composite materials: morphological, chemical and cytotoxic evaluation.

Hydroxyapatite (HA) and HA-alumina (HA/Al2O3) composites, with Al2O3 contents of 5, 10, 20, and 30%, were synthesized using a wet precipitation method and sintered at 900 and 1300 degrees C. We investigated the effect of sintering temperature and relative concentration of HA and Al2O3 on the chemical composition, surface morphology, and cytotoxicity of the composite powders. The XRD results show that in the 1300 degrees C composites, HA partially decomposed into CaO which combined with Al2O3 to form different calcium aluminates. For the 900 degrees C composites the CaO phase was not detected, though a Ca/P ratio larger than 1.67 measured by XPS suggests that CaO was present in trace amounts. SEM-EDX analysis indicated that the HA microstructure was affected by the sintering temperature, and this HA is present on the surface of Al2O3 particles. The cytotoxicity of the composites was assessed indirectly using the MTT assay. The short-term effect of leachables was quantified by exposing a L929 mouse fibroblast cell line to the degradation products released by the composites after immersion in the cell culture medium. Degradation products were less toxic to L-929 at lower extract concentrations (10, 50%) than at 100% concentration. Cell viability was also influenced by leachable size.

Accelerated fatigue behavior and mechano-physical characterizations of in vitro physiological simulation of nitinol stents.

In this study, we have provided an experimental evaluation of the fatigue behavior of the nitinol (NiTi) endovascular device (peripheral stent). The accelerated fatigue tests were performed using arterial conditions, which mimicked actual physiological conditions. Natural, rubber latex-tubing materials were used to simulate human arteries. The equipment design and the test parameters used allowed for the simulation of a compliant artery and the application of circumferential forces to the device.The stent compliance values were good indicators for tracking the time evolution of fatigue behavior. Moreover, the analyses of changes on the surface morphology and on the chemical composition were used to establish a relationship between surface characteristics and peripheral stent response during 400 million cycles, which is equivalent to 10 yrs of human life. In order to determine the influence of the accelerated fatigue, an evaluation of both mechanical and surface characteristics was carried out before and after testing using the following tests and methods, respectively: radial hoop testing (RH), scanning electron microscope analysis (SEM), auger electron spectroscopy (AES), atomic absorption spectroscopy (AAS), and X-ray photoelectron spectroscopy (XPS). Under these experimental conditions, the studies have shown that after 400 million cycles, the tested stents did not demonstrate any mechanical failure. Moreover, the surface did not undergo any changes in its chemical composition. However, we did observe an increase in roughness and signs of pitting corrosion.

Bioperformance of shape memory alloy single crystals.

Shape memory alloys (SMA) represent a large family of alloys that show unique characteristics. They have been exploited in several fields for diverse applications. For the last 20 years, these alloys and more particularly Ni-Ti alloys have revolutionized the field of metallic biomaterials. Applications in the biomedical area are multiple and these materials improve significantly the quality of the diagnostics, treatments and surgeries. To our knowledge, most devices are made of SMAs in the polycrystalline form. Nevertheless, the single crystal form shows several promising advantages especially concerning its mechanical performances. In this paper we describe the advantages, advances and limits of using different SMA single crystals for biomedical applications, including biocompatibility and corrosion resistance. We also discuss the low response time of classical thermal SMAs as well as the new advances in research on magnetic SMA single crystals.

Surface characterization of Streptococcus mutans biofilms grown on polyethylene and beta-titanium.

In this study, we considered the biofilms as a surface, characterizing them using instruments for surface analyses, environmental microscopy, IR-spectroscopy (ATR-mode) and goniometry of the contact angle. The bacteria that formed the biofilms were grown on two different supports: beta-titanium alloy (beta-Ti) and polyethylene (PE). Environmental microscopy allowed the observation of biofilms in situ and in their hydrated state. On the metallic support, the biofilm quickly adhered and formed a dense structure with micro-colonies, but on the PE a thinner biofilm layer was observed covering a large surface area of the support. IR-spectroscopy is another effective method to detect the biofilm quickly and in situ, without pre-treating the sur-face. Nevertheless, problems with the overlapping of the characteristic bands on the spectra are frequent between the biofilm and PE. Finally, we compared the surface energy (SE) of the supports before and after biofilm formation. Our results indicate that the SE of the supports depends on the sterilization method, and that the SE of the biofilms varies depending on the support and the sterilization method. The biofilm on the beta-Ti had the highest SE, and as mentioned above, microscopic images showed a higher roughness on its surface.

