Optimized Electrophoretic Deposition of Chitosan/Mesoporous Glass Nanoparticles with Gentamicin on Titanium Implants: Enhancing Hemocompatibility and Antibacterial Activity.

Titanium-based implants have long been studied and used for applications in bone tissue engineering, thanks to their outstanding mechanical properties and appropriate biocompatibility. However, many implants struggle with osseointegration and attachment and can be vulnerable to the development of infections. In this work, we have developed a composite coating via electrophoretic deposition, which is both bioactive and antibacterial. Mesoporous bioactive glass particles with gentamicin were electrophoretically deposited onto a titanium substrate. In order to validate the hypothesis that the quantity of particles in the coatings is sufficiently high and uniform in each deposition process, an easy-to-use image processing algorithm was designed to minimize human dependence and ensure reproducible results. The addition of loaded mesoporous particles did not affect the good adhesion of the coating to the substrate although roughness was clearly enhanced. After 7 days of immersion, the composite coatings were almost dissolved and released, but phosphate-related compounds started to nucleate at the surface. With a simple and low-cost technique like electrophoretic deposition, and optimized stir and suspension times, we were able to synthesize a hemocompatible coating that significantly improves the antibacterial activity when compared to the bare substrate for both Gram-positive and Gram-negative bacteria.

Melaleuca armillaris Essential Oil as an Antibacterial Agent: The Use of Mesoporous Bioactive Glass Nanoparticles as Drug Carrier.

Bioactive glasses have been proposed for bone tissue engineering due to their excellent biocompatibility and osteo-inductive behaviour. The generation of mesoporous bioactive glass (nano) particles adds a high surface area for the dissolution and release of bioactive ions, and the possibility to load them with different drugs for antibacterial purposes. Essential oils (EO) are an interesting resource for alternative medical therapy, providing antimicrobial compounds that come from organic/natural resources like aromatic plants. Also, a biological polymer, such as chitosan, could be used to control the release of active agents from mesoporous bioactive glass (MBG) loaded particles. This work presents MBG particles with nominal composition (in mol) 60% SiO2, 30% CaO and 10% P2O5, loaded with essential oil of Melaleuca armillaris, which contains 1,8-cineol as the main active component, with an inhibitory in vitro activity against several bacterial species. Also, co-loading with a broad-spectrum antibiotic, namely gentamicin, was investigated. The MBG particles were found to be of around 300nm in diameter and to exhibit highly porous open structure. The release of EO from the particles reached 72% of the initial content after the first 24 h, and 80% at 48 h of immersion in phosphate buffered solution. Also, the MBG particles with EO and EO-gentamicin loading presented in vitro apatite formation after 7 days of immersion in simulated body fluid. The antibacterial tests indicated that the main effect, after 24 h of contact with the bacteria, was reached either for the MBG EO or MBG EO-gentamicin particles against E. coli, while the effect against S. aureus was less marked. The results indicate that MBG particles are highly bioactive with the tested composition and loaded with EO of Melaleuca armillaris. The EO, also combined with gentamicin, acts as an antibacterial agent but with different efficacy depending on the bacteria type.

Short-Term In Vivo Response to Anodized Magnesium Alloy as a Biodegradable Material for Bone Fracture Fixation Devices.

Biodegradable materials based on magnesium alloys have a huge potential for bone fracture fixation devices due to their adequate mechanical properties and biocompatibility. However, their fast degradation and the consequent liberation of hydrogen gas at the initial stages of implantation is the major limitation for their use. In this study, the AZ91D magnesium alloy was surface treated by an environment-friendly, nontoxic, and low-cost anodizing process and the early in vivo response was studied in a rat transcortical model. Adequate maturation of woven bone around implants-detected at day 7 post implantation-to lamellar bone was observed from day 15. Lamellar bone after 15 and 30 days of implantation presented similar volume, mineralization pattern, mineral to protein content, and estimated bone maturity between anodized AZ91D and polylactic acid (control) implants. Histology observation showed neither release of hydrogen bubbles in the region closed to the anodized AZ91D implant nor systemic effects on liver, kidney, and spleen. Thus, anodizing of AZ91D in the conditions stated here induced an adequate short-term in vivo response, which postulates their use as potential biodegradable fracture fixation devices for bone healing.

Functional behavior of chitosan/gelatin/silica-gentamicin coatings by electrophoretic deposition on surgical grade stainless steel.