Bioactive hydroxyapatite coatings on polymer composites for orthopedic implants.

Hydroxyapatite [HA, Ca10(PO4)6(OH)2] coatings on polymer composite substrates were investigated for their bioactivity and their physicochemical and mechanical characteristics. HA holds key characteristics for use in orthopedic applications, such as for coating of the femoral stem in a hip replacement device. The plasma-spray technique was used to project HA onto a carbon fiber/polyamide 12 composite substrate. The resulting HA coatings exhibited mechanical adhesion as high as 23 MPa, depending on the surface treatment of the composite substrate. The purpose of this investigation was to evaluate the bioactivity of an HA-coated composite substrate. HA- coated samples have been immersed in simulated body fluid (SBF) and maintained within a shaker bath for periods of 1, 7, 14, 21, and 28 days at 37 degrees C. Scanning electron microscopy, energy dispersive X-ray spectroscopy, and X-ray diffraction techniques were performed on the samples before and after immersion into SBF. SBF was analyzed using inductively coupled plasma atomic emission spectrometry for element concentration and evaluation of the solution's purity. SBF conditioning led to the deposition of crystalline HA onto the surface of the coatings. The calcium-to-phosphorous ratios of initial HA coating and of newly deposited HA were respectively 1.72 and 1.65, close to the HA theoretical calcium/phosphorous value of 1.67. Results demonstrated that bioactive HA coatings were produced by plasma spraying, because SBF conditioning induced newly formed HA with high crystallinity. Mechanical adhesion of the HA coatings was not significantly affected upon SBF conditioning.

Interjoint coordination in lower limbs in patients with a rupture of the anterior cruciate ligament of the knee joint.

Previous studies of movement kinematics in patients with a ruptured anterior cruciate ligament (ACL) have focused on changes in angular displacement in a single joint, usually flexion/extension of the knee. In the present study, we investigated the effect of an ACL injury on the overall limb interjoint coordination. We asked healthy and chronic ACL-deficient male subjects to perform eight types of movements: forward squats, backward squats, sideways squats, squats on one leg, going up a step, going down a step, walking three steps, and stepping in place. Depending on the movement concerned, we applied principal component (PC) analysis to 3 or 4 degrees of freedom (DFs): thigh flexion/extension, knee flexion/extension, ankle flexion/extension, thigh abduction/adduction. The first three DFs were investigated in all movements. PC analysis identifies linear combinations of DFs. Movements with a fixed ratio between DFs are thus described by only one PC or synergy. PCs were computed for the entire movement as well as for the period of time when the foot was in contact with the ground. For both the control and the injured groups, two synergies (PC vectors) usually accounted for more than 95% of the DFs' angular excursions. It was possible to describe 95-99% of some movements using only one synergy. Compared to control subjects, injured subjects employed different synergies for going up a step, walking three steps, squatting sideways, and squatting forward, both in the injured and uninjured legs. Those movements may thus be more indicative of injury than other movements. Although ACL-deficiency did not increase asymmetry (angle between the PCs of the same movement performed on the right and the left sides), this result is not conclusive because of the comparatively low number of subjects who participated in the study. However, the finding that synergies in both legs of patients were different from those in control subjects for going up a step and walking three steps suggests that interjoint coordination was affected for both legs, so that the asymmetry index might have been preserved despite the injury. There was also a relationship between the asymmetry index for squatting on one leg, squatting forward, walking three steps and some of the outcomes of the knee injury and osteoarthritis outcome score (pain, symptoms, activities of daily living, sport and recreation function, and knee-related quality of life). This suggests that significant differences in the asymmetry index could be obtained if more severely-injured patients participated in this study. It is possible that subjects compensated for their mechanical deficiencies by modifying muscle activation patterns. Synergies were not only modified in injured subjects, but also rearranged: the percentage of movement explained by the first PC was different for the injured and/or uninjured legs of patients, as compared to the legs of the control group, for going up a step, going down a step, walking three steps, and squatting forward. We concluded that the analysis of interjoint coordination may be efficient in characterizing motor deficits in people with knee injuries.

Adapting MRI systems to propel and guide microdevices in the human blood circulatory system.