Metals are used in several orthopedic applications as fixation elements for the stabilization of fractures or as prostheses. One of the most common orthopedic biomaterials in many developing countries is surgical grade stainless steel (SS). However, its use as permanent implant in orthopedic surgery is conditioned due to its limited corrosion resistance in physiological media, lack of osseointegration, and absence of antibacterial effect. The aim of this work is to generate a degradable coating with antibacterial properties for stainless steel to be used in implants/medical devices. The coating is composed of a biopolymer/silica-gentamicin nanoparticles composite obtained by electrophoretic deposition (EPD) on surgical grade stainless steel plates. The coating surface was characterized by microscopic examination, and in vitro performance was evaluated after immersion in phosphate-buffered saline (PBS) solution, simulated body fluid (SBF), and cell culture medium, to analyze coating degradation, antibiotics release, cell attachment (ST-2 stromal cells), and antibacterial (Escherichia coli and Staphylococcus aureus) properties. EPD coatings were uniform and covered homogeneously the surface of the SS substrate. Also the distribution of silica-gentamicin nanoparticles was homogeneous on the coated area. The degradation of the chitosan-gelatin coatings was evident after 7 days of immersion. The gentamicin release led to excellent antibacterial behavior at 24 h, meanwhile the cell proliferation (at 7 days culture) was not inhibited. The results show that the coating system exhibits promising behavior which could contribute to prevent hospital infections at early implantation times.

In vitro and in vivo characterization of anodised zirconium as a potential material for biomedical applications.

In vitro studies offer the insights for the understanding of the mechanisms at the tissue-implant interface that will provide an effective functioning in vivo. The good biocompatibility of zirconium makes a good candidate for biomedical applications and the attractive in vivo performance is mainly due to the presence of a protective oxide layer. The aim of this study is to evaluate by in vitro and in vivo approach, the influence of surface modification achieved by anodisation at 30 and 60V on zirconium implants on the first steps of the osseointegration process. In this study cell attachment, proliferation and morphology of mouse myoblast C2C12-GFP and in mouse osteoprogenitor MC3T3-E1 cells was evaluated. Also, together with the immune system response, osteoclast differentiation and morphology with RAW 264.7 murine cell line were analysed. It was found that anodisation treatment at 60V enhanced cell spreading and the osteoblastic and osteoclastic cells morphology, showing a strong dependence on the surface characteristics. In vivo tests were performed in a rat femur osteotomy model. Dynamical and static histological and histomorphometric analyses were developed 15 and 30days after surgery. Newly formed bone around Zr60V implants showed a continuous newly compact and homogeneous bone just 15 after surgery, as judged by the enhanced thickness and mineralization rate. The results indicate that anodising treatment at 60V could be an effective improvement in the osseointegration of zirconium by stimulating adhesion, proliferation, morphology, new bone thickness and bone mineral apposition, making zirconium an emerging candidate material for biomedical applications.

Calculation of cancellous bone elastic properties with the polarization-based FFT iterative scheme.

The Fast Fourier Transform-based method, originally introduced by Moulinec and Suquet in 1994 has gained popularity for computing homogenized properties of composites. In this work, the method is used for the computational homogenization of the elastic properties of cancellous bone. To the authors' knowledge, this is the first study where the Fast Fourier Transform scheme is applied to bone mechanics. The performance of the method is analyzed for artificial and natural bone samples of 2 species: bovine femoral heads and implanted femurs of Hokkaido rats. Model geometries are constructed using data from X-ray tomographies, and the bone tissue elastic properties are measured using microindentation and nanoindentation tests. Computed results are in excellent agreement with those available in the literature. The study shows the suitability of the method to accurately estimate the fully anisotropic elastic response of cancellous bone. Guidelines are provided for the construction of the models and the setting of the algorithm.

A comparative study of zirconium and titanium implants in rat: osseointegration and bone material quality.

Permanent metal implants are widely used in human medical treatments and orthopedics, for example as hip joint replacements. They are commonly made of titanium alloys and beyond the optimization of this established material, it is also essential to explore alternative implant materials in view of improved osseointegration. The aim of our study was to characterize the implant performance of zirconium in comparison to titanium implants. Zirconium implants have been characterized in a previous study concerning material properties and surface characteristics in vitro, such as oxide layer thickness and surface roughness. In the present study, we compare bone material quality around zirconium and titanium implants in terms of osseointegration and therefore characterized bone material properties in a rat model using a multi-method approach. We used light and electron microscopy, micro Raman spectroscopy, micro X-ray fluorescence and X-ray scattering techniques to investigate the osseointegration in terms of compositional and structural properties of the newly formed bone. Regarding the mineralization level, the mineral composition, and the alignment and order of the mineral particles, our results show that the maturity of the newly formed bone after 8 weeks of implantation is already very high. In conclusion, the bone material quality obtained for zirconium implants is at least as good as for titanium. It seems that the zirconium implants can be a good candidate for using as permanent metal prosthesis for orthopedic treatments.

Can anodised zirconium implants stimulate bone formation? Preliminary study in rat model.