Magnetic resonance imaging (MRI) systems are widely used to gather noninvasively images of the interior of the human body. This paper suggests that an MRI system can be seen beyond being just a tool for imaging purpose but one that can propel and guide special microdevices in the human body to perform specific medical tasks. More specifically, an MRI system can potentially be used to image the region of interest, propel a microdevice through the generation of magnetic gradients, determine the location of the device, compute the corrective actions through feedback control algorithms and adjust the generation of the magnetic gradients accordingly to navigate such a microdevice in a preplanned path. This paper presents an introductory description of the proposed techniques, the main issues to consider, and some preliminary data indicating the validity of this approach.

Computer based method for the three-dimensional kinematic analysis of combined posterior cruciate ligament and postero-lateral complex reconstructions on cadaver knees.

The aim of this study is to evaluate the effect of combined posterior cruciate ligament (PCL) and postero-lateral corner (PLC) reconstruction on laxity and three-dimensional kinematics of cadaver knees. We performed anatomical double bundle PCL reconstruction, and functional one bundle 'over-the-bottom' PCL reconstruction combined with one type of PLC reconstruction, running from the postero-lateral tibia to an isometric point near the lateral epicondyle of the femur. Our results showed that combined reconstruction was necessary to restore rotatory laxity. PLC reconstruction, according to the technique described, invariably created a shift towards internal rotation of the kinematic curves, compared to the intact knee.

Comparison of two methods for reconstruction of the posterior cruciate ligament using a computer based method: quantitative evaluation of laxity, three-dimensional kinematics and ligament deformation measurement in cadaver knees.

The aim of this paper is to present a biomechanical comparison of two different methods for reconstruction of the posterior cruciate ligament in cadaver knees. We used an original computer-based method allowing precise calculation of three-dimensional (3D) knee kinematic parameters as well as the estimation of combined graft deformation (elongation-flexion-torsion). After isolated posterior cruciate ligament (PCL) dissection, double bundle and 'over-the-bottom' methods were performed successively on each knee using synthetic polyester ligaments. The effect of pre-tensioning was tested with the 'over-the-bottom' method. antero-posterior (A-P) and rotational laxity as well as 3D kinematics were recorded and analysed. Our computer based method allowed us to show that both reconstruction methods were equivalent in restoring A-P and rotational laxity as well as kinematic curves. Combined deformation of the prostheses was equivalent for both ligaments.

Computer-based method for the 3-D kinematic analysis of posterior cruciate ligament and postero-lateral corner lesions.

Posterior cruciate ligament (PCL) rupture, whether or not combined with postero-lateral corner (PLC) tears, are more often diagnosed today thanks to improved imaging techniques. However, due to the lack of reliable instrumentation to quantitatively evaluate the knee, much is still unknown about the function of these ligamentous structures. The aim of this paper is to present results on the effect of progressive resection of the PCL and PLC on knee laxity and 3-D knee kinematics. The results show that 3-D movement analysis is important and complements laxity measurements by helping to interpret the complex alteration of knee function.

Effect of surface treatment of NiTi alloy on its corrosion behavior in Hanks' solution.

The effect of surface modification of NiTi shape memory alloy on its corrosion behavior in Hanks' solution was determined. The near-equi-atomic super elastic NiTi (Ni 55.8 wt %) alloy used for this study was provided by Memry USA. The surfaces of heat-treated samples were modified by mechanical polishing (MP), electropolishing (EP), and electropolishing followed by chemical passivation (CP). As-heat-treated samples with straw-colored oxide finishes (SCO) and blue-colored oxide finishes (BO) also were included in the study. Surface analysis was performed using auger electron spectroscopy (AES), atomic force microscopy (AFM), and contact angle measurements (CAM). It was shown that surface roughness increased in the order CP < EP < SCO < BO < MP. The nickel release within the five groups of NiTi samples, as determined by atomic absorption spectrophotometry, reduced in time over the measured period. The level of Ni ions released over a 25-day immersion period was highest in the SCO sample (0.002 microg/day). This Ni level is negligible compared with the daily intake of Ni in an ordinary diet. The auger electron spectroscopy (AES) analyses indicated that before immersion in Hanks' physiologic solution, the main surface composition of all the samples was titanium and nickel, with a small amount of oxygen, carbon, and sulphur as contaminants. And the surface oxide thickness of the different samples increased in the order CP < EP < MP < BO < SCO. On the other hand, for the electrodes treated under the same conditions, the mean breakdown potential value decreased in the order BO > MP > CP > EP > SCO while the corrosion current density and rate increased in the order CP < SCO < EP < BO < MP.

Polyethylene wear characteristics in vivo and in a knee stimulator.

Validation of a wear simulator requires that the device produce a similar type and amount of wear and particles of a comparable morphology as occurs clinically. Using techniques previously established to compare polyethylene particles from hip simulators to those from retrieved tissues, particles isolated from six revised posterior stabilized knee replacements were characterized and compared to particles generated from the same knee design worn in a knee simulator. The particles produced in the knee simulator were of comparable size but had less variability in their form factor compared to the particles produced in vivo. Comparable wear features were seen on the articulating surfaces in both groups. These results indicate that this knee joint simulator is able to reproduce a baseline type of wear that is similar to that in vivo and should encourage further use of this device to better understand knee component wear and function.

Safety of plasma-based sterilization: surface modifications of polymeric medical devices induced by Sterrad and Plazlyte processes.

Plasma-based sterilization is a promising alternative to the use of pure ethylene oxide (EO), for low-temperature clinical sterilization of medical instruments and devices. However, few studies have been published that evaluate its safety in terms of possible damage to materials, particularly polymers. The objective of this work was to evaluate polymer surface modifications induced by commercial plasma-based sterilizers, in comparison with pure EO: Samples from 5 polymer-based devices were subjected to 1, 5, and 10 sterilization cycles by Sterrad-100, Plazlyte, and pure EO. Surface analysis was carried out by X-ray photoelectron spectroscopy (XPS), dynamic contact angle measurements (DCA), and scanning electron microscopy (SEM). Surface oxidation and wettability changes were observed on all samples sterilized by plasma-based techniques, the degree of modifications depending on the sterilizer (Sterrad, Plazlyte) and the type of polymer. Drastic changes of surface appearance were also observed by SEM on PVC samples sterilized by Plazlyte and by pure EO. Possible repercussions on safety are discussed.

Characteristics of porous nickel-titanium alloys for medical applications.

We investigate the behavior of NiTi porous alloys, possessing the property of shape memory, by using different characterization methods XPS, Auger, DSC and SEM. The study mainly focuses on the determination of porosity, surface characteristics and the phase transformation. In the case of porous material the biomechanical compatibility is closely related to the internal structure and porosity distribution. To describe appropriately the influence of the properties of NiTi on the memory shape, two types of materials provided by different sources has been analyzed. Despite the fact that both materials present different pores size, they exhibit an open and interconnected porosity. Our measurements show that the temperature of the inception of the martensite-austenite phase transition occurs at 60 degrees C, which is by 20 degrees C greater than the body temperature. Moreover, we show that the surface characteristics can be greatly influenced by heat treatment. Furthermore, we observe that the R-phase occurs only for one of the used materials after its heat treatment. The correlation between the composition and the other characteristics measured has been found.

Low-temperature sterilization using gas plasmas: a review of the experiments and an analysis of the inactivation mechanisms.

Utilizing an ionized gas (plasma) to achieve sterilization is an alternative to conventional sterilization means as far as sterilization of heat-sensitive materials and innocuity of sterilizing agents are concerned. The literature on plasma sterilization is reviewed. A major issue of plasma sterilization is the respective roles of UV photons and reactive species such as atomic and radicals. Insight into this matter is obtained by analyzing the survival curves of microorganisms. In contrast to classical sterilization where such plots show a unique straight line, plasma sterilization yields survival diagrams with two or three different linear segments. Three basic mechanisms are involved in the plasma inactivation of microorganisms: (A) direct destruction by UV irradiation of the genetic material of microorganisms; (B) erosion of the microorganisms atom by atom, through intrinsic photodesorption by UV irradiation to form volatile compounds combining atoms intrinsic to the microorganisms; (C) erosion of the microorganisms, atom by atom, through etching to form volatile compounds as a result of slow combustion using oxygen atoms or radicals emanating from the plasma. In some cases, etching is further activated by UV photons, increasing the elimination rate of microorganisms. These mechanisms make plasma sterilization totally different from classical sterilization techniques and suggest its use to inactivate nonconventional infectious agents such as the abnormal prions.