The mechanical properties and good biocompatibility of zirconium and some of its alloys make these materials good candidates for biomedical applications. The attractive in vivo performance of zirconium is mainly due to the presence of a protective oxide layer. In this preliminary study, the surface of pure zirconium modified by anodisation in acidic media at low potentials to enhance its barrier protection given by the oxides and osseointegration. Bare, commercially pure zirconium cylinders were compared to samples anodised at 30 V through electrochemical tests and scanning electron microscopy (SEM). For both conditions, in vivo tests were performed in a rat tibial osteotomy model. The histological features and fluorochrome-labelling changes of newly bone formed around the implants were evaluated on the non-decalcified sections 63 days after surgery. Electrochemical tests and SEM images show that the anodisation treatment increases the barrier effect over the material and the in vivo tests show continuous newly formed bone around the implant with a different amount of osteocytes in their lacunae depending on the region. There was no significant change in bone thickness around either kind of implant but the anodised samples had a significantly higher mineral apposition, suggesting that the anodisation treatment stimulates and assists the osseointegration process. We conclude that anodisation treatment at 30 V can stimulate the implant fixation in a rat model, making zirconium a strong candidate material for permanent implants.

Bone quality around bioactive silica-based coated stainless steel implants: analysis by micro-Raman, XRF and XAS techniques.

Surface modification of surgical stainless steel implants by sol gel coatings has been proposed as a tool to generate a surface that besides being protective could also create a "bioactive" interface to generate a natural bonding between the metal surface and the existing bone. The aim of this work is to analyze the quality and bone formation around hybrid bioactive coatings containing glass-ceramic particles, made by sol-gel process on 316L stainless steel used as permanent implant in terms of mineralization, calcium content and bone maturity with micro Raman, X-ray microfluorescence and X-ray absorption techniques. Uncoated implants seem to generate a thin bone layer at the beginning of osseointegration process and then this layer being separated from the surface with time. The hybrid coatings without glass-ceramic particles generate new bone around implants, with high concentration of Ca and P at the implant/tissue interface. This fact seems to be related with the presence of silica nanoparticles in the layer. The addition of bioactive particles promotes and enhances the bone quality with a homogeneous Ca and P content and a low rate of beta carbonate substitution and crystallinity, similar to young and mechanical resistant bone.

Improving the osteointegration and bone-implant interface by incorporation of bioactive particles in sol-gel coatings of stainless steel implants.

In this study, we report a hybrid organic-inorganic TEOS-MTES (tetraethylorthosilicate-methyltriethoxysilane) sol-gel-made coating as a potential solution to improve the in vivo performance of AISI 316L stainless steel, which is used as permanent bone implant material. These coatings act as barriers for ion migration, promoting the bioactivity of the implant surface. The addition of SiO(2) colloidal particles to the TEOS-MTES sol (10 or 30 mol.%) leads to thicker films and also acts as a film reinforcement. Also, the addition of bioactive glass-ceramic particles is considered responsible for enhancing osseointegration. In vitro assays for bioactivity in simulated body fluid showed the presence of crystalline hydroxyapatite (HA) crystals on the surface of the double coating with 10mol.% SiO(2) samples on stainless steel after 30 days of immersion. The HA crystal lattice parameters are slightly different from stoichiometric HA. In vivo implantation experiments were carried out in a rat model to observe the osteointegration of the coated implants. The coatings promote the development of newly formed bone in the periphery of the implant, in both the remodellation zone and the marrow zone. The quality of the newly formed bone was assessed for mechanical and structural integrity by nanoindentation and small-angle X-ray scattering experiments. The different amount of colloidal silica present in the inner layer of the coating slightly affects the material quality of the newly formed bone but the nanoindentation results reveal that the lower amount of silica in the coating leads to mechanical properties similar to cortical bone.

Modificacion superficial de aleaciones base cobalto para uso en cirugia ortopedica / Surface modification of cobalt base alloys for surgical devices

Las aleaciones que se usan en ortopedia tiene en comun la formacion de una pelicula pasiva que disminuye su velocidad de corrosion. La migracion de iones metalicos puede causar una respuesta adversa en el huesped y desarrollar una cascada de eventos que pueden culminar con la perdida del implante. Los metales a su vez, son incapaces de generar union natural al tejido oseo sin cementacion o fijacion externa. En este trabajo se presenta la modificacion superificial de implantes de aleacion de CrCoMo (F75) por medio de recubrimientos obtenidos por la tecnica de sol-gel con el agregado de particulas de vidrio o vitroceramico bioactivo del sistema CaO-SiO2-P2O5 con el doble proposito de proteccion contra la corrosion y generacion de union natural con el tejido oseo (resumen truncado)

Modificacion superficial de aleaciones base cobalto para uso en cirugia ortopedica / Surface modification of cobalt base alloys for surgical devices

Las aleaciones que se usan en ortopedia tiene en comun la formacion de una pelicula pasiva que disminuye su velocidad de corrosion. La migracion de iones metalicos puede causar una respuesta adversa en el huesped y desarrollar una cascada de eventos que pueden culminar con la perdida del implante. Los metales a su vez, son incapaces de generar union natural al tejido oseo sin cementacion o fijacion externa. En este trabajo se presenta la modificacion superificial de implantes de aleacion de CrCoMo (F75) por medio de recubrimientos obtenidos por la tecnica de sol-gel con el agregado de particulas de vidrio o vitroceramico bioactivo del sistema CaO-SiO2-P2O5 con el doble proposito de proteccion contra la corrosion y generacion de union natural con el tejido oseo (resumen truncado